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Title:
ESSENTIAL AND IMPORTANT GENES OF PSEUDOMONAS AEROGINOSA AND THE USE THEREOF TO DESIGN OR IDENTIFY ANTIBACTERIAL AGENTS
Document Type and Number:
WIPO Patent Application WO/2003/089572
Kind Code:
A2
Abstract:
The invention includes a database of candidate essential genes in Pseudomonas aeruginosa, as well as otherwise important genes that, when mutated, lead to a growth attenuated phenotype. Such genes and mutants of such genes are important for identifying antibacterial agents suitable for treating and preventing Pseudomonas aeruginosa infections. The invention includes methods for confirming the essentially or importance of candidate genes, as well as methods for utilizing those genes to screen for new antibacterial drugs. The invention also includes the antibacterial agents identified using the disclosed methods, as well as methods of using the same for treating and preventing Pseudomonas infection.

Inventors:
Folger Bruce, Kim (9756 Lakeshore Blvd, NE Seattle, WA, 98115, US)
Warrener, Paul (2803 N.W. 67th Street, Seattle, WA, 98117, US)
Hou, Kevin (18123 N.W. Village Park Drive, Issaquah, WA, 98027, US)
Application Number:
PCT/US2002/035518
Publication Date:
October 30, 2003
Filing Date:
November 05, 2002
Export Citation:
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Assignee:
CHIRON CORPORATION (4560 Horton Street, Emeryville, CA, 94608-2916, US)
Folger Bruce, Kim (9756 Lakeshore Blvd, NE Seattle, WA, 98115, US)
Warrener, Paul (2803 N.W. 67th Street, Seattle, WA, 98117, US)
Hou, Kevin (18123 N.W. Village Park Drive, Issaquah, WA, 98027, US)
International Classes:
C07K14/21; G06F19/00; A61K38/00; (IPC1-7): C12N/
Attorney, Agent or Firm:
Hale, Rebecca M. (Chiron Corporation, Intellectual Property-R338 P. O. Box 809, Emeryville CA, 94662-8097, US)
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Claims:
What is claimed:
1. An isolated nucleic acid molecule comprising a nucleic acid sequence encoding a polypeptide having at least 80% sequence identity to a polypeptide encoded by a nucleic acid sequence selected from the group consisting of the Pseudomonas aeruginosa open reading frames (ORFs) listed in Table 1.
2. The isolated nucleic acid molecule of claim wherein the sequence encodes a polypeptide having at least 90% sequence identity to said nucleic acid sequence.
3. The isolated nucleic acid sequence of claim 1 wherein the sequence encodes a polypeptide having at least 95% sequence identity to said nucleic acid sequence.
4. An isolated nucleic acid molecule comprising a nucleic acid sequence encoding a polypeptide having at least 80% sequence identity to a polypeptide encoded by an essential or important nucleic acid sequence selected from the group consisting of the Pseudomonas aeruginosa open reading frames (ORFs) listed in Table 1, wherein said essential or important nucleic acid sequence is identified as being essential or important by integration knockout coupled with extrachromosomal complementation.
5. The isolated nucleic acid sequence of claim 4 wherein the sequence encodes a polypeptide having at least 90% sequence identity to said essential polypeptide.
6. The isolated nucleic acid sequence of claim 5 wherein the sequence encodes a polypeptide having at least 95% sequence identity to said essential polypeptide.
7. An isolated nucleic acid molecule comprising a nucleic acid sequence encoding a polypeptide having at least 80% sequence identity to a polypeptide encoded by an essential or important nucleic acid sequence selected from the group consisting of the Pseudomonas aeruginosa open reading frames (ORFs) listed in Table 1, wherein said essential or important nucleic acid sequence is identified as being essential by integration of a regulatable promoter into the gene.
8. The isolated nucleic acid sequence of claim 7 which encodes a polypeptide having at least 90% sequence identity to said polypeptide.
9. The isolated nucleic acid sequence of claim 8 which encodes a polypeptide having at least 95% sequence identity to said polypeptide.
10. A method of screening for an antibacterial agent, comprising determining whether a test compound is active against the bacterial gene of claim 1.
11. A method of screening for an antibacterial agent, comprising determining whether a test compound is active against the bacterial gene of claim 2.
12. A method of screening for an antibacterial agent, comprising determining whether a test compound is active against the bacterial gene of claim 3.
13. A method of screening for an antibacterial agent, comprising determining whether a test compound is active against the protein encoded by the bacterial gene of claim 1.
14. A method of screening for an antibacterial agent, comprising determining whether a test compound is active against the protein encoded by the bacterial gene of claim 2.
15. A method of screening for an antibacterial agent, comprising determining whether a test compound is active against the protein encoded by the bacterial gene of claim 3.
16. A method of screening for an antibacterial agent, comprising determining whether a test compound is active against the essential or important bacterial gene of claim 4.
17. A method of screening for an antibacterial agent, comprising determining whether a test compound is active against the essential or important bacterial gene of claim 5.
18. A method of screening for an antibacterial agent, comprising determining whether a test compound is active against the essential or important bacterial gene of claim 6.
19. A method of screening for an antibacterial agent, comprising determining whether a test compound is active against the protein encoded by the essential or important bacterial gene of claim 4.
20. A method of screening for an antibacterial agent, comprising determining whether a test compound is active against the protein encoded by the essential or important bacterial gene of claim 5.
21. A method of screening for an antibacterial agent, comprising determining whether a test compound is active against the protein encoded by the essential or important bacterial gene of claim 6.
22. A method of screening for an antibacterial agent, comprising determining whether a test compound is active against the essential or important bacterial gene of claim 7.
23. A method of screening for an antibacterial agent, comprising determining whether a test compound is active against the essential or important bacterial gene of claim 8.
24. A method of screening for an antibacterial agent, comprising determining whether a test compound is active against the essential or important bacterial gene of claim 9.
25. A method of screening for an antibacterial agent, comprising determining whether a test compound is active against the protein encoded by the essential or important bacterial gene of claim 7.
26. A method of screening for an antibacterial agent, comprising determining whether a test compound is active against the essential or important bacterial gene of claim 8.
27. A method of screening for an antibacterial agent, comprising determining whether a test compound is active against the essential or important bacterial gene of claim 9.
28. The method of claim 13, comprising the steps of : a) contacting said protein or a biologically active fragment thereof with a test compound; and b) determining whether said test compound binds to said protein or said fragment ; wherein binding of said test compound to said polypeptide or said fragment is indicative that said test compound is an antibacterial agent.
29. The method of claim 14, comprising the steps of : a) contacting said protein or a biologically active fragment thereof with a test compound; and b) determining whether said test compound binds to said protein or said fragment; wherein binding of said test compound to said polypeptide or said fragment is indicative that said test compound is an antibacterial agent.
30. The method of claim 15, comprising the steps of : a) contacting said protein or a biologically active fragment thereof with a test compound; and b) determining whether said test compound binds to said protein or said fragment; wherein binding of said test compound to said polypeptide or said fragment is indicative that said test compound is an antibacterial agent.
31. The method of claim 19, comprising the steps of : a) contacting said protein or a biologically active fragment thereof with a test compound; and b) determining whether said test compound binds to said protein or said fragment; wherein binding of said test compound to said polypeptide or said fragment is indicative that said test compound is an antibacterial agent.
32. The method of claim 20, comprising the steps of : a) contacting said protein or a biologically active fragment thereof with a test compound; and b) determining whether said test compound binds to said protein or said fragment; wherein binding of said test compound to said polypeptide or said fragment is indicative that said test compound is an antibacterial agent.
33. The method of claim 21, comprising the steps of : a) contacting said protein or a biologically active fragment thereof with a test compound; and b) determining whether said test compound binds to said protein or said fragment; wherein binding of said test compound to said polypeptide or said fragment is indicative that said test compound is an antibacterial agent.
34. The method of claim 25, comprising the steps of : a) contacting said protein or a biologically active fragment thereof with a test compound; and b) determining whether said test compound binds to said protein or said fragment; wherein binding of said test compound to said polypeptide or said fragment is indicative that said test compound is an antibacterial agent.
35. The method of claim 26, comprising the steps of : a) contacting said protein or a biologically active fragment thereof with a test compound; and b) determining whether said test compound binds to said protein or said fragment; wherein binding of said test compound to said polypeptide or said fragment is indicative that said test compound is an antibacterial agent.
36. The method of claim 27, comprising the steps of : a) contacting said protein or a biologically active fragment thereof with a test compound; and b) determining whether said test compound binds to said protein or said fragment; wherein binding of said test compound to said polypeptide or said fragment is indicative that said test compound is an antibacterial agent.
37. A method for evaluating a test agent for inhibition of expression of the gene of claim 1, comprising: a) contacting a cell expressing said gene with said agent; and b) determining the amount or level of expression of said essential gene in said sample.
38. A method for evaluating a test agent for inhibition of expression of the gene of claim 2, comprising: a) contacting a cell expressing said gene with said agent; and b) determining the amount or level of expression of said essential gene in said sample.
39. A method for evaluating a test agent for inhibition of expression of the gene of claim 3, comprising: a) contacting a cell expressing said gene with said agent; and b) determining the amount or level of expression of said essential gene in said sample.
40. A method for evaluating a test agent for inhibition of expression of the essential or important gene of claim 4, comprising: a) contacting a cell expressing said essential or important gene with said agent; and b) determining the amount or level of expression of said essential or important gene in said sample.
41. A method for evaluating a test agent for inhibition of expression of the gene of claim 5, comprising: a) contacting a cell expressing said gene with said agent; and b) determining the amount or level of expression of said essential gene in said sample.
42. A method for evaluating a test agent for inhibition of expression of the gene of claim 6, comprising: a) contacting a cell expressing said gene with said agent; and b) determining the amount or level of expression of said essential gene in said sample.
43. A method for evaluating a test agent for inhibition of expression of the essential or important gene of claim 7, comprising: a) contacting a cell expressing said essential or important gene with said agent; and b) determining the amount or level of expression of said essential or important gene in said sample.
44. A method for evaluating a test agent for inhibition of expression of the gene of claim 8, comprising: a) contacting a cell expressing said gene with said agent; and b) determining the amount or level of expression of said essential gene in said sample.
45. A method for evaluating a test agent for inhibition of expression of the gene of claim 9, comprising: a) contacting a cell expressing said gene with said agent; and b) determining the amount or level of expression of said essential gene in said sample.
46. The method of claim 37, wherein said level of expression is measured by measuring the amount of expression product in said cell relative to a cell that has not been contacted with said agent.
47. A method for evaluating a test agent for inhibition of expression of the gene of claim 38, comprising: a) contacting a cell expressing said gene with said agent; and b) determining the amount or level of expression of said essential gene in said sample.
48. A method for evaluating a test agent for inhibition of expression of the gene of claim 39, comprising: a) contacting a cell expressing said gene with said agent; and b) determining the amount or level of expression of said essential gene in said sample.
49. The method of claim 37, wherein said level of expression is measured by measuring the level of expression of a gene fusion to said gene relative to a cell containing said gene fusion that has not been contacted with said agent.
50. The method of claim 38, wherein said level of expression is measured by measuring the level of expression of a gene fusion to said gene relative to a cell containing said gene fusion that has not been contacted with said agent.
51. The method of claim 39, wherein said level of expression is measured by measuring the level of expression of a gene fusion to said gene relative to a cell containing said gene fusion that has not been contacted with said agent.
52. The method of claim 37, wherein said level of expression is measured by measuring the level of expression of a protein fusion to said gene relative to a cell containing said protein fusion that has not been contacted with said agent.
53. The method of claim 38, wherein said level of expression is measured by measuring the level of expression of a gene fusion to said gene relative to a cell containing said gene fusion that has not been contacted with said agent.
54. The method of claim 39, wherein said level of expression is measured by measuring the level of expression of a gene fusion to said gene relative to a cell containing said gene fusion that has not been contacted with said agent.
55. A method for evaluating an potential antibacterial agent, comprising the steps of : a) providing a bacterial strain comprising a mutant form of the gene of claim 1, wherein said mutant form of the gene confers a growth conditional or attenuated growth phenotype; b) contacting bacteria of said bacterial strain with said test compound in semipermissive or permissive growth conditions; and c) determining whether the growth of said bacterial strain comprising said mutant form of a gene is reduced in the presence of said test compound to a greater extent than a comparison bacteria comprising a normal form of said gene.
56. A method for evaluating an potential antibacterial agent, comprising the steps of : a) providing a bacterial strain comprising a mutant form of the gene of claim 2, wherein said mutant form of the gene confers a growth conditional or attenuated growth phenotype; b) contacting bacteria of said bacterial strain with said test compound in semipermissive or permissive growth conditions; and c) determining whether the growth of said bacterial strain comprising said mutant form of a gene is reduced in the presence of said test compound to a greater extent than a comparison bacteria comprising a normal form of said gene.
57. A method for evaluating an potential antibacterial agent, comprising the steps of : a) providing a bacterial strain comprising a mutant form of the gene of claim 3, wherein said mutant form of the gene confers a growth conditional or attenuated growth phenotype; b) contacting bacteria of said bacterial strain with said test compound in semipermissive or permissive growth conditions; and c) determining whether the growth of said bacterial strain comprising said mutant form of a gene is reduced in the presence of said test compound to a greater extent than a comparison bacteria comprising a normal form of said gene.
58. A library of nucleic acid sequences consisting essentially of nucleic acid sequences having at least 80% protein sequence identity to a nucleic acid sequence selected from the group consisting of the Pseudomonas aeruginosa open reading frames (ORFs) listed in Table 1, wherein said library of nucleic acid sequences is employed to identify essential genes in Pseudomonas.
59. The library of 58 wherein said nucleic acid sequences encode proteins having at least 90% sequence identity to the open reading frames in Table 1.
60. The library of 58 wherein said nucleic acid sequences encode proteins having at least 95% sequence identity to the open reading frames in Table 1.
61. A map of at least about 10,000 to about 14,000 transposon insertions in the genome of Pseudomonas aeruginosa, wherein said map is useful for identifying genes that are essential or important for survival of said Pseudomonas aeruginosa.
62. A vector comprising a promoter operably linked to the nucleic acid sequence of claim 1.
63. A vector comprising a promoter operably linked to the nucleic acid sequence of claim 2.
64. A vector comprising a promoter operably linked to the nucleic acid sequence of claim 3.
65. The vector of claim 62, wherein said promoter is active in Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus, Hemophilus influenzae, Neisseria gonorrhea, Klebsiella pneumoniae, and Stfeptocooci.
66. The vector of claim 63, wherein said promoter is active in Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus, Hemophilus influenzae, Neisseria gonorrhea, Klebsiellapneumoniae, and Streptocooci.
67. The vector of claim 64, wherein said promoter is active in Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus, Hemophilus influenzae, Neisseria gonorrhea, Klebsiellapneumoniae, and Streptocooci.
68. A host cell comprising the vector of claim 65.
69. A host cell comprising the vector of claim 66.
70. A host cell comprising the vector of claim 67.
71. A fragment of the nucleic acid of claim 1,2 or 3 said fragment comprising at least 10, at least 20, at least 25, at least 30, or at least 50 consecutive bases of said nucleic acid.
72. A protein having at least about 80% sequence identity to the protein encoded by the nucleic acid of claim 1,2 or 3.
73. A protein having at least about 80% sequence identity to the protein encoded by the nucleic acid of claim 4,5 or 6.
74. A protein having at least about 80% sequence identity to the protein encoded by the nucleic acid of claim 7,8 or 9.
75. An antibody or antibody fragment capable of specifically binding the protein of claim 72.
76. An antibody or antibody fragment capable of specifically binding the protein of claim 73.
77. An antibody or antibody fragment capable of specifically binding the protein of claim 74.
78. An agent identified as having antibacterial activity by any of the methods of claims 1057.
79. A method for inhibiting the growth or survival of Pseudomonas aeruginosa comprising contacting said bacteria with an agent identified by a method as set forth in any one of claims 1057 so as to inhibit growth or survival.
80. A pharmaceutical composition comprising an agent according to claim 78.
81. A method for treating a patient having a Pseudomonas aeruginosa infection, comprising administering to said patient an amount of an agent according to claim 78 effective to reduce or inhibit growth or survival of said Pseudomonas aeruginosa.
82. A method of protecting a patient against a Pseudomonas aeruginosa infection, comprising administering to said patient an amount of an agent according to claim 78 effective to prevent said patient from acquiring a Pseudomonas aeruginosa infection.
83. The isolated nucleic acid molecule of claim 4,5 or 6, wherein said nucleic acid contains an essential gene selected from the group consisting of Pseudomonas aeruginosa uppS, ispB and metK.
84. The isolated nucleic acid molecule of claim 4,5 or 6, wherein said nucleic acid comprises the Pseudomonas aeruginosa ispA gene.
85. The nucleic acid library of claim 58,59 or 60, wherein said map is in electronic form.
86. The library of claim 85, wherein said electronic form is selected from the group consisting of magnetic storage media, such as a floppy disc, a hard disc storage medium, and a magnetic tape; optical storage media such as CDROM; electrical storage media such as RAM and ROM; hybrids of these categories such as magnetic/optical storage media; computer readable forms such as a word processing text file, database format, searchable files, executable files and search program software.
87. The transposon insertion map of claim 61, wherein said map is in electronic form.
88. The map of claim 85, wherein said electronic form is selected from the group consisting of magnetic storage media, such as a floppy disc, a hard disc storage medium, and a magnetic tape; optical storage media such as CDROM; electrical storage media such as RAM and ROM; hybrids of these categories such as magnetic/optical storage media; computer readable forms such as a word processing text file, database format, searchable files, executable files and search program software.
89. A method for identifying a library of putative essential or important genes using a High Throughput Transposon Insertion Database (HTTIM), comprising: (a) mutagenizing a bacterial genome with a transposon such that individual cells containing at least one transposon insertion are isolated; (b) collecting and mapping said at least one transposon insertion in each individual cell so as to form a database of transposon insertion sites, or an HTTIM; (c) comparing said database of transposon insertion sites with a database comprising the genomic sequence of the bacterium to identify open reading frames in said genomic sequence database that are not disrupted by a transposon insertion; (d) forming a library from said putative essential or important genes that are not disrupted by a transposon.
90. The method of claim 89, wherein said bacteria is P. aeruginosa or S aureus.
91. The method of claim 89, wherein said transposon inserts randomly into the target genome.
92. The method of claim 91, wherein said transposon is Tn5.
93. The method of claim 91, wherein said HTTIM comprises at least about 5000 transposon insertion sites.
94. The method of claim 91, wherein said HTTIM comprises at least about 10000 transposon insertion sites.
95. The method of claim 91, wherein said HTTIM comprises at least about 13000 transposon insertion sites.
96. The library of putative essential or important genes identified by the method of claim 91, wherein said library comprises at most about 3000 genes.
97. The library of putative essential or important genes identified by the method of claim 91, wherein said library comprises at most about 2500 genes.
98. The library of putative essential or important genes identified by the method of claim 91, wherein said library comprises at most about 2000 genes.
99. The library of putative essential or important genes identified by the method of claim 91, wherein said library comprises at most about 1700 genes.
100. The method of claim 91, further comprising a statistical calculation for identifying putative essential or important genes.
101. The method of claim 100, further comprising the statistical method applied herein.
102. The method of claim 91, further comprising a physical mutagenesis experiment in order to verify essential or important genes.
103. The method of claim 102, wherein said physical mutagenesis comprises knocking out a putative essential or important gene or creating a promoter swap mutant.
104. An essential or important gene identified by the method of claim 102.
105. An antibacterial agent that targets the gene of claim 104, or the gene product encoded by said gene.
106. A pharmaceutical composition comprising said antibacterial agent of claim 105.
107. A method of identifying a nucleic acid motif associated with a Pseudomonas aeruginosa infection, comprising screening the library of claim 58, 59 or 60 for conserved nucleic acid fragments.
Description:
ESSENTIAL AND IMPORTANT GENES OF PSEUDOMONAS AERUGINOSA AND THE USE THEREOF TO DESIGN OR IDENTIFY ANTIBACTERIAL AGENTS Cross Reference Related to Related Applications [0001] This application relates to U. S. Provisional Serial No. 60/372,095, filed on April 15,2002 and which is incorporated in their entirety by reference herein.

Field of Invention [0002] The present invention relates to the identification of essential and important genes in Pseudomonas aeruginosa, and the use thereof in screening assays and diagnostic methods to identify, evaluate or design antibacterial agents useful for the treatment of Pseudomonas infections. Such agents are particularly useful in preventing and treating opportunistic infections in immunocompromised individuals and for treating and preventing pulmonary infections in patients having cystic fibrosis disease.

Also disclosed is a Bayessian statistical model that may be utilized to increase the statistical confidence that any given gene identified using the disclosed methodology is essential.

Background of Invention [0003] Pseudomonas aeruginosa is a versatile Gram-negative bacterium that is able to adapt to and thrive in many ecological niches, from water and soil to plant and animal tissues. The bacterium is capable of utilizing a wide range of organic compounds as food sources, thus giving it an exceptional ability to colonize ecological niches where nutrients are limited, such as soil, marshes and coastal marine habitats.

Hardalo, C. & Edberg, S. C. Pseudomonas aeruginosa : assessment of risk from drinking water. Crit. Rev. Microbiol. 23,47-75 (1997). It also forms biofilms on wet surfaces such as those of rocks and soil. Costerton, J. W. , Stewart, P. S. & Greenberg, E. P. Bacterial biofilms: a common cause of persistent infections. Science 284,1318- 1322 (1999). Ahearn, D. G. , Borazjani, R. N. , Simmons, R. B. & Gabriel, M. M.

Primary adhesion of Pseudomonas aeruginosa to inanimate surfaces including biomaterials. Methods Enzymol. 310,551-557 (1999). Analysis of the P. aeruginosa genome has identified genes involved in locomotion, attachment, transport and utilization of nutrients, antibiotic efflux, and two component and other regulatory systems involved in sensing and responding to environmental changes. Because its natural habitat is the soil, where it exposed to bacilli, actinomycetes and molds, it has developed resistance to a variety of their naturally-occurring antibiotics.

[0004] The emergence of P. aeruginosa as a major opportunistic human pathogen during the past century may be a consequence of its resistance to the antibiotics and disinfectants that eliminate other environmental bacteria. P. aeruginosa is now a significant source of bacteraemia in burn victims, urinary-tract infections in catheterized patients, and hospital-acquired pneumonia in patients on respirators.

Bodey, G. P. , Bolivar, R. , Fainstein, V. & Jadeja, L. Infections caused by Pseudomonas aeruginosa. Rev. Infect. Dis. 5,279-313 (1983). It is also the predominant cause of morbidity and mortality in cystic fibrosis patients, whose abnormal airway epithelia allow long-term colonization of the lungs by P. aeruginosa. Thus, people with cystic fibrosis, burn victims, individuals with cancer and AIDS, and patients requiring extensive stays in intensive care units are particularly at risk of disease resulting from P. aeruginosa infection. P. aeruginosa is also a cause of a variety of different disorders including septicemia, urinary tract infections, pneumonia and chronic lung infections, endocarditis, dermatitis, osteochondritis, ear and eye infections, bone and joint infections, gastrointestinal infections and skin and soft tissue infections, including wound infections, pyoderma and dermatitis.

[0005] Cystic fibrosis is one of the most common fatal genetic disorders in the United States, affecting about 30,000 individuals. A comparable number of people in Europe also have CF. It is most prevalent in the Caucasian population, occurring in one of every 3,300 live births. The gene involved in cystic fibrosis was identified in 1989 and codes for a protein called the cystic fibrosis transmembrane conductance regulator (CFTR). This protein, normally produced in a number of tissues throughout the body, regulates the movement of salt and water in and out of these cells. One hallmark of CF is the presence of a thick mucus secretion that clogs the bronchial tubes in the lungs and plugs the exit passages from pancreas and intestines, leading to loss of function of these organs and resulting in a predisposition toward chronic bacterial infections.

Pseudomonas aeruginosa, having a propensity to live in warm, wet environments, is a particular problem for CF patients, whose lungs typically become colonized (inhabited long-term) by P. aeruginosa before their lOth birthday. Although antibiotics can decrease the frequency and duration of these attacks, resistant bacteria are quick to develop and the bacteria are never completely eradicated from the lung. More effective antibiotics are necessary for improving lung function and quality of life for CF patients for extended time periods.

[0006] Pseudomonas aeruginosa is notorious for its resistance to antibiotics and is, therefore, a particularly dangerous and dreaded pathogen. Todor, K. 2000 Pseudomonas aeruginosa, University of Wisconsin-Madison, http://www. bact. wisc. edu/microtextbook/disease/ pseudomonas. html, available on April 25,2001. The permeability barrier afforded by its outer membrane LPS also contributes to its natural antibiotic resistance, as do the presence of two antibiotic resistance plasmids, both R-factors and RTFs, which are commonly transferred between cells by the bacterial processes of transduction and conjugation. Only a few antibiotics are effective against Pseudomonas, including tobramyocin (TOBI; Chiron), fluoroquinolone, gentamicin and imipenem, and even these antibiotics are not effective against all strains.

[0007] Pseudomonas aeruginosa disease generally begins with some alteration or circumvention of normal host defenses and may involve several different virulence determinants. Todor, 2000, supra. The ultimate Pseudomonas infection may be seen as composed of three distinct stages: (1) bacterial attachment and colonization; (2) local invasion; (3) disseminated systemic disease. Particular bacterial determinants of virulence mediate each of these stages and are ultimately responsible for the characteristic syndromes that accompany the disease. For instance, Pseudomonas utilize fimbriae or pili to adhere to the epithelial cells, apparently via binding to specific galactose or mannose or sialic acid receptors on epithelial cells. Fimbrial adherence may be an important step in Pseudomonas keratitis and urinary tract infections, as well as infections of the respiratory tract. Mucoid strains, which produce an a exopolysaccharide (alginate) have an additional or alternative adhesin which attaches to the tracheobronchial mucin (N-acetylglucosamine). Therefore, mucoid strains of P. aeruginosa are commonly seen in lung infections.

[0008] The ability of P. aeruginosa to invade tissues depends upon its resistance to phagocytosis and the host immune defenses, and the extracellular enzymes and toxins that break down physical barriers and otherwise contribute to bacterial invasion.

Todor, 2000, supra. For instance, Pseudomonas elastase cleaves collagen, IgG, IgA, and complement, and also lyses fibronectin to expose receptors for bacterial attachment on the mucosa of the lung. Alkaline protease interferes with fibrin formation and lyses fibrin. Together, elastase and alkaline protease destroy the ground substance of the cornea and other supporting structures composed of fibrin and elastin. Elastase and alkaline protease together are also reported to cause the inactivation of gamma Interferon (IFN) and Tumor Necrosis Factor (TNF).

[0009] P. aeruginosa produces three other soluble proteins involved in invasion, including a cytotoxin (MW 25,000) and two hemolysins. Todor, 2000, supra. The cytotoxin is a pore-forming protein originally named leukocidin because of its effect on neutrophils, but it appears to be cytotoxic for most eukaryotic cells. Of the two hemolysins, one is a phospholipase and the other is a lecithinase. They appear to act synergistically to break down lipids and lecithin. The cytotoxin and hemolysins contribute to invasion through their cytotoxic effects on eukaryotic cells.

[0010] Pseudomonas aeruginosa also produces two extracellular protein toxins, Exoenzyme S and Exotoxin A. Exoenzyme S may act to impair the function of phagocytic cells in the bloodstream and internal organs to prepare for invasion by P. aeruginosa, and is typically produced by bacteria growing in burned tissue. Exotoxin A is partially identical to diphtheria toxin, and exhibits a necrotizing activity at the site of bacterial colonization and is thereby thought to contribute to the colonization process. Indirect evidence involving the role of exotoxin A in disease is seen in the increased chance of survival in patients with Pseudomonas septicemia that is correlated with the titer of anti-exotoxin A antibodies in the serum.

[0011] While therapeutic measures aimed at any of the above virulence factors may help to slow the progression of an infection and may be useful in combined therapeutic regimens, given the variety of virulence factors of P. aeruginosa, antibacterial agents that inhibit growing bacteria by interacting with essential genes and essential gene products are necessary. Although, this is not to say that genes encoding virulence factors would not be essential to survival in particular niches or environments, emphasizing the importance of screening for gene essentiality in various pathogenic environments. See, e. g., Coulter et al., 1998, Staphylococcus aureus genetic loci impacting growth and survival in multiple infection environments, Mol. Microbiol.

30 (2): 393-404. However, as P. aeruginosa becomes more and more resistant to existing antibacterial agents, new compounds are required.

[0012] Indeed, reports of bacterial strains resistant to the most powerful known antibiotics are becoming more common, signaling that new antibiotics are needed for all bacteria, not only P. aeruginosa. For instance, the United States Center for Disease Control recently announced that one of the most powerful known antibiotics, vancomycin, was unable to treat an infection of Staphylococcus aureus (staph), an organism commonly found in the environment and responsible for many nosocomial infections. If this trend continues, some have warned that we could return to a time when a common bacterial infection is a life threatening matter. See Zyskind et al. , WO 00/44906, published August 3,2000.

[0013] Historically, however, the identification of new antibacterial drugs has been painstaking and laborious with no guarantee of success. Traditional methods involve blindly and randomly testing potential drug candidate molecules, with the hopes that one might be effective. Today, the average cost to discover and develop a new drug is nearly $500 million, and the average time is 15 years from laboratory to patient. New identification and screening methods that shorten and improve this process are much needed.

[0014] A newly emerging technique for identifying new antibacterial agents is to first identify gene sequences and proteins required for the proliferation of bacteria, or "essential"genes and proteins, and then conduct a biochemical and structural analysis of that target gene or protein in order to derive compounds that interact with the target.

Such methodology employs molecular modeling techniques, combinatorial chemistry and other means to design candidate drugs, and offers a more directed alternative to merely screening random compounds with the hope that one might be suitable for a particular bacterium.

[0015] Nevertheless, even this preferred approach presents obstacles including the identification of essential genes and proteins, and the design of new assays for the genes thus identified in order to efficiently screen candidate compounds. Several groups have proposed systems for the identification of essential genes. For instance, Zyskind and colleagues propose a method of identifying essential genes in Escherichia coli by subcloning a library of E. coli nucleic acid sequences into an inducible expression vector, introducing the vectors into a population of E. coli cells, isolating those vectors that, upon activation and expression, negatively impact the growth of the E. coli cell, and characterizing the nucleic acid sequences and open reading frames contained on the subclones identified. See WO 00/44906, herein incorporated by reference. The disadvantage of this method is that the overexpression of nonessential genes can also negatively impact the cell, particularly the overexpression of membrane proteins and sugar transport proteins that are not necessary for growth where alternative carbon sources exist. Such proteins typically become trapped in membrane export systems when the cell is overloaded, and would be identified by this methodology. See Muller, FEMS Microbiol. Lett. 1999 Jul 1; 176 (1) : 219-27.

[0016] Another group proposes the identification of growth conditional mutants, and more specifically temperature sensitive (ts) mutants, as a means to identify essential genes in Staphylococcus aureus. See Benton et al. , U. S. Patent 6,037, 123, issued March 14,2000, herein incorporated by reference. Each gene is identified by isolating recombinant bacteria derived from growth conditional mutant strains, i. e. , following introduction of a vector containing a library of nucleic acid sequences, which would grow under non-permissive conditions but which were not revertants. These recombinant bacteria were found to contain DNA inserts that encoded wild type gene products that replaced the function of the mutated gene under non-permissive growth conditions. By this method, Benton and colleagues were able to identify 38 loci on the S aureus chromosome, each consisting of at least one essential gene.

[00171 The disadvantages of this method are first, the chemical employed to induce mutagenesis (diethyl sulfate, DES) is capable of causing several mutations in the same cell, thereby complicating interpretation of the results. Second, the method is particularly labor intensive in that one must painstakingly analyze replica plates of individual colonies grown at permissive and non-permissive temperatures, where replica plates include both mutant and non-mutant cells. Thus, employing the appropriate level of mutagen to achieve a balance between minimizing the number of non-mutant colonies one must screen in order to identify one mutant, while at the same time avoiding multiple mutations in the same cell, may be an arduous task.

[0018] Another group has proposed a transposon mutagenesis system for identifying essential genes called"GAMBIT" ("genomic analysis and mapping by in vitro transposition"), and has used the system to identify essential genes first in the gram positive bacteria Haemophilus influenzae and Streptococcuspneumoniae, and more recently in Pseudomonas aeruginosa. See Akerley et al., Systematic identification of essential genes by In vitro mariner mutagenesis, Proc. Natl. Acad. Sci USA 95 (15): 8927-32; Wong and Mekalanos, 2000, Proc. Natl. Acad. Sci. USA 97 (18): 10191-96 ; and Mekalanos et al. , U. S. Patent No. 6,207, 384, issued March 27, 2001, herein incorporated by reference. GAMBIT involves first isolating and purifying specific genomic segments of approximately 10 kilobases using extended-length PCR, and creating a high density transposon insertion map of the isolated region using Himarl transposon mutagenesis. The transposon insertions are then transferred to the chromosome following transformation of the bacteria with the transposon containing vectors, and selection for the antibiotic resistance marker on the transposon. The position of each transposon insertion with respect to a given PCR primer is then determined by genetic footprinting, i. e. , by amplifying sub-PCR products using one of the original PCR primers and a primer that recognizes an internal site in the Himarl transposon. By analyzing the length of PCR fragments thus identified, it is possible to identify regions that are devoid of transposon insertions, thereby signaling regions that might contain essential genes.

[0019] While the GAMBIT method is a good technique for looking at a small region of the genome for essential genes, it would be extremely labor intensive to use this method for analyzing the entire genome. This is particularly true for P. aeruginosa, whose genome (-6 megabases) is about 70% greater in size than the H. influenzae genome (-1. 8 megabases). Furthermore, GAMBIT would not be readily applicable to use in organisms that are less recombinogenic than H. influenzae. Indeed, while the H. influenzae genome contains about 1700 protein coding genes, P. aeruginosa contains about 5570. According to U. S. Patent 6,207, 384, one would need to clone and mutagenize the 6 million base pair genome of P. aeruginosa in 10,000 base pair fragments, isolating and characterizing 400-800 mutants per 10,000 base pair fragment.

Generating 6 X 105 mutants and characterizing them via PCR on gels would require a significant investment of labor, materials and time.

[0020] Another group at Abbott Laboratories has proposed a genome scanning method for identification of putative essential genes in H. influenzae, whereby random transposon insertions are mapped and analyzed to identify open reading frames containing no insertion in order to identify putative essential genes. Reich et al. , 1999, Genome Scanning in Haemophilus influenzae for Identification of Essential Genes, J.

Bacteriol. 181 (16): 4961-68. However, even though transposon insertions were isolated that spanned the whole genome, the authors employed a genomic footprinting technique similar to that used in GAMBIT to map insertions in a short contiguous region of the chromosome. The method further employs the methods of mutation exclusion and zero time analysis in order to monitor the fate of individual insertions after transformation in growing culture, which looks at individual insertions on a case- by-case basis. Again, such techniques would be extremely labor-intensive for the P. aeruginosa genome, which is 70% larger than the genome of H. influenzae.

[0021] Wong and Mekalanos also proposed identifying essential genes in P. aeruginosa by starting with the knowledge of three essential genes in H. influenzae and using genetic footprint analysis to determine if the homologues of these genes are essential in P. aeruginosa. Of three homologues tested, only one was unable to accommodate a transposon insertion. See Wong and Mekalanos, supra. Such results underscore the fact that a gene that is shown to be essential in one species will not necessarily be essential in another, given that some gene products may fulfill different functional roles in different species. Furthermore, given the larger coding capacity of the P. aeruginosa genome relative to that of other bacteria, it would not be surprising for P. aeruginosa to possess an increase in redundant gene functions, thereby decreasing the actual number of essential genes, and making them more difficult to identify.

10022] Another method is entitled Transposon Mediated Differential Hybridisation (TMDH), which is disclosed in WO 01/07651, herein incorporated by reference. This method entails (i) providing a library of transposon mutants of the target organism; (ii) isolating polynucleotide sequences from the library which flank inserted transposons; (iii) hybridising said polynucleotide sequences with a polynucleotide library from said organism; and (iv) identifying a polynucleotide in the polynucleotide library to which said polynucleotide sequences do not hybridise in order to identify an essential gene of the organism. However, the problem with this methodology is that it has a high propensity to lead to false positives, and many essential genes will be missed.

Furthermore, the method does not yield any detailed information regarding the loci disrupted by transposons, or whether they were hit more than once.

[0023] Thus, there is a great need for more efficient methods to identify essential genes, particularly in P. aeruginosa, so that new antibacterial agents may be designed therefrom for use in treatment of P. aeruginosa infections.

Summary of Invention [0024] The present inventors have devised a database of potential essential or otherwise important genes in P. aeruginosa, which may be used to verify essentiality and design antibacterial agents active against the targets thus identified. In particular, the inventors have isolated and mapped a library of at least about 5,000 to at least about 14,000 transposon insertions in the genome of P. aeruginosa, and more preferably a library of at least about 8000 to at least about 14,000 transposon insertions, and even more preferably a library of at least about 10,000 to at least about 14,000 transposon insertions, using the recently published P. aeruginosa gene sequence. The map thus generated was used to form a database of approximately 1500 to 3000 open reading frames, or more preferably about 1500 to 2000 open reading frames, for which no transposon insertions could be obtained, each of which possibly represents an essential gene required for growth and proliferation of P. aeruginosa on rich media, or an important gene, the mutation of which results in an attenuated growth mutant. Also disclosed is a Bayessian statistical model that may be utilized to increase the statistical confidence that any given gene identified using the disclosed methodology is essential.

[0025] Thus, one aspect of the invention is a database of putative essential or otherwise important genes, defined by the absence of transposon insertions in those genes in a High Throughput Transposon Insertion Map (HTTIM) database comprising about 10,000 to about 14,000 transposon insertions in the genome of Pseudomonas aeruginosa. Minimally, such a database comprises approximately 1800 open reading frames (ORFs), each of which may be further tested for essentiality using a variety of tests disclosed herein. However, predictions of essentiality or importance may be bolstered based on length of the ORF and predicted function and other statistical factors, thereby providing for more narrow databases of putative essential genes. Thus, the invention also includes databases that are more narrow and comprise only those genes for which essentiality or importance may be predicted with at least an 80% confidence level, and include at least about 850 to about 875 genes. The invention also includes databases assigned a confidence level of about 85% and including at least about 675 to about 700 genes. The invention further includes databases assigned a confidence level of about 90% including at least about 475 to about 500 genes.

Further, the invention includes databases assigned a confidence level of about 95% and including at least about 200 to 250 genes.

[0026] The transposon insertion map and database of putative essential or otherwise important open reading frames (ORFs) obtained may be used to confirm the essentiality or importance of genes, for example by integration knock outs in the presence of chromosomal complementation or by integration and activation of a regulatable promoter. An"essential"gene is one that cannot be"knocked out, "i. e. for which null mutants having complete absence of the gene product are not viable. This does not mean, however, that such genes could not tolerate point mutations or truncations that preserve sufficient gene product function so as to enable cell growth and survival.

Essential genes are to be distinguished from"important"genes, which are also included in the present invention, in that a"knock out"of an important gene does not lead to cell death but rather results in an attenuated growth mutant. Such genes may be included in the database of open reading frames not hit by random transposon mutagenesis as described herein, because attenuated growth colonies may be significantly smaller than the average P. aeruginosa colony and may have been overlooked when transposon insertion mutants were picked to generate the high throughput transposon insertion database (HTTIM).

[0027] Nevertheless, important gene products may interact with or regulate other genes, gene products or cellular processes that are essential, thereby making such gene products appropriate targets for drug design. Moreover, most drugs don't effectively kill all the pathogenic bacteria in the body ; rather, they kill or growth attenuate a portion of the bacteria, empowering the immune system to target the remainder.

Hence, important genes that, when targeted with an antibacterial agent, result in attenuated growth, are also targets for the antibacterial drugs of the present invention.

[0028] The invention also includes a database of attenuated growth mutants identified from the HTTIM transposon database. The genes marked by such mutations are of the same class of importance as the"important"genes identified in the no-hit database of genes, except that the growth attenuated nature of such transposon mutants was discovered at the transposon mutagenesis stage, rather than at the stage where essentiality is tested via targeted knock out. Thus, genes that when mutated confer attenuated growth may be identified from two sources: (1) from the library of open reading frames that did not receive a transposon insertion during HTTIM but were subsequently identified as an important gene when essentiality was tested via knock out and/or promoter swap strategies, and (2) from the HTTIM database itself when in the process of accumulating transposon insertion mutants it was observed that a particular insertion conferred an attenuated growth phenotype.

[0029] Such attenuated mutants grow more slowly than wild type, and may grow more slowly due to reduced expression of an essential gene, i. e. , transposon is in gene that regulates expression of an essential gene, or due to expression of a truncated form of an essential gene, i. e. , transposon is in the essential gene itself and leads to expression of a truncated mRNA. For example, mutants that show a higher drug susceptibility could be the result of insertions in a gene that potentiates resistance, such an efflux pump, or due to reduced expression of essential genes involved in the mechanism of action of the drug. Expression of mutated forms of essential and important genes may make the cell more susceptible to compounds that inhibit that particular gene or gene product, and may allow the identification of antibacterial agents with greater sensitivity. Furthermore, screening in whole cells overcomes the potential problems of uptake and efflux that are sometimes an issue for compounds identified via enzyme-based assays.

[0030] The essential and important genes of the invention may be used to design, screen for and evaluate potential antibacterial agents for the purpose of developing new treatments for P. aeruginosa infection. Antibacterial agents identified according to the invention may have activity against the gene or against the corresponding gene product or metabolic pathways requiring the gene product. For instance, antibacterial agents according to the invention may include antisense nucleic acids or regulatory proteins that bind to open reading frames, to upstream polar sequences or to promoters that drive expression of the genes encoded by such open reading frames. Active agents according to the invention may also include antibodies or proteins that bind to proteins encoded by open reading frames, or to transcriptional or translational regulators of such genes or proteins, or to binding partners of such proteins. Agents may also be chemical compounds designed following molecular modeling of essential gene products according to the invention, or mutant proteins designed therefrom that compete with the essential wild type protein for reactive cell components or for interacting nutrients, as well as agents from random chemical.

[0031] The present invention therefore includes methods and assays for identifying antibacterial agents having specificity for the essential or important open reading frames identified, or to genes and proteins that interact with such open reading frames or the products encoded thereby. Once essential and important open reading frames are identified, antibacterial agents may be identified using the assays and methods described herein, or by any suitable assay. Such assays may vary depending on the function delineated for each essential locus, as would be apparent to those of skill in the art. For instance, enzyme assays may be designed based on the predicted function of essential and important genes in order to define classes of inhibitors to be tested. Also, random chemical libraries may be screened for activity against the isolated genes or gene products. Cell lines may be designed or isolated that demonstrate reduced expression of essential genes, thereby providing a sensitive screening tool for inhibitors that effect the activity of that gene or gene product as it functions in the cell. Such cell lines may be devised from cells having transposon insertions that lead to attenuated growth, or may be constructed by the promoter swap techniques described herein, by using a regulatable promoter that can be used to increase gene expression, allowing for confirmation of target specificity. Here, the minimal inhibitory concentration of the inhibitor is directly related to the expression level of the target gene, such that under low expression, an attenuated growth cell is more susceptible to an inhibitor than the wild type strain, and as you raise the expression level, the minimum inhibitory concentration (MIC) increases. The MIC shift will be consistent when the inhibitor acts on the regulated target.

[0032] Active agents and compounds can be formulated into pharmaceutical compounds and compositions, effective for treating and preventing Pseudomonas infections in accordance with the methods of the invention. Such therapy will be particularly useful in the hospital setting for preventing and treating nosocomial infections, and for administering to cystic fibrosis patients to improve lung function and quality of life. Depending on the activity of the essential or important gene targeted, such agents could also be useful in treating all types of Pseudomonas infections ranging from bacteraemia and septicemia, urinary-tract infections, pneumonia and chronic lung infections, burn infections, cancer, AIDS, endocarditis, dermatitis, osteochondritis, ear and eye infections, bone and joint infections, gastrointestinal infections and skin and soft tissue infections, including wound infections, pyoderma and dermatitis. Further, the invention provides pharmaceutical compositions appropriate for use in methods of treating bacterial infections described above.

Brief Description of the Drawings [0033] Figure 1. Depiction of a single crossover recombination event resulting in integration of a plasmid into the bacterial chromosome. Isolation of such recombinants indicates that the targeted gene is not essential.

[0034] Figure 2. Single crossover and integration of a plasmid resulting in the replacement of a wild type promoter with a regulatable promoter.

[0035] Figure 3. Depiction of the'promoter swap'strategy, using transformation of pBEMlO into P. aeruginosa in order to replace the IpxC promoter with the arabinose araBAD promoter, thereby allowing modulation of its lpxC expression by the use of a simple sugar, arabinose.

[0036] Figure 4. Graph showing the susceptibility or non-susceptibility of various E. coli and P. aeruginosa strains to the inhibitor L161, 240.

[0037] Figure 5. Graph depicting the effect of tetracycline and L161, 240 on the growth of P. aeruginosa strain PA01 with and without polymixin permeabilization.

[0038] Figure 6. Sensitivity of various E. coli and P. aeruginosa strains to inhibitor L161, 240 following promoter swap and transformation with vector expressing E. coli LpxC or P. aeruginosa lpxC. E. coli"swaps"refer to P. aeruginosa containing a vector comprising E. coli lpxC, and"PA swaps"refer to P. aeruginosa containing a vector comprising P. aeruginosa lpxC.

[0039] Figure 7. Graph illustrating ORF coverage by Tn5 achieved in High- Throughput Transposon Insertion Mapping (HTTIM), wherein 30% of the genes in the genome are candidate essential genes where ORF size is not taken into account in predicting essentiality.

[0040] Figure 8. Graph depicting the probability of identifying an essential gene given no transposon insertion, as a function of gene size.

[0041] Figure 9. A circular map of the P. aeruginosa genome showing distribution of transposon insertion sites constituting a HTTIM of the invention, and demonstrating the random nature of the transposon employed. The length of the bars radiating outward from the center of the circular map reflect the number of transposon insertions per non- overlapping kilobase.

[0042] Figure 10. Histogram depicting the number of ORFs in the P. aeruginosa genome of (a) up to 4000 base pairs and (b) from 4000 up to 16884 base pairs.

[0043] Figure 11. Graph showing likelihood and accumulative likelihood gains.

[0044] Figure 12. Trajectory of the algorithm projected in a subspace spanned by two gene sizes. The x-axis represents genes of sizes 151-160 DNA base-pairs and y-axis represents genes of sizes 171-180 DNA base-pairs. Here n= (2, 1) and M= (7,9). The median gene size of each group is used as the gene size. At iteration number 66, the likelihood gain is maximum in the direction of increasing the number of nonessential genes by one for genes with size 171-180 DNA base-pairs. At iteration number 443, the largest likelihood gain is obtained in the direction of increasing one nonessential genes for genes of sizes 151-160 DNA base pair. At any point, moving backwards has a negative likelihood gain.

[0045] Figure 13. More trajectories of the searching algorithm projected in different subspaces.

[0046] Figure 14. Plot of likelihood for different initial values.

[0047] Figure 15. Trajectories of the algorithm with different starting values projected in the subspace spanned by two gene sizes: 1101-1150 DNA base-pairs for X-axis and 921-930 DNA base-pairs for y-axis.

[0048] Figure 16. Top: (A) The top line is M, number of genes, the bottom line is ñ, the number of genes with at least one observed insertion; the line in the middle is N, the number of estimated nonessential genes. For demonstration purpose, a cubic spline smooth is applied to the data. Bottom: Histogram of resamples of r (B) and (C).

[0049] Figure 17. Plot of RIM,. The doted line is the value of lv ; M ; and the solid line is a moving average smooth.

[0050] The essential and important open reading frames identified in the present invention were originally part of a library of putative nucleic acid sequences generated from P. aeruginosa strains PA01 and PAK. See Table 1. Nevertheless, it is expected that the genes identified will also be essential or important in related P. aeruginosa strains as well as other Pseudomonas species, given the low sequence diversity that exists between P. aeruginosa strains of widely diverse environments and the pronounced structural and functional homology of gene products. See, e. g., Spangenberg et al. , 1998, Structural and functional implications of sequence diversity of Pseudomonas aeruginosa genes oriT, ampC andfliC, Electrophoresis 19 (4): 545-50; Ruimy et al., 2001, Genetic diversity of Pseudomonas aeruginosa strains isolated from ventilated patients with nosocomial pneumonia, cancer patients, bacteremia, and environmental water, Infect. Immun. 69 (1) : 584-8. For instance, comparative sequencing of several P. aeruginosa genes from several environmental and clinical isolates revealed the sequence diversity to be about one order of magnitude lower than in comparable housekeeping genes from Salmonella. See Kiewitz and Tummler, 2000, Sequence diversity of Pseudomonas aeruginosa : impact on population structure and genome evolution, J. Bacteriol. 182 (11): 3125-35. Thus, it is expected that agents identified as antibacterial based on their interaction with genes or gene products of P. aeruginosa PA01 or PAK will be broadly applicable as antibacterial agents against a variety of Pseudomonas species as well as other bacteria including but not limited to Escherichia, Hemophilus, Vibrio, Borrelia, Enterococcus, Heliobacter, Legionella, Mycobacterium, Mycoplasma, Neisseria, Staphylococcus, Streptococcus, etc.

[0051] Thus, the present invention encompasses an isolated nucleic acid molecule comprising a nucleic acid sequence encoding a polypeptide having at least 80% sequence identity to a polypeptide encoded by a nucleic acid sequence selected from the group consisting of the Pseudomonas aeruginosa open reading frames (ORFs) listed in Table 1. More preferably, the present invention encompasses an isolated nucleic acid molecule comprising a nucleic acid sequence encoding a polypeptide having at least about 85 to 90% sequence identity to a polypeptide encoded by a nucleic acid sequence selected from the group consisting of the Pseudomonas aeruginosa open reading frames (ORFs) listed in Table 1. Even more preferably, the present invention encompasses an isolated nucleic acid molecule comprising a nucleic acid sequence encoding a polypeptide having at least about 90 to about 95% sequence identity to a polypeptide encoded by a nucleic acid sequence selected from the group consisting of the Pseudomonas aeruginosa open reading frames (ORFs) listed in Table 1.

[0052] In particular, the invention encompasses isolated nucleic acid molecules comprising nucleic acid sequences encoding polypeptides having at least 80% sequence identity, or more preferably at least about 85 to 90 to 95% identity, to a polypeptide encoded by an essential or important nucleic acid sequence selected from the group consisting of the Pseudomonas aeruginosa open reading frames (ORFs) listed in Table 1, wherein essentiality or importance of said nucleic acid sequence is determined by integration knock-out coupled with extra-chromosomal complementation. Likewise, the invention encompasses isolated nucleic acid molecules comprising nucleic acid sequences encoding polypeptides having at least 80% sequence identity, or more preferably at least about 85 to 90 to 95% identity, to a polypeptide encoded by an essential nucleic acid sequence selected from the group consisting of the Pseudomonas aeruginosa open reading frames (ORFs) listed in Table 1, wherein essentiality or importance of said nucleic acid sequence is determined by integration of a regulatable promoter into the gene, or via any other suitable method.

[0053] In one embodiment, the polynucleotides of the invention are recombinant.

Recombinant polynucleotides of the invention include proteins of genomic, cDNA, semisynthetic, or synthetic origin, which, by virtue of its origin or manipulation (1) is not associated with all or a portion of a polynucleotide with which it is associated in nature; (2) is linked to a polynucleotide other than that to which it is linked in nature; or (3) does not occur in nature.

[0054] Given that the library of nucleic acid sequences encompassed in Table 1 provides an unprecedented tool useful for the identification of essential and otherwise important genes in Pseudonsonas and the construction and isolation of attentuated mutants, the present invention includes a library of nucleic acid sequences consisting essentially of nucleic acid sequences having at least 70% sequence identity, or more preferably at least about 80 to 90 to 95% identity, to a nucleic acid sequence selected from the group consisting of the Pseudomonas aeruginosa open reading frames (ORFs) listed in Table 1, wherein said library of nucleic acid sequences is employed to identify essential or otherwise important genes or to construct or isolate attenuated mutants in Pseudomonas.

[0055] Also encompassed in the invention is a map of at least about 10,000 to about 14,000 transposon insertions in the genome of Pseudomonas aeruginosa (High- Throughput Transposon Insertion Database or HTTIM), wherein said map is useful for identifying genes that are essential or important for survival of said Pseudomonas aeruginosa, i. e. , by permitting the generation of a database of open reading frames that do not contain a transposon insertion. Figure 9 contains a circular map of the P. aeruginosa genome depicting 12,000 to 13,000 transposon insertion sites constituting a HTTIM of the invention, and demonstrates the random nature of the transposon employed. The length of the bars radiating outward from the center of the circular map reflect the number of transposon insertions per non-overlapping kilobase. Table 3 contains a list of 13,515 specific Tn5 transposon insertion sites generated in either PAK or PA01, with the 473 mutants 12516-13043 being identified as attenuated for growth.

[0056] Thus, the databases and libraries disclosed herein may be used to formulate useful subsets of these libraries and databases. Accordingly, the invention includes subsets of the databases and libraries disclosed. For instance, mutants 12516-13043 are identified as attenuated for growth and as such, the genes in this subset could be useful drug targets. Accordingly, this group of 473 mutants from the HTTIM database of 13,515 transposon hits provides a useful subset database for comparing homologies with essential genes of other organisms, for computer modeling of potential antibacterial agents, etc. A particularly useful database subset is one containing essential genes from P. aeruginosa that are also identified as essential in other Gram negative or Gram positive bacteria. Indeed, genes that have essential homologs in other bugs are likely to provide useful targets for broad spectrum antibacterial agents, i. e., agents that have broad spectrum activity as an antibacterial agent. Genes in the putative essential or important gene database have already been identified via BLAST or other database analyses, and constitute an exemplary subset database of the present invention. See Table 4.

[0057] Further, the databases and subset databases of the present invention may also be used as comparative tools with other like databases or database subsets to identify broad spectrum. For instance, particularly envisioned is an embodiment wherein the database of putative essential and important genes identified in P. aeruginosa is cross- referenced with a similar database formed from S. aureus, wherein homologues present in both databases signal a potential target for a broad spectrum antibacterial agent.

Cross-referencing between P. åeruginosa and S. aureus in particular will identify antibacterial targets for identifying broad spectrum antibiotics active against both Gram negative and Gram positive bacteria. However, databases derived from any bacteria could be employed in such comparisons, as well as databases formed from yeast, fungi, mycoplasma, and other potential pathogens.

[0058] Also encompassed in the invention is the use of essential and important genes and the corresponding proteins expressed thereto in the design of vaccines for eliciting prophylactic or therapeutic immune responses against Pseudomonas aeruginosa.

[0059] Such vaccines will typically comprise a Pseudomonas aeruginosa protein antigen or fragment or variant thereof encoded by an essential gene. Additionally, such antigens will preferably be a protein expressed on the surface of the bacteria.

[0060] Such vaccines will typically comprise a Pseudomonas aeruginosa protein antigen or fragment or derivative thereof encoded by an essential or important gene.

Preferably, the protein antigen expressed from a recombinant polynucleotide.

[0061] Where the invention is directed to a fragment of a protein encoded by an essential or important gene, said fragment is preferably at least 8 to 12 amino acids long, and even more preferably at least about 20 to 30 amino acids long. Preferably, the fragment comprises either a B cell or a T cell epitope.

[0062] Where the invention is directed to a derivative of a protein encoded by an essential or important gene, said derivative contains one or more amino acid substitutions, additions or deletions. Preferably, the amino acid substitutions are conservative amino acid replacements. Conservative amino acid replacements are those that take place within a family of amino acids that are related in their side chains.

Genetically encoded amino acids are generally divided into four families: (1) acidic = aspartate, glutamate; (2) basic = lysine, arginine, histidine ; (3) non-polar = alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan; and (4) uncharged polar = glycine, asparagine, glutamine, cystine, serine, threonine, tyrosine.

Phenylalanine, tryptophan, and tyrosine are sometimes classified jointly as aromatic amino acids. For example, it is reasonably predictable that an isolated replacement of a leucine with an isoleucine or valine, an asparate with glutamate, a threonine with a serine, or a similar conservative replacement of an amino acid with a structurally related amino acid will not have a major effect on the biological activity. Polypeptide molecules having substantially the same amino acid sequence as the protein by possessing minor amino acid substitutions that do not substantially affect the functional aspects are encompassed with the scope of derivatives of the proteins of the invention.

[0063] The polypeptide fragment or derivative is preferably immunologically identifiable with the polypeptide encoded by the essential or important gene. The polypeptide fragment or derivative is preferably immunogenic and is able to cause a humoral and/or cellular immune response, either alone or when linked to a carrier, in the presence or absence of an adjuvant. The polypeptide fragment or derivative may be fused to or incorporated into another polypeptide sequence. This other polypeptide sequence may include one or more other proteins, fragments or derivatives thereof encoded by an essential or important gene. The other polypeptide sequence may also include a polypeptide sequence which allows for presentation of the polypeptide fragment or derivative.

[0064] Accordingly, the present invention encompasses an isolated polypeptide and fragments and derivatives thereof, wherein said polypeptide has at least 80% sequence identity to a polypeptide encoded by a nucleic acid sequence selected from the group consisting of the Pseudomonas aeruginosa open reading frames (ORFs) listed in Table 1. More preferably, the present invention encompasses an isolated polypeptide and fragments and derivatives thereof, wherein said polypeptide has at least about 85 to 90% sequence identity to a polypeptide encoded by a nucleic acid sequence selected from the group consisting of the Pseudomonas aeruginosa open reading frames (ORFs) listed in Table 1. Even more preferably, the present invention encompasses an isolated polypeptide and fragments and derivatives thereof, wherein said polypeptide has at least about 90% to about 95% sequence identity to a polypeptide encoded by a nucleic acid sequence selected from the group consisting of the Pseudomonas aeruginosa open reading frames (ORFs) listed in Table 1.

[0065] In particular, the invention encompasses isolated polypeptides and fragments and derivatives thereof, wherein said polypeptides have at least 80% sequence identity, or more preferably at least about 85 to 90 to 95% identity, to a polypeptide encoded by an essential or important nucleic acid sequence selected from the group consisting of the Pseudomonas aeruginosa open reading frames (ORFs) listed in Table 1, wherein the essentiality or importance of said nucleic acid sequence is determined by integration knock-out couple with extra-chromosomal complementation. Likewise, the invention encompasses isolated polypeptides and fragments and derivatives thereof, wherein said polypeptides have at least 80% sequence identify, or more preferably at least about 85 to 90 to 95% identity, to a polypeptide encoded by an essential nucleic acid sequence selected from the group consisting of the Pseudomonas aeruginosa open reading frames (ORFs) listed in Table 1, wherein essentiality or importance of said nucleic acid sequence is determined by integration of a regulatable promoter into the gene, or via any other suitable method.

[0066] Also encompassed in the invention are therapeutic and prophylactic vaccines that comprise ligands that specifically bind antigens encoded by essential or important genes identified according to the invention, for use in, for instance, passive immunization. Preferred ligands are antibodies and antibody fragments that specifically bind the antigen encoded by the essential gene. Such antibodies may be polyclonal or monoclonal. Types of antibodies and antibody fragments include by way of examples murine antibodies, chimeric, antibodies, humanized antibodies, Fab fragments, Fab2 fragments and human antibodies and scFv's. Methods for producing antibodies and antibody fragments by recombinant and non-recombinant methods are well known to those skilled in the art. In some embodiments the antigen used in such passive immunization may be attached to a cytotoxic moiety, e. g. , a radionuclide or other agent that is cytotoxic against the bacteria.

[0067] Further encompassed within the scope of the invention are cells or viral vectors that express on their surface a Pseudomonas aeruginosa essential gene, fragment or variant identified according to the invention.

[0068] In the case of prophylactic vaccines, the vaccine will comprise an immunogenic composition comprising a prophylactically effective amount of an antigen, antibody, cells or vector expressing an antigen encoded by an essential or important gene and will be formulated such that upon administration it elicits a protective immune response. In the case of therapeutic vaccines, the vaccine will comprise an immunogenic composition comprising a therapeutically effective amount of an antigen, antibody, cells or vectors expressing an antigen encoded by an essential or important gene and will be formulated such that upon administration it elicits a therapeutic immune response. Dosage effective amounts of prophylactic and therapeutic vaccines will be determined by known methods and will typically vary from about 0.00001 g/kg body weight to about 5-10 g/kg body weight.

[0069] The immunogenic compositions of the invention can be administered by known methods, i. e. , mucosally or parenterally.

[0070] Suitable routes of mucosal administration include oral, intranasal (IN), intragastric, pulmonary, intestinal, rectal, ocular, and vaginal routes. Preferably, mucosal administration is oral or intranasal.

[0071] Where mucosal administration is used, the immunogenic composition is preferably adapted for mucosal administration. For instance, where the composition is administered orally, it may be in the form of tablets or capsules (optionally enteric- coated), liquid, transgenic plants, etc. Where the composition is administered intranasally, it may be in the form of a nasal spray, nasal drops, gel or powder. Where the antigen composition is adapted for mucosal administration, it may further be formulated such that the antigen remains stable, for instance by the use of carriers and excipients.

[0072] The immunogenic compositions of the invention can further comprise a mucosal adjuvant. Mucosal adjuvants suitable for use in the invention include (a) E. coli heat-labile enterotoxin ("LT"), or detoxified mutants thereof, such as the K63 or R72 mutants; (B) cholera toxin ("CT"), or detoxified mutants thereof; or (C) microparticles (i. e., aparticle of slOOnm to ~15011m in diameter, more preferably ~200nm to ~3011m in diameter, and most preferably ~500nm to ~1011m in diameter) formed from materials that are biodegradable and non-toxic (e. g. a poly (a-hydroxy acid), a polyhydroxybutyric acid, a polyorthoester, a polyanhydride, a polycaprolactone etc.) ; (D) a polyoxyethylene ether or a polyoxyethylene ester (see International patent application WO 99/52549); (E) a polyoxyethylene sorbitan ester surfactant in combination with an octoxynol (see International patent application WO 01/21207) or a polyoxyethylene alkyl ether or ester surfactant in combination with at least one additional non-ionic surfactant such as an octoxynol (see International patent application WO 01/21152) ; (F) chitosan (e. g. International patent application WO 99/27960) and (G) an immunostimulatory oligonucleotide (e. g. a CpG oligonucleotide) and a saponin (see International patent application WO 00/62800). Other mucosal adjuvants are also available (e. g. see chapter 7 of Vaccine design: the subunit and adjuvant aproach, eds. Powell & Newman, Plenum Press 1995 (ISBN 0-306-44867-X).

[0073] Mutants of LT are preferred mucosal adjuvants, in particular the"K63"and "R72"mutants (e. g. see International patent application WO 98/18928), as these result in an enhanced immune response.

[0073] Microparticles are also preferred mucosal adjuvants. These are preferably derived from a poly (a-hydroxy acid), in particular, from a poly (lactide) ("PLA"), a copolymer of D, L-lactide and glycolide or glycolic acid, such as a poly (D, L-lactide-co- glycolide) ("PLG"or"PLGA"), or a copolymer of D, L-lactide and caprolactone. The microparticles may be derived from any of various polymeric starting materials which have a variety of molecular weights and, in the case of the copolymers such as PLG, a variety of lactide: glycolide ratios, the selection of which will be largely a matter of choice, depending in part on the coadministered antigen.

[0074] Antigen may be entrapped within the microparticles, or may be adsorbed to them.

[0075] Entrapment within PLG microparticles is preferred. PLG microparticles are discussed in further detail in Morris et al. , (1994), Vaccine, 12: 5-11, in chapter 13 of Mucosal Vaccines, eds. Kiyono et al., Academic Press 1996 (ISBN 012410587), and in chapters 16 & 18 of Vaccine design : the sgbunit and adjuvant aproach, eds. Powell & Newman, Plenum Press 1995 (ISBN 0-306-44867-X).

[0076] LT mutants may advantageously be used in combination with microparticle- entrapped antigen, resulting in significantly enhanced immune responses.

[0077] Suitable routes of parenteral administration include intramuscular (IM), subcutaneous, intravenous, intraperitoneal, intradermal, transcutaneous, and transdermal (see e. g., International patent application WO 98/20734) routes, as well as delivery to the interstitial space of a tissue.

[0078] The immunogenic compositions of the invention may be adapted for parenteral administration (e. g. , in the form of an injectable, which will typically be sterile and pyrogen-free).

[0079] The immunogenic composition may further comprise a parenteral adjuvant.

Parenteral adjuvants suitable for use in the invention include: (A) aluminum compounds (e. g. aluminum hydroxide, aluminum phosphate, aluminum hydroxyphosphate, oxyhydroxide, orthophosphate, sulfate etc. (e. g. see chapters 8 & 9 of Vaccine design : the subunit and adjuvant aproach, eds. Powell & Newman, Plenum Press 1995 (ISBN 0-306-44867-X) (hereinafter"Yaccine design"), or mixtures of different aluminum compounds, with the compounds taking any suitable form (e. g. gel, crystalline, amorphous etc.), and with adsorption being preferred; (B) MF59 (5% Squalene, 0. 5% Tween 80, and 0. 5% Span 85, formulated into submicron particles using a microfluidizer) (see Chapter 10 of Vaccine design ; see also International patent application WO 90/14837); (C) liposomes (see Chapters 13 and 14 of Vaccine design), (D) ISCOMs (see Chapter 23 of Vaccine design) ; (E) SAF, containing 10% Squalane, 0.4% Tween 80, 5% pluronic-block polymer L121, and thr-MDP, either microfluidized into a submicron emulsion or vortexed to generate a larger particle size emulsion (see Chapter 12 of Vaccine design) ; (F) RibiTM adjuvant system (RAS), (Ribi Immunochem) containing 2% Squalene, 0.2% Tween 80, and one or more bacterial cell wall components from the group consisting of monophosphorylipid A (MPL), trehalose dimycolate (TDM), and cell wall skeleton (CWS), preferably MPL + CWS (DetoxTM) ; (G) saponin adjuvants, such as QuilA or QS21 (see Chapter 22 of Vaccine design), also known as Stimulodrm ; (H) ISCOMs, which may be devoid of additional detergent (International patent application WO 00/07621); (1) complete Freund's adjuvant (CFA) and incomplete Freund's adjuvant (IFA); (J) cytokines, such as interleukins (e. g. IL-1, IL-2, IL-4, IL-5, IL-6, IL-7, IL-12, etc.), interferons (e. g. interferon-y), macrophage colony stimulating factor, tumor necrosis factor, etc. (see Chapters 27 & 28 of Vaccine design) ; (K) microparticles (see above); (L) monophosphoryl lipid A (MPL) or 3-O- deacylated MPL (3dMPL) (e. g. chapter 21 of Vaccine design) ; (M) combinations of 3dMPL with, for example, QS21 and/or oil-in-water emulsions (European patent applications 0835318,0735898 and 0761231); (N) oligonucleotides comprising CpG motifs (see Krieg (2000) Vaccine, 19: 618-622; Krieg (2001) Curr. Opin. Mol. Ther., 2001,3 : 15-24; WO 96/02555, WO 98/16247, WO 98/18810, WO 98/40100, WO 98/55495, WO 98/37919 and WO 98/52581, etc. ) i. e. containing at least one CG dinucleotide, with 5-methylcytosine optionally being used in place of cytosine ; (O) a polyoxyethylene ether or a polyoxyethylene ester (International patent application WO 99/52549); (P) a polyoxyethylene sorbitan ester surfactant in combination with an octoxynol (International patent application WO 01/21207) or a polyoxyethylene alkyl ether or ester surfactant in combination with at least one additional non-ionic surfactant such as an octoxynol (International patent application WO 01/21152) ; (Q) an immunostimulatory oligonucleotide (e. g. a CpG oligonucleotide) and a saponin (International patent application WO 00/62800); (R) an immunostimulant and a particle of metal salt (International patent application WO 00/23105); (S) a saponin and an oil- in-water emulsion (International patent application WO 99/11241); (T) a saponin (e. g.

QS21) + 3dMPL + IL-12 (optionally + a sterol) (International patent application WO 98/57659); and (U) other substances that act as immunostimulating agents to enhance the effectiveness of the composition (e. g. see Chapter 7 of Vaccine design).

[0080] Aluminium compounds and MF59 are preferred adjuvants for parenteral use.

[0081] The immunognic compositions of the invention may be administered in a single dose, or as part of an administration regime. The regime may include priming and boosting doses, which may be administered mucosally, parenterally, or various combinations thereof.

[0082] In some instances the vaccines of the invention may comprise several antigens, fragments or variants encoded by essential genes identified according to the invention. Alternatively, the vaccine may further comprise antigens identified by other methods, or specific to other bacteria, e. g. , in order to provide multivalent vaccines.

[0083] With respect to libraries according to the invention, a library of polynucleotides or a library of transposon insertion sites is a collection of sequence information, which information is provided in either biochemical form (e. g. , as a collection of polynucleotide molecules), or in electronic form (e. g. , as a collection of polynucleotide sequences stored in a computer-readable form, as in a computer system and/or as part of a computer program). The sequence information of the polynucleotides can be used in a variety of ways, for instance as a resource for gene discovery, i. e. , for identifying and verifying essential and important genes in P. aeruginosa, or for identifying essential or important homologues in other genera or species. A polynucleotide sequence in a library can be a polynucleotide that represents an mRNA, polypeptide, or other gene product encoded by the polynucleotide, and accordingly such a polynucleotide library could be used to formulate corresponding RNA or amino acid libraries according to the sequences of the library members.

[0084] The nucleotide sequence information of the library can be embodied in any suitable form, e. g. , electronic or biochemical forms. For example, a library of sequence information embodied in electronic form comprises an accessible computer data file (or, in biochemical form, a collection of nucleic acid molecules) that contains the representative nucleotide sequences of essential and important genes and/or insertion mutants that are differentially expressed (e. g., attenuated growth mutants). Other combinations and comparisons of cells affected by various diseases or stages of disease will be readily apparent to the ordinarily skilled artisan. Biochemical embodiments of the library include a collection of nucleic acids that have the sequences of the genes or transposon insertion sites in the library, where the nucleic acids can correspond to the entire gene in the library or to a fragment thereof, as described in greater detail below.

[0085] The polynucleotide libraries of the subject invention generally comprise sequence information of a plurality of polynucleotide sequences, where at least one of the polynucleotides has a sequence of any of the sequences in Tables 1-3. By plurality is meant at least 2, usually at least 3 and can include up to all of the sequences included in these tables. The length and number of polynucleotides in the library will vary with the nature of the library, e. g., if the library is an oligonucleotide array, a cDNA array, a computer database of the sequence information, etc.

[0086] Where the library is an electronic library, the nucleic acid sequence information can be present in a variety of media. "Media"refers to a manufacture, other than an isolated nucleic acid molecule, that contains the sequence information of the present invention. Such a manufacture provides the genome sequence or a subset thereof in a form that can be examined by means not directly applicable to the sequence as it exists in a nucleic acid. For example, the nucleotide sequence of the present invention, e. g. the nucleic acid sequences of any of the polynucleotides of Tables 1-3, can be recorded on computer readable media, e. g. any medium that can be read and accessed directly by a computer. Such media include, but are not limited to: magnetic storage media, such as a floppy disc, a hard disc storage medium, and a magnetic tape; optical storage media such as CD-ROM; electrical storage media such as RAM and ROM; and hybrids of these categories such as magnetic/optical storage media. One of skill in the art can readily appreciate how any of the presently known computer readable mediums can be used to create a manufacture comprising a recording of the present sequence information. "Recorded"refers to a process for storing information on computer readable medium, using any such methods as known in the art. Any convenient data storage structure can be chosen, based on the means used to access the stored information. A variety of data processor programs and formats can be used for storage, e. g. word processing text file, database format, etc. In addition to the sequence information, electronic versions of the libraries of the invention can be provided in conjunction or connection with other computer-readable information and/or other types of computer-readable files (e. g., searchable files, executable files, etc, including, but not87 limited to, for example, search program software, etc.).

[0087] By providing the nucleotide sequence in computer readable form, the information can be accessed for a variety of purposes. Computer software to access sequence information is publicly available. For example, the gapped BLAST (Altschul et al. Nucleic Acids Res. (1997) 25 : 3389-3402) and BLAZE (Brutlag et al. Comp.

Chem. (1993) 17: 203) search algorithms on a Sybase system can be used to identify open reading frames (ORFs) within the genome that contain homology to ORFs from other organisms.

[0088] As used herein, "a computer-based system"refers to the hardware means, software means, and data storage means used to analyze the nucleotide sequence information of the present invention. The minimum hardware of the computer-based systems of the present invention comprises a central processing unit (CPU), input means, output means, and data storage means. A skilled artisan can readily appreciate that any one of the currently available computer-based system are suitable for use in the present invention. The data storage means can comprise any manufacture comprising a recording of the present sequence information as described above, or a memory access means that can access such a manufacture.

[0089]"Search means"refers to one or more programs implemented on the computer-based system, to compare a target sequence or target structural motif, or expression levels of a polynucleotide in a sample, with the stored sequence information.

Search means can be used to identify fragments or regions of the genome that match a particular target sequence or target motif. A variety of known algorithms are publicly known and commercially available, e. g. MacPattern (EMBL), BLASTN and BLASTX (NCBI). A"target sequence"can be any polynucleotide or amino acid sequence of six or more contiguous nucleotides or two or more amino acids, preferably from about 10 to 100 amino acids or from about 30 to 300 nucleotides. A variety of comparing means can be used to accomplish comparison of sequence information from a sample (e. g., to analyze target sequences, target motifs, or relative expression levels) with the data storage means. A skilled artisan can readily recognize that any one of the publicly available homology search programs can be used as the search means for the computer based systems of the present invention to accomplish comparison of target sequences and motifs. Computer programs to analyze expression levels in a sample and in controls are also known in the art.

[0090] A"target structural motif, "or"target motif, "refers to any rationally selected sequence or combination of sequences in which the sequence (s) are chosen based on a three-dimensional configuration that is formed upon the folding of the target motif, or on consensus sequences of regulatory or active sites. There are a variety of target motifs known in the art. Protein target motifs include, but arc not limited to, enzyme active sites and signal sequences. Nucleic acid target motifs include, but are not limited to, hairpin structures, promoter sequences and other expression elements such as binding sites for transcription factors.

[0091] The present invention encompasses the use of the library of essential and important genes to search for polynucleotide and amino acid sequences in common among the essential and important genes. Such identified sequences can be used to design and develop antibacterial agents and vaccines against Pseudomonas aeruginosa.

[0092] A variety of structural formats for the input and output means can be used to input and output the information in the computer-based systems of the present invention. One format for an output means ranks the relative expression levels of different polynucleotides. Such presentation provides a skilled artisan with a ranking of relative expression levels to determine a gene expression profile.

[0093] As discussed above, the"library"as used herein also encompasses biochemical libraries of the polynucleotides of Tables 1-3, e. g., collections of nucleic acids representing the provided polynucleotides. The biochemical libraries can take a variety of forms, e. g., a solution of cDNAs, a pattern of probe nucleic acids stably associated with a surface of a solid support (i. e., an array) and the like. Of particular interest are nucleic acid arrays in which one or more of the sequences of Tables 1-3 is represented on the array. By"array"is meant a an article of manufacture that has at least a substrate with at least two distinct nucleic acid targets on one of its surfaces, where the number of distinct nucleic acids can be considerably higher, typically being at least 10 nt, usually at least 20 nt and often at least 25 nt. A variety of different array formats have been developed and are known to those of skill in the art. The arrays of the subject invention find use in a variety of applications, including gene expression analysis, drug screening, mutation analysis and the like, as disclosed in the above-listed exemplary patent documents.

[0094] In addition to the above nucleic acid libraries, analogous libraries of polypeptides are also provided, where the polypeptides of the library will represent at least a portion of the polypeptides encoded by a gene corresponding to one or more of the sequences in Tables 1-3.

[0095]"Identity"as it is used in the present invention should be distinguished from "homology"or"homologous. "In the context of the coding sequences and genes of this invention, "homologous"refers to genes whose expression results in expression products which have a combination of amino acid sequence similarity (or base sequence similarity for transcript products) and functional equivalence, and are therefore homologous genes. In general such genes also have a high level of DNA sequence similarity (i. e. , greater than 80% identity when such sequences are identified among members of the same genus, but lower when these similarities are noted across bacterial genera), but are not identical. Relationships across bacterial genera between homologous genes are more easily identified at the polypeptide (i. e. , the gene product) rather than the DNA level. The combination of functional equivalence and sequence similarity means that if one gene is useful, e. g. , as a target for an antibacterial agent, or for screening for such agents, then the homologous gene is probably also useful, but may not react in the same manner or to the same degree to the activity of a specific antibacterial agent.

[0096] Nevertheless, the identification of one such gene serves to identify a homologous gene through the same relationships as indicated above, and can serve as a starting point to determine whether the homologous gene is also essential, whether it responds to the same antibacterial agents, etc. Typically, such homologous genes are found in other bacterial species, especially, but not restricted to, closely related species.

Due to the DNA sequence similarity, homologous genes are often identified by hybridizing with probes from the initially identified gene under hybridizing conditions that allow stable binding under appropriately stringent conditions. For instance, nucleic acids having sequence similarity are detected by hybridization under low stringency conditions, for example, at 50°C and 10XSSC (0.9 M saline/0.09 M sodium citrate) and remain bound when subjected to washing at 55°C in 1XSSC. Sequence identity can be determined by hybridization under stringent conditions, for example, at 50°C or higher and 0. 1XSSC (9 mM saline/0.9 mM sodium citrate). Hybridization methods and conditions are well known in the art, see, e. g., USPN 5,707, 829. Nucleic acids that are substantially identical to the provided polynucleotide sequences, e. g. allelic variants, genetically altered versions of the gene, etc., bind to the provided polynucleotide sequences under stringent hybridization conditions. By using probes, particularly labeled probes of DNA sequences, one can isolate homologous or related or substantially identical genes. The equivalent function of the product is then verified using appropriate biological and/or biochemical assays.

[0097] Using such hybridization technique for the identification of homologous genes, it will be possible to screen other species of bacteria, particularly other genera of gram negative pathogenic bacteria although gram positive bacteria may also be screened, to determine if any essential or important gene identified herein has a homologue in that particular genus of bacteria. If so, such gene could be cloned and isolated for essentiality in the particular genus, and further tested for sensitivity or susceptibility to the antibacterial agents and inhibitors identified herein. Specific genera of bacteria particularly appropriate for hybridization screening for the presence of homologues of essential and important genes include Escherichia, Hemophilus, Vibrio, Borrelia, Enterococcus, Heliobacter, Legionella, Mycobacterium, Mycoplasma, Neisseria, Staphylococcus, Streptococcus, etc.

[0098] "Identity, "on the other hand, is gauged from the starting point of complete homology. Thereafter, identity may be described in terms of percentages according to the number of base changes in the DNA sequence taking into account any gaps. For purposes of the present invention, variants of the invention have a sequence identity greater than at least about 65%, preferably at least about 75%, more preferably at least about 85%, and can be greater than at least about 90% or more as determined by the Smith-Waterman homology search algorithm as implemented in MPSRCH program (Oxford Molecular). A preferred method of calculating percent identity is the Smith- Waterman algorithm, using the following. Global DNA sequence identity must be greater than 65% as determined by the Smith-Waterman homology search algorithm as implemented in MPSRCH program (Oxford Molecular) using an affine gap search with the following search parameters: gap open penalty, 12; and gap extension penalty, 1.

[0099] Amino acid sequence variants are also included in the invention. Preferably, naturally or non-naturally occurring protein variants have amino acid sequences which are at least 85%, 90%, or 95% identical to the amino acid sequences identified herein, or to a shorter portion of these sequences. More preferably, the molecules are 98% or 99% identical. Percent sequence identity is determined using the Smith-Waterman homology search algorithm using an affine gap search with a gap open penalty of 12 and a gap extension penalty of 2, BLOSUM matrix of 62. The Smith-Watennan homology search algorithm is taught in Smith and Waterman, Adv. Appl. Math. (1981) 2: 482-489.

[0100] Also included in the invention are fragments of the nucleic acid sequences and amino acid sequences identified herein, as well as RNAs and RNA fragments corresponding to the DNA sequences disclosed. Such nucleic acid fragments are at least about 10 nucleotides, more preferably at least about 20 to 25 nucleotides, and more preferably at least about 50 to 100 nucleotides, and can include any fragment or variant of a fragment. Such nucleic acid fragments may be used as probes for identifying similar or substantially identical or identical nucleic acid sequences in other genera, or as tools in constructing nucleic acid vectors for knock out and promoter swap experiments. Such amino acid fragments are at least about four amino acids in length, more preferably at least about 8 to 12 amino acids in length, and more preferably at least about 20 to 30 amino acids in length, and may be used as agonists or antagonists to test binding interactions of the proteins disclosed herein, or alternatively as immunogens to isolate antibodies that recognize and bind to specific epitopes of a target protein.

[0101] Once a gene is identified as being essential or important for Pseudomonas growth on rich media or in any specific environment, the invention also encompasses the identification of antibacterial agents that have specific activity against the essential or important genes or their gene products or the biochemical pathways in which they are involved. In this context, the term"biochemical pathway"refers to a connected series of biochemical reactions normally occurring in a cell, or more broadly a cellular event such as cellular division or DNA replication. Typically, the steps in such a biochemical pathway act in a coordinated fashion to produce a specific product or products or to produce some other particular biochemical action. Such a biochemical pathway requires the expression product of a gene if the absence of that expression product either directly or indirectly prevents the completion of one or more steps in that pathway, thereby preventing or significantly reducing the production of one or more normal products or effects of that pathway.

[0102] Thus, an agent specifically inhibits such a biochemical pathway requiring the expression product of a particular gene if the presence of the agent stops or substantially reduces the completion of the series of steps in that pathway. Such an agent, may, but does not necessarily, act directly on the expression product of that particular gene. An"expression product"of a gene means that, in a bacterial cell of interest, the gene is transcribed to form RNA molecules. For those genes that are transcribed into mRNAs, the mRNA is translated to form polypeptides. More generally, in this context, "expressed"means that a gene product is formed at the biological level that would normally have the relevant biological activity (i. e. , RNA or polypeptide level).

[0103] Thus, the invention includes a method of screening for an antibacterial agent, comprising determining whether a test compound is active against an essential or important bacterial gene identified by the methods herein. The invention also includes a method of screening for an antibacterial agent, comprising determining whether a test compound is active against a protein encoded by an essential bacterial gene identified herein, or active to inhibit the biochemical pathway that involves said protein. The term"antibacterial agent"refers to both naturally occurring antibiotics produced by microorganisms to suppress the growth of other microorganisms, and agents synthesized or modified in the laboratory which have either bactericidal or bacteriostatic activity. An"active"agent in this context will inhibit the growth of P. aeruginosa and possibly related species. The term"inhibiting the growth"indicates that the rate of increase in the numbers of a population of a particular bacterium is reduced. Thus, the term includes situations in which the bacterial population increases but at a reduced rate, as well as situations where the growth of the population is stopped, as well as situations where the numbers of the bacteria in the population are reduced or the population even eliminated. If an enzyme activity assay is used to screen for inhibitors, one can make modifications in uptake/efflux, solubility, half life, etc. to compounds in order to correlate enzyme inhibition with growth inhibition.

[0104] Assays may include any suitable method and may be expected to vary on the type of essential gene or protein involved. For instance, one embodiment is a method comprising the steps of : a) contacting said protein or a biologically active fragment thereof with a test compound; and b) determining whether said test compound binds to said essential gene product or protein or fragment of said protein; wherein binding of said test compound to said polypeptide or said fragment is indicative that said test compound is an antibacterial agent. It is quite common in identifying antibacterial agents, to assay for binding of a compound to a particular polypeptide where binding is an indication of a compound which is active to modulate the activity of the polypeptide. Binding may be determined by any means according to the agent tested and techniques known in the art. l0105] Also, agents that inhibit binding of two proteins or polypeptides may also be identified, for instance using a yeast two-hybrid system. Such a system will entail cloning the genes encoding each protein and expressing each in a reporter cell system such that interaction between the two proteins is monitored by observing the expression of a reporter gene. For instance, cDNAs cloned in a yeast two-hybrid expression system (Chien et al. (1991) Proc. Natl. Acad. Sci. (U. S. A. ) 88: 9578; Zervos et al.

(1993) Cell 72: 233) can be used to identify other cDNAs encoding proteins that interact with the protein encoded by the first, thereby produce expression of the GAL4- dependent reporter gene. Thereafter, cells expressing both proteins leading to expression of the reporter gene are used to screen for agents that interact with either protein, or the gene encoding either protein. Such systems are well known in the art and are well within the realm of ordinary skill.

[0106] Another embodiment is a method for evaluating a test agent for inhibition of expression of an essential gene identified according to the methods herein, comprising: a) contacting a cell expressing said essential gene with said agent; and b) determining the amount or level of expression of said essential gene in said sample.

[0107] The exact determination method will be expected to vary depending on the characteristics of the expression product as would be readily apparent to one of ordinary skill in the art. Such methods can include, for example, antibody binding methods, enzymatic activity determinations, and substrate analog binding assays. Such level of expression could be monitored by monitoring the level of the product of the essential gene in the cell, i. e. , by SDS-PAGE, or by colorimetric assays using, for example, a lacZ gene or protein fusion and detection on media using X-Gal or spectrophotometric detection.

[0108] When such fusions are employed, fusions may be designed using the chromosomal gene so long as the fusion does not disrupt the function of the essential gene, i. e. , as with a gene fusion where lacZ is inserted just downstream of the essential gene and is expressed from the same promoter as the essential gene. Alternatively, one could employ an extrachromosomal fusion construct whereby the wild type chromosomal copy of the gene is not disrupted. In this case, one could employ a protein fusion, i. e. , where a portion of lacZ sufficient to be detected with a colorimetric test is fused in frame with the coding region of the essential gene such that a fusion protein is obtained. Other detectable or measurable proteins commonly used in the art may be used as an alternative to lacZ, for instance, phoA, Lux/luciferase, etc.

[0109] Another method of the invention for evaluating an potential antibacterial agent, comprises the steps of : a) providing a bacterial strain comprising a mutant or normal form of the essential or important gene, wherein said mutant form of the gene confers a growth conditional phenotype; b) contacting bacteria of said bacterial strain with a test compound in semi- permissive or permissive growth conditions; and c) determining whether the growth of said bacterial strain comprising said mutant form of a gene is reduced in the presence of said test compound to a greater extent than a comparison bacteria comprising a normal form of said gene.

[0110] In this context, a"mutant form"of a gene is a gene which has been altered, either naturally or artificially, changing the base sequence of the gene, which results in a change in the amino acid sequence of an encoded polypeptide. The change in the base sequence may be of several different types, including changes of one or more bases for different bases, small deletions, and small insertions. Mutations may also include transposon insertions that lead to attenuated activity, i. e. , by resulting in expression of a truncated protein. By contrast, a normal form of a gene is a form commonly found in a natural population of a bacterial strain. Commonly a single form of a gene will predominate in natural populations. In general, such a gene is suitable as a normal form of a gene, however, other forms which provide similar functional characteristics may also be used as a normal gene. In particular, a normal form of a gene does not confer a growth conditional phenotype on the bacterial strain having that gene, while a mutant form of a gene suitable for use in these methods does provide such a growth conditional phenotype.

[0111] As used in the present disclosure, the term"growth conditional phenotype" indicates that a bacterial strain having such a phenotype exhibits a significantly greater difference in growth rates in response to a change in one or more of the culture parameters than an otherwise similar strain not having a growth conditional phenotype.

Typically, a growth conditional phenotype is described with respect to a single growth culture parameter, such as temperature. Thus, a temperature (or heat-sensitive) mutant (i. e. , a bacterial strain having a heat-sensitive phenotype) exhibits significantly reduced growth, and preferably no growth, under non-permissive temperature conditions as compared to growth under permissive conditions. In addition, such mutants preferably also show intermediate growth rates at intermediate, or semi-permissive, temperatures.

Similar responses also result from the appropriate growth changes for other types of growth conditional phenotypes. A growth conditional phenotype can also be conferred by cloning an essential or important gene behind a regulatable promoter, for instance, a promoter that is only active, or only leads to transcription, under particular environmental conditions or in response to a specific environmental stimulus. Such growth conditional promoter mutants may be isolated according to the promoter swap strategies described herein.

[0112]"Semi-permissive conditions"are conditions in which the relevant culture parameter for a particular growth conditional phenotype is intermediate between permissive conditions and non-permissive conditions. Consequently, in semi- permissive conditions the bacteria having a growth conditional phenotype will exhibit growth rates intermediate between those shown in permissive conditions and non- permissive conditions. In general, such intermediate growth rate is due to a mutant cellular component which is partially functional under semi-permissive conditions, essentially fully functional under permissive conditions, and is non-functional or has very low function under non-permissive conditions, where the level of function of that component is related to the growth rate of the bacteria.

[0113] The term"method of screening"means that the method is suitable, and is typically used, for testing for a particular property or effect in a large number of compounds. Therefore, the method requires only a small amount of time for each compound tested; typically more than one compound may be tested simultaneously (as in a 96-well microtiter plate, or in a series of replica plates), and preferably significant portions of the procedure can be automated. "Method of screening"also refers to determining a set of different properties or effects of one compound simultaneously.

[0114] Because the essential and important genes identified herein can be readily isolated and the genes cloned into a variety of vectors known in the art, the invention also encompasses vectors comprising the nucleic acid sequences, open reading frames and genes of the invention, as well as host cells containing such vectors. Because the essential genes identified herein can be readily isolated and the encoded gene products expressed by routine methods, the invention also provides the polypeptides encoded by those genes, as well as genes having at least about 50%, or more preferably about 60%, or more preferably about 70%, or more preferably about 80%, or more preferably about 90%, or most preferably about 95% protein sequence identity.

[0115] Thus, by identifying certain essential and/or important genes, this invention provides a method of screening for an antibacterial agent by contacting a polypeptide encoded by one of the identified essential or important genes, or a biologically active fragment of such a polypeptide, with a test compound, and determining whether the test compound binds to the polypeptide or polypeptide fragment. In addition, to simple binding determinations, the invention provides a method for identifying or evaluating an agent active on one of the identified essential genes. The method involves contacting a sample containing an expression product of one of the identified genes with the known or potential agent, and determining the amount or level of activity of the expression product in the sample.

[0116] In particular, antibodies to essential and important gene products are anticipated to be suitable diagnostic binding and antibacterial agents. Thus, antibodies to the proteins encoded by the essential and important genes identified by the methods described herein are also included in the invention. Such antibodies may be isolated according to well known techniques in the art, i. e. , Kohler and Milstein for monoclonal antibodies. Also included are polyclonal antibodies and antibody fragments such as Fv, Fab and Fab2 fragments, as well as chimeric and humanized antibodies, and human antibodies, i. e. , made using a Xeno mouse.

[0117] In a further aspect, this invention provides a method of diagnosing the presence of a bacterial strain having one of the genes identified above, by probing with an oligonucleotide at least 15 nucleotides in length, which specifically hybridizes to a nucleotide sequence which is the same as or complementary to the sequence of one of the bacterial genes identified above. In some cases, it is practical to detect the presence of a particular bacterial strain by direct hybridization of a labeled oligonucleotide to the particular gene. In other cases, it is preferable to first amplify the gene or a portion of the gene before hybridizing labeled oligonucleotides to those amplified copies.

[0118] In a related aspect, this invention provides a method of diagnosing the presence of a bacterial strain by specifically detecting the presence of the transcriptional or translational product of the gene. Typically, a transcriptional (RNA) product is detected by hybridizing a labeled RNA or DNA probe to the transcript.

Detection of a specific translational (protein) product can be performed by a variety of different tests depending on the specific protein product. Examples would be binding of the product by specific labeled antibodies and, in some cases, detection of a specific reaction involving the protein product. Diagnostic assays find particular use in assaying tissue and fluid samples of patients suspect of having a Pseudomonas infection.

[0119] Antibacterial agents identified according to the methods of the invention may be employed in pharmaceutical compositions. Such compositions may be administered to patients in order to treat an infection by or involving P. aeruginosa, either alone or in combination with secondary agents targeted at, for instance virulence factors of P. aeruginosa, or other bacteria that may be present in addition to P. aeruginosa. In this context, the term"administration"or"administering"refers to a method of giving a dosage of an antibacterial pharmaceutical composition to a mammal, where the method is, e. g. , topical, oral, intranasal, inhaled, intravenous, transdermal, intraperitoneal, or intramuscular. The preferred method of administration can vary depending on various factors, e. g. , the components of the pharmaceutical composition, the site of the potential or actual bacterial infection, the bacterium involved, and the severity of an actual bacterial infection.

[0120] As used above and throughout this application, "hybridize"has its usual meaning from molecular biology. It refers to the formation of a base-paired interaction between nucleotide polymers. The presence of base pairing implies that at least an appreciable fraction of the nucleotides in each of two nucleotide sequences are complementary to the other according to the usual base pairing rules. The exact fraction of the nucleotides which must be complementary in order to obtain stable hybridization will vary with a number of factors, including nucleotide sequence, salt concentration of the solution, temperature, and pH.

[0121] The term, "DNA molecule", should be understood to refer to a linear polymer of deoxyribonucleotides, as well as to the linear polymer, base-paired with its complementary strand, forming double-strand DNA (dsDNA). The term is used as equivalent to"DNA chain"or"a DNA"or"DNA polymer"or"DNA sequence", so this description of the term meaning applies to those terms also. The term does not necessarily imply that the specified"DNA molecule"is a discrete entity with no bonding with other entities. The specified DNA molecule may have H-bonding interactions with other DNA molecules, as well as a variety of interactions with other molecules, including RNA molecules. In addition, the specified DNA molecule may be covalently linked in a longer DNA chain at one, or both ends. Any such DNA molecule can be identified in a variety of ways, including, by its particular nucleotide sequence, by its ability to base pair under stringent conditions with another DNA or RNA molecule having a specified sequence, or by a method of isolation which includes hybridization under stringent conditions with another DNA or RNA molecule having a specified sequence.

[0122] References to a"portion"of a DNA or RNA chain mean a linear chain which has a nucleotide sequence which is the same as a sequential subset of the sequence of the chain to which the portion refers. Such a subset may contain all of the sequence of the primary chain or may contain only a shorter sequence. The subset will contain at least 15 bases in a single strand. However, by"same"is meant"substantially the same" ; deletions, additions, or substitutions of specific nucleotides of the sequence, or a combination of these changes, which affect a small percentage of the full sequence will still leave the sequences substantially the same. Preferably this percentage of change will be less than 20%, more preferably less than 10%, and even more preferably less than 3%. "Same"is therefore distinguished from"identical" ; for identical sequences there cannot be any difference in nucleotide sequences.

[0123] As used in reference to nucleotide sequences, "complementary"has its usual meaning from molecular biology. Two nucleotide sequences or strands are complementary if they have sequences that would allow base pairing between the strands according to the usual pairing rules. This does not require that the strands would necessarily base pair at every nucleotide ; two sequences can still be complementary with a low level of base mismatch such as that created by deletion, addition, or substitution of one or a few (up to 5 in a linear chain of 25 bases) nucleotides, or a combination of such changes.

[0124] Other embodiments of the invention will be immediately envisaged by those of skill in the art upon reading the methods and examples to follow. Such examples are merely illustrative of the invention, and should not be construed as limiting the scope of the invention in any way.

Methodology Generation of Transposon Library [0125] Transposon insertions were generated using an improved transposon system for P. aeruginosa that utilizes a mini-Tn5-type transposon on a delivery vector that does not replicate in Pseudomonas. The delivery vector contains a modified transposase gene with three amino acid substitutions that have been shown to increase the frequency of Tn5 insertions. Weinreich et al. , 1994, Evidence that cis preference of the Tn5 transposase is caused by nonproductive multimerization, Genes Dev. 8 (19): 2363-74. The Tn5 transposase was placed under control of a lac promoter and the complete transposable element was minimized to 1.7 kilobases in length, including a tetracycline resistance marker and transcription terminator to prevent read-through into the genome. The transposon vector is delivered to P. aeruginosa via conjugation from a suitable E. coli host (e. g. SM103w). Following conjugation, transposon mutants are selected by resistance to tetracycline conferred by the trasnposable element.

[0126] Libraries were created in both P. aeruginosa PAK and PA01. The average diversity of the libraries created using this strategy is estimated to be-40, 000 to -50, 000 independent mutants per conjugation. Care is taken to minimize passage of each transposon conjugation before plating for mutant selection in an effort to minimize the potential for siblings, i. e. , by stopping the conjugation after sufficient time for a single round of conjugation events.

High-Throughput Transposon Insertion Mapping (HTTIM) [0127] Precise transposon insertion sites were determined by an anchored, semi- random PCR method for amplification of the transposase/genome junction region.

O'Toole and Kolter, 1998, Initiation of biofilm formation in Pseudomonasfluorescens WCS365 proceeds via multiple, convergent signaling pathways: a genetic analysis, Mol. Microbiol. 28 (3): 449-61. The technique, HTTIM, uses both Tn5 specific and semi-random primers with conserved primer tails. A small aliquot of transposon mutant liquid culture is used as a template and amplification of a fragment containing an insertion site is achieved in a two-step process. The PCR product is then sequenced and the insertion site is entered into an Oracle database for analysis. To date, more than 10,000 to 14,000 insertions have been mapped, each insertion representing the disruption of a gene or intergenic region that is not essential for survival on rich media.

[0128] With every insertion added to the map, the regions of the genome containing essential genes, and particularly those containing operons containing essential genes (because of potential polar effects of insertions in upstream genes), begin to become apparent because these regions will not be able to accommodate transposon insertions.

Table 1 shows a listing of the open reading frames identified as existing between transposon insertions, as well as an indication of whether the gene has homologues that have been identified in other bacteria pursuant to BLAST sequence database analysis.

Open reading frames were tentatively assigned names prior to being identified pursuant to HTTIM analysis, as disclosed in the Pseudomonas genome project, and reported in Stover et al., Complete genome sequence of Pseudomonas aeruginosa PAO1, an opportunistic pathogen, August 21,2000, Nature 406: 959-964, herein incorporated by reference in its entirety.

[0128] For instance, the predicted ORFs were examined individually for (1) identity with known genes of P. aeruginosa with sequences deposited in GenBank, (2) similarity with well-characterized genes from other bacteria, or (3) presence of known functional motifs (see http://www. pseudomonas. com for complete list). In each case the literature was searched to ensure that the proteins encoded by the homologous genes were functionally characterized to avoid the perpetuation of poorly supported functional assignments. In addition, 61 researchers who were members of the P. aeruginosa research community or had experience in particular aspects of bacterial physiology were enlisted for the Pseudomonas Community Annotation Project (PseudoCAP) to provide expert assistance and confirmatory information in the genome project for the analysis of identified ORFs and assigned functions.

[0129] The genome project was able to assign a functional class to 54.2% of ORFs.

As in other bacterial genomes, a large proportion of the genome (45.8% of ORFs) consists of genes for which no function could be determined or proposed (confidence level 4). Of these, nearly a third (769 ORFs) possess homology to genes of unknown function predicted in other bacterial genomes, and the remainder (32% of ORFs) do not have strong homology with any reported sequence. The 372 ORFs from the entire genome analysis that are known P. aeruginosa genes with demonstrated functions (confidence level 1) are primarily genes encoding lipopolysaccharide biosynthetic enzymes, virulence factors, such as exoenzymes and the systems that secrete them, and proteins involved in motility and adhesion. ORFs with strong homology to genes in other organisms with demonstrated functions (confidence level 2; 1,059 ORFs) include those required for DNA replication, protein synthesis, cell-wall biosynthesis and intermediary metabolism.

[0130] The ORFs that provided the most new information about P. aeruginosa biology via the genome annotation were those that could be assigned a probable function on the basis of similarity to established sequence motifs, but could not be assigned a definite name (confidence level 3; 1,590 ORFs). Most of these genes encode products that are in one of three functional classes: putative enzymes (405 genes), transcriptional regulators (341 genes) or transporters of small molecules (408 genes). In some cases genomic context provided additional information, allowing us to identify loci that appear to encode systems such as metabolic pathways and secretion systems, although the substrates for such systems could not be identified. The system for assigning name and putative function to each essential or important gene was gleaned from the Pseudomonas genome project data already available.

Statistical Analysis of Putative Essential and Important Genes [0131] The open reading frames listed in Table 1 are also presented in Table 2, wherein the ORFs are listed in order of length of base pairs from longest to shortest.

Also listed in Table 2 is the probability of essentiality assigned to each of the open reading frames. Probability correlates with length of the ORF, such that the longer the ORF, the higher the probability of hitting the ORF in a random transposon mutagenesis experiment, and the higher the confidence level that the ORF represents an essential or an important gene given that no transposon insertions therein were isolated. Statistical confidence levels in essentiality or importance can help narrow the focus in the screening of specific genes, thereby shortening the verification process and the subsequent identification of antibacterial agents specific for that gene or gene product.

Thus, one of the benefits of the HTTIM approach is that it is a quantitative approach that lends itself well to statistical analysis.

[0132] The High-Throughput Transposon Insertion Mapping (HTTIM) strategy utilizes a transposon, which is a small, mobile DNA element that randomly inserts into the chromosome. Although HTTIM was performed using a Tn5 transposon, any transposon may be employed so long as its insertion into the chromosome is random, i. e. , devoid of hot spots. Reznikoff, W. S. , 1993, The Tn5 transposon, Annu. Rev.

Microbiol. 47: 945-63. Although the Tn5 derivative employed here contained a modified transposase gene with three amino acid substitutions that have been shown to increase the frequency of Tn5 insertions (see supra), the frequency of insertion is generally quite low. For instance, mutants with even one insertion occur at a rate of only 1 in 105 or 106 bacteria, and must be specifically selected from a background of cells with no insertions. Because the frequency of a single insertion is so low, the frequency of a double insertion is so low as to be insignificant.

[0133] When the transposon insertion disrupts one of the 5570 genes in the Pseudomonas genome, the function of that gene is lost. If the disrupted gene is essential for growth, the transposon insertion mutant dies and cannot be characterized.

If the transposon disrupts a gene that is non-essential, the mutant survives, grows and the transposon insertion site is mapped. By examining the insertion sites of a large number of transposon mutants, all, of the non-essential P. aeruginosa genes can be identified, and by implication, all of the essential genes may be identified as well.

Characterization of over 13,000 transposon insertions revealed insertions in 3890 genes and resulted in an even distribution of insertions across the entire length of the genome.

The remaining 1658 genes, in which a transposon insertion has never been observed, are candidates of essential genes (30%). See Figure 7, showing a graph illustrating ORF coverage by Tn5 achieved in High-Throughput Transposon Insertion Mapping (HTTIM), wherein 30% of the genes in the genome are candidate essential genes where ORF size is not taken into account in predicting essentiality.

[0134] Because insertion of the transposon used here into the chromosome was proposed to be random, it was possible that some of the 1658 genes that did not receive a transposon insertion were simply not hit by random chance. One cannot truly know that a transposon has no hot spots and is entirely random until the data is analyzed, and the data here confirmed that the Tn5 derivative employed underwent random insertion in P. aeruginosa. Thus, the chance that a gene will not be hit by the transposon as a matter of random chance increases as the length of the gene decreases, particularly for very small genes (< 600 base pairs). See Figure 8, Probability of Being an Essential Gene Given No Hit. Thus, by deleting smaller ORFs (< 600 base pairs) in which there is a lower confidence in essentiality, the probability of essentiality goes up while the number of predicted essential genes decreases. Further, the curve in the graph depicted in Figure 8 should level off faster. Thus, in predicting the essentiality of genes from the HTTIM candidate set, the closer one can come to a probability of 1.0 as depicted in Figure 8, the higher the confidence level of essentiality that can be assigned to each gene in the candidate subset. For a representation of the number of ORFs of various lengths in P. aeruginosa, see the histogram in Figure 9.

[0135] A Bayessian statistical model for truncated counting data was applied to the candidate essential gene set, and permitted a determination that 16 to 17 percent of P. aeruginosa genes are essential. Such a model may therefore be utilized to increase the statistical confidence that a given gene in the candidate subset is essential. An exemplary statistical model is provided in Example 1.

Physical Methods for Target Gene Validation [0136] While the above methodology and the database of putative essential and important gene candidates established thereby is believed to be superior to existing methods with regard to the quantity of experimentation required to identify essential and important genes in Pseudomonas aeruginosa and the degree of confidence conferred, it should be understood that the methodology described herein can be incorporated into combined protocols with technology known in the art. For instance, the methods for verifying essentiality disclose in WO 01/07651, herein incorporated by reference in its entirety, would be useful as a secondary method to be utilized in combination with the methods described in this disclosure. Alternatively or additionally, one of several approaches may be used to determine whether a particular gene is essential (absolutely required for survival on rich medium) or important (the absence of which results in attenuated growth) to P. aeruginosa.

Integration Knockouts [0137] This is the simplest and most rapid strategy. PCR is used to amplify a small (200-500 base pairs) portion of the coding sequence, or open reading frame (ORF) of the gene of interest. This gene fragment must be centrally located within the ORF--it cannot include either termini of the gene's coding region. This fragment is cloned into a plasmid vector that can replicate in E. coli, but not in Pseudomonas. The vector used should have a drug resistance marker that is suitable for selection in Pseudomonas, and an origin for conjugal transfer. This feature allows the plasmid to be transferred by conjugation from a suitable E. coli donor strain to a Pseudomonas strain when the two are co-cultured under the appropriate conditions.

[0138] Following conjugation the co-cultured mixture is harvested and plated on media which selects against the E. coli donor and for Pseudomonas which contain the plasmid. Since the plasmid is incapable of extra-chromosomal replication in Pseudomonas, colonies that arise are the result of homologous recombination between the Pseudomonas chromosome and the cloned gene fragment on the plasmid. This is referred to as single-crossover recombination ; a single recombination event takes place between the plasmid and the chromosome. The result is integration of the plasmid into the bacterial chromosome and disruption of the gene from which the fragment was amplified (Fig. 1).

[0139] Variations of this approach are possible. For instance, one could clone out the entire locus and isolate transposon insertion mutants in E coli using known techniques, i. e. , by transposition from the E. coli genome, selecting plasmid insertions by mobilizing the vector into a recipient cell that does not contain the transposon or the antibiotic resistance marker encoded by the transposon, and screening the plasmid for insertions in the cloned gene. Thereafter, a similar assay could be performed by screening for double crossover events in P. aeruginosa that result in recombination of the transposon into the chromosomal locus from a suicide vector.

[0140) Integration of the plasmid or other insertion at the locus can be confirmed by a relatively rapid PCR-based screen of recombinant colonies. The advantage of this strategy, particularly the plasmid single crossover strategy, is that it requires only amplification of a short stretch of DNA followed by a single cloning step before recombination experiments can be performed. The disadvantage is that if the target gene is essential, no recombinants can be obtained. Failure to obtain recombinants as proof of essentiality is pretty thin evidence. However, if a gene is in fact non-essential, this method will demonstrate that quickly.

Integration Knockouts with Extra-chromosomal Complementation [0141] This variation of the above method provides more convincing data when the target gene is essential. It employs the same type of non-replicating integration plasmid described above, but recombinations are performed in strains already carrying a second copy of the target gene on an extra-chromosomal plasmid. This second copy can then supply the essential function when the chromosomal copy is disrupted. If disruptions can only be obtained when a complementing plasmid is present and not when a control plasmid is present, this is rather strong evidence that the target gene is essential. The advantage of this method is that you obtain colonies even when your gene is essential.

The disadvantage is that construction and sequencing of the complementation plasmid takes additional time.

Integration with a Regulatable Promoter (Promoter Swap) [0142] This approach also involves selecting for chromosomal integration of non- replicating plasmids via homologous recombination. However, the design of the integrating plasmid is different. In this case, the N-terminal coding sequence (300-500 base pairs) of the target gene is PCR amplified and cloned into a vector downstream of a regulatable promoter, i. e., a lac promoter, which is inducible in the presence of IPTG, or an arabinose promoter (pABD), inducible in the presence of arabinose. The activity of the promoter can be modulated by the presence of a specific inducer molecule. The plasmid is conjugated into Pseudomonas and integration selected for under conditions where the regulatable promoter is active. The resulting chromosomal integration replaces the target gene's natural promoter with the regulatable promoter from the plasmid (Fig. 2). If the target gene is essential, recombinants can only survive when the inducer molecule is present in their growth media to stimulate gene expression. If the gene is non-essential, the recombinant's growth is independent of the addition of the inducer. The advantage of this strategy is that it requires only amplification of a short stretch of DNA followed by a single cloning step before recombination experiments can be performed.

Examples : Essential Genes Identified Example 1: A Bayessian Statistical Model for Increasing Statistical Confidence of Essentiality [01431 When the Tn5 transposon inserts into the Pseudomonas DNA, one of three things happen: 1) The insertion disrupts a nonessential gene. The cell survives to be characterized and the location of the insertion is determined. 2) The insertion disrupts an essential gene. The cell does not survive and the insertion site is not determined. 3) The insertion is in an intergenic region (between genes) and no information is gained.

Genes with identified insertions are nonessential genes. However, genes without identified insertions could be essential genes or nonessential genes with zero transposon insertion. To determine the number of essential genes, we have developed a multivariate Bayession model for truncated Poisson data and applied it to the Pseudomonas genome data set. A likelihood gain based searching algorithm was developed to obtain maximum likelihood estimates. The property of the algorithm was studied. Different approaches were compared for both multivariate and univariate approaches.

A. Structure of the Data and Preliminary Considerations [0144] A transposon Tn5 insertion mutagenesis library was constructed in Pseudomonas aeruginosa strains PAK and PAO1. Mutants were randomly picked and their genomic insertion site sequence determined through polymerase chain reaction (PCR) and automated DNA sequencing. BLASTN analysis of transposon/genome junction sequences was used to map the location of the insertions relative to the completed strain PAO1 genome sequence. More than 20,000 mutants were analyzed which resulted in 12,219 independent insertions being mapped. In order to identify essential genes, transposon insertion sites were analyzed with respect to the protein- encoding genes in this organism. A data set consists of the ID of genes, their length in DNA base-pairs, and the number of transposon insertions were obtained from experiments. The data set consists of 5570 genes with 881 different sizes ranging from 72 to 16884 DNA base-pairs. The distribution of the gene sizes are extremely skewed to the right with majority of the genes being smaller than 2000 DNA base-pairs as shown in Figure 10.

[0145] A randomly selected subset of the data is shown in Table 4, where 8 is gene size, x is the observed number of transposon insertions. Insertions to essential genes are not observable since the insertion mutants can not survive for characterization when the transposon is inserted into an essential gene. Therefore, a gene with zero observed transposon insertions can either be an essential gene or a nonessential gene with zero transposon insertion. Consequently, the count of transposon insertions x is truncated with the truncation region being a single element {0}.

Table 4: A sample of the gene data set Genes 6 x <BR> <BR> 298 1359 3 4047 618 0 1170 735 1 4953 1044 1 5526 213 0 4624 1707 4 5069 426 3 [0146] Since the insertion into the chromosome of Pseudomonas aeruginosa is random (Reznikoff WS. 1993), and the probability of receiving an insertion for a given gene is proportional to its size measured in DNA base-pairs, the number of transposon insertions into a gene is distributed as truncated Poisson with parameter k6, where S is the size of the gene and k is an unknown parameter, which is independent of gene size.

B. A Bayessian Model [0147] Let R be a measurable subset of the probability space Q such that a random variable X is observable only if X##\R. In this example, no observations can be obtained from essential genes, whereas only nonzero observations can be obtained from nonessential genes, the set R consists of a single element {0}.

1. a. One Gene Size [0148] Assume all genes in a genome have same size, 8, and let N be the number of nonessential genes in this genome. Then the observations Xi, X2,..., XN from the N nonessential genes are i. i. d. Poisson (S b), of which, all observations of value zero are truncated. The product ?,. 8 indicates that the probability of a gene receiving an insertion is proportional to its size.

[0149] Let denote the subset of all nonzero observations. Then this subset composes a random sample of size n from a truncated Poisson distribution whose distribution function can be written as [0150] Let q l-e~Ay denote the probability that an observation from Poisson (###) is not truncated, and let p =1-q=-##. Then, conditional on the parameters n and N, the likelihood function of the joint distribution of {x ;, xi,..., xn} can be written as [0151] Let S=X1*+X2*+###+Xn* (3. 3) denote the sum of all nonzero observations and notice that n follows a binomial distribution B (N, q). The likelihood function of the joint distribution of {n,X1*,X2*,###,Xn*}, conditional on the parameter N, can be obtained as [0152] The Bayesian model consists of the conditional model (2.4) and a prior distribution of the parameter N. Assuming N, the number of nonessential genes, is binomial B (M, y), where M is the total number of genes of size 8, which is known, and y is the portion of nonessential genes which is unknown and is independent of gene size, we can write the likelihood function of the joint distribution of {n,N,X1*,X2*,###,Xn*} as [0153] This is the likelihood function of n nonzero observations from M genes of the same size 8, of which N genes are nonessential. It is easy to see that (3.5) is proportional to the likelihood function of the posterior distribution of N given observations n and S.

2. b. Multiple Gene Sizes [0154] For a given genome consists of genes of different sizes, let #=(#1,#2,###,#g)T denote the vector of g different gene sizes, and let M= (M1,M2,###,Mg)T the vector of known numbers of total genes, # = (N1,N2,###,Ng)T the unknown numbers of nonessential genes, the numbers of nonzero observations from the nonessential genes, and #= (S1,S2,###,Sg)T the sums of nonzero observations, as defined in (3.3).

[0155] The likelihood function of the joint distribution of can be written as where #*# is the L1 norm of a vector, and [0156] Let 3 = ln (L). Then up to an additive constant, the log likelihood function of the joint distribution of {#,#,#} can be written as where (ra, N) are the parameters of interests. The vector # is defined on {ni # Ni # Mi; = 1,2,###,g} and 3 (7, AR) is proportional to the likelihood function of the posterior distribution of N given ñ and s.

[0157] When g is large, say, in the order of hundreds as in the situation we are dealing with in this paper, obtaining the maximum likelihood (ML) estimate of Ñ= (N1,N2,###,Ng)T from (3.7) in such a high dimensional parameter space is a very difficult task both theoretically and computationally. In the next section, we will present a stepwise, maximum likelihood gain based method to obtain the ML estimation.

C. ML ESTIMATION OF PARAMETERS [0158] For any #=(N1,N2,###,Ng)T, it is easy to verify using (3.7) that the ML estimations of the parameters y and k are r=IIÑII/IIMII (4.1) and #=###/(#T##) (4.2) respectively. Substituting (4.1) and (4.2) for y and X in (3.7), we have [0159] For 1#i# g, define <BR> <BR> <BR> <BR> #i#*(#)=#*(#+#i)-##(#) (4.4)<BR> <BR> <BR> <BR> <BR> <BR> for any # # {ni # Ni < Mi, nj # Nj # Mj: i # j}. In equation (4.4), #@ = =(0,###0,1,0,###,0)T with 1 at the i position. For notational purpose, let #(k)=k#ln(k)+(###-k)#in(###-k) (4.5) for 11 ñ ## k ###. Then, (3.4) can be written as [0160] To obtain ML estimation of Ñ, we define an operator, ED, between the observed vector ñ and any integer k with 0 # k ## ##-### as follows: <BR> <BR> <BR> <BR> <BR> <BR> K$0=K,<BR> <BR> <BR> <BR> ##1={#+#i:#$i#*(#)##j#*(#) for all j#i}, and (4.7)<BR> <BR> <BR> <BR> ##k=(##(k-1))#1 for k # 2.

We also define a likelihood-gain function G with G (0) =0 and <BR> <BR> <BR> <BR> <BR> <BR> G (k) = #*(##k)-##(##(k-1)) (4.8)<BR> <BR> <BR> <BR> <BR> <BR> <BR> <BR> <BR> <BR> <BR> <BR> <BR> <BR> <BR> <BR> for 1#k####-###.

[0161] Using this likelihood-gain function, we can search the ML estimation for Ñ as follows: 1. Start with the observation ii as the initial estimate of #, and denote it as No.

2. For each gene size 8 ; with n ; < Mus, il 2,..., g, calculate a likelihood difference #i#*(#0)=#*(#0+#i)-#*(#0) by set Ni0 = ni+1 and NJ = nj for all jgi.

3. Update the initial values #0 by setting N° = N° + 1 such that #i#*(#0)=max{#j#*(#0), j=1,2,###,g}. This maximum likelihood difference is the likelihood gain defined in (4.8).

4. Repeat the process until it converges. By convergence we mean that either the estimated number of nonessential genes equals to the number of genes in each size group or when increasing the number of nonessential genes in any size groups will result in a loss of likelihood.

[0162] This algorithm searches the ML estimator in a high dimensional space (881 in our study) along a path such that at each iteration, it moves in a direction (that is, increases the number of nonessential genes in this size group by one) along which the likelihood gain is maximum among all possible directions. Because the searching algorithm prohibits reversal of previous moves at any later iteration, it moves towards the ML estimator along the shortest path with the deepest ascending (maximum likelihood gain) at each step. Table 5 and Figures 11 and 12 show the values of likelihood gains in each iteration. With very few exceptions where the monotonous is violated only at the fourth or fifth decimal places that probably can be attributed to rounding errors, the likelihood gain is a monotonously decreasing function.

Table 5. A Sample of Likelihood Gains at Each Iteration Iteration id 6 M n N (i) G (i) 1 28 210 13 2 3 2. 67559 2 60 306 14 3 4 2.41082 3 44 258 14 5 6 2.34388 4 63 315 15 5 6 2.29243 ... ... ... ... ... ... ...

18 32 222 7 1 2 2.05160 19 81 369 11 2 3 2.05166 ... ... ... ... ... ... ...

774 122 492 12 8 11 0.00692 775 266 924 16 14 15 0.00544 776 85 381 14 3 11 0.00531 The following three theorems show that the estimates obtained through the above algorithm are indeed the maximum likelihood estimates.

THEOREM 1: if then G (1) >0.

Proof : IfG (1) <0, thenby (4.5), #i#*(#)#0 for all 1#i#g, which leads to C (###+1)-#(###)-####in(1+#i/(#T##))+ln(Mi-ni)#0 Using the facts that (1+1/x)x<e, (1+1/x)x+1>e, and (1-1/x)x-1>e-1 for any x>0, we obtain This is contradictory to condition (4.9).

For g=l, (4.9) becomes ln (n) > (X1+X2+###+Xn)/n. Hence, when the mean of the observed transposon insertions is less than the log of the number of nonzero observations, the vector n can not be the ML estimator of Ñ and there must be truncated observations from nonessential genes.

THEOREM 2: foralli&num j (4. 10) Proof : By definition in (4.5), for any 0<x<###. Hence #(###+1)-#(###) is an increase function of ###. Using this result, we have <BR> <BR> <BR> <BR> <BR> <BR> A, #*(#)-#i#*(#-#j)=<BR> <BR> <BR> <BR> <BR> <BR> (#(###+1)-#(###))-(#(###)-#(###-1))<BR> <BR> <BR> <BR> <BR> <BR> -####ln(1_#i/(#T##))+####ln(1+#i/(#T##-#j))<BR> THEOREM 3: Under (4.9), for any 1#j#g and 1#k#K*, with K* = max{k*#0:G(k)#0 for all 0 # k # k*}, <BR> <BR> <BR> <BR> <BR> <BR> if # = ##k -#j#{nj#Nj#Mj}, then<BR> <BR> <BR> <BR> <BR> <BR> <BR> <BR> <BR> <BR> <BR> <BR> <BR> <BR> <BR> <BR> <BR> <BR> <BR> 3 (##k)>#*(##k-#j) (4.11) Proof : This is obviously true when k=l. Assume (3.11) is true for integers 1,2,..., k.

For integer k+1, we have <BR> <BR> <BR> <BR> <BR> <BR> #*(##(k+1)-#j)-#*(##k)<BR> <BR> <BR> <BR> <BR> =[#*(##(k+1)-#j)-#*(##k-#j)]+<BR> <BR> <BR> <BR> <BR> <BR> [#8(##k-#j)-#*(##k)] < [3* (Me (k+1)-#j)-#*(##k-#j)] By theorem 2, #(##(k+1)-#j)-#*(##k-#j)<#*(##(k+1))-#(##k) Therefore #*(##(k+1))>#*(##(k+1)-#j) Combining theorems 1-3, we obtain THEOREM 4: If the likelihood function defined in (3.7) has an unique solution, the ML estimator of N is: #=##K* (4.12) [0163] Theorem 3 guarantees that the trajectory of the searching algorithm follows the shortest path in the sense that a reversal of a previous move (that is, removal of a previously added nonessential gene of any gene size) at any later state will result in a loss of likelihood. This property is illustrated in Figure 4 which shows the trajectory of the searching algorithm projected in a subspace spanned by two different gene sizes.

For illustration purpose, genes are grouped into 143 groups by grouping genes with similar sizes together to increase the length of the trajectory. As indicated in the plot, at any state, moving backwards in any direction results in a loss of likelihood. Figure 13 shows more trajectories projected in different subspaces.

[0164] Now we need to demonstrate that the likelihood function (3.7), which is defined in a high dimensional discrete space, has an unique solution. This can be established if the same estimations are obtained from different initial values. Since the initial values can be any value between the observation ñ and the total number of genes M, we need to extend the searching algorithm (4.7) as follows: For any initial value {N'' : nj<Ni°<Mj fori= 1, 2,---, g} and any integer k with <BR> <BR> <BR> <BR> #0#0=#0,<BR> <BR> <BR> <BR> <BR> 0#k####-##0# such that #j0#l={30~#i:#i#*(#0)##f#*(#0) for all j#i}, and (4.13)<BR> <BR> <BR> <BR> <BR> #0#k=(#0#(k-1))#1 for k#2.

The likelihood gain function is extended similarly as G (0) =0 and <BR> <BR> <BR> <BR> <BR> <BR> G (k)=#*(#0#k)-#*(#0#(k-1)) (4.14)<BR> <BR> <BR> <BR> <BR> <BR> <BR> <BR> <BR> <BR> <BR> <BR> <BR> <BR> <BR> <BR> <BR> <BR> <BR> for 1#k####-##0#.

[0165] Algorithm (4.13) preserves all the properties of algorithm (4.7) and it searches the ML estimator the same way as that of algorithm (4.7) with two exceptions. Unlike algorithm (4.7), which uses ñ as initial values of N and at each iteration, the number of nonessential genes is increased by one in gene groups of size bi to find the maximum likelihood gain, this algorithm uses ? as initial values of N which can be greater than the ML estimator. Therefore, at each iteration, the number of nonessential genes in a group with size ces ; can be either increased or decreased by one such that the likelihood gain is maximum.

[0166] Randomly selected initial values ? were used for data with grouped gene sizes and data with exact gene sizes. The estimations of all parameters are exactly the same and the final likelihood for all initial values ? are exactly the same as indicated in Figure 14, which plots twenty seven different initial values ouf ?. The line in the far left represents the likelihood when Ñ° = n, and the lines in the middle are randomly selected. Figure 15 is the trajectory projected into a subspace spanned by two gene sizes. Each circle represents the projection of a different initial value Ñ°. Regardless of the initial values, the trajectories all converge to the ML estimator.

D. ANALYSIS OF PSEUDOMONAS AERUGINOSA DATA 1. Multivariate Model with Exact Gene Sizes [0167] The data considered here consist of observations from 5570 genes in 881 different sizes, ranging from 72 to 16884 DNA base-pairs. Distribution of gene size is severely skewed to the right as indicated in Figure 10. For many sizes, especially for sizes smaller than 200 or greater than 2000 DNA base-pairs, there is only one gene in a given size and the observation of transposon insertions for small genes are usually truncated. Since all genes are modeled simultaneously in a single model with a prior y enforcing the essentialness of a gene being independent of its size, the sparseness of the data does not impose limitations on the computation. However, as discussed in the next section, the prior may play a dominating role for small genes where data are sparse.

The estimations of y and A, together with the 95 percent BCa confidence intervals are presented in Table 6 and the estimation of Ñ is presented in Figure 16.

Table 6. Parameter Estimation of y and Estimate Bias SE BCa Confidence Intervals y 0. 8434 3. 942x10-3 9. 893x10-3 (0.818, 0.859) 2. 547x10-3-1. 027x10-5 4. 392x10-5 (0.00247, 0.00264) Here the bias and standard error are estimated with bootstrap 2. Multivariate Model With Grouped Gene Sizes [0168] The prior y plays an important role in enforcing the fact that the essentialness of a gene is independent of its size. It also made possible to estimate the number of essential genes where data are very sparse. However, for small genes where data are extremely sparse, the prior y becomes the dominating source of information. In order to moderate the dominance of the prior on small genes with sparse observations, we grouped the genes into 143 groups according to their sizes, using the median size of each group as the gene size. Table 7 is a sample of estimated JV based on grouped and exact gene sizes. In the table, m is the number of unique sizes in each group ; N1 is estimated using grouped data and N2 is estimated using ungrouped data.

Table 7: Estimated N with Grouped and Exact Gene Sizes Gene size m M x N Ni N2 [72,120] 6 7 3 2 6 7 (120,150] 4 7 3 2 6 7 (150,160] 3 7 2 2 6 7 (160,170] 2 8 0 0 7 7 (170,180] 3 9 1 1 8 8 (180,190] 3 9 1 1 8 8 (190,200] 3 12 4 4 11 11 (200,210] 4 27 7 7 23 24 (210,220] 3 19 7 5 16 17 ... ... ... ... ... ... ... ...

[0169] We see that here N2 # N1. However, this is true only for data in the above tabl where the ungrouped data are extremely sparse and most of the data are truncated. The estimated proportion of non-essential genes, y, is actually larger for grouped data which is presented in Table 8. Grouping genes with similar sizes reduces the sparseness of the data and consequently, the dominance of the prior. Another obvious advantage of grouping is dimension reduction of the parameter space, and therefore, drastic reduction of computation time. Of cause, such grouping introduces another source of variation, and the algorithm could be unrobust against different grouping. In our study, however, different grouping resulted only in slight difference in estimates.

3. Conditional Maximum Likelihood Estimates For a given gene size Aj, the likelihood function (3.4) can be written differently as [0170] Here f (. ,. ) is defined in (3.1), and xj,1*,xj,2*,###,xj,nj* are the nj nonzero observations from Nj genes of size 8j. Assume there are g different gene sizes, the likelihood function can be written as with [0171] Assuming the number of observations nj for each gene size 8j being fixed, we can obtain the conditional maximum likelihood estimate of X by maximize L2 as nj<BR> Equation (5.3) reduces to equation (4.2) if we estimate nj by Nj= .<BR> <P> 1-e-##j The proportion of truncated nonessential genes can be calculated as p=P (x=O} nonessential) = F26dF (a). (5. 4) n [0172] Here Q is the set of nonessential genes, which can be approximated by the set of all untruncated genes.

Therefore, # =n/M+ # (5.5) Estimations from the three approaches are very similar as shown in Tabe 8. If the primary interest is to estimate X and y, the conditional MLE approach has the advantage of simplicity. However, in estimating #, this approach omitted information of M, and y is estimated separately after k is estimated. Another obvious limitation of this approach is that it can only estimate ###, the total number of nonessential genes by 11 ñ 11/Y. The estimation of Ñ by jilp is not reasonable because even though y is independent of gene size, we can not assume the proportion of non-essential genes in different sizes being the same as shown in Figure 17.

Table 8. Estimates of y and k with the Three Approaches Estimates Bias SE 95% BCa Confidence intervals Multivariate Model with Y 0.843 3. 942x10-3 9. 893x10-3 Exact Gene Sizes (0. 818,0. 859) X 2. 547x10-3-1. 024xlO-5 4. 320x10-5 (2.473, 2.642) x10-3 Multivariate Model with Grouped Gene Sizes y 0.853 7. 221xlO 8. 051x10-3 (0.835, 0.867) 2. 524x10'3 2. 803x10-6 4. 063x10-5 (2.451, 2.610) x10-3 Conditional Maximum Likelihood Estimates y 0.828-7. 621x10-5 7. 273x10-3 (0.815, 0.843) 2. 539x10-3 9. 713x10-7 4. 058x10-5 (2.455, 2.618) x10-5 E. DISCUSSION OF ONE DIMENTSIONAL CASE [0173] When the model does not depend on gene size, which can happen for example, when we study a subset of genes with a fixed size, or in other settings where the distribution is identical, model (2.6) reduces to (2.5). Blumenthal, Dayhiya, and Gross (1978) studies estimations of complete sample size from an incomplete Poisson sample using conditional, unconditional, and modified maximum likelihood functions.

The modified likelihood estimation weights the likelihood function and maximizes it. This approach is similar to providing priors to ? and N. Table 9 presents four types of estimations of N using data randomly selected from the 143 grouped genes. Here M and n are number of genes and number of genes with at least one observed transposon insertions. Nm-b is a subset of Ni in Table 7, which is estimated using model (2.6) with grouped data; Nb is estimated with model (2.5) ; Nc and Nu are conditional and unconditional estimates of N as described in Blumenthal., Dayhiya, and Gross (1978).

Table 9: Comparison of Estimations with Different Methods Gene size M n Nm-b Nb Nu Ne [72,120] 7 2 6 2 3 2 (400-410] 44 31 40 37 36 36 (430-440] 46 22 38 25 25 26 (470-480] 80 42 66 57 56 57 (500-510] 54 30 45 39 39 39 (610-620] 47 29 39 33 33 34 (640-650] 54 35 45 44 43 43 (710-720] 50 35 42 41 41 41 (750-760] 56 37 46 39 46 40 (770-780] 61 43 51 53 52 52 (910-920] 60 47 52 53 53 52 (980-990] 57 45 49 52 51 51 (1050-1100] 137 107 115 117 115 117 (1200-1250] 129 100 106 106 106 106 (1400-1450] 121 110 111 112 112 112 (2100-2150] 23 20 20 20 20 20 [0174] We see that the estimations from the three univariate models are very similar.

For fairly large genes, estimations from the multivariate model are similar to those of the univariate models. However, for small genes with high truncation rate, estimations from the multivariate model are larger than estimations from the univariate models. In the univariate models, only the information related to a particular gene size is used and the estimations are obtained separately for each gene size. This approach tends to underestimate N for small genes with sparse observations. The multivariate model uses a prior to enforce the fact that the essentialness of a gene is independent of its size and maximizes the likelihood jointly for all genes. Therefore, it alleviates the underestimation of N for small genes with high truncation rate.

Example 2: IpxC [0175] Lipid A constitutes the outer layer of the outer membranes of gram-negative bacteria and is essential for bacterial growth. This makes all the enzymes involved in the biosynthesis of this molecule essential for bacterial growth, and therefore ideal targets for drug design. A series of synthetic molecules was previously identified that inhibited the first committed step in lipid A biosynthesis. Onishi H. R. , B. A. Pelak, L.

S. Gerckens, L. L. Silver, F. M Kahan, M-H Chen, A. A. Patchett, S. M. Galloway, S.

A. Hyland, M. S. Anderson, and C. R. H. Raetz. 1996. Science. 274: 980-982. This step is catalyzed by a unique deacetylase (UDP-3-O-[R-3-hydroxymyristoyl]-GlcNAc deacetylase), LpxC.

[01761 UDP-3-0- [R-3-hydroxymyristoyl]-GlcNAc deacetylase (LpxC) is a deacetylase that catalyzes the first committed step of lipopolysaccharide (LPS) biosynthesis in gram negative bacteria. This is the second step following the first acylation of N-Acetylglucosamine (GlcNAc). This enzyme functions to deacetylate the UDP-3-0-[R-3-hydroxymyristoyl]-GlcNAc. This step was shown to be essential for growth in E. coli wherein a point mutant (envol) expresses an LpxC protein that has reduced activity. Beall B. and J. Lutkenhaus, 1987. Sequence analysis, transcriptional organization, and insertional mutagenesis of the envA gene of Escherichia coli. J.

Bacteriol. 169: 5408-5415. A 30% reduction in the amount of LPS on the cell wall of such mutants results in hypersensitivity to antibiotics. Attempts to create null mutants in lpxC were unsuccessful in a number of pathogenic bacteria, indicating that inhibitors of LpxC would be effective antibiotics for a number of gram negative organisms.

[0177] Previously identified inhibitors are chiral hydroxamic acids that had unique hydrophobic aromatic moieties, and were suspected to bind a metal in the active site of the deacetylase. The most potent inhibitor, L-161,240, displayed a minimal inhibitory concentration of about 1 microgram per milliliter against E. coli, caused three logs of bacterial killing in 4 hours, and cured mice infected with a lethal intraperitoneal dose of E. coli. Considering the very high degree of homology between the E. coli and P. aeruginosa enzymes, it was initially presumed that an inhibitor of the E. coli enzyme might also inhibit the P. aeruginosa enzyme. However, this molecule inhibited LpxC from P. aeruginosa only at very high concentrations, and even then it did so poorly. It had no effect on bacterial growth in this organism. Thus, there was some question as to whether the IpxC homologue had the same function in P. aeruginosa, and whether it was essential to P. aeruginosa given its decreased sensitivity to the L, 161,240 inhibitor.

[0178] Nevertheless, P. aeruginosa IpxC was one nucleic acid identified as being unable to accommodate a transposon insertion in the library depicted in Table 1 (PA4406). To test the essentiality of P. aeruginosa lpxC, we first tested the sensitivity of P. aeruginosa transformants expressing E. coli LpxC following a"promoter swap" integration. Using this technique, we completely shut off expression of the native P. aeruginosa lpxC, while expressing only the E. coli enzyme encoded on a plasmid. This strategy resulted in a P. aeruginosa mutant that was more sensitive to L-161,240. This suggested that the E coli IpxC gene was substituting for the function of the P. aeruginosa gene, and moreover, that there were no duplicate functional homologues in P. aeruginosa that were active in the absence of lpxC.

[01791 Materials. Pseudomonas aeruginosa PAO1 was grown at 37°C in Luria- Bertani (LB) broth (Difco) or plated on sheep blood agar (Remel). Tetracycline at 100 u. g/ml in LB media was used to maintain the selection of the integrated plasmid pBEMlO in PAO1. LB broth or agar with 10 ug/ml of tetracycline was used for <BR> <BR> <BR> <BR> growing E. coli DH5a (Gibco BRL) and E. coli S-17 transformants. Plasmids pPS72 and pBADHisB were from Promega and Invitrogen, respectively. EDTA, bis-tri buffer, sucrose, arabinose, and DMSO were purchased from Sigma as Ultrapure agents. Yeast extract and Tryptone were obtained from Difco. Restriction enzymes, and T4 DNA Ligase, and their reaction buffers were from New England Biolabs. Polymixin B nonapeptide was from Sigma. The antibiotics, tetracycline, ampicillin, carbenicillin, gentamicin, and kanamycin were all purchased from Sigma. DNA and deduced amino acid information were analyzed using a family of programs included in the Dnastar package. BLASTP was used to search for amino acid'similarities among a host of protein databases available on-line through the National Library of Medicine (USA).

Altschul, T. F. , W. Gish, W. Miller, E. W. Myers, and D. J. Lipman. 1990. Basic local alignment tool. J. Mol. Biol. 215: 403-410.

[0180] DNA manipulations. Standard recombinant DNA procedures were used.

Sambrook, J. , E. F. Fritsch, and T. Maniatis. 1989. Molecular cloning : a Laboratory Manual, 2nd Edition. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory.

Primers were designed to the N-and C-terminal regions of the E. coli or P. aeruginosa lpxC gene that encompassed only the coding region and included NdeI and EcoRI restriction sites for subsequent cloning. For the E. coli gene the primers were (5'- GGGAATTCCATATGATCAAACAAAGGACACTTAAACGT-3'and 5'- CCGGAATTCTTATGCCAGTACAGCTGAAGGCGCT-3') and for P. aeruginosa gene they were (5'- GGGAATTCCATATGATGATCAAACAACGCACCTTGAAGAACAT-3'and 5'- CCGGAATTCCTACACTGCCGCCGCCGGGCGCATATAG-3'). These primers were used in a polymerase chain reaction (PCR) containing either P. aeruginosa genomic DNA (10-50 pg) or plasmid pKD6 containing the E. coli lpxc gene (1. 0 ug) as template (Sorensen, P. G. , J. Lutkenhaus, K. Young, S. S. Eveland, M. S. Anderson, and C. R. H. Raetz. 1996. Regulation of UDP-3-O- [R-hydroxymyristoyl]-N- acetylglucosamine deacetylase in Escherichia coli. The second enzymatic step of lipid A biosynthesis. J. Biol. Chem. 271 (42): 25898-25905). The lpxC genes were amplified using Pwo DNA polymerase (Roche) in a 100 p^1 reaction mixture containing 200 uM concentration of each dNTP and 0.5 gM concentration of each primer for 30 cycles (94°C denaturation, 55°C annealing, and 72°C polymerization (according to the manufacturer's instructions). The PCR products were purified with the Qiaquick PCR Purification Kit from Qiagen (according to the manufacturer's instructions) and digested with NdeI and EcoRI restriction enzymes at sites introduced by the primer sequences. Bands of the correct sizes predicted for the LpxC genes were separated by gel electrophoresis, and the excised DNA purified using the Qiaquick Gel Extraction Kit from Qiagen (according to the manufacturer's instructions). The purified DNA was ligated into the T7 expression vector (Studier, F. W. , A. H. Rosenberg, J. J. Dunn, and J.

W. Dubendorff. 1990. Use of T7 RNA polymerase to direct expression of cloned genes.

Methods Enzymol. 185: 60-89) pET21b (Novagen), that had been cut in the multiple cloning site with the same enzymes, transformed into DH5a and plated on LB agar containing ampicillin (250 pg/ml). The resulting clones had their DNA sequenced to confirm the fidelity of the PCR reactions before it could be transferred into the expression strain. Subcloning of these fragments into other vectors was carried out as needed for expression in various backgrounds. These included pEXl 8T (cbR) for allelic exchange mutagenesis in P. aeruginosa (Schweizer, H. P and T. T. Hoang. 1995.

An improved system for gene replacement and xylE fusion analysis in Pseudomonas aeruginosa. Gene 158 (1) : 15-22), pDNl9 (tetR) for low copy number complementation of E. coli JBK-1 (Nunn, D. , S. Bergman, and S. Lory. 1990. Products of three accessory genes, pilB, pilC, andpilD, are required for biogenesis of Pseudomonas aeruginosa pili. J. Bacteriol. 172 (6): 2911-2919), and pUCP30T (gmR) for P. aeruginosa'promoter swap'mutant complementation (Schweizer, H. P. , T. R.

Classen, and T. Hoang. 1996. Improved methods for gene analysis and expression in Pseudomonas. In: Nakazawa, T. , K Furukawa, D. Haas, S. Silver. (Eds. ) Molecular Biology of Pseudomonas. American Society for Microbiology, Washington, DC. pp.

229-237).

[0181] Construction of pBEMlO and'promoter swap'mutagenesis. Plasmid pPW101 was made by ligating oriT, the region that encodes conjugative plasmid transfer, into pSP72 (Promega). oriT had been amplified from plasmid pEXlOOT (Schweizer and Hoang, 1995, supra) with an introduction of an Ndel and an AatII restriction sites. To create the lpxC'promoter swap'vector, pBEMlO, the following different DNA pieces were amplified and sequentially ligated into pPW101. These included the tetracycline resistance marker (tetR) from plasmid pUCP26 (Olsen, R. H., G. DeBusscher and R. R. McCombie. 1982. Development of broad-host-range vectors and gene banks: self-cloning of the Pseudomonas aeruginosa PAO chromosome. J.

Bacteriol. 150: 60-69), the araBAD promoter from the plasmid pBAD HisB (Invitrogen) with an altered ribosome binding site (rbs) (Guzman, L. M. , D. Belin, M.

J. Carson, and J. Beckwith. 1995. Tight regulation, modulation, and high-level expression by vectors containing the arabinose pBAD promoter. J Bacteriol. 177 (14): 4121-4130), the araC gene, also from pBAD HisB (Lee, N. 1980. Molecular aspects of ara regulation. In The Operon. J. H. Miller and W. S. Reznikoff, eds. Cold Spring Harbor, NY. Cold Spring Harbor Laboratory, pp. 389-410 ; and Schleif, R. S.

1992. DNA looping. Ann. Rev. Biochem. 61: 199-223), and the first 340 base pairs of the P. aeruginosa LpxC gene. The tetR marker was amplified using a forward primer that introduced a BglII site (5'-AGATCTCAAGGGTTGGTTTGCGCA-3') and a reverse primer that introduced an EcoRI site (5'- GAATTCTAATTCTCATGTTTGACA-3'). The araBAD promoter and araC gene were amplified as one piece from the pBAD HisB vector. The forward primer introduced an Mol site (5'-CTCGAGGCATGCATAATGTGCCTGTC-3') and the reverse primer introduced a HintII site (5'- AAGCTTCTCCTGTTAGCCCAAAAAAACG-3'). The rbs was altered from its original AGGAG to CTTCT. The following primer set was used to make these changes and introduced an upstream BssHII site (5'- GCGCGCGGACGAAAGTAAACCCAC TGG-3') and a downstream HindJII site (5'- AAGCTTATTCAGAAGGTTAGCCCAAAA AAACGGG-3'). The first 340 bases of PAO1 IpxC were amplified from PAO1 genomic DNA. The forward primer introduced a HindII site (5'-AAGCTTATGATCAAACAACGCACCTT-3') and the reverse primer introduced anXbaI site (5'-TCTAGAAGCGCTGCCATCCATGATCGG-3').

These pieces were then ligated into pPWIOl to form the final product, pBEMlO, which was used for the'promoter swap'mutagenesis of lpxC. The'promoter swap'scheme is a homologous recombination strategy, whereby transformation of pBEMlO into P. aeruginosa removed the native lpxC promoter and placed the tightly regulated araBAD promoter upstream of the chromosomal copy of lpxC, allowing modulation of its expression by the use of a simple sugar, arabinose (Figure 3). In the absence of arabinose the IpxC was effectively shut off, and expression was inducible by addition of arabinose. Such mutants were selected in the presence of arabinose, and if lpxC is essential, these mutants would not be viable in media that is not supplemented with arabinose, but fully capable of growth in the presence of arabinose.

[0182] Growth curves. Bacterial cultures were prepared by diluting stationary phase overnight cultures to an OD600 of 0. 1 in 5 ml of LB. The inhibitor, L-161,240, was resuspended in DMSO to a final concentration of lOmg/ml and added to the bacterial cultures to a final concentration of 50 vug/ml or 10 pLg/ml. In the samples without inhibitor, DMSO was added to keep the final concentration of DMSO equivalent between samples. The cultures were incubated with shaking and 0.8 ml was taken for OD600 readings over the course of the experiment. DH5a, PA01, and PA0200 (Schweizer, H. P. 1998. Intrinsic resistance to inhibitors of fatty acid biosynthesis in Pseudomonas aeruginosa is due to efflux: application of a novel technique for generation of unmarked chromosomal mutations for the study of efflux systems.

Antimicrob. Agents Chemother. 42: 394-398) were all grown at 37°C. In the cases where temperature sensitive JBK strains were being assayed, the cultures were grown at 42° C for both the overnight and the time course cultures.

[0183] Outer membrane permeabilization. Polymixin B nonapeptide (Sigma) was prepared as a suspension in DMSO at 3 mg/ml final concentration. Erythromycin and Tetracycline were resuspended in DMSO to a final concentration of 250 mg/ml and 125 mg/ml, respectively. L-161,240 was prepared as above in DMSO to a final concentration of 1 Omg/ml. These DMSO antibiotic solutions were individually added to LB to the appropriate final concentration and mixed. Polymixin B nonapeptide was then added to the appropriate samples and mixed. DMSO was added to each sample to keep the final concentration of DMSO equivalent between samples. A stationary phase overnight culture of PA01 was added to each sample to bring the final concentration to 0.1 OD600. Samples were removed for OD600 determinations every 1-2 hours for 6.5 hours and the data from these time points were plotted.

[0184] MIC determinations for'promoter swapped'mutants. Single colonies of DHSa, PA01 and each promoter swap strain were picked and grown in LB at 37°C with shaking for approximately 4 hours. Assuming that an OD600 reading of 1.0 is equivalent to 109 cells/ml, dilutions were made of all cultures to 5x105 cells/ml. 200 of each diluted culture was added to each well where a two-fold serial dilution of inhibitor had been placed. The 96-well plates were incubated at 37°C overnight and their OD600 determined using the Spectramax Plus (Molecular Devices) plate reader.

[0185] To confirm the effect of the arabinose-sensitive promoter in regulating the LpxC expression in the swapped mutants, MIC determinations were performed as above, except that arabinose was added to induce expression of the chromosomal locus and override the effects of the plasmid borne lpxC. In this case the stationary-phase overnight bacterial culture was diluted to 5x105 cells/ml in LB containing Arabinose to a final concentration of 0.2% (a 20% stock made up in water).

RESULTS AND DISCUSSION: [0186] Homology between the E. coli and the P. aeruginosa LpxC enzymes. Using protein analysis software, this study and others have compared the deduced amino acid sequence of LpxC from both E. coli and P. aeruginosa (Hyland, S. A. , S. S. Eveland, and M. S. Anderson. 1997. Cloning, expression, and Purification of UDP-3-O-Acyl- GlcNAc Deacetylase from Pseudomonas aeruginosa : a metalloamidase of the lipid A biosynthesis pathway. J. Bacteriol. 179 (6): 2029-2037). This comparison revealed 82% similarity and 57% identity shared between the two sequences. This homology was found over the entire length of the protein sequence (data not shown). Significant homology with other known acetyl-or acyltransferases was not found, suggesting that LpxC is unique among acetyltranferases. The two proteins also share a total of five fully conserved Histidine residues that are presumed to be responsible for the zinc metal cofactor coordination. It was therefore expected that an inhibitor that functions by chelating the metal cofactor away would affect both enzymes similarly.

[0187] LpxC is essential for growth in P. aeruginosa. Since the hydroxamate inhibitor was effective in preventing growth of E. coli, but completely ineffective against P. aeruginosa, there was a possibility that LpxC was not essential in P. aeruginosa This could be as a result of the presence of another enzyme that catalyzed a similar function. If that were the case, elimination of the LpxC function should be possible without inhibiting bacterial growth. A thorough analysis of the P. aeruginosa genome sequence revealed only one LpxC homologue. An attempt to disrupt the function of this LpxC homologue was made by conjugating wild type PAO1 with a suicide vector (pEXl 8T) carrying lpxC whose BamHI-SalI fragment had been replaced with a gentamicin cassette. However, P. aeruginosa null mutants could not be established by this method. In several attempts a few gentamicin resistant trans- conjugants were obtained, but in all these cases allelic replacement of the chromosomal LpxC by the defective copy had not occurred. Instead, a gene duplication had occurred, placing the suicide vector and the disrupted copy next to the wild type allele (data not shown). This could be demonstrated by the carbenicillin resistance and sucrose sensitivity acquired by these trans-conjugants, both of which are encoded on the suicide vector. These data indicated a strong negative selection for the sought after disruption of IpxC suggesting that lpxC is essential for growth. To confirm this, an experiment was carried out whereby the trans-conjugants were transformed with either lpxC on a low copy, replicating vector, or vector alone. In 100% of lpxC transformants, resolution of the gene duplication as demonstrated by the loss of carbenicillin resistance and sucrose sensitivity was observed, as opposed to no such resolution among those transformed with vector alone. These results suggested that the wild type genomic allele could be disrupted if a functional copy was present on the transforming plasmid.

10187] In another attempt at demonstrating essentiality of LpxC in P. aeruginosa, the 'promoter swap'strategy as described in materials and methods was carried out.

'Promoter swapped'pseudomonas mutants were fully capable of growth in the presence of arabinose when the arabinose sensitive lpxC promoter was turned on, but completely incapable of growth in the absence of this inducer. This further confirmed that in P. aeruginosa, just as in E. coli, LpxC is essential for growth.

[0188] E. coli expressing LpxC from P. aeruginosa is more resistant to L-161, 240. The E. coli strain JBK-l/pKD6 contains the chromosomal lpxC gene disrupted with a kan element and a wild type copy of E. coli lpxC on the temperature-sensitive replicon pKD6. The strain was constructed as described by Sorensen et al., 1996. Since lpxC is essential for growth, this strain is not viable at 42° C because the functional copy is on the temperature sensitive replicon. Transforming JBK-l/pKD6 with IpxC from either E. coli or P. aeruginosa on a non-temperature-sensitive replicon (pKD19, TetR) and selecting at 42°C, produced transformants that were viable at 42°C, tetracycline resistant, and kanamycin sensitive. This result indicated that lpxC from P. aeruginosa could be expressed in the E. coli background, and was capable of substituting for the missing chromosomal copy. An unexpected result was that whereas the JBK-1 carrying the LpxC copy from E. coli was still sensitive to killing by a slightly higher concentration of L-161, 240, the JBK-1 carrying the LpxC copy from P. aeruginosa was resistant to up to 50, ug/ml, about 50 times above the MIC of the wild type organisms (data not shown). This suggested that the P. aeruginosa enzyme was uniquely resistant to this inhibitor. It also meant that this resistance was the reason for the failure to inhibit growth of P. aeruginosa, and not reduced permeability, or efflux or modification of drug by the pseudomonal enzymes. This, in turn, suggests that a program designed to search for inhibitors for the pseudomonal enzyme should be based on screening directly on that enzyme, and not the surrogate enzyme from E. coli.

[0189] L-161,240 is a substrate for the major drug efflux pump of P. aeruginosa.

The completed P. aeruginosa genome reveals genes for at least nine homologous, multicomponent, multidrug efflux systems (Stover et al. , 2000, Complete genome sequence of Pseudomonas aeruginosa PAO1, an opportunistic pathogen, Nature 406: 959-64). However the only one that is constitutively expressed to a high degree in the wild type strains is MexAB-OprM (Kohler, T. , M. Michea-Hamzehpour, and U. Henze.

1997. Characterization of MexE-MexF-OprN, a positively regulated multidrug efflux system of Pseudomonas aeruginosa. Mol. Microbiol. 23: 345-354). Therefore, mutants of this efflux system can be used to evaluate the consequences of diminished efflux pump activity. These mutants would be expected to be highly sensitive to a number of antibiotics. Such a mutant, PAO 200, has been isolated (Schweizer, 1998, supra), and whereas it shows a higher level of sensitivity to a number of antibiotics (Westbrock- Wadman, S. D. R. Sherman, M. J. Hickey, S. N. Coulter, Y. Q. Zhu, P. Warrener, L. Y.

Nguyen, R. M. Shawar, K. R. Folger, and C. K. Stover. 1999. Characterization of a Pseudomonas aeruginosa Efflux Pump Contributing to Aminoglycoside Impermeability. Antimicrobial Agents and Chemotherapy. 43 (12): 2975-2983), it was not more sensitive to L-161,240 (Figure 4). This suggests that this drug compound is not a substrate for this efflux system in P. aeruginosa.

[0190] P. aeruginosa is not less permeable to L-161,240. Low permeability of the outer membrane is a major contributing factor to the observed high levels of intrinsic drug resistance in P. aeruginosa (Nikaido, H. 1998. The role of outer membrane and efflux pumps in the resistance of gram-negative bacteria. Can we improve access? Drug Resistance Updates. 1: 93-98). This low permeability is due to the fact that P. aeruginosa lacks the homolog of the relatively efficient, trimeric porins like OmpF. P. aeruginosa has, instead, OprF, the OmpA homolog, which produces channels only when it is folded into a rare conformation, and only a small fraction of these channels occurs in the open conformation. As is usually the case with P. aeruginosa it was assumed that the reason L-161,240 was ineffective against P. aeruginosa was the lack of permeability of the outer membrane to this inhibitor. Polymixin B nonapeptide (PMBN), a derivative of Polymixin B that lacks the fatty acid tail, is capable of binding to the polyanionic LPS molecules and disrupting the bilayer structure, thus increasing the permeability of the outer membrane. PMBN has been used this way to permeabilize the outer membrane of many gram-negative bacteria (Vaara, M. and T. Vaara. 1983.

Sensitization of gram-negative bacteria to antibiotics and complement by a nontoxic oligopeptide. Nature 303: 526-528), including P. aeruginosa (Vilianen, P. and M.

Vaara, 1984. Susceptibility of gram-negative bacteria to polymixin B nonapeptide.

Antimicro Agents Chemother. 25: 701-705) and effectively sensitize them to lipophilic antibiotics. Unlike the acylated polymixin B, PMBN is not cidal. In order to determine the effect of outer membrane exclusion of L-161, 240, we exposed P. aeruginosa to PMBN in combination with L-161,240, and with other lipophilic antibiotics as positive controls. Whereas PMBN lowered the MIC of tetracycline for P. aeruginosa more than 16 fold, the sensitivity towards L-161, 240 remained unchanged (Figure 5). This, together with the E. coli expression data indicated that permeability was not a major factor causing the inability of L-161,240 to inhibit pseudomonal growth.

[0191] P. aeruginosa expressing only E. coli LpxC is more sensitive to L161-240 than wild type. Using the'promoter swap'technique as described in the methods, it was possible to replace expression from the wild type chromosomal copy of P. aeruginosa lpxC, with expression solely from a plasmid borne copy. For this experiment,'promoter swapped'P. aeruginosa was transformed with either vector containing P. aeruginosa lpxC ("PA Swap &num 1"), or vector containing E. coli lpxC ("PA Swap &num 2'). The transformants were then exposed to various concentrations of L- 161,240 for MIC determination. Transformants expressing the E. coli enzyme only were much more sensitive to the inhibitor compared to organisms expressing the P. aeruginosa enzyme (Figure 6). These transformants were sensitive enough to be comparable with the sensitivity seen in E. coli. Since the validity of this observation relied on the un-induced arabinose-sensitive promoter to shut down expression from the chromosomal copy of lpxC, it was necessary to demonstrate how effectively this happens. To do that, MIC determinations were performed as above, except that arabinose was added to induce expression of the chromosomal locus. For this experiment stationary-phase overnight bacterial cultures were diluted to 5x105 cells/ml in LB containing 0.2% arabinose. In this case all the transformants, regardless of what gene the vector contained, were resistant to killing due to the expression of the chromosomal copy of P. aeruginosa IpxC. This confirmed that certain intrinsic properties of the P. aeruginosa enzyme are resistant to inhibition by this hydroxamate inhibitor. It also confirmed that neither reduced uptake, efflux, nor modification of the inhibitor play a significant role in this observed resistance. Considering the very high similarity between the two enzymes, this finding was not expected.

[0192] But on further examination and analysis of existing data, it was possible to recognize some inherent differences that might explain this finding. Whereas both these enzymes share five conserved Histidine residues, the E. coli enzyme has two more Histidines that have no counterparts in the P. aeruginosa enzyme. This is an important difference because these residues are probably involved in the metal cofactor coordination. It was also observed earlier that whereas the E. coli enzyme is not sensitive to EDTA, the P. aeruginosa enzyme was significantly inhibited by as little as 2 u. M EDTA. Evidence that the E. coli enzyme is also a metalloenzyme is that the envol mutation, which has one of the conserved Histidines (His 19) replaced by a Serine, is sensitive to EDTA. It was because of these observations that these investigators suggested that the E. coli enzyme has a more stably bound metal than that of the EnvAl mutant protein, and thus it is less accessible to EDTA than the wild type P. aeruginosa enzyme. These observations suggest that the Histidine'patch'that is involved in the metal coordination is not similar between the two enzymes. It is conceivable therefore that since the inhibitor works by chelating the metal cofactor away from the enzyme, each'patch'has unique features that result in disparate reactivities towards the inhibitor. It is also important to consider the findings of Wyckoff et al. , 1998. Hydrocarbon rulers in UDP-N-acetylglucosamine acyltransferases. J. Biol. Chem. 273 (49): 32369-32372. These investigators found that LpxA, the first enzyme of lipid A biosynthesis, is very selective for the length of its acyl donor substrates. Whereas E. coli LpxA prefers R-3-hydroxymyristoyl-ACP to R- 3-hydroxydecanoyl-ACP, P. aeruginosa LpxA prefers the opposite. The products of the LpxA reaction therefore differ in the carbon chain length of their lipid moieties between the two bacteria. Since the product of the LpxA reaction is the substrate of the LpxC reaction, this observation suggests that the two LpxCs would have substrate binding pockets of different sizes to accommodate the different size substrate. That would, in turn, suggest that inhibitors that have to occupy that active site would be unique for each enzyme.

Examples 3-7: ispA, ispB, uppS, aroC, aroK, and metK [0193] Several more candidate genes from the HTTIM gene database were tested for essentiality using a single crossover knock-out strategy. The Pseudomonas genes targeted for knocking out were ispA, ispB, uppS, metK, aroC, and aroK. To attempt knock-outs, regions of about 300 bp were cloned into the vector pPW120. These regions were selected so that known active site residues (or highly conserved residues likely to be essential for enzyme function) would be separated after generation of a single-crossover knock-out. The regions were (numbering from the start codon): ispA, 283-594; ispB, 319-610, uppS, 103-402 ; metK, 415-732; aroC, 385-684; aroK, 175- 375.

[0194] The pPW120 vector carries an E. coli origin of replication, but not a Pseudomonas origin of replication, making it a suicide vector. It also carries an origin of conjugal transfer and antibiotic resistance genes for tetracycline and ampicillin. An E. coli donor strain (SM10) carrying the pPW120 knockout constructs was incubated with Pseudomonas strain PAO1 to allow conjugal transfer, and recombinants were selected by plating onto media containing tetracycline at 100 pg/mL and chloramphenicol at 10 j. g/mL. Pseudomonas recombinants will be resistant to this antibiotic mixture while wild-type PAO1 and the E. coli donor strain will be sensitive.

Aromatic amino acid recombinants (aroC and aroK) were then tested for auxotrophy by plating onto minimal media with and without phenylalanine, tryptophan, tyrosine, and folic acid at 100 jug/mL while maintaining tetracycline selection. The genes ispB, uppS and metK did not yield recombinants, demonstrating that they are essential genes in all media conditions, while ispA yielded slow-growing recombinants (suggesting that this gene may nevertheless be an"important"gene according to the invention).

[0195] For ispA, ispB, uppS, and metK, the conjugation procedure was also done in the presence of the complementing plasmid pBAD/HisP. This plasmid has both E. coli and Pseudomonas origins of replication, an antibiotic resistance gene for carbenicillin, and an arabinose-inducible copy of the full-length wild-type gene. In this way, recombinants with the chromosomal copies of ispA, ispB, uppS, and metK knocked out could be isolated since the vector copy would provide complementation.

[0196] The genes ispB, uppS, and metK are novel with regard to P. aeruginosa. The gene ispB (PA4569, ranging from 5116864 to 5117832 in the genome), has 67% similarity/52% identity to IspB in E. coli, and was assigned to the function class concerned with biosynthesis of cofactors, protein groups and carriers, and energy metabolism, with a confidence level of 2. It is thought to be involved in the pathway of ubiquinone biosynthesis.

[0197] The gene uppS (PA3652, ranging from 4091654 to 4090899), coding for undecaprenyl pyrophosphate synthetase, has 69% similarity/57% identity to the uppS gene in E. coli, and was assigned to the function class involved in biosynthesis of cofactors, protein groups and carriers, cell wall and capsule, with a confidence level of 2. It is separated by one gene (casa) from dxr, which is involved in the synthesis of isopentenyl diphosphate, a precursor of undecaprenol phosphate.

[0198] The gene metK (PA0546, ranging from 604896 to 603706) had never been characterized in P. aeruginosa, although it is 82% similar/72% identical to MetK in E. coli. The gene encodes methionine adenosyltransferase (adomet synthetase) which is involved specifically in methionine metabolism, and was originally assigned to a function class of amino acid biosynthesis and metabolism and central intermediate metabolism with a confidence level of 2.

Example 8: rrF [0199] The essentiality of the P. aeruginosa rrF (PA3653, ranging from 4092227 to 4091670) gene was tested using the promoter swap methodology disclosed herein. The N-terminus region (position 1-327) of the gene encoding the ribosome recycling factor (frr) was cloned into the plasmid vector pBEM10. A single crossover was constructed as described above for lpxC. Recombinants were unable to grow in the absence of arabinose, confirming the essentiality of this gene. The rrf gene encodes ribosome recycling factor, alternatively known as ribosome releasing factor, assigned to the functional class pertaining to translation, post-translational modification and degradation with a confidence level of 1. Although this gene was previously known in Pseudomonas aeruginosa, confirming the essentiality of known genes using the methods disclosed herein will reveal new utilities for such genes as targets for the identification and design of new antibacterial drugs. _- .", < _ _. w ;,,. , :. s s ! ! t' ' :-. a . fy : ; J'w : . aa'g . . r, r.. . s Y ...,..,.. .. .. d.. t 'Y'r .. Pid. n,.. e., ,' !", : s.. t, w/. y. - : 'PA0001 j 39J 9 tchrompsonjep) icationmiatpr protein ! : _P_A0004 9945822DNA g _rase subunit B, YrB : PA0006 9945824 conserved hypothetica) proteinj ! j/3sD""' 'PAOOOS""'9945826Jg) ycy)-tRNA synthefase beta chain) g) yS''"""""' '650-'9945827 fycyi-tRNAsynthetaseatpha chan ! g) yQ J---'"--"--- iPA0011J 45830 probabte'"'"""'"-.. - ! pA0015 JIMJlX. P° ! lst ! pr, .... L.. L. L'I. L" "J"II 1..--I"" ; PA0022 9945841 conserved hypotheticalNprotein , wyrdC ^ 'PA0026 9945846 hypothetical protein ! PAID033 9945854 hypothetical protein 'PA0035 9949945856 t ptophan synthase alpha chain _ trpA ! PA0038 9945859 hypothetical protein PA039__ T_9_945860 hypothetical protein 9945864 h pothetical protein ! PA0047 9945869 hypothetical pot (Lin (PA0052""'"9945875 hypothetica) protein ! "] LA005L 9945876 hypothetical protein t PA0054 9945877 conserved hypothetica) protein ! yjitJ PA0055 9945878 hypothetical protein H' ; PA0058 9945881 hypothetical protein PA0059 9945882 osmoticaOy inducibfe proteinpsmC ! osmC 1pua0060 9945883 conservpot in 'iPA0061 9945884 hypothetical protein , _. ... .. _. _......., _. __.... _. _.,. .., _. ...... iPA0065 9945889 hypothetical protein _945892 hKpotefii protein JPA0070"9945894 hypothetica ! proteinJ PA0070 9945894 hypothetical protein PA0080 9945905 hypothetical protein PA0082 9945907 hypothetical protein PA0094 9945920 hypothetical protein ip JPA0105 9945932 cytochrome c oxidase, subunit) !'pxB co !""' ! _. _ ; __ 0P994593_ _y9oh- ; 9 45936 h pothetical rotein ; 9945938 h pothetical protein _- , PA0113 9945940 probable cytochrome c oxidase assembly factc Y' . _... __. _. PA011414 9945941 conserved hypothetical protein Pua0115 9945942 conserved hypothetical rotein ; PA01_16 9945944 hypothetical protein i ; 3 PA0119 9945947 probable dicarboxylate transporter, _ 91P.-1. 9945948 j probabie transcriptipna) reguiatorH......... [I......'...... J........ 1111] 124 9945952 hypothetical protein PA0124 yp p m PA0125 9945953 hypothetical protein j PA0128 9945956 conserved hypothetica) protein ! "jphnA, 9945960 hypothetical protein . PA0133 1 9945962 probable transcriptional requlator iPA0135 9945964 hypothetical protein 9945969 alkyl hydroperoxide reductase subunit C'amp 'iPA0143" ; 9945973 prpbabtenudepsidehydrofase...... 11.."""" !". 1-.... 1...-1 ' ! PA0145 9945975'h 'jPA0149"J 9945980 iprobabiesigma-70 factor. ECFbfamity""'""""'J : PA01] 54 9945985'protocatechu3, 4-dioxqenase, aipha subURpcaG'--------............ i PA0159 9945991 ! probabie transcriptionaijregutator"'"","""'"""'"""''" : P_AO_1_67 ; _99_4_5999p_robable transcrip_tionai regula#or.... m, _M... _.. _.... _.........' 'PA0161 9945993 hypothetical protein y_-_. __. . _.a __. _. _. ___. _____.. _ PA0170""19946003 J hypothetical protein""""""""""" :"'------------'.-....--.--. j JPA0171'""'"9946004 J hypotheticai protein'"IL'''1 IjJIII""'""-"---"----i PAO 182 9946016 ! ! probable short-chain dehydrogenase asa . . _.. __.. _... _.. ___. __ : _. _. . _. _.. _____. __. _... _. _____. _. _.. ____. _..... __. _. ___. ___.. ___. _____. __.. __., _. __...... _... _. _. _.. _. _. ___.. _. _........ _...... _......... __... _...... _.... _..... _. _. _.. ___., 99460181 prob _oTponent of ABC trani3 PA0184 able ATP-binLirIQ PA0187 994602lihyp, in _. _. _w : _______. _..... _.. ___. _. _, .. PA0200 9946035 hypothetical protein LPA0202. 99460371 probable amidase PA0203 . °'-. ___.. robable bindin rot __ 9946038 f p g p ein component of ABC tr ; ; PA0204 9946039 probable permease of A_BC transporter f , PA020_5 _ :99946040, probable permease of ABC transporter _ PA0207 ! 9946042 proba. btetranscriptionatregujatpr"""'j''""j PA0209 9946045, conserved hyp thetical prote. in mdcb , PA0211 99460471 malonate decarboxylase beta subunit 1 mdcd PA021_3 __ ; Y999460491 hypothetical protein i, much PA0216 99460521 probable lmadm PA0233 j _ 9946071 probable transcriptional regulator PA0236 bable transcriptional regulator PA0238 1 99460761 hypothetical protein {PA0243"'9946082 J probabie transcriptionai regmator L-*" TL1---II--LIII LIIJ . PA0244'")'9946083 hypothetical protein"''T'J'"'L----I---. J iPA0244 I 9946083i hypothetical protein PA0245 3-dehydroquinate dehydratase aroQ2 laroD2 PA0250 9946090 ! conserved hypothetical PA0258" ! 9946098 jT) ypotheticai protein'"j'") PA0258 1 9946098 hypothetical protein 9946101 hypothetical protein P 264 9946105 hefical-ro-tein PA0273 99461151 probable MFS transporter 1PA0279 9946122 probabie transcriptionai regutatprj l5IIIIIIIIJ IPA0280 9946123 sulfate transport protein ic_ ______. _. _. ____. __..... _ ! PA0284 ; 9946127 hypothetical protein- _ 3 __ j ; PA0309 9946155 ! hypothetica ! protein J"'f , PA0311 9946157 hypothetical protein ! PA0320"T 9946167j conserved hypothetical protein iPA0332"T 9946180 hypotheticat protein] 'IL- PA0332 : w99461801 h othetical protein _ _ E iPA0332---646 ! 8bi hypothetical protein PA0339' (9946187 hypothetica) proJ.. L. II"'J"L--II-.-.-1 PA0341 J 9946190 protipoproteindiacytg ! ycery) transferase Jigt umpA PA0341y 'g946191 thymidylate synthase, _ y_ v PA0342 th A ; PA0350 j 9946200 dihydrofolate reductase olA tmrA __ PA0358 _j 9946208 hypothetical rotein _ __ ____ ___ ___ PA0362 9946213 ferredoxin [4Fe-4S) ifdxl ! PA0363 9946214iphpsphopantetheineadeny) y) trans lE hypothetical protein iPAa370__.. _.-.---9946222'. conserved hypothetical protein---....... . wHH-. __. _. __....... _.. _iyhhF... _. __...... __.. __. _...... __. _.. _.. _. _... _. _.... _. ! ; PA0373 9946225 signal recog nition particle receptor FtsY ftsy E PA0376 9946228 ! jsi'gma factor PA0377 9946229'hypothetical protein PA0380 9946232 ! conserved lpoth ! ical .......... PA0384 _g946237 . . - -_.... _.... _.... _.... _. _. _.. _.. _..... _. ___.... __.. ;'y.... __.. _^__. YPothetical protein __. _. _.. _. ____ _. __... _ __. ___ __. _. _.. ___. _.. PA0402 99462561aspartate carbamoyltransferase pyrb 9946257'transcriptional regulator PyrR pyrr PA04. 10. 4.--......,. !--. 9946258i conserved hypothetical protein iPA0405 i 9946259 ! conserved hypothetical protein IEPA101407 99462621glutathione synthetase lgshb PA0416 9946271 probable transcriptional regulator ichpd PA0416] 9946271 jprobabte franscriptionaf reguiatorL-J. 1-IIItchpD ["-"-"-- PA0422 1 994Q278 conserved hypothetical protein PA0427 9946283 outer membrane protein OprM precursor zou PA0433 9946290 hypothetical PA0442 9946300 hypothetical protein PA0443 1 9946301'probable transporter 46304 probable transposase PA0446 J 9946305 conserved hypothettca) protein ! PA0448'. _. !99946307 probable transcriptioiial _. PA0453 9946312 hypothetical protein PA0456 j 9946316 probabte coid-shock proteint"j r yp. e_n PA0466 i 9946327 hypothetical protein JITT'llj PA0475 9946337 probable transcriptional requlator --AO4-77-- 9946339 probable transcriptional regulator PA0477'9946339 p J j PA0479 9946341 probabie transcriptiqnai regutator L--" PA0488"1 9946351 conserved hypothetical protein Im PA0489] 9946352 probabie phosphoribosyt transferase j PA0490 9946353 hypothetical protein PA0493 1 9946356yi p''quiring _ _ PA049able biotm-retuinng enz me Pari 9946365 8-amino-7-oxon'IIILJJ -A050-2 9946366 probable biotin biosynthesis protein b_ioH bioH PA0503j 9946367 probabie biotin synthesis protein BioCJ [biqC J PA0504 9946368 dethiobiotin synthase _ T ! bioD j PA0505 9946369 hypothetical protein PA0514 9946379 heme d1 biosynthesis protein Girl PA0527 s^946393 transcriptional regulator Dnr __A_ adnr _ ___ __ _ PA0531 j 9946397 probabfe gfutamine amidotransferase. jI j PA0535 9946402 ! probable transcriptional regulator PA0540 j 9946407) hypothetical protein PA0542 ; 9946409 e conserved hypothetical rotein ; _-1 C PA0543 99464101 hypothetical protein------ PA0544 T 9946411h othetical roten I-, _ PA0546 9946414 methionine adenosyitransferase metK PA05SO 9946418 conserved hypotheticai protein ygbM posphoglyycerate kinase pg k 9946422 ! hypothetical protein [PA0555r 9946424'fructose-1, 6-bfsphosph fda . J. .'. cbbA, cfxB s ; PA0559 9946428iconserved hypothetical protein tyhiN, PA0565---"-6946-4341 conserved h pothetical protein PA0565 9946434} conserved hypothetical protein iyqaE PA0570 §-946-4401 hypothetical rotein PA0570''9946440 jhyppthetica) protein"I"'.'.... I "'".----. PA0571-------5 : Z6-4=-4-1-.,'hypothetical protein ', PA0574 ... . _. _. _.. 9946444i hypothetical protein. PA0578 9946449 !, conserved hypothetical protein PA0579""''9946450) 308 ribosoma) protein21"""IPsU"'"' : , PA0580 9946451 jo-siaiogjycoprotein endopeptidase 92EL YSJD.. ; PA0582 9946453 ! dihydroneopterin aldolase fo I B PA0585 99464561. hypothetical protein PA0589 9946461 ! conserved hypothetical protein 9 p PA0591 conserved hypothetical protein PA0593 j 9946465 pyridoxai phosphate biosynthetic protem PdxAjpdxA j PA0594 ; 9946466 peptidyl-prolyl cis-trans isomerase SurA sura PA0595 ! 9946467 organic solvent tolerance protein OstAprecurpstAirnp ! } PA0607 ! 9946481 ribuiose-phosphate 3-epimeraserpe dod ! PA0610 9946484 ttranscriptiona ! reguiator PrtN jprtN ! 1 PA0611"9946485jtranscriptiona) regutator PrtR"fprtR"'''"T PA0613---9946-487 hypothetical protein PA0614 r 9946488 hypothetical protein PA0617 9946492 j probable bacteriophage protein ! PA0626 9946501 j hypothetical protein ! PA0627 9946502 conserved hypothetica) prote ! n... jj PA0617 9946492 probable bacteriophage protteiin I PA0627 ; _- 9946502consehrveld hyp t_ein.-p othetical rotem_ -PA06-30-"'F----§-946-505 rotein PA0632 9946507 hypothetical protein___,- PA0635 9946511 hypothetical protein ! PA0643""1"9946519 hypothetica) proteinJJIIIIIIL j ! PA0644 j'9946520 hypothetica) protein"'j" PA0643 9946519 hypothetical protein _ _ _ _ __ _ PA0644 99465201 hypothetical protein PA0647 99465241 hypothetical protein PA0648 99465251 hypothetical protein IPA0652 9946529 1 transcriptional regulator Vfr 1 vfr ! f PA0653 i _994_6530 conserved hypothetica_I protein ; --_, y fA PA0655-. M. j 9946532 h potheticalrolp tein_--, , .. _ y-. _.. _ , PA0680 9946538 h othetical prote'in ; 'PA0661_ I 9946539 conserved hypothetical protein _ W _, _ _" PA0665 i 9946543 ! cpnserved hypothetica) protein...... YR PA0678 9946558 probabte type)) secretion system protein PA0679'"'9946559 hYpothetica) protein""jhxcP '" PA0680_ s 9946560 p_robable type II secretion system prote ! n hxcV PA0684 ; 99probable type II secretion syterg rqtein hxcz PA0686"'9946566 probable type)) secretion system prote ! n' hxcR 994687 994656i probable type II secretion system protem ; PA06_8 9946570 hypothetical protein y _ PA0697 9946579 hypotheticat protein.. PA0698) 9946580 hpothetical protein s -----_-_-y ' h othet'_p PA0700 9 46582 yp ical rotem f_ JPA0702"J 9946585 j hypothetica) protein"L-JI. JHI'lL- !- 'PA0702 9946585'hypothetical protein . PA0705 9946588 ; probable glycosyl transferas .. __. __ _. . _ 9A PA0708'9946591 probable transcriptiona ! regulator"j'......--.---. ! ! PA0709 9946592 hypothetical protein ! PA0710'"""9946593 iactoyig ! utathione ! yase'','g'o6'" __.... _.. . _.. _.. _..-.. . , _PAO_712'_ 9946595 i hypothetical protein 'PA0714 99465981 hypothetical protein ! PA0716"9946600 hypothetica ! protein'""I"1L-..-'J """---- P _716 _9946600 hypothetic_al protein_ _... ___. ____ _.. __ :.. _. _.. m. "'.. ___. __. _ _.. _. _ !. PA0717"9946601 hypothetica ! protein of bacteriophagePflj"''"' PA0720""9946604 hejixdestabiiizing protein of bacteriophagePf ; PA0728 99466136probable bacteriophage integrase N w 1 _ __. _. _... __ _ 1PA0729 9946614 ! hypothetical protein PA0730 9946615 probable transferase 99466 ip PA0733 9946618 probabtepseudoundyiatesynthasej rsuA PA0734 9946619 hypothetical protein Pua0738 ! 9946624 conserved hypothetica ! protein !' ! FPA-07472-9946628 hypothetical protein j PA0759 ! 9946647 conserved hypothetical protein ----§946-651 anti-sigma factor Muta Pua0767 j 9946655 GTP-binding protein Liera ......... PA0768--6661signal peptidase I __. _. __., __. ei SPASE.. I _. _. _... ______. _... PA0771 9946660IGTP-binding protein Ers liera PA0776 9946665 hypothetical protein YP P. _ PA0778 9946, 667 hypothetical protein........ PA0786 9946676 probable transporter PA0787 9946677 hypothetical protein PA0790 fuzz 9946681 hypothetical rotein PA0802 9946694 hypothetical protein PA0805 99466971 hypothetical protein PA0808 99467011 hypothetical protein PA0815 99467081 probable transcriptional regulator PA0818 in PA0822 99467161 hypothetical protein PA0822 ! 9946716 hypothetica) protein) j i PA0825 othetical protein P, hy., __.... _....,.. __ PA0829 1. 9946723 probable hydrolase PA0837 9946732 peptidyI-prolyl cis-trans isomerase SlyD,,. islyd IPA0850 9946747 hypothetical protein PA0851; 9946748 hypothetical protein PA0853 j 9946750 ! probabieoxidoreductaseT" ' PA0857 j 9946754 j morphogene protein Bols bols I PA0862 _ ; 9946760, hypothetical protein PA0867 1 9946766 hypothetical protein.........-------- PA0868"9946766 conserved hypotheticai protein''"'"""ya&J'""'"""' PA0869 9946767 D-alanyi-D-alanine-endopeptidase rvpbpG i ^_ _T PA0871 9946770Epterin-4-alpha-carbinolamine dehydratase phhB 1 PA0874 9946773 hypothetical protein PA0880 9946779 probable ring-cieaving diqxygenase't ! ! PA0894 99467941 hypothetical protein iPA0900 i 9946801 l hypothetical protein PA0903 9946804 ! alan 1-tRNAsynthetase : alas s : PA0904'9946806iaspartatek ! nase alpha iPA0905 9946807 jcarbon storage regulator .. .'9srA''<'--- PAt) 906 9946808 j probabie transcriptiona ! f7egu ! ato -.-.-.......- g u 1 ; to r iPA0908 9946810 : protein PA0913"'""9946815 ! probable Mg transporter MgtE'"'"""" ? rngtE'.-.-.----..- ; PA 468251 hypothetical protein __. _ __'. f-__. r j __ ; ,.. _. _. : __. ___. __. __.. _______ . ___, ___.. ____.. ; IPAO_927 _ _."., _9946831 D lactate dehydroge_nase__ (fermentatme..""".,-,"_ cldhA w IPA0932- _m 9946836jcysteine synthase B_-, __ H s. jPA0937 ; 9946842iconserved hypothetical protein' 3yaiL [PA0939''""'j9946844 hypothetica ! prote ! r . L . I ! I ! {PA0944, 9849 pjhosphobpsytamHoimid iP [N ! *" ! [PA0945 99465-0'lphosphoribosylaminoimidazole svnthetase ipurm PA0947 i 9946853 conserved hypotheticat protein ! -P-0953 9946859 ! probable thioredoxin PA0954 f 9946860 probab) eacy) phosphatase ! '"J PA0956 i 9946862 ! Lprolyi-tRNA synthetase PA0960 i 9946867 hypothetical protein PA0962 j946869 probable dna-binding stress protein ! ? A0969 9946876 ToQ protein J to) Q''"'J PA0970 46877 ToIR protein tolR 9946878TolA rotem PA0971 _ p'tolA PA0971--9-946-878, TolA protein tola . . J _ .. Pua0973 9946880 outer membrane protein OprL precursor PA0976 j'-9946884 conserved hypothetical protein PA0978 9946886 conserved hypothetical protein PA0979 9946887 jconserved hypothetica) proteinIj. ! IL---' PA0980_ _ 9946888 hypothetical protein ,. _. _... _.. _. __. PA0981 9946889 hypothetical protein PA0983 9946891 conserved hypothetical protein ---946-893 1. p .... ,. .... », JPA0986j 9946894) conserved hypothetical protein conserved hypothetical protein_. __. PA0991 9946900 1 hypothetical protein PA0993 9946902 probabie pi) i assembiy chaperone PA1000 9946909, hypothetical prote_in__ _ ( !'_. __.... _ PA1008 9946918 bacterioferritin comigratory protein loci Pua1010) 9946920 dihydrodipicolinate synthase dapA ; PA1012 1 9946922 conserved hypothetical protein PA1013 9946923 phosphoribosyiaminoimidazote-succjnocarboxurC' ! PA1021 _j 9946932 ; probable enoyl-CoA hydratase/isomerase j PA1021 9946932 probable enoyl-CoA hvdratase/isomerase ! PA1026 9946938, h pothetical protein PA1035 9946948 hypothetical protein PA1039 ! 9946952 conserved hypotheticai proteinL-.-. L' ychJ ! PA1049 9946963 pyridoxine S'-phosphate oxidasepdxHj PA1055 9946969 conserved hypotheticat protein lphac PA1063 ; 99478 hypothetical protein _ y _ _ _ _ __ ! ! PA1068 j 9946983 probabie heat shock protein (hsp9p fami ! y)"'"'J'') 'PA1076) 9946992 hypothetical protein yPA1088 ! 9947004 ; hypothetical protein _. _.. _..... _.. _ _....... s (PA1089 "9947005 jconserved hypothetica) protein ''..'. 1'"1'.''HL ---.--- IPA1095 9947012'hypothetical protein iflis = r_.. . ___ t,"o_com o_nent s_ensor A1098-". 9947015 ;-P _. _ _..... _.....' rPA1_102.. __ 994701 g ____-_. _. _ _. _. fliG _.. __ iPA-1105 . ... __9947022flaellar protein FIiJ _, _. _rv. _ 1 PAl 106 9947023 hypothetical protein PA1107. _, _... _. __9947025 ; conse_rve_d hypothetical protein........ _... _... _......... ____... _M_.. _Y___. _^... _. _. _. _. __.... _. _. __ iPA1114 a__ etical rotein v _ _ : PA1118 9947037 ! hypothetical protein , PA1120 9947039conserved hypothetical protein yfiN__ _, __ PA1122 _w,... _. ____. __ 9947041 amble peptide deformylase _y , . _ .. . _.. . _.. _v f def, pdf, fus PA1125 ! _9947044 ; probable cobalamin biosynthetic protein M', ^ ; cobB__ PAI 129 1 9947049 probable fosfomycin resistance Rrottein PA1134 '9947054 hypothetical protein' _,. _ ;...... __ .. ___. _. __ PA1135 9947055 conserved hypothetical protein PA1138_ I, _ 9947058probable transcriptional reculator _ y _ ! _ PA1145 i 9947066 pro6able transcriptional re ulator i I-i3'A 114-9 9947071 hypothetical protein PA1151 9947073 pyocin S2 immunity protein _. -i5Al-l 9947076 conserved h potheti6al protein, _. PA1155'9947077 ! ribonucleoside reductase, small chain nrdB _ PA1156 _ 9947078ribonucleoside reductase, large chain ; nrdA _ , PA1129 9947049 j probabie fosfomycin resistance protein'j PA1134 ! 9947054) hypotheticaf protein ! PA1135 J 9947055 j conserved hypotheticat protein ! ysdU PA1138 J9947058 probabie transcriptionai reguiator. [ . PA1145'9947066 probabie transcriptionai regutator j F PA1149 f*"994707T hypothetical proteinHL-.-. "- PA1151 T'9947073 pyocin S2 immunity proteintl'ILt'-.-PA1154 1 9947076 conserved hypothetical proteinJ"'PA1155 9947077 ribonudeoside reductase, smat) chainmrdB PA11569947078 ribonucteoside reductase, targe chainJjirdA J PA1157 J 9947080 probabie two-component response regujator ; ! PA1159"'p'9947082 probabie coid-shock protein JlLjI. LJI. T-. ! JLL-. LLLLL ! PA1160 9947083 hypothetical protein VAif-I 9947 inyl-diaminopimelate desucsin PA1164 ! 9947087 conserved hypothetical protein [J... PA1165 r'9947088 hypothetical protein PA1167 j 9947091 hypothetical protein ° PA1167 9947091 hypothetical protein IPA1172 1 9947096 ;, cytochrome c-type protein NapC __napC ' PA1173 _9947097 cytochrome c-type protein NapB precursor, _eapB PA1176 r 9947100 ferredoxin protein NapF spF ! ! ) PA1177 j 9947101 peripiasmic nitrate reductase protein NapE jnapE'JJ . p p.. _ _...... P_ _ _ p _. p .... ___. _.. Pua1183 9947108 C4-dicarboxylate transport i-,--, 0947119 hypothetical protein , PA1203 9947130 hypothetical protein PA1204 ! 9947131 conserved hypothetical protein ^ __ ! yieF' _ _, , I Pua1206 9947133 hypotheticai protein ____ _., __, __. _.. __. _. ____. _.. ___ _. __. _. ___. .. ____. _ PA1213 9947141, hypothetical protein PA1213 9947141 hypothetical protein. JILjIIII IL-JIIIII--L-J PA1215 3 hypothetical rotein PA1216 9947144'hypothetical protein PA1217 9947145 probable 2-isopropylmalate synthase.. __.... _. _'__...... _.. __. _.. _. _i. ___. __.. _.... _.. _... _____. _. __. p 9947147 hypothetical protein IPA1223 (9947152 probable transcriptional reguiator ; PA1224 9947153'probable NAD (P) H dehydrogenase _ ; Pua1228 9947157 hypothetical protein PA1230 1--'947-159'hypothetical protein 9947162 hypothetical protein 9947167 ! probable multidrug resistance efflux pump [PA1250 9947181. ! aikaiineproteinaseinh ! b ! torApri . I... lapr !"' 'PA1261 9947193 t probable transcriptional regulator , PAi269 9947202'Drobable tra t lator 'PA1280'9947214 ! hypothetical protein fi cobC PA1285'9947220 j probabie transcriptionat regulator j"" ;""'"'""" s_ i tional re PAl 298 9947234 ! conserved hypothetical protein 'PA1300 9947236'. probable sigma-70 factor, ECF subfamily _...,........ _. _.. _......... _.. : PA1306. 99472433probable HIT family protein, y S PAl 306 3 j probable HIT family protein 6. 1' 9947243 prpbabje H) T fami ! y protein"JI"'"""""" 947252 li-Al 315 pro bable, transcri ptional requlator ßPA1321'9947259icvtochrome o ubiquinol oxidase protein CyoElcyoE 3PA1323 9947261 {Ihypothetical protein probable transcriptional re uliatior PAl 328 9947267 9947270 conserved hypothetical protein PA1342 9947282 probable binding protein component of ABC tr z PA1348'9947289ihypothetical protein PA1349 9947290 conserved hypothetical protein PA1349 ! 9947290jconserved hypothetica) protein"T''"I-.- ! PA ! 35CL_ 9947291 hypothetical protein PA1352 9947293 conserved hypothetical protein PA1353 9947295 hypothetical protein PA1355 9947297 hypothetical protein 9947300 hypothetical protein IPA1362 t'9947304 hypothetical protein PA1362 9947304 hypothetical protein PA1364 ! 9947306 probable transmembrane sensor _ i --Al 366 9947309 hypothetical protein IP 1369 9947312 hypothetical protein IPA1369 9947312 hvpothetical protein B g PA1371 9947314 hypothetical protein Yp PAl 372 1 9947315 hypothetical protein | PA1372 9947315 1 hypothetical protein _ __ _ _ _ I PA1375 9947319 1 erythronate-4-phosphate dehydrogenase pdxB _ I 9947319jeythronate-4-phosphate dehydrogenase lpdxb IPA1377 9947321 tconserved hypotheticai protein çyhhY iPA1378 9947322'hypothetical protein |PA1379 probabie short-chain dehydrog_ase'Ç __ ___ 99473401 hypothetical protein IPA1397 9947343 tt rmguiator} ç _ X PA1398 J"9947344 hypothetica ! protein J'1'''J PA1398 9947344, h othetcal rotem f s YP p PA1<1 9947344 hypothetical protein t PA1404 9947351 hypothetical protein | PA1404 W_ H _'i'_ ~_ PA1409 ! 9947356 ace Ipol amine aminohydrolase aphA | PA1426 _ 994737_5 hypotheticai protein-_ _. _ , _ i tPA1427 l 99473761hypothetical protein I , PA1426 9947375 ! hypothetical protein iPA1427 1 99473761 hypothetical protein PA1431 99473801 regulatory protein RsaL rsal I PA1442 ! w __ W §oliL ____*__ PA1427)'9947376'hypotheticai protein111--..-J... I JL'j ifs PAl 442 b9473931conserved hypothetical protein PA1454 9947406 flagellar synthesis regulator FieN PA1456 9947408 two-component response regulator CheY PA1461 9947413 probable c emotaxis protein motb PA1462 9947414 probabie. plasmid partitioning protein PA1464 9947417, probable purine-binding chemotaxis protein J cheW iPA1465 99474181 hypothetical protein 'PA1468 9947421 ! hypothetical protein , PA1472 | 9947425iconserveu_ypotnetlcal protein.,'. Y . _.. __.... _ w fRS 994742 heme"exporterprot-------.....--. PAl 476 9947429' ! heme exporter protein CcmB : ccmb tlO ; cycw ; helb bPA1477 9947431 Iheme exporter protein CcmC, ccmC. pfcyt1 cycZ heiC sPA1478. 9947432i, hypothetical protein ; pfcyt2 ccmD cycX helD.' PA 1480 : 9947434 cytochrome C-type biogenesis protein CcmF ; ccmF cycK ; cc ! 1 JL-'9947435 cytochrome C biogenesis protein CcmG ccmG dsbE iPA1482 ; 9947436Jcytochrome C-type biog_nes_prote, n Ccm tccmH _ c12 cycL _ CPA1488 ; 9947442 ! hypothetical protein '. PA1489 9947443. hvDothetical protein fPA1492'--99474471 hypothetical protein sPA1496 t 99tassium channel,, 9~,, _ _ t PA1504 9947460 i probable transcriptional regulator 1 s 9947464l, hypothetical protein S t ical rotem I PA1. 509, 9947465. hypothetlcal proteln PA1. 509 9947465 ! hypothetical protein PA1514 ! 9947471 conserved hypothetica) protein < IybbT w s PA1517 __ 9947474 conserved hypothetical protein PA1518 1 99474753btical protein S PAl 518 9947475 ! conserved hypothetical protein PA1526 1 9947484Sprobable transcriptional regulator. o_ PA1528, 9947486 cell division protein ZipA PA1529 J !'SdZ. DNAiigaseEII dnaL ! jgAJ' PAl 532 9947490 DNA polymerase subunits gamma and tau dnax PA1533 1 9947491', conserved hypothetical protein S _I PA1535, 9947494 probable acyl-CoA dehydrogenase. PA1539 ! _ 9947498, hypothetical protein I, EAI 54Q 9947499 1 conserved hypothetical protein PA1541"r 99475p0robab ! edruge i. IIL-L-- PA1548 9947508 consenced hypothetical protein I'fixS FPAL 5-48 1 9947508 1 conserved hypothetical protein fixs PA1551 j 9947511 probable ferredoxin í nxe PAl 555 9947515 Probable cvtochrome c Ifixp CCOP 9947519 hypothetical protein PAl 558 PAl 559 1 9947520, hypothetical protein PA1560} 9947521 thypothetical protein _'_S PA1564 i 9947526 conserved hypothetical protein PA1568_ ; 9947530 ; conserved hypothetical protein PAl 571 9947533 ! hyp, in i IPA1582 99475451succinate dehydrogenase (D subunit) t_ I, > PA1582 9947545t e (D subunit) sdhd PA1583 9947546isuccinate dehvdroaenase (A subunlt) ssanA I PA1583 9947546Ssuccinatedehydrogenase (A subunit) sdhA"" PA1587 i 9947550. 1ipoamide dehydrogenase-glc llpau, ipaA u-CoA synthetase beta chain suc PA1589 1 9947553 | succinVl-CoA svnthetase alpha chain, sucD PA1591 9947555 ; h othetical rotein -vir= PA1592..... f_.. 9947556_ _othe_tical pro_tein,. _. ____... ___... _ _....... __... _.. _...... _..... __ PA1591 j 99475551 hypothetical protein PA1592 9947556 1 hypothetical protein PA1595 9947559 ii hypothetical protein { m. r | A _ PA1610 9947559 hypothetical protein PA1610 9947576 beta-hydroxydecanoyi-ACP dehy . irise IP A 1 9947584 1 conserved hypothetical protein lybdb PA1619 t 5. 90 j conserved hypothet ! capot : 9947589'probable hydrolase _. _..... _. ____. ____... _... __.-.9_. _______. _. .. ; _.. __.. __.... __. _. __.. _...... _....... _. _....... _..... _.. _- : _.... _... _. _.......... _.......... _..... _. ___..... _.... _. _........ _....... _......... __.. _. ___... : ; PA1624 ^.. _ 9947591 ; hypothetical proten _. _. _.. _. ___. __. _. _... ____... __..... _... 'PA1632 99476005KdpF protein kdpF'. 3PÂ1635 9947603. potassium-transporting ÅTPase C chaln Kdpi_ s, atkC iPA1638 R 9947606Econserved hypothetical protein <, yneH ÆPÂ1641 ; _ 99476104. hypothetical protein __ _ _,'__ _ __ _ I PA1641'. _PA1657947627 ; conserved hypothetical_p_ro_te__in_ _"_, m"^x. _,''" --"-T-. T-.. _____. _.. _. fPA1641 99476101, hypothetical protein V PA1666'99476371 hypothetical protein i, si IPÂ1673 1 9947645Shypothetical protein S , 3PA1674'9947646 GTP cyclohydrolas-e I precursor, folE2 IEA 16 L3 9947645 hypothetical protein PA1675"'9947647 j conserved hypotheticarprotein)'""""j'"""'j PA1676 i 9947648thypothetical protein 3 PA1687, 9947660 spermidine synthase spee PA1690 9947663 transiocation protejn in type 111 secretion pscU IPA1691 1 9947664 translocation protein in type III secretion psct PA1696 3 9947669 trans ocation protein in t e ifl secretion psc0 ; PA1696 9947669 translocation protein in type III secretion PSCO PA1698 3 9947672 outer membrane Protein POPiN pOplN PA1701 9947675 conserv in type III secre ; PA1702 E 9947676 conserved h pothetical protein in type I I I sec j pcr4 -r j l f PA1705 99476791 regulator in type III secretion. pcru | 3 PA1710, 3 99476843exoenzyme S synthesis proteiX exsC PA1711 f 9947685 hypothetical protein PA1718 9947693 type III export protein PscE _ pscE _S _ ____ PA1719 j 9947694 type III export protein PscF pscg PA1720 9947695 type) !) export protein PscG _ pscG v PA1722 9947697 type))) export protein Psc) i. PscL.'... III. PA1732 w 9947708 conserved h othetical rotem YP p _... _. _. _. . _.. _ PA1732 9947708 conserved hypothetical protein _'_ _* PA1743 ! 99477201hypothetical protein 3j PA1743 9947720 1, hypothetical protein PA1745 99477221 hypothetical protein PA1750 9947728 phospho-2-dehydro-3-deheptonate aldotas, -, _rj-- PA1757 W I 99477_354homoserine_k_inase ; thrH i_ PA1757 I 9947735 ! homoserine kinase PA1772 1 9947752 probabfemethyftransferase""menG""''' 3iPA1777 ii 99477571outer membrane protein uprF precursor ioprr | IPA1780 9947760 assimilatory nitrite reductase small subunit inird nase IPA1783 99477641 nitrate transporter lnasa nast PA1787 99477 aconitatehydratase2 . HLJspn B 'j '9947768 t aconitate hydratase 2 _ acnB PA1787 ' 'PA1790 j 994. 7772 Yhypothetical protein _ _ : _ __ 3PA1792, 99477741conserved hypothetical protein [ybbF PA1794 ! 9947776 gtutaminy)-tRNA synthetase'""" ! ghS"f'""""''"" PA1795 i 9947777|cysteinyl-tRNA synthetase cysS t E gena<,, TolD t PA1796 = _9947778C5', 10-methylene-tetrahydrofolate dehydrog_ena f ID Y--_ __ _ PA1803 9947786 Lon protease () on capR deg muc JopA PA1815 9947800 ribonuclease H rnha [E : 1816 9947801, DNA polymerase lll,, epsilon chain dn@Q imutd (PA1817 99478021 hypothetical protein i. PA1820 i 9947805 ! sodium/proton antiporter NhaB : nhaB 3 __... __. _.. _......... _. _....... . PA1825_ 9_ p P_. t ____....... __.. ___ PA1830'9947816i, hyethetical protein PA1835 y uhyp het p :-,--_-"...-. _... __. _.. _. __.. ___. __.. _. _..... .. _.,.. _.. __... __.. _. _.. i hypothetical protein ; PA1837-. Y. __.. 9947823 ; hypoth_etical protein _-'m. _.. _____. _.. , _.... __. _ _ _ _. ; h- othe _.... _. _. _ IPA1842 9947829 ! hypothetical protein iiPA1845 9947832 hypothetical protein _ e. _. _. _ 9947835 ; conserved. hypothetical protein .......... PA1855 1 99478433hvpothetical protein PA1859. 9947848iprobable transcriptional regulator i 1 ---------- PA1867 T 9947857Lypothetical protein 3 tjxphA . _. PA1869 9947859 probable acyl carrier protein PA1872 9947862 hypothetical protein PA1882 9947873 probable transporter ii PA1882' !"'9947873 probable transporter, [. JIIT*') PA1883 : 9947874 (probable NADH-ubiquinone/plastoquinflne oxo " PA1883 PA1884 99478751 probable transcriptional reg lator 'PA1885 1 9947876 conserved hypothetical protein PA1892 1 9947884 hypothetical protein PA1894 9947886 hypothetical protein PA1895 9947887 hypothetical protein PA1896 99478 8 hypothetical protein PA1897 1 hypothetical protein PA1897 ; PA1900 i 9947893 probable phenazine biosynthesis protein i zB2 PA1905 i probable pyridoxamine 5'-phosphate oxldasw ipnzz PA1911 9947904 probable transmembrane sensor PA1912 99479051 probable si ma-70 factor, ECF subfamily PA1914 9947907 ! conserved hypothetical rotein hvn PA1917 t 9947910|hypothetical protein t iJ PA1924 1 9947918 hypothetical protein PA1 92i5 A ~ x t PA1928 zu 9947923Jiribosomal protein alanine acetyltransferase frimJ __ PA1929 1 9947924 hypothetical protein PA1936 9947932 hypothetical protein A1937 9947933 conserved hypothetical. protein PA1938 9947934 conserved hypothetical protein PA1939 9947935 hypothetical protein I PA1951 9947949 hypothetical protein l PA1952 9947950 hypothetical protein PA1955 9947953 hypotheticai rotp ein ; _ 1PA1956 1 9947953 hypothetical protein PA1962"* ! 9947960 conserved hypothetical protein PA1963 9947962 hypothetical protein PA1965 9947964 hypothetical protein PA1967 9947966 hypothetical protein l s PA1968 1 9947967|hypothetical protein _.. _--S A1 9947974ihpthetical rrtem... ___. ____-.. ___= PA1974 9947974'hypothetical protein g "_.. __ PA1980 i 99479801probable two-comDonent response_tor { ; PA1985 ! 9947986 ; pyrroloquinoline quinone biosynthesis protein i, pqqA : ; .. _........ _. _ __... _ _... _.. _... __. ___..,..... __.. _. _., _ PA1986 : 9947987i. pyrroloajuinoline quinone biosynthesis protein tqB' 1PA1988 9947989rroloquinolme quinone biosynthesis protein tqqD si PA1994'_ 9947996 1 hypothetical protein _ __ ç _ i __ _ _ H95 i, 99479971 hypothetical protein A 9 i 9 tÇcis träñs isor i äse C1 ppj ... _ __ _---_+ __S-- '_ ce I CoA acet Itransferase B PA2002 99480041conserved hypothetical protein i iatoE PA2007 99480101 maleylacetoacetate isomerase ; maia iPA20l6 994801 sfpTobabie transcript')-.....--.--... PA2013''9948016tprobable en l-ydratase/isomerase i ; menB PA2013'9948016 probabte enoyJ-CoA hydratase/isomerase ! J"JmenB---- PA2015'9948019 probabte acy !-CoA dehydrogenase'"11111. . ! PA2016 probable transcriptional PA2017"'9948021 hypotheticaf protein"--"--"-''' PA2021'i 9948025 hypothetical protein i PA2026 1 9948031 wls=r ed hz het e n __1 __ yfeH i PA2029 L 9948034 hypothetical protein 9....... PA2031 948036 h pothetical protein PA2034 9948039 hypothetical protein_________ I PA2037 9948043 hypothetical protein PA2042 i 9948048 probable transporter (membrane subunit) yg, jU PA2051 1 9948058 probable transmembrane sensor i ; PA2051 9948058 probable transmemirane sensor PA2052 Jl9948059 cyanate tyasejcynS"' pA2060'999948068 probable permease of ABC transporter PA2062 9948071 probabie pyridoxai-phosphate dependent enzy ; ! PA2066 9948075 hypothetical protein . _. _ __-.. _... _. _. _... ...., _, _.. __ PA2071 9948081 eion aton factor G _ fusA2 PA2073 ! 9948083 probable transporter (membráne subunit) $ ; PA2074 F 9948084 hypothetical protein__ PA28086 hypothetical protem PA2080 1 99480911 hypothetical protein _ _ PA2090 9948102 ! hypothetical protein rter ^. ... . _ ... .... _. . J'.. _. l PA2095 9948108 hypothetical iPA2097 9948110iprobable flavin-binding monooxygenase i l I PA2101 ! 9948114 conserved hypothetica) proteinl ! l....... . i PA2103 9°. 9948117 ; probable mol bdopterin biosynthesis proteinN' -_m-W moe8y . PA2105 bable acetyltransferase 5 99481195 _~_ co__ 99481191 pro P PA21 10 9948124 1hypothetical protein PA211 I 994813_3i06-methylguanine-DNA methyltransferase ada PA2119 a) coho ! dehydrogenase (Zn-dependent) adh PA2120 i 99481351hypothetical protein g L p 99481381 probable transcriptional A2123 lat...... PA2131 {9948147ihypothetical protein _ PA2136 1 994818153 hypothetical protein ; u ( PA2142 9948159 probabte short-chain dehydrogenaseY yhxC T PA2143 1 9948160 1 hypothetical protein T 4 PA2143 9948160 hypothetical protein PA2146 9948164 conserved hypothetical protein _y {Pm7 <9protein __] __ L, _ _____. _--- 99-48175 i hypothetical . _.. j _. _... ! PA2166""""'"9948186 hypotheticai protein J"------- - hypothetical rotein , PA2170'"J.""' 9948190 ! hypotheticat protean"""' ;""'"'"'"""-'-"-"'"-"-" _, pothetical protem °-.. ___.. _. __. __. _. _____ ____... .,. Y. __. _. _... ___.. __. _. _ ___... __. _... __. _..- PA2167'v r. w. 9948187 ; hypotheticalrotein-T-. Y-_. _.... _... .. _... _... _..... _.. __. _ww... _. _.. _. _. _. _.. _.,. _... _ _. _.... _.,... _,., _.. . f-PA2171 99481911 yR2Lhetical protein 9948194 hypothetical . 2PA2167 994Ent glwt PA2170 : 99481<ypothetical protein 217i'94 91ihZp theticalproten PA2174 9948194'hypothetical protein'. tPA2175 PA2174 protein_. _. __.. : P 182 _. : 994 Yp 'p.-.-. _. __... _. __. __ ;-. V _. %. . __. _____. _.. _.. __......... _.. _... __.. _ 'PA2182. 9948203 hypothetical protein iPA2183 9948204 hypothetical protein I proteln PA2185 ; 9948206 hypotheticai protein PA2187 ___-rv9948208 hypothetical protein. _. _... _, . 4 V _ _ PA21'87-9948208 ! hypothetical protein A2190 9948211 conserved hypothetical'protein 9948214 conserved hypothetical.. protein PA2196 j 9948218iprobable transcriptional regulator PA2197 9948218 probable transcriptional regulator PA2205 ì 9948228 jhypothetical protein {_ S PA2207 9948230 hypothetical protein PA2207 9948230 hypothetica) proteinJj PA2211 9948234 conserved hypothetical protein PA2214 9948238 probable MFS transporter PA2219 9948243 membrane protein OpdE opte ! PA2220 probable transcriptional regulator oprm PA2221 j 9948245 conserved hypothetical protein _ _ _ PA2222t 9948247 hypothetical protein JL-.'L. JII""'I'L.--ir'''' ! PA2223 1 9948248 h_.-_. _~wo__~__ XPA2224 9948249 hypothetical protein _. S PA2225- 9948250 hypotheticai pr_otein_ PA2225 99482501 hypothetical protein PA2226 i 99482511hypothetical protein __ ? __, ___ 9948252 probable transcriptional reguliatior PA2228 i 9948253 hypothetical protein _ _ i _I _ FP-A2228 9948253 hypothetical protein PA2234 r 9948259 probabte exopotysaccharide transporter ! ; PA2242 ; 99948268 hypothetical protein ___ _ _ l PA2242 9948268'h othetical protein JPA2251 9948278 hypothetica ! proteinJ j" PA2253 i 9948280 ; iL-asparacinase I lanse _ _ __ f PA2257 j 1_ vcD _, _ PA2258 9948285 [transcriptional regulator PtxR ptxr PA2260 1 99482881 hypothetical protein IPA2280 f 99483091conserved hypothetical protein H _ jlPA2282 $<sça M 5'I PA2282 9948312 h pothetical protein w {",, t., j [, PA2293 1 99483241hypothetical protein. j_ _ _. __'. _ o. _____ ! 1PA2292 9948323 hypothetical protein PA2293 9948324 hypothetical protein PA2295 9948326 probable permease of ABC transporter 1"j' PA2297'"} 9948328 probabteferredoxinT".... LlI'*' IPA2298. 99483291probable oxidoreductase PA2303 99483351h pothetical p ! PA2311 9948344 ! hypothetical protein '. PA2316 f >, robable transcriptional regulator. 2 fPA2329F'9948364, probab ! eATP-bindingcom""T"------"----'' ; PA2329_,. _, _. ____ 83 p b dtng po 'PA2331 9_4836 h1Usthet > DBtY n __ _ , PA2336'. 9948371 : hypothetical protein : t PA2331 9948366 hypothetical protein : _PA2336.. ___.. _........ __994837_1 hypothetical protein '"''... __ _, __u.. _. _... _.. _.... ___ __. . ___-.. ___w.... _. _..,. _ _. _.. __. __.... ____.. _. _ _. _. p __. , PA2338 9948374 i probable binding protein component of A m, mtle PA2347 9948384 ß hypothetical protein : PA2347 9948384 ! hypothetical protein "PA2347""'"'"9948384hypothet) ca) protein-------------j--- PA2349-9948386, conserved hypothetical protein sPA2351. 9948388. probable permease of ABC transporter PA2365, _9948403, conserved hy othetrcai _ rotein . PA2367 9948406lhypothetical protein. | PA2368 l 9948407. hypothetical protein I tPA2370 9948409} hypothetcal protein IPA2372 l 9948411 ! hypothetical proteln _ IPA2375 n 99484141hypothetical protein'| T PA2383 _i_ 9948423 jprobable transcriptional regulator 9948411 hypothetical protein _... __. _.. _.. ^ PA2411"j 9948455 probabte thioesterasei 'J PA2411 __ 9948455 probable thioesterase . w 1 __. ; _, PA2406 t wypothetical protein _ t S PA2411'9948455 probåble thioeste_ase} PA2412, 9948463 ! conserved hypothetical protein _t PA2428 994846 ypothetical protein E PA2422 9948473 hypothetical protein PA2427 t 99484731hypothetical protein PA2428 9948474 hypothetical tPA2429 1 9948475 ! hypothetical protein IPA2434 99484 pothetical protein PA2441 1 99484881h othetical protein PA2441, 9948488 hYpothetical protein PA2446) 9948494 giycine cleavage system protein H2 jgcvH2 j ! PA2451 1 9948499 hypothetical protein |PA2451 <, hypothetical protein _ _ _ Pua2455 ! 9948502 jhypothetical protein _ _ E_. _,. _. _ _. | _ ___ _ _-. PA2456 protein) "''' PA2455 ; 9948504 hYpothetical protein , PA2456 1 9948505 ! hypothetical protein PA2459 9948508, hypothetical protein i _ J tPA2460.,., 3 S lPA2461 9948510jhypothetical protein i'_ 4 SPA2464 9948514 j hypotheticai proteinJl.. JIIjJ PA2467 994851 7 ; probable transmembrane sensor ç __ PA2469 9948519'probable transcriptional regulator 'PA2464 -P 99485171 probable transmembrane sensor yp P _ 9948519 1 probable transcriptional regulator PA2474 1 99485251 hypothetical protein PA2475 1 9948526 probable cytochrome P450 . __. __. _ _. .. _ p. _ p 'jPA2490 t 9948542 ! conserved hypothetical protein X, jydbB 9948539 hypothetical rotein jxPA2492 i'99485441transcriptional regulator MexT,. mexT 0 j PA2496 _ I v _ _. i.....'o_-~--~----5 ; PA 1_.. . 9948554 hypothetical pro_tein_ a r 1_p_o ein = (PA2492--9948544 transcriptional regulator Mex. mexT iPA2501 99485541 hypothetical protein s _____ _,, _,, _,, _ ___ _ _. !...... ___. _. _. _. __. _. __ _.,,, _ _ 9 _ _ _ _ _ __,, _ _x". _~_ _, _...... I''''''''''''''--}- '''"-'__-__ __ _. ____c_a_ _. ___. _ PA2515'9948569 ! cis-1 2-ydroxvcvclohexa-3, 4-dienecarboxylivlL 'PA2517 9948571 itoluate 112-dioxygenase beta subunit, xylY PA2521.. . w 994857_6R_ND_div_a_len_t me_ta_I c_ation_e_fflux mem_br_ane fu, czcB . y---w.-Vyu.. -'".. , PA2536 99 93 S probable phosphatidate cyt y yltransferase __ _, yn i3 _ _ iPA2536 9948593 probable phosphatidate cltriladylyltransfe 5PA2538 9948595t, hypothetical protein ___ _ __ __ __ _ __ _ _ iPA2538 9948595 ! hypothetical protein h PA2544 9948602 E hypothetical protein IPA2549. 9948608conserved hypothetical protein I EygiT jPA2551'7''9948610jprobab) etJIIIIIjI""'""" PA2552 9948611 jprobabte acy !-CoAjjehydrogenase jacdB ! ; PA2551 9948610Eprobable transcriptional ret E I PA2552 ; 9948611 ! proba_e acyi-Co dehydrogenase, 9, _, _,"",,, d _, « ~,,,, ~ acci,,, 3 w, 9"_,, i ìPA2553 9948612*probable acyl-CoAthiolase t PA251 99486101 nscriptional requlator PA2577' ["'9948639 probab) etranscnptiona) regu ! ator-j--. PA95-77 9948639 probable transcriptional regulator PA2584, _ 9948647 CDP-diacylglycerol--glycerol-3-phosphate 3-pi IpgsA E PA2591'. 9948655sprobable transcriptional reguiator _ I I T PA2602'"r"9948667 hypothetica) protein i. IILIIL-ZLi-111-"' H ! I PA2605 $'9948670 w _, ~ E ~, _ yheN _S_ m PA2606 i 9948671 conserveu typotSnetlcal protein EE yheM PA2607 iZ _ 9948672iconserved hypothetical protein _,,,,,,,,,,,,,,,,"","",,,, T,,,,,, _,,,",,, _ __ _ _ _ _ e PA260_7_.. 9948672 ; conserved hypOthetical protein 9948671 ! conserved hypothetical protein serS I PA2612.'9948677 seryt-tRNA synthetase serSj PA2614 T 9948680Zperiplasmic chaperone LOIA JOIA 9948681 cell division protein FtsK ftsK PA2617 . .-. 99486831euc I/phenrlalan I-t-, RNA-proteintransfera_se aat ---w-_. __. .. __ PÁ2619 T 99486851initiation factor Z jnfA i PA2621 1_99486871conservedhypotheticalpr tein __ t_ I_ __ _ PA2626 9948693jtRNAmethy ! transferae ! l [ [ ! l'='........ JItas.) L'Ej,..., J _... _... s... __. . _. _. y , P i _9948696 aden losuccinate I ase _ __ _ PA2638 E 9948706, NADH dehydrogenase I chain B nuoB î _r |jo * PA2641 j"'9948709JNADH dehydrogenase) chain FL-Jl !"°IIIIIlT] PA2645 ! 9948714JNADH dehydrogenase i chain J inu_oJ_ _ _ I PA2646 i 9948715iNADH dehydrogenase I chain K ìnuoK I | kPA2658 (9948_72_8 hypothetica) protein PA2658 1 9948728 hypothetical protein PA266 34 hypothetical protein PA2687 t 9948738tconse ved hypothetical protein __<_ _ PA2688 ì 9948739fhvpothetical protein PA26-ro8 9948739 hypothetical protein iPA2673 99487441 probable type If secretion system protein <". jhptV"j PA2674 j 9948745 probable type 11 secretion system protein ihplu PA2675 9948746|probable typwm_rotein _ t_ _ A, k iPA2678 9948749tprobable permease of ABC-2 transporter J 4 UB2681 _ 9948753jprobable transc ltgulator _., _~~_ » L-o iPA2B83 9948755|probable senne ! threonne de dratasel degrac tdcB _ _ t. Y. .. ___ ___ _ _ _ IPA2-689 _ _ 9948762 1 hypotheticai protein __ _ _ ; _ _'_ _. _. ~ __1 iPA2689 99487621 hypothetical protein IPA2690 9948763 probable transposase j, ioredoxin j e iPA2708 ; 9948780 ; hypothetical proten I ßPA2703} 99487771hypow D) iPA2706 l 9948780ihypothetical protein. t __ t PÄ27i5 lY 9948790 jprobable ferredoxin 0 'PA2719 9948795 ! hypothetical protein 9948796 hypothetical protein ! iPA2721 9948797 ! hvpothetical protein e 22 9948798 h othetical roten PA2723 _y _ical rot_ein ; ,,, _ _----"''""'''"'""'""''"'''""'"'' ? ? PA2726 ? 9948802 ? probable radical activating enzyme _i_ {PA2730 9948807 hypothetical protein ? JPA2722"'9948798 hypotheticat protein """"''.....""'j''"-. ..-.....-.-..-.....-.... A2733 9948810 i ical proten . PA2731 9948808 ? hypothetical protein J _^ _ . 4 ; conserved hypot_het' ; ' .". _....... _. _.... tPA2733 9948810 ?, conserved hypotheticai eroteln _ X _ fi v 994881 1 w ? ? PA2736 9948814 ! hypothetical protein PA2737 99488151 conserved hypothetical protein .......... PA2737 ! 9948815 cqnseryed hypothetica) protein T'I ! PA2738"i 9948816jintegration host factor, a ! pha subumt'*TrnA j.---.--.-..--.-. PA2739,.. __9948817lphenylalanyl-tRNA synthetase, betasubunit__-'pheT v ; F__T..- --.. ____. ____ ..........-- =,'PA2740 _ 9948818 phenylalanyl-tRNA synthetase, alpha-subunit phe_S iPA2741 ; 994_8819 50S ribosomal protein L20 rpIT PA2742 r 9948820 J 503 ribosoma ! protein L35jpmj ! J P PA2743 itiation factor IF-3 infc PA2744 i 99488221threonyl-tRNA svnthetase thrb PA2749 1 99488281DNA-specific endonuclease I endA _. < PA2753 99488 : al protein PA2756 99488. 35 hypothetical protein PA2759 9948839 hypothetical protein EPA2759 3 9948839 hvpothetical protein PA2762 1'9948842 hypothetical protein oo PA2763 9948843 hypothetical protein PA2767 QoARRA7 r) rnbable enovi-CoA hydratase/isomerase PA2768 99488481 hypothetical protein PA2769 9948849 hypothetical protein _2948855 YPO PA2775'1'9948856 hypothetical protein'""""""'"' !---.--.------ PA2780 9948861|hypothetical protein ___ I _____ PA2781 99488621 hypothetical protein 'PA2782 1 9948863 1 hypothetical protein liPA2784 4i 9948866 hypothetical protein PA2785 9948867 conserved hypothetical protein PA2786 9948868 hypothetical protein -PA2792 9948874 hypothetical protein PA2794 1 9948877 hypothetical protein PA2797 ! 9948880 hypothetical protein , PA2799 L 9948882 hypothetical protein ~ PA2800 9948883 conserved hypotheticai protein ! JvacJ J PA2803 t 9948886 hypothetical protein PA2805 9948888 hypothetical protein PA2807 9948891 hypothetical protein {PA2808 i 9948892 ! hypothetical protein _,, yf _ tf PA2808 9948892 hypothetical protein PA2818 9948902 hypothetical-protein PA2819 1 9948903 hypothetical protein PA2827 9948912 conserved hypothetical protein 4 __. yeaA PA2829 9948914 hypothetical protein , PA2831 1 9948917 conservedpothetical protein __ iPA2832'. 99489181thiopurine methyltransferase tpm SpAS839'9948925 ! conserved hypothetical protein'iY9tD.. __ _ X f PA2843 _ 9948930 ; probable aldolase _ {i PA2845 994eSn {PA2851"''9948938itrans) ationefongation'"'"'''""'" ; g'''''"j"-".................. ! PA2852"'""9948939) hypothetica) protein I. . Il.. j"""'""' si PA2852 9948939 i hypotheticai protein sPA2853 _ 9948941.'outer membrane lipoprotein Oprl precursor, oprl {, [PA2859r 9948947 ! transcription elongation factor GreB" ; greB'1"'j PA2859'9948947itranscription elongation factor GreB greB'ij a, PA2863'9948_1I ase modulator p otein'ilipi. __ PA2868 . 99489571 h othetical roten J = _. 99489571hypothetical protein __ _ _ __ _ __ _ _ _ ! tRA2874 _'9948963 I hyp + , 3PA2876 1 9948966, orotidine 5'-phosphate decarboxylase'ipyrF iPA2877 9948967i probable transcriptional requlator IPA2879 i 99489691 probable transcriptional regulator PA2883 T ; 9948973 hypothetical protein - _ _ _ _ _____.. __ PA2894 9948985 hypothetical protein PA2898 9948990 hypothetical protein PA29_01 9948993hypothetical prote_in., _ _ _ PA2901, 9948993, 3hypothetical protein I I PA2915 _a 9949008 hypothetical protein PA2915 i 9949008 hypothetical protein 3j. 1 3 PA2922 9949008 hypothetical protein .. __-. _..-. PA2928-49023 h othetica) rotem E othetical ro#en _ PA29 1 9 9949030 hypothetical protein 9949031 hy othetical protein PA2937 IP P _ _ _ PA2940 probable acyl-CoA thiolase,.. PA2949 9949046 probable lipase PA2949 L<ffl o~-- PA2951 9949048 electron transfer flavoprotein alpha-subuniHiF PA2952 9949049 electron transfer flavoprotein beta-subunit etfb PA2953 L 9949050jelectrontransferflavoprotein-ubiquinoneOxidO. _ PA2960 9949058 type 4 fibrin ! biogenesis protein PiIZ pilZ PA2961 i 9949059iDNA polymerase 111, delta prime subunit IholB PA2962 i 99490601thymidylate kinase _'tmk Ft 9949061 |conserved hvpothëtical protein i'wG li 9 conserved hypothetical protein lyceg PA2966 9949064 acyl carrier protein sPA2967 r 9949065 ; 3-oxoacyl-[acyi-carrier-protein] reductase ifabG PA2968 99490B6 B _ <_ i PA2970 9949069 50S ribosomal protein L32 I rpmf PA2971 f 9949070 ! conserved hypothetica) protein11111111" ! yc' PA2975 1 9949074 ! ribosomal large subunit pseudouridine syntha¢. rluC IyceC PA2977 9949076 UDP-N-acetyipyruYqytgfucosamine reductase murB ! PA :) sine protein phosphatase liptpA Ij gj t PA2979 ì 9949078 tj 3-deoxy-manno-octulosonate cytidylyltransfera kdsB f tjPA2980' 9949079iconserved hypothetical protein W ycaR . __. __. _..-.. _------. * PA2980 erved h pothetical protein ycar PA2982 9949081 conserved hypothetical protein i ij PA2983' 9949082 P probable #oIQ-type transport proten PA2985 tj 99490851hypothetical protein PA 9949085 hypothetical protein PA2986 949086 conserved hypothetical protein , PA2987 9949087 probable ATP-binding'component of ABC tran, 's lycfv ! PA2988', 9949088 i, oon_rW ! PA2989 1 9949089 hypothetical-protein iPA2991 9949091 ;, soluble pyridine nucleotide transhydrogenase Isth,. . p p. . _ :.. ___.. __ ___. ___. ___z 'PA2992 9949092thypothetical protein''. , PA2996'9949096iNa+-translocating NADH : uniquinone oxidoredl, anqrD sPA3001""'9949102 j probabje giyceraidehyde-3-phosphate dehydrc'....-.-.--.--.-.-.- PA3004 PA3004 9949105i probable nudeoside phosphoryfase ['J PA3009y_ u_9949111 hpothetical, pr_ote_m PA3009 9949111'i hypothetical protein iPA3011 9949113 ! DNAtopoisomerase I ítOp ; iR ----------------- protein ! PA3012 9949114 ! h pothetical protein PA3017 99491201, con, served hypothetical protein [PA3022'"9949125 hypotheticatprotein'j J' { PA3021 pothetical protein PA3024 1 9949127, probable carbohydrate kinase,,,",,,, _ _L. _ _ _ _} ____. __. _ __ _ _ _ _. ______. _. r ip 9949127 probable carbohydrate kinase "PA3030 9949134 probable molybdopterin-Quanine dinucleotide b moba PA3033 9949137 hypothefical protein w ; r PA3036 t 9949140ihypothetical protein % -fi PA3040 t 9949145 conserved hypothetical protein_ __ _ yqjD I _ __ .-. _..... PA3041 9949146 ! hypothetical protein --'9949147 hypothetical protein Put3042 PA3049, 9949155 ribosome moduiation factor rmf. _-., _ _ PA______... ---9949157h pothetical protein _... _. ____. _. _.. _........ _.... __. __. _. _. . _. __. ___. . _____.... _. __ '_. _. PA3067 # 9949174iprobable transcriptional regulator PA308_1 _, 9949189 ! conserved hypothetical protein y [ PA3051 9949157 hypothetical protein'I PA3081 1 9949189 ! conserved hypothetical protein _/_.... _ _ _| PA3088 9949197 conserved hypotheticai protein ijYfl§ ! PA3085 9949193 hypothetical protein PA3093, _ 994 0 ypothetical protein _'_ PA3089 1 949198. pothetical protein PA3093 ; 9949202 MebM, _ _ __. ~ __ _ ; PA3095 9949205 general secretion pathway protein M xcpZ PA3096 J 9949206 genera) secretion pathway protein L XCPY PA3100 1 9949210 general secretion pathway protein HxcpUpddB'"''''"'' ! PA3103' ! 9949213 general secretion pathway protein E xcpR PA3103 99492131general secretion pathway protein E L _ 9949221 thypothet __ __ _ _ w _ _ PA3112 1 9949221 hypothetical protein PA3117 1 9949229 aspartate semialdehvde dehvoroqenase asa PA3117 1 994929r se PA3123 ; 9949235 conserved hypothetical protein PA3140 9949254, hypothetical protein PA3140 i 9949254 jhypothetical protein PA3142 9 ___9256 hypothetical protein PA3144 9949258 hypothetical protein PA3145 ! 9949259 glycosyltransferase WbpL lwbpL_ - ; 'PA3146'9949260 ! probable NAD-dependent epimerase/oenyurat3vDpK 1 { tPA3147 í 9949261 probable glycosyl transferase WbpJ LwbpJ l _t PA3148 9949262 probable UDP-N-acetylqlucosamine 2-epimerE twpi PA3149 4 9< o-i i ! PA3150 9949264 LPS biosynthesis protein WbpG IwbpG v PA3151 9949266 ; imidazolegcerol-phosphate synthase, cyclase : hisF2 PA3152"9949267jgiutamine amidotransferase'"ThisH2"' iPA3153 t 994926810-antigen translocase lwzx IwbpF, rfbX PA3154 9949269 B-band 0-antigen poiymerase wzy rfc j PA3155 9949270 probabte aminotransferase WbpEwbpE 9. PA3156 1 s= _pD, wbpD j _J PA3155 9-492-701 probable aminotransferas WbiLE pE PA3157Y 9949272 probabie acetyitransferase . . L-- J"wbpCj ! PA3158'r 9949273 j probabte oxidoreductase WpbS. LUbpB" [..'.. ! , PA3159 99492745probable UD_glucose/GDP-mannose ciehy_re NbpA >_ _ _ _ __., P. __ Y_ __ 'PA3160 9949276iO-antigetth guiator'wzz Scld, rol i'PA3161 : 9949277iintegration hostfactor beta subunit''himD EPA3162 : 9949278i30S ribosomal protein Su rusa PA3163""""""9949279 jcytidyfate kinase"""""""''] cmk"" T----------- . - T. . PA3167, 9949283 ! 3-phosphoserine aminotransferase iserC iPA3168 9949285 ! DNAgyrasesubunitA tgyrA £'. PA317 1 < 9949288 1 3-demethylubiquinone-9 3-methyltransferase, ubiG jPA3178"""'9949296\hypbtheticaj prote ! n"jJ'-"-"--T"'"""""" ; i_.... ___ __ _. _ _. _ ! PA318fj 9949299 2-keto-3-deo -..---,.....---.... ....... j f PA3185 9949303 hypothetical protein. t_ T ? 9949314 glyceralidaehvde 3-phosphate dehydrogenase tgapA ij Y _.. _. __ conserved hypothetical protein PA3203 9949323 hypothetical protein PA3207 9949327 hypothetical protein PA3211 9949331 probable permease of ABC transporter PA3220 F 9949341 probabte transcriptiona) regufator ! """-"--'--"'- } PA3227 ! 9949348 peptidyt-protytcis-transisomeraseAfppiA j cyHIL--."' [PA3230 1 99493521conserved hypothetical protein PA3232 t 9949354 ! probable nuclease PA3237 _'9949359 hypothetical protein'_. _ _ 1 _. _ _ PA3242 9949365 probable lauroyl acyltransferase PA3245 9949368 cell division topological specificity factor MinE minE PA3246 9949369ipseudouridine synthase RIuA rluA {yabO PA3249J 9949372 probabietranscriptipna) JIIIL. 11."1 PA3255 9949379 hypothetical protein PA3260 ! 99493841probable transcriptional regulator PA3266 9949391 'cold acclimation protein B capB csp _ _S PA3273 99493981 hypothetical protein PA3274 1 99493991 hypothetical protein PA3275 9949401 3conserved hypothetical protein 3 IynfA PA3278 99494041 hypothetical protein PA3280 j 9949406) outer membrane porin OprO precursor _opr0 , _ _ PA3281 9949407 h pothetical protein onserved hypothetical protein PA3287 99494141c pezs 94as m I'PA3289 1 9949416 hypothetical PA3291 9949418 hypothetical protein PA3291 9949419 hypothetical protein IPA3298 i 9949426ihypothetical protein I 1 3 PA3298 9949426, hypothetical protein PA3302 99494301 conserved hypothetical protein ) g 9949438 conserved hypothetical protein PA3312 ! 9949441 í probable 3-hydroxyisobutyrate dehYdrogenase ; I' PA3314 ! 9949443lprobable ATP-binding component of ABC tran (; [i, PA3315 9949444 probable permease of ABC transporter PA3317 T 9949447 hypothetica) proteinJ j IPA3318 1 9949448ihypothetical protein'} PA3320 9949450 hypothetical protein PA332G ! 9949457 probable Clp-Pfamily ATP-dependent protease s_-, T ipA3330 i 9949461 ! probable thortwgenase 1 3 PA3332 3 9949463'conserved hypothetical protein S i X 334 j 9949465 probab) eacy) arrierprptejn i. I.... I... f" !. ......"J 3PA3338 1 9949470E. hypothetical protein S i PA333_8 _ 9949473WnEgwegulator >, _. Ws~WW___. _____.-_. _ *WW _ _------------s a PA3341 m_ 99494_73 ! probable transcriptional regulator 'PA3347 9949480 hypothetical rotein JPA3341 ! 9949473 probabie transcriptionairegufator " ;"--- __ _ y..--_. _ __. ntPA3348 9949481ipr babi emotaxisproteinmethyitransferaseN. cheR1 PA3351"'9949484 hypotheticai protein'r I LiT T''". I Jf ! g [. 1""""I" PA3353'""'9949486) hypotheticat protein'J !'""-"--'--'-'--- : 1PA3353 9949486 hypothetical protein ypothetical protein , PA3354 9949487 h iPA3360 9949494 probable secretion prot-,,, n p__A3367 _ H ___ 9949502lhypotheticalproten _. . _ _M, ., . cydcA _.. _... _.......' PA3368 9949503 probable acetyltransferase 9, 67., H__, _9949502 LyS9titet ~ww M _ _w w W*_ www_w tw_ _ ww oeYwC _A_ _ _ w _w _ _s PA3370 994950 rotein _.. _ PA3370'. 9949505 hvpotheticalproteln e s PA3371'9949506 hypothetical protein i t PA3380 Z1-"1 f"3S49506hypothetjcaf protein riLJ"...--.-.. II--........... . I'"J"'-..-----. PA3380 ! 9949515 conserved hypothetica ! protein"" phnG"'J PA3384 ! 9949519 ATP-binding component of ABC phosphonatesphnC _ w_s PA3390 í 9949525'hypothetical protein S PA3390 9949525, hypothetical protein _ _ #.. ,. _ _. r. PA3395 9949531 NosY protein inosy PA3396 1 9949532 NosL protein lnosl PA3397 9949534 ferredoxin--NADP+ reductase _ _ Eev PA3403 i 9949540 hypothetical protein . PA3407 9949545 heme acquisition protein HasAp hast iyebG PA3413 9949551 conserved h othetical rotem. _ PA3414 9949552 hypothetical protein t r PA3416 99495541probable pyruvate dehydrogenase E1 compon PA3432 99495721 hypothetical protein PA3433 f 9949573 probabte transcnptionat regufator {ywb)"''"n . IJ PA3433 W=guiator _wol lYWblwww__ s PA3434 9949574 probable transposase PA3435 9949575 conserved hypothetical protein mioc PA3438 9949578 GTP cyclohydrolase I precursor [folE1.........,,,", f PA3439")''9949579 d-erythro-7. 8-dihydroneopterintriphosphate'''Jfo) X'T J ., probable permease of ABC transporter PA3444 ! 99495850conserved hypothetical protein fi ? ssuD PA3445 9949586 conserved hypothetical protein PA3446 9949587, conserved hypothetical protein IssuE s PA3450 1 9949591 m __ 9fL_H PA3450 ! 9949592 hypothetical protein f I 1 v . ~ PA3460 1 9949602 probableacetyltransferase PA3470 fì 9949614 m PA3472 9949616, hypothetical protein PA3472 9949616 h pothetical protein E PA3477 1 9949621 transcriptional regulator RhIR jrhlR _ PA3482 9949627 metKionyl-tRNA synthetase lmetg PA3488 9949634 hypothetical protein } PA3489'J 9949635 conserved hypotheticai protein ! mfA" ? PA3492, hypotÇi~ __, __.,, ~ irnfD PA3494""9949640 conserved hypothetical protein"J'J') rnfE " ! |W t Fiease 111--r-=~ PA3494 9949640 conserved hypothetical protein rnfe PA3495 y ___ 99496 endonuclease III _. _, ns th __. T_. __, _-_ ____. __ PA3496 9949642 hypothetical protein PA3501 ° 9949647 hypothetical protein iPA3502 hypothetical protein ! PA3505 9949652 hypothetical protein PA3512 j 9949659 probabie permease of ABC transporter PA3519 t _ mypotheticai protein _e : PA3520 ! 9949668jhypothetical PA3523 PA3523 9949671 ; probable RND efflux membrane fusion protein spa3528 9949677, ribonuc-iease T ; rnt EPA3530-9949679 ! conserved hypothetical protein _ ; tifd _A3533 9949682 jconser ed hypo hetical protein _ 4 _, y xnD __..... __... _... _. _, __w _... _... _. : PA3542y-. v. ,. 9949693 ; alinate bios nthesis protein AIg44^ .,. T.. . ."M_r ; alg44"". y_".,..,.. _T.. _.. _. w PA3550 9949702, alginate o-acetyltransferase A gF ____ ga 9r __ _ _,,,, _,, _, _, wo ; tPA3558 9949710ihypothetical protein fi, ; iPA3566 9949719, conserved hypothetica', prote ! n _ _, lycnE ! PA3570" J9949724 methytmabnate-serniatdehydedehYdrpgenasem'---' j-'j 1PA3572 99497261 hypothetical protein ) PA3575 ! 9949729 ihypothetica ! protein, J' J1""I, ! : ; PA3575, 9949729 I hyt_H PA3578""'9949732 conserved hypothetical protein"I...... L.... Li..." J" ! 1"'."''J" ! rPA3588 1_9949744|probable acyl-CoA thiolase ___ , PA3600 ì, 99497561conserve_ hypothetical protein __ i _ __ Irpl36 ~ IPA3600 9949756 ! conserved hypothetical protein rpl36. 1PA3601 9949757 ! conserved hypothetical protein X PA3605. 9949762 l hypothetìcal protein _ Q _ _ _ L_ PA3605 9949762 ; hypothetical protein . ------.-.. . ! PA3605--99497621 hypo ! hefical erot2in _PA3_606_, _ y etical rotem_ _ _ _ PA3609' 9949766 poiYamine transportprotein P ! po" F LIIIL-----I ! r b94976-61 polya mine transport p teinPotC-po PA3611 99497681 hypothetical protein PA3616 1 9949774 conserved hypothetical protein _ _, _____ recX __ __ e-| PA3617 i, 99497751RecA protein irece Hi « ffi PA3627 1 9949785, conserved hypothetical protein _ _ ygbB____ i PA3632 9949791 conserved hypothetical protein _ yedF tPA363_3 j _9949792c_onserved h py othetical prfltein _ _ . . iygbP PA3634 9949793, 1 conserved hypothetical protein g PA3635 99497_94enolase eno _ _ _ _ _ xyp p PA3636_ _.. 99497952_deh dro 3 deo _ hos hooctonate aldolase kd_s_A _ . i PA3637'''9949796 CTP synthase''."'""""pyrG""' !'""""""""1 PA3637 9949796 CTP synthase. pyru PA3638 9949797 conserved hypothetical protein. __ PA3639 I 9949798 acetyl-coenzyme A carboxylase carboxyl transRaccA S r | n v PA3640 99498001 DNA polymerase 111, alpha chain -----LdnaE C -w z W ; PA3644 9949804 B glucosamin yç l pXA,. _ _ PA3645 9949805 (3R)-hydroxymyristoyacyt carrier p sefA IPA3646 i 9949806iUDP-3-0-[3-hydroxylauroyl] glucosamine N-atpxD firA omsA,' ! PA3647 9949807 probable outer membrane protein precursor j''j <' PA3648 9949808 probable. outer membrane protein j iPA3648 i 9949808 probable outer membrane prote n I I = _. v 'PA3650 ! 994981111-deoxy-d-xylulose 5-phosphate reductolsomeaxr * PA3651 j 9949812 phosphatidatecytidy) y) transferasecdsA"j PA3652 9949813 undecaprenyt pyrophosphatesynthetase. uppSyaeS J r PA3653 9949814 ribosomerecyciing factorfrr rrf] PA3654 994981i5liuridylate kinase ipyrH t PA3655 9949816 n factor Ts tsf PA3656""''9949817J30S ribosoma) protein S2"-""'""''"''sB"""'" PA3657 j 9949818 methionine aminopeptidase ! map i j 'PA366_2'9949824 hy othetical protein', v 1PA3662 9949824 hypothetical protein PA3666 9949828 tetrahydrodipicotinate succinytaseJPJD ! _. _... _ jPA3671 l ; 9949833, probab1e permease O _ ! PA3674" ! 9949837 hypotheticai proteinIjmi.'L---LL-- i IPA3678 1 9949841 probable transcriptional regulator m. _.. ___. ____. __....... _... _. _........ _. ___. _.. _.. __. _. ___. _.. . __. __. _.... _____. ______... _... _. _... ___. _...... _.... _. _... _. _. _.. __..... _.... _. _. _.... _.. _.......,. _... :.. _. __.... _... _..,......., __..... _. _... _... _.... _... __......... _.. ) PA3684""1" 9949848 hypothetica) protein LlIl..-.-. II--... IIIIII"... . JlI''"'''"TII"'l ! ; PA3685 9949849iconserved hypothetical protein iPA3685 9949849 ! conserved hypothetical protein 'PA36 : conserved hypothetical proten_, ; _ ide cham release factor 2 fp ; __. JPA3704"'9949868J probable chemotaxis sensor/effector fusion PA3704 99 49885 h othe_tical rotei_n _ _ 9949885 hypothetical protein ; PA3725 _ 9949891 ; single-stranded-DNA-sp_ecific_exon_uc_le_ase ReaecJ u Pua3719 Yp'p' P_A3730 : 994989 1h othetical roten __ _ : PA3731. ; . 994989_8 ; c_onserved hypothetical protein, _. WW. _. _V_. _. _.. . _. f PA3733 T... _.. ___... _.. ___9949900hypothetical proteini _, H ; -.. __. _^____.. _... _____. i Pua3731 PA3742 910 50S ribosomal protein L19 rpIS PA3743 ; tRNA (guanine--methyltransferase trmD. ' 9949911 N1) PA3744 ! 9949912 16S rRNA processing protein rimM PA3745 1 99499l3i3OS ribosomal protein S16 PA3746 (9949914 signat recognition partide protein FfhffhJ" PA3752 9949921 hypothetical protein PA3754 1 9949923 hypothetical protein PA3756 1 9949925 1 hypothetical protein yIfK PA3759 9949928 probable aminotransferase PA3765 ; 9949935 hypothetical protein ___ _ PA3767 9949937 conserved hypothetical protein PA3769 9949940 GMP synthase iQuaA PA3773 1 99499441 hypothetical protein PA3776 j 99499471 probabie transcriptiona ! reguiatqr !' PA3777" 9949948 exodeoxyribonuctease Vi) iarge subunit"*3eA PA3782" 9949954 probable transcriptionai regulatori--.---------.-. -. -...- PA3784499599i hypothetical protein __ I _ PA3785 99499571 conserved hypothetical protein PA3787 ; y ical rotein '. PA3788 99499601 hypothetical hypothetical protein PA3796 99499691 PA3803 (9949976, conserved hypothetical protein gcpE IPA3803 1 99499761 conserved hypothetical protein PA3805 i 9949978 t e 4 fimbnal bio enesis rotem PiIF i iIF PA3806 ; 9949979 conserved hypoth_etic_al protein 3 y B PA3806 9949979 conserved hypothetical protein PA3808 9949989 2conserved hypothetical protein-. ""_ _ ; ___'yfhJ PA3809 9949983ferredoxin [2Fe-2S] y fdx_2 _ y ^- _PA3810 I 9949984 heat shock rotein HscA _ I hscA_ _ PA3811 9949985 heat shock protein HscB' ; hsc_l3 , y, PA3812 9949986 probable iron-binding protein iscA."IIIJ'scA"J PA3813 9949987 probable iron-binding protein loci issu PA3816) 9949989 conserved hypothetical protein A3821 J 9949995 secretion protein Secs used ! PA3822 9949996 conserved hypothetical protein ; PA3827 ; 9950_002 conserved hypothetical protein .""____T.... _..... r_lgQ .... __--__ ilPA3827 i 9950002,, conserved hypothetical protein ; PA3828 ! conserved hypothetical__, p ! PA3829 pothetical protein iPA3834 99500û9. valyl-tRNA synthetase, valS PA3834 9950009. valyltRNP, snthetase ,. w m_. _. ___. _.. _. _.. _, aa (S.. _.... _ 1PA3843 9950019 hypothetical protein PA3840 : 9950016 conserved hypothetical protein. IybiN ßPA3843 9950019. hypothetical protein. {' PA_ . hypothetical _, 9950028 ! hypothetical protein I PA3854'9950031 5 hypothetical protein PA3856' ! 9950033ihypothetica [protein'1.'". !"'*' PA3857""T''"9950034Jconservedhypotheticarprot"]"'"""'""""''" " PA3857v. ;. .. 9950034 ; conserved ^M PA3859', 9950037'probable carboxylesterase PA3867 {9950046X, probable DNA invertase | PA3868'9950047 1 hypothetical protein % W cal rotem ; _. _. f......... _... __9950048-Ip----__p.-. _. ___.. _...-__. _. _..... ___. __.. ___r_.. PA3869_ _ h othetical rotem _ _ _ _. __ ! PA3876'j''9950056 nitrite extrusion protein 2jnarK2 1. TIII'I'' PA3884 r 995JOwP t PA3884")'9950064 hypothetica) protein _ _ . .' PA3884 1 9960064 hypothetical protein 9950066 hypothetical protein PA3888 9950069 probable permease of ABC transporter PA3890 9950071 probabie permease of ABC transpprter ! L.--I PA3891''9950072 probabie ATP-binding componentpf ABC tran1 9950073, conserved hypothetical p PA3905 j 9950088 hypothetica) protein ! j PA3905 l 9950088 hypothetical protein _ _ ! PA3905 _i, _ 9950088|hypothetical protein ____ _ l _ _. PA3906 : 9950089, hypothetical protein ì PA3906 9950089 ! 1 hypothetical protein A3911 9950094 ! conserved hypothetical protein PA3916 9950100) mo ! ybdopterin converting factor, large subunitiao PA3917 9950101 molybdopterin convertinq factor small subunitlmoaD PA3918 9950102 molybdopterin biosynthetic protein C ImoaC __ _-J PA3936 j 99501221probable permease of ABC taurine transporter, tauC . PA3960 r 9950149') hypothetica) protein ! -M t m hyp iPA3962' 9950151 hypothetical protein PA3965 9950154 probable transcriptional regulator PA, 3, 9-, 6, 7, 9950156 hypothetical protein PA3969 9950158 conserved hypothetical protein PA3973 9950163 probable transcriptional regulator I ! iaior 9. Y.. . _ pothefical protein oX ! PA3981 9950172 conserved hypothetical protein wi ybeZ PA3982 995017 served hypothetical protein PA3984 j 9950175 ! app) ipoprotein N-acy) transferaseiint cutE (' 8wI 99'0177 B et __ ___ PA3987, 9950178 jieucyl-tRNA synthetase, ileuS PA3987 99501781leucyl-tRNA synthetase PA3988) 9950179 j hypothetical protein ! j' JPA3989'f"9950180 JDNA poiymerase iii, delta subunit''"''ihoiA'"'. .. 1. 1I. J PA3990* r 9950182 conserved hypothetical protein : J PA3993 9950185 probable transposase PA3996 9950188 fipoatesynthasesfipAj) p ! PA3998 9950190 1 conserved hypothetical protein ybed -'' rodA mrdB PA4002 9 9950194 rod shape-determining protein [PA4005"j 9950197jconserved hypothetica) protein jybeBJ lPA4006 1 9950198, hypothetical protein _ __ w ; PA4008 9950201 iprobable hydrolase,... _......... ...... _........ _.... _...... __.. _. __. _....... __..... _. ___... ... __..... _.. _ _.... __. __. _. _.. __. _ _ _. ___. _____. 5 ; h othetical PA401 8. 9950211 hy, pothetical protein 5 PA4012 9960205'hypothetical protein PA4019 9950212'probable aromatic acid xylose _ S PM028 9950222. hypothetical protein sPA4029_ 9950224 ; conserved hypothetical proten _ : __ FdedA _ aPA4031 9950226 inorganic pyrophosphatase _ _ppa _ _tp_ __, _, _,,, _, ! PA4033 9950228'hypothetical protein IPA4037 9950232 probable AT- nding 1-K4-043 9950239 i geranyltranstransferase 1 ispa F. T. t PA4047'9950243iGTP cyclohydrolase 11, ribA L I PA4043 9950240 ; 1 deoxyxylulorse 5 phos __ i is A F PA4047_ _,'99502_43GT_P cyclohydrolase II V. w. ... _. _____ _=rib T_ PA4050 H _i-_H _99502461phosphatidrlgl cerophosphatase_A _ _pg_pA _ PA4051 9950247 thiamine monophosphate kinasefthiLL--I-L-- p {PM051''_ 9950247 CEPase IthiL t ____ _ PA4052''9950248 NusB protein'nusBssyB"' PA4052 9950248 NusB protein inusb ssyb PA4053 4 9950249 6, 7-dimethyl-8-ribityllumazine _y hase 0 ribE _ ribH PA4055 4 ; 9950252. riboflavin s nthase alpha chain V_ nbC _ y ribB ~ 0f77w777777_o777977 _777 PA4056 9950253 ribof ! avin-specificdeaminase/reductase ribD ribGj PA4056 9950253 riboflavin-specific deaminase/reductase ribg p 9950254 conserved hypothetical protein ybaD PA4059 950256 hypothetical protein ; PA4060 | _ 9950257 j hypothetical protein _. PA4063 r 9950260ihvpothetical protein v PA4064'9950261 j probable ATP-binding component of ABC tran, oH PM068 1. 9950266iprobableepimerase 1 F { PA4076 PA4076 y ; -9950274 j hypothet_ical, protein--,---_.. ___ PA4076 99502741 hypothetical protein PA4077 r 9950275 Jprobabte transcriptionai reguiator ! 11111 ! PA4083 9950282 probable pili assembly chaperone __ PA4097 9950298 probable alcohol dehydrogenase (Z_-depende _"."""*.. Y, d. L,,, _, » _ »., _.,,,,,,"__ 4 PA4099 9950300 hypothetical protein PA4104 1 9950305 conserved hypothetical protein PA4107 9950309 hypothetical protein PA4114 9950317 spermidine acetyitransferase.'. jb ! tD j iPA4121 t 9 0324jcon_er ed-yEgt-rotein _~ PA4122 1 E PA4122) 9950325 conserved hypotheticat protein j J ! PA4125 I 9950329 5-carboxymethyl-2-hydroxymuconate isomera ;p; hpcD PA4134 9950339, _hypothetical rotem PA4141 99503461 hypothetical protein PA4149 9950355, conserved hypothetical protein'acoX PA4151 J 9950357 acetoincataboiism protein AcoBjacoB ! PA4157 L 9950364 probable transcriptional regulator IPA4164 1 99503721hvDothetical Drotein,, I ,'jPA4167 i 9950375, probable oxidoreductase __~ yafB _____', PA4169 ! 9950377 conserved hyp ! in I w w}'--'~ PA4170 9950378 hypothetical protein iPA4171 probable protease PA4174't 9950383 probab ! e transcriptiona) reguiator. 11 PA4176 j 9950385 eptidyi-proiyi cis-trans isomerase C2, ppiC2 PA4181 1 9950390, hypothetical protein PA4182 9950391 !, hypothetical protein PA4183 i 9950392 ii hypothetical protein 'PA4190, 9950400, probable FAD-dependentmonooxYgenase, X, E Qerase ___. _. _. __.. __..... m.. __... ___. _. _. __. _____.. ____. _______. _.. ____. _. __.. _. _.... _. ___... ___... _. _... : PA42l6' 7 9950423jprobab ! ephenazmeb ! osynthesfsp""'"""' PA4211 9950424'. probable phenazine bios nthesis protein , PA4212 9950425iphenazine biosynthesis protein PhzC 'PA4215 able phenazine biosynthesis protein t tphzF1 _.... _ _ ;.. _-. _. _ i PA4216 99504301 probable ridoxamine 5'-phosphate oxidase I s 7 p ìPM230 : 9950446isalicylate biosynthesis protein PchB jpchB jPA4232'9950448'j singie-stranded DNA-binding protein jssb s'-.-.--- IPA4239 9950456130S ribosomal protein S4 ìPA4238, 99504551DNA-directed RNA polvmerase alpha chain rpoA W PA4240 9950457130S ribosomal protein S4 rpsD t { lpA4240 9950457t, 30S ribosomal protein S11"''7'\JrpsK''"i'"""""*" ! _...... _..,. _. _________-_. __. ... , _____. lPA4242. 9950459150S ribosomal Drotein L36 rpmJ S PA4243 ! 9950460 secretion protein SecYsecYpr ! A""' ! ° __. __ PA4244 9950461150S ribosomal protein L15 __ _ _trplO PA4245"T 9950462 50S ribosomai protein L30" rpmD''"1'F'T'-"ZL IPA4246 , 9950463130S ribosomal protein S5 rpsE t PA4246 9950463130S ribosomal protein S5 rpse PA4247. 995046150S ribosomal protein Ll 8 rpIR PA4248"j"9950465 50S ribosoma ! protein L6 7piF !-- j PA4250 9950466 30S ribosomal protein S14 rpsN PA4250 9950467 30S ribosomal protein Sl. 4 rpsn PA4251 l 9950468|50S ribosomal protein L5 rpiE j J PA4251 PA4253" !"9950470 50S ribosomai protein L14 rplN PA4254 j 9950471 30S ribosomal protein L14 pl ! PA4254 9950471 30S ribosomal protein S17 rpsq PA4255 l 9950472 50S ribosomal protein L29 rpmC PA4256 9950473 5 S ribosomal protein L16 _ rpiP _ _ __............. PA4257 _ii 9950474 30S ribosomal protein S3 rpsC _PA4258 E 9950475 50S ribosomal protein L22 rplV-1 . c.-_ PA4259 I, 9950476 30S ribosomal protein S19 rpsS __ ; PA4260 1 9950477 50S ribosomal protein L2 rplB _.. l. __. __. . _ PA4259 ! 9950476 30S ribosomai protein S19 rpsS j PA4261 9950478 50S ribosomal protein L23 I rplw or PA4263 1 9950480 50S ribosomal protein L3 1p C __ tPA4264 i 9950482 j30S ribosomal protein S10 2rPsJ..... 1,,-2 pA4267 9950485 30S r p PA4267 l 9950485130S ribosomal protein S7 IrpsG 2 Q PA4268 f 9950486 30S ribosomal protein S12 _jrpsL L PA4269 i 995048is DNA-directed RNA polymerase beta* chain irpoC IPA4270, i 9950488jDNA-directedRNApolymerasebetachain jrpoB---L--- PA4271 44E 9950490i50S ribosomal Drotein L7/L12 IrplL PA4 9950490 50S ribosomal protein L7/L12 rpil iPA4273 1 9950492|50S ribosomal protein L1 lirplA PA4272--9950491 50S ribosomat protein L11 _ _ M. ^. rplK_ _ rA4275 i 9950494Xtranscriptionantjterminationprotein NusG 2nusG] 'PA4276 i 9950495 secretion protein SecEsecE prJG ! PA4276 1 99504951 secretion protein 2 ; IPA4279 1 99504981 hypothetical protein PA4296 9950518 probable two-component response reguiator' l PA4298 2 9950520 i hYPotggt _ 2 w 'PA4299 l 9950521 Ihypothetical protein-2 2 2pA4305 1 9950527 ! hypothetical protein : {,, _ ' ; PA4305 hypothetical protein PA4 06 995052 h ot e cã r te ñ 'PA4314'9950538 fo ywofolatedeformylase purU1_ '_PA4314 T » _995_0538_eformyltetrahydrofolate d_ef_ormylase,,. m .. _ : purU1".,. ^. » _ w wy." »., ".,. y.,",.. x_. _.. hypothetical protein i PA4324. 9950548 i hypothetical protein 'PA4324 m 9950548 hypothetical protein ip PA4329 995055zt ; pyruvate kinase 11 1 pyka pyk-11 iPA4330 i 9950555iprobable enoyl-CoA hy ? dratase/isomerase t', 'PA4341.. 9950567.'probable transcriptional regulator __. __ _ _ ePM345 9950572ihypothetical protein ? - PA4348 9950575) conserved hYpothetica ! protein i [pA4349""9950576 ! hYpothetica ! protein'IIL1-ILj lLjL--I'-I 11111- ! 'PA4345 JPA4354 ; 9950581 conserved hypothetica ! protein i." ! PA4357) 9950584 conserved hypothetica ! protein "JyhgG''J ! .... ___ _ _. _.. .. _ ___ _. _.. __... ___... PA4359'J 9950586) conserved hypotheticat proteinTjfeoA ! =PA4366 j 9950594'superoxide dismutase . sodB , V v ; IPA4350 PA4373) 9950602 hypothetical protein JJ PA4359 9950586 1 conserved hypothetical protein ifeoa tPA4373 __ 9950602 hypotheuca protein _ __ __ i______, _ PA4383 j 9950613 conserved hypotheticalJcrcB ! PA4383 !"5"F i'1F __ _. ___W r_Mrr PA4377 I 9950607 ! h othetical protein PA4388. 9950616 GroES protein'"groES jinoRB"I ! ß--- w- PA4388 99506181 hypothetical protein PA4389 9950619 probabie short-chain dehydrogenase' ! ! PA4392 9950623 ! conserved hypothetical protein PA4395 9950626 conserved hypothetical protein PA4403"r 9950635 secretion protein SecA ! secA ! PA4405 9950637ih pothetical protein PA4406 3 9950638|UDP-3-0-acyl-N-acetylglucosamine dea_ylaiipxC _ 3envA asmB_ r.. Ir. 7 _ PA4407 J 9950639 jcell divislon proteln FtsZ ltSL WA4408 T 9950640Scell division protein FtsA < _ _ --- PA4408 99506401 cell division protein FtsQ jftsQ PA44p9 9950641 jceit division protein FtsQ ftsQTJ {PA4411 9950643 JUDP-N-acetyimuramate-aianineiigase murCjj PA4411' JPA4412 9950644} UDP-N-acetyigiucosamine--N-acetytmurarny1- ! j , PA4411 murc PA4413 f 9950645tcell division protein FtsW {ffsW PA4414 ! 9950646 UDP-N-acet Imuramo lalanine--D-lutamate I murD . _" -__.... _... ___. _y Y 9 __ __. s ! pA4415 9950647 phospho-N-acetytmuramoy)-pentapeptide4ranra} ORFY . _. _. __.... ____ _____.. . tY Y Y 9 y _ g 'H".. _,. .... s... _. _ PA4416 9950648 UDP-N-ace Imuramo lalan I-D-lutam _I 2 murF"_ j __,-". _,... __ ; PM417 _ UDP-N : acetylmuramoylalanyl gMmapE T. s PA4418 9950650 penicillin-bindin protein 3 ftsl pbpB ! PA4418, 9950650 penicillin-binding protein 3 ipb-pB 'PA4419 9950651'cell division protein FtsL _ ftsL _ _ _ _. _ tPA4420 __ 9950652$conserved hypotheticai protein _ w m W yabC ylxA __ ; | PA4421 T m---yabB___ _4 PA4424 f 9950657tconserved hvpothetical protein I yraN YPO Imraw yab ! pA44249950657jqnserved hypotheticai proteint yraNj 4425*""') 9950658 prpbabie phosphoheptoseisomerase yraOj . g _,... ....., j., . __ _, » ».. (PA4427 w_. _. 9950660 ; strin en_t starvation proten_B sspB _. PA4428 9950661 Istringent starvation protein A _ sspA _ _ ssp pog_ {PA4430'99506631probable cvtochrome b PA4432 9950665 30S ribosomal protein S9 _ irpsl .. _----.- PA4433 9950666 SOS ribosqmai protein L13'irpiM PA4436 1 9950670t. probable transcriptional regulator (PA4438"9950672 conserved hypotheticai protein ! ; PA4439M : _ 9950673=tryptophanyl-tRNA synthetase . __M_ ; trpS IPA4439 _ ! 9950673'Etryptophanyl-tRNA synthetase_ _ m _ t__ ___ PA444b 74 ! hypothetical protein , PA4450"''J''''99'50685] UDPacetyu'""" : PA4450 ^ _"9_95_0685°UDP-N-ace Iglucosamine 1-carboxymnyltransmurA ___r, w PA4452'9950687 ! conserved hvpothetical protein. $ 9950687 ! conserved hypothetical protein . PA4453 9950688'conserved hypothetical protein .......... tPA4455 9950690 probable permease of ABC transporter,'VrbE fPA4457'99506931conserved hypothetical proteinL-1J1 jyrbHkpsF""""""""" [PA4459. 9950695 iconserved hypothetica) protein j'jyrbK' iPA4459. 9950695iconserved hypothetical protein E, yrbK ; _. _ _ _ _.... _.... _. _... _ _..... .. ___l_. _.. _..... _. _ 1PA4461 9950697 l probable ATP-bindinp, 2omponent of ABC tran'3 lyhbg } pA4462' j"'9950698 jRNApoiymerase sigma-54 factor ! lE° ! iJl !. l [J"""" PA4462 i X w v f rpoN 3 ntrA PA4463'99506991conserved hypothetical protein-Y lbH RA4464 9950700 j nitrogen regufatory HA protein jjtsN'"''J'--- --'950-700"nitrogen regulatoEK il [A protein ptsn iPA4466, 9950702lDrobable phosphorvl carrier proteln f fS PA4471 L 9950707 hypothetical protein f fa'gA p'reC i I PA4471 9950707 1 hypothetical protein a I (P-g protem M m_r PA4480 9950718 rod shape-determining protein MreC mreC--j PA4482 9950720 Glu-tRNA (Gln) amidotransferase subunit C _ gavu ? tPA4483 9950721 G u-tRNA (Gln) amidotransferase subunit A gatA _ ~_5 __ PA4482 JPA4484'9950722 JG) u-tRNA (Gin) amidotransferase subunit B i gata _PA4_485. 9950723iconserved hypothetical protein PA4492 X 9950730|conserved hypothetical protein _ _ _L PA4499 9 9950738 probable transcriptiona) regulator ; PA4507 9950747"hypothetical protein PA4524 9950766'nicotinate-nucleotide pyrophosphorylase naau J PA4525 1 99507673type 4 fimbrial precursor PiiA ~ p A__,,, _~ o__K,. _-g to_ o _ * r PA4526 3 99507683tvpe 4 flmbrial biogKeMB Pl PA4527 i 9950770 stillframeshlft yHftlt p C_ _s_. _t PA4529 _ 9950772 conserved h othetical rotein i _ , YP P PA4530 99507701 ty PA4537 1 9950780|hypothetical protein __L 3 PA4537 9950780 hypothetical protein IPA4544. 9950788 pseudouridine synthase 1 riud yfil jPM547_ i 9950791thvo-componentres oBegulatorPilR __3pllR_ _3 PA4552'""''9950797Jty pe 4 fimbriaibiogenes protein Pi) W iPA4552 9950797'type 4 fimbrial biogenesis protein PilX tpllA g 3 ? W <~3~29--sl 3 PA4657< 9950802 j LytB protein ! IvtB _ < 3PA4559 3 9950804fprolig ? oprotein signal peptidase 3. 151> 3 s PA4560 3 99508051isoleucyl-tRNA synthetase liles PA4561'9950806 {ribof) avi ribFij PA4563, 9950809130S ribosomal protein S20 rpsT IPA4564 ç 9950810iconserved hypothetic, al protein I ÇscreA Ç3 PM565'j 99508111glutamate5-kinåse __ leror _ I _. ooi PA4565-_ 9950811 _lutamate 5 Wnase , _roB. T.,.. j IPA4567 9950812 ! GTP-binding protein Obg''tobg"jU. I' 99508131505 ribosomal protein L27 rIlmA i PA4567 ; PA4568. j.. _. 99508_14 50 S ribosomal protein L21_ PA4569 9950815 joctaprenyi-diphosphatejsyntnasepB ! ce) tPA4574 ? 3 9950821 Xconserved hypothetical protein ___, _ __ _yL < PA4575'9950822 hypothetica) protein j"'" !' PA4577 9950824 hypothetical protein 1' 4m} 99508241 hypothetical protein j'.'LJ'l-------I IPA4586 t 99508341hvDothetical Drotein i I ? PA4577 9950824 hypothetical protein r rPA4603 t 9950853 ! hypothetical protein JPA4610 j995086Thypotheticaiprotein'.... IL...... i. J"---- i PA461 1 9950862i hypothetical protein tPA4617 9950868. . conserved hypothetical protein ì íygjo PA4630 9950883 j hypothetical protein 'hypothetical pro in . PA4637 9950891.'hypothetical protein'. iPA4638 9950892'i hypothetical protein , PA4638 9950892 X hvpothetical protein i PA4642 9950895 S hypothetical protein . ___... g950895 ical protein ". _.. ___.. _. 'iPA4642 9950895i hypothetical protein PA4646'9950ituracil phosphoribosyltransferase lup. p PA4649 hypothetical protein PA4649 PA4655 ; 9950909iferrochelatase _ -hemH rv s f _ : _. _____. _.. _... _ _.. __.. __ _. _ PA4662 ! 9950917 g ! utamate racemase mur)..--.----- PA4663,'9950918 molybdopterin biosynthesis MoeB prot, iln moeb chIN f PA4665 9950920 E peptide chain release factor 1 f prfA rf1 PA4666 1 9950921, giutamyl tRNA reductase _ XhemA hem1 ; glUtR_ 9 PA4669 99509241isopenten I mono hosphate kinase ipk ychB PA4669 9950924 ! isopentenyl monophosphate kinase ipk PA4670 §1 99509251ribose-phosphate pyrophosphokinase prs prsA PA4671 9950927 probabie ribosoma) protein L25 IP ! Y.. ! P p -y,,. PA4672 9950928 peptidy-tRNA hydrolase _ {pth PA4672-9950928 peptidyi-tRNA hydrol6se pth PA4674 j 9950930 conserved hypothetical protein PA4676 able carbonic y PA4679 1 9950935 hypothetical protein f PA4679 9950935 hypothetical protein __ .. _.. _. ... . __. PA4693 9950950 phosphatidytserine synthase pssA j : PA4696 9950953 acetoiactate synthase targe subunitHM''''"i PA4697"9950955 hypotheticai protein"'j j PA4698"1'"'9950956fhypotheticat protein''"'------Y-----------yqcCj''"'"""'"'""""""J PA4699 j 9950957) hypotheticai protein L---JLJIL----- PM699 S 99509571hypothetical protein _ S PA4702 1 9950960 hypothetical protein =,- PA4 69 8 995095, 61 hypothetical protein PA4711 1 9950970 hy omcalprotein I _ _ H' PA4699 9950957 hypothetical protein IPA4728 I 9950988 2-amino-4-hydroxy-6-hydroxymethyldihydroptefolK PA4711 1 99509701 hypothetical protein li5A4728 i 9950988 2-amino-4-hydroxy-6-hydroxymethyldihydroptdfolK tPA4731 t__ 9950992 aspartate 1-ciecarboxylase precursor e~_ PA4732 j 9950993 g) ucose-6-phosphate isomerase pg) ! PA4737 i 9950998 hypotheticai protein T' ! PA4737 9950998 i hypothetical protein PA4739 9950999 conserved hypothetical protein yjbj PA4740 99 9951002 [polyribonucleotide nucleotidyltransferase pnp PA4741 9951003 9951003ì30S ribosomal protein S15'rpsu S PA4744 J 995106 transtation initiation factor) F-2" f) nfB"'"'J] iPA4745 r 99S1007 N utiiization substance protein A (nusA''""""" j PAA745 9951007 N utilization substance protein A PA4746 9951008 conserved hypothetical protein _y PA4747 j 9951009 secretion protein SecG tPA4748 1. 99510101triosephosphate isomerase ItpiA _ Itpi _~, _ PA4749 9951011 phosphogiucosamine mutase gimtVi yhbF, mrsA j PA4750 ! 9951012 dihydropteroate synthase"'Ifoip dhpS"J [PA4752'995105 ceii division protein FtsJ'ftsJ S sPA4753 t 9951016>conserved hypothetical protein S _tyhbY,,,", __,,,,",,",",,"_, _. _. _u PA4757. 9951020'conserved h othetical protein '_PA4759_^. _... _, __. 9951022dihydrodipico_linat_e_reductase. T _..,.... . _........ <dapB... _,....-. _........ _. _.... _. heat shock roten Gr E E rot in fur ,, p g. . _.. _...... _.' ___ ___. __, ______ , PA4765 9951029iouter membrane lipoprotein OmIAN_. _. _. _.. _. _. __. mom_IA, T_ . _ ! °PrX_. __... __... ___.. _... _.. _. ___.. _ PA4767'9951031) conserved hypothetical protein iPA4773 9951038 hypothetical protein JPA4776 9951041] probab) etwo-componentresponse regutator L. . ! !'MZs" Jstor I '. m." ! yH.'l'1""11" protein S PA4782 _'9951047,'hypotheticai protein X f ___ , A4788 v-r IPA4789 9951055, conserved hypothetical protein v f PA4790 ! 9951056 conserved hypothetical protein f smtA PA4792 9951058 conserved hypothetical protein PA4797 99510641 probable transposese PA4802 {9951069 hypothetical protein', _ jPA4809l""r""9951077 FdhE protein. U'Hr III'IIIIIIII). fdhI"II"tH IIIIIIir"IIIIIII'. IPA4809, 9951077iFdhE protein ffdhE PA4823 1 _9951 0911 hypothetical protein PA4813 | 9951080flipase LipC _ e9_ PA4823 1 9951091 hypothetical protein _ PA4826 9951095 hypothetical protein PA4828 9951097 conserved hypothe'ti'c'in Z PA4831 9951100 probable transcriptional regulator PA4841 9951111 conserved hypothetical protein PA4847 i 9951 118 biotin carboxyl carrier protein (BCCP) $accB fabE , PA4848 1 99511191biotin carboxylase _ {, accC _ L IPA4850, 9951121 tribosomal protein L11 methvltransferase IprmA 9951121 ribosomal protein L11 methyltransferase rprmA PA4853m. 9951124jDNA-binding protein Fis _____ fis __Hw_ } PA4861 j 99511 33 I probable ATP-binding component of ABC tranS _ t . _. _. ___. Pua4864 PA4864 9951137iurease accessory protein lurpr) PA4866't 99511391conserved hypothetical protein s I _ ______ __ _' ; ureC PA4868 9951141 grease alpha subumt a_..,. _. ; p 9951141 urease alpha subunit-- !-Gre-C PA4874 9951148 conserved hypotheticai proteinj jpsjF PA4871 9951144 ! hypothetical protein T » 874 JmiF _ PA4875 PA4878 i 99511521 probable transcriptional regulator i PA4885 $ 9951159itwo-componentresponse_TiriiR TPA4887 r 9951161 {probable MFS transporter T PA4890 9951165 conserved hypothetical protein lyljc m i 99511671urease accessory protein UreF PA4894 9951169 hypothetical protein iPA4895 1 99511701probable transmembrane sensor g} * 99511701 probable transme IPA4906 9951182 probable transcriptional rilalatol iPA49-08 9951185 hypothetical PA4916.-..... ____. g951_-. 193 i hypothetical protein iPA4920 1 9951198 ! NH3-dependent NÄD synthetase ;'nadE A4923 9951201 conserved hypothetical protein a 1PA4925 9951203 conserved hypothetical protein : PA4926 9951204 conserved hypothetical protein ! pA4931"* ! 9951210 repticative DNA helicase jPA4934"j"9951213 30S ribosomai protein S18''"JrpsR'j.-1-J'I ! 'PA4935 g 9951214t30S ribosoma'l protein S6'rpsF} nbosomal proten SB.. _.-.- ! PM938, 9951218iadenylosuccinate synthetase ipurA ripa4940 PA4940 r 9951220jconserved hypotheticat protein' ylsT L----j-. 'PA4944': 9951224iconserved hypothetical protein'hfq PA4945 9951225 J delta 2-isopenter) ! yipyrophosphate transferase miaA"" !--.--- PA4948 9951228, conserved hypothetical protein PM952. 9951233 jconserved hypothetical protein _, yjeQ PM956 9951237, thiosulfate sulfurtransferase _ _, rnc A _ __, ___"_., _______ PA4961 9951243ihypothetical protein PA4962 9951244 conserved hypothetical protein LyJIIILJII PA4964 9951246 topoisomerase) Vsubunit A JB-.'" PA4964 99512461topoisomerase IV subunit A _PA_4_ _99512471 hypothetical protein. r I i ____ __i___. S. __, # __ _. ___,. A : PM966 i 9951248lhypothetical protein PA4966 i 9951248 hypothetical protein PA4967 9951249 topoisomerase IV subunit B 1'parE IPA4969 9951252 conserved hypothetical PA4972 99512551 hypothetical protein jPA4980''9951263jprobabte enoyt-CoA hydratase/fsomerase !'''' PA4988 ! 9E : =sonic-acid (KDu tranwaaA {kdtA PA4990 i 9951275lSMR multidrug efflux transporter _ H ___ -S tv s PA4991 t 9951276 hypothetical protein 1PA4992 hypothetical protein PM992 9951277 hypothetical protein'_, _1 _ _"__-__ __. _ _. i'A4998 - 9951283conserved hypothetical protein ; __..... _ PA5006 S 9951292) hypothetica) protein'*") J PA5007 i 99512922hypothetical protein _ PA5007 i wapQ InaA __ PA5008 i 9951294, hypothetical protein __ s watwaaX PA5008 1PA5009 9951295ilipopolysaccharide core biosynthesis protein aaP rfap PA50 10, 9951296 J ; UDP-glucose : (heptosyi) _LPS< waaG _ rfaG PA5011 9951297tjheptosyltransferase i twaas a p Y ^' m ___... _ _.. lpA5012 1 9951298 heptosvltransferase 11 iwaaF rfaF L PA5028 9951317 conserved hypo ri --A5032 robable transcriptional reguliatior---T-- PA5033 99513221 hypothetical protein JPA5034 j 9951323 uroporphyrinogendecarboxyiase smj... j. [PA5039r'9951329) shikimatekinase''''TaroK''" ! i PA5044 i, 99513341type 4 fimbrial biogenesis protein PilM'jpilM},,, _ PA5050 J'995134Tprimosomat protein N'JpnA' ! PA5051'"T"9951342 arginyt-tRNA synthetase""argS"""""""""""i PA5052 : 9951343 hypothetical protein _ ru-A j 9951353 conserved hypotheticai proteinj phajmH ! PA5063 1 99513551ubiquinone biosynthesis methyltransferase UD] UDlt J PA5064 1) rotein PA5065 9951357jconserved hypotheticat protein' ! lTI) aarFytgRi PA5067 j 9951360 ! phosphoribosyATPpyrophosphphydro L----IL- ! PA5068 i 9951361 translocation protein TatA....... "jmttAyigT....] 5PA5071 7 995_4iconserved hypothetical protein t _ _ PA5072 9951365 probabte chemotaxis transducer J PA5081 i 9951375 hypotheticat protein ! PA5085 j 9951379 probabte transcriptiona) regufator 1 e7PA5072 7 9951365|probablechemotaxis transducer PA5072 9951365 probable chemotaxis transducer 7PAS081 7 995-375 jhypothetical protein PA5085 99513791 probable transcriptional regulator iPA5110 _ ! 9951406 ! fructose-1, 6-bisetgt __ __bp _ Icrr r iPA511j9'J9951417jgjutamireynthetase"'igtnA "''T. ! IPA5116 i 99514131probab, etranscriptio ~ _ ~ iPA511g. .'. 9951417 ; glutaminssynthetase __ _.. w.. _^.. 3.. gInA _. _-,. ,. __. . ... . -... _. _. _. _. ___. _. __. _.... ___. __......... _.... ___...... _____.. __..... _ _.... _ __. _.. _.,...., il_A5128 _ 9951427 ìsecretion protein SecB, secB ; PA5129...... _ _995_1428Mlutaredoxin____ _ ........ _. 9 ! W. _ _.. _. _... _.... _.... _.... _.. _.. _ _. _., a 'PA5130. 9951429i. conservedMttrotein'"e. _ 'PA5 3i t 9 51430'p osphogdycer mutase, P9. m,,, _. _ Y !, b,. 0 9951429 ! conserved hypothetical protein ! PA511'9951430) phosphog) yceratemutase pgm' ; yib0 fA51-3'-2'** ^. __.. _..,. ... 9951431 fhpothetical rotem _, __.. __.... __ _... _.. _. _ _.. _. ___.. N_ PA5142 :'9951442'gjutamine amidotransferase ... I'lHI--r--"-Y- ; PA5144 9951444 ; hypothetical protein _ _ __ ' JPA5148"9951448iconser hypotheticai proteir),...... yggX'" _ __4 _9514... _. ____.. _. ; PA5144 9951444 ! hypothetical protein PA5161 9951463 dTDP-D-glucose 4, 6-dehydratase ! rm B rfbb PA5162 {9951464 dTDP-4-dehydrorhamnose reductase, rmlD irfbD PA5163, 9951465iglucose-1-phosphate thyMlyyitra_sferase lrrnlA'rfbA _ PA5164"I"'''9951466 dTDP-4-dehydrorhamnose 3, 5-epim''" PA5173"r 9951476 carbamate kinase) arcC"") PA5176 _. _.. _ __ econserved h othetical rotein yrfE {PA5178 1 9951481. conserved hypothetical protein tPA5182 1 9951486rhypothetical protein PA5187 9951491, 1 probable acyl-oA dehydrognase PA5190 9951495 probable nitroreductase IPA5195 9951500 probable heat shock. rotein rfrh PA5221-"---F-'"--9951529 probable FAD-depe6dent monooxy enase ivisc PA5222 ; 9951530 hypothetical protein PA5224 9951532. aminopeptidase P lpepp PA5225 9951533 hypothetical protein PA5227 y95 _9951535 conserved hypothetical protein_y_V ; --w.. _. _. _. _ygfE W_ _. _. _. _. _.. ___. ___. __. _. __ PA5229, 9951537 conserved hypothetical protein i, 9951537 conserved hypothetical protein _P_A_5_239'9951548 transcri tion termination factor Rho r_ho .. _. i__. ____ .. p _ __. _.. PA5240 1 9951549 thioredoxin i trxa PA5246 j 9951556 conserved hypotheticai protein iyigj) PA5247 1 9951557|conserved hypothetical protein yail PA5259, 9951570 uroporphyrinogen-lll synthetase _ _ _4hemD _ ____. __, __ _ PA5260 = gen deaminase _, hemC HopE___ PA5276 9951589 jtipopeptideLppL precursor) tppLJJ 'PA5276 j 9951589Semg eE iiPPL_ 2 PA5276, 9951589 lip, , pP p, _p PA5278 vgwldiaminopimelate epimerase HpF S............... ! Y. gB ig M w--r PA5296 ; 9951611 IATP-dependent DNA helicase Rep rep IPA5300 j 9951616icytochrome c5 _, icycB, _. ___. _. __,. _ {PA5303 1 9951619. conserved hvpothetical protein fiPA5316, Mm M PA5316 9951633, 0 ribosomal protein L28 'r mB , I PA5319 9951636 DNA repair protein RadC _ r adC PA5319j 9951636 DNA repair protein RadC"'] 7adC' J PA5319 9951636, DNA repair protein RadC lradc tPA5321 1 9951638 jdeoxyuridine 5'-triphosphate nucleotidohy olEdut _ _ i _ _I Y p p Y _ 4 __ ; PA5325 9951643 hypothetical protein wl 9951646|probable cytochrome c (mono-heme type) PA533 9961648 hypothetical protein v g i r PA5331 1 9951649lorotate phosDhoribosvltransferase ìpyrE I _i PA53339951652iConserved hypotheticai protein"i ' ! I- P A-99516531ribonuclease PH rp------- h. othetical iPA5347 9951667 ! hypothetical protein iPA5339 99516581conserved hyEothetical protein iPA5350 9951670 i rubredoxin JPA5350""9951670 kubredoxin""""" J"J""'-------" ! PA5351""""''995167l1rubredoxin"J 11111111'''L Umn'-""- iPA5358 9951678) 4-hyd roxybenzoate-octaprenynransferase J'ubiA , PA5351 : 9951671 rubredoxin ! PA5364 9951685) probable two-component responseregutator J""""""" PA5381 9951704ihypotefical protein IPA5385 9517091 hypothetical protein PA5381 9951704 ; hypothetical protein.'i i ; PA5385 : 9951709ifMbe X _ __ 'iPA5390 99517141 able peptidic bond hydrolase i'iPA5396, 9951721 ihypothetical protein if_'A 03 ; 99517291 probable transcnptional regulator.,., . _., _",.. _". _.. -T. __. _. __. ___. _. _..... _.. _... _.... _. _. _ PA5404 1 9951730i, hypothetical protein ___ ;, __ ì_. _.,,, _ _,,,.,, _ iPA5406 i 9951732 hvpothetical protein f --------------------- 1PA5406. _- :--995173 : hpothefical protein JPA5406"""9951732 hypothetical protein'"'"') ILL-I11-"1- PA5417 I PA5417 9951744) sarcosineoxidase delta subunit (soxD j PA5408 9951734 ! hypothetical protein PA5460 | 99517921hypothetical protein. 1 _, PA5417 1-788 1h pothetical protein PA5457 PA5469 ! conserved hypothetical protem f PA5470, f 9951802, jprobable peptide chåin release factor, Sprrt PA5480 1 9951813fhypothetical protein _ _ PA5482 9951816, l hypothetical protein PA5482 9951816 hypothetical protein PA5503 f 9951839 probabieATP-binding'component of ABCt' !"..... ! PA5526 9951864 ypothetical proten i < PA5529, 9951867probable sodnm/roton anti orter _ __. __, _... _.. _ PA5529 . _, __m. _.. _ ; 9951871 hypothetical '9951873 hypothetical protein PA5534 __ PA5543-T951-882 hypothetical protein PA5549 ! __9951889jglucosamine--fructose-6-phosphate aminotrarigl_mS 9 PA5552 1 9951892 1 glucosamine-1-phosphate acetyltransferase/'Imu ca PA5553 1 9951893, (ATP synthase epsilonchain u G PA5554 j 9951894 ATP synthase beta chain atpd uncd papb PA5555 ! 9951895ATP synthase gamma chain atpGuncGpapC... J PA5556'T 9951896 JATP synthase alpha chain--"'-'''''- atpA"uncApapA"j r. i I x b J PA5557 9951897 ATP s nthase delta chain atpH ncH papE PA5558 9951898 ; ATP synthase B chain atpF fflF papF PA5559 J 9951899 atp synthase C chain atpE uncEpapH J PA5560 i ? 99519001ATP synthase A chain _ _, ~ un pK sPA5561 1 9951901 IATP svnthase protein I ? axpl ? uncl i iPA5562 ! 9951903 chromosome partitioning protein SppOJ ! spoOJ J jPA5566 f 9951907lhypothetical protein, f IPA5566 1 99519071 hypothetical prot ! iln IP 9951909 ! conserved hypothetical protein id 1PA5569--9519101ribonuclease P protein component I rnpa 1 9951911150S ribosomal protein L34 IP@e V DNA replication, recombination, modifica 483 2027 DNA replication, recombination, modificat 4275; 6695 Hypothetical, unclassified, unknown 833 7803 : Translationl post-translational modificatiofn 124i 10434. Translation, post-translational modificatioi 12488 ; 10434 Translation, post-translational modificatioh 13435j_12488. Cell wall/LPS/cap§ule 14235ii 15122 : Hypothetical, unciassified, unknown 7217 1 S900j Hypothetical, unclassified, unknown 24001 Hypothetical, unclassified, unknown 27646j 28632 Hypothetical unclassified, unknown 36270 5 35905 Amino acid biosynthesis and_metabolism l _ __ _378_93--37087 i I__.. _... . __. .. ; Hypothetical, unclassifiedl unknown _ ! 40190j_ 404054. Hypothetical, unclassified, unknown 40190 40405i Hypothetical, unclassified, unknown 40589 40816i Hypothetical, unclassified, unknown 56546 56941 ; Hypothetical, unclassified, unknown 61879 62388J Hypothetical, unclassified, unknown 68616 681881 ! Hypothfetical, unclassifiedl unknown _ 69272i 695261 Hypothetical, unclassified, unknown 70091 C43 s f Hypothetical, unclassified, unknown 70636 70130 Hypothetical, unclassified, unknown 72880 73384 :. Adaptation, protection 73468 73923 t Hypothetical, unclassified, unknown 74034 74267 Hypothetical, unc ! assified, unknown 74715 74279 ! Hypothetical, unclassified, unknown 78097 77432 ' Hypothetical, unclassified, unknown 80752f 81027i Hypothetical, unclassified, unknown 811161 821741 Hypothetical, unclassified, unknown 83318 82404- Hypothetical, unclassified, unknown 98218 97754. Hypothetical, unclassified, unknown I 100124 101158i Hypothetical, unclassified, unknown 115045 1146111 Hypothetical, unclassified, unknown 1201641 Hypothetical, unclassified, unknown 121346 1222661, Energy metabolism 127378 1285021 Hypothetical, unclassified, unknown 131792 13158 Hypothetical, unclassified, unknown132577 33155j Energy metabolism 134319i 135233i Hypothetical, unclassified, unknown 1352591 135894j Hypothetical, unclassified, unknown 136386 135934 Hypothetical, unclassified, unknown i 136518gf 136991 Transport of small molecules 138818 140167 Transcriptional regulators 140218 140902f iHypothetical, unclassified unknown | 143848 143567a H pothetical, unclassified, unknown 1 4072 143845 Hypothetical, unclassified, unknown 145542--1-458-831 Hypothetical, unclassified, unknown 1 1494251 149138, Transcriptional regulators E 150906 151823 ; Hypothetical, unclassified, unknown 153696 153836 Adaptation, protection 158199158762 Nucleotide biosynthesis and metabolism iL 164415 ; Hypothetical unclassified, unknown 165737i 165219i , Transcriptional regulators r 16936iL i69906 zCarbon compound catabolism'1755D33 176108 ; Transcriptional regulators, 1827681 183706 www4 ~ Hypotheticall unclassified, unknown_. _ 184287 184439' Transcriptional regulators'J 191697 192362 H pothetical, unclassified, unknown 1 194179i 193799 HYpothetical unciassified, unknown, 19474831 194206' Putative enzymes)"207071 207823 Carbon compound catabolism ! 209533 j 207923 i transport of sma ! ! motecuiesj 210460J 209621 Hypothetical, unclassified, unknown j 2 214634 Hypothetical, unclassified, unknown, HãF 215512' Hypothetical unclassified unknown i 229738 229526, Putative enzymes, 232000 2305433 Transport of small molecules j 233100 232066'. Transport of small molecules j 233932 2331231 Transport of small molecules 1 234849, 2339293 Transcriptional regulators 236218 2371111 Hypothetical, unclassified, unknown 238896 239777 j Carbon compound catabolism 240071 240934 Hypothetical, unclassified, unknown 241753 2424455 Transport of small molecules 243841 244605i Transcriptional regulators 262557 263498 Transcriptional regulators 266616 267395 Hypothetical, unclassified, unknown 268704 2695191 Transcri regulators. 275772 276440 Hypothetical, unclassified, unknown 277334 276480 Amino acid biosynthesis and metabolism 277777 277331 i Hypothetical, unclassified, unknown 282757 282323 Hypothetical unclassified unknown 283553 282912 ! H othetical, unclassified, unknown-i 289390 2892051 v i Hypothetical, unclassified, unknown !, 293304 291154 Hypothetical, unclassified, unknown 293798 293301) Hypothetical, unclassified, unknown 299497 299081' Transport of small molecules 309092 307878' Transcriptional regulators 313227 3139251 Transport of small molecules 31 49Z71 313938' Hypothetical, unclassified, unknown 318148 317966 ! i 31814a 3179661 Hypothetical, unclassified, unknown 350841 350089 ! Hypothetical, unclassified, unknown 352164 351610 Hypothetical, unclassified, unknown 359982 360332J Enerqy metabolism i 371833 3711621 Hypothetical, unclassified, unknown 373725 3741921 Hypothetical, unclassified, unknown 378096 378575 Hypotheticat. uncfassified, unknown 382792 382037 ìFattY acid and phosPhöiiPid métaboiism 383727 3w w S Nucleotide biosynthesis and metabolism 384733 385527 Biosynthesis of cofactors, prosthetic grou 393308 393814 Hypothetical, unclassified, unknown 402020 402598 Energy metabolism 406498 406247' S s Central intermediary metabolism 407098 ! Hypothetical, unclassified, unknown 413654 SHYpotheticall unclassified, unknown ! 414529i 413933, -F Protein secretion/export 4175271 418894 ! Transcriptiona) reguiators'420683J 421537J Hypothetical, unclassified, unknown i 422207 : IHYpothetical unclassified, unknown, 423460J 423660 ', Hypothetical, unclassifiedl unknown 1 4271201 426863 i Hypothetical, unc ! ass ! fied, unknown 439991 ! 440395 ; ! Nucleotide biosynthesis and metabolism 445691 ! 444687j iTranscriptiona) reg uiators"446227 j 445715j . _. _ 9 t ~ Hypothetical, unclassified, unknown"446773 ! 446339 j , Hypothetical, unclassified, unknown i 4473421 446773 ; Biosynthesis of cofactors, prosthetic grou 449384 448431) Motility & Attachment 453239 4541141 Transcriptional regulators 463079 463873 ; Hypothetical, unclassified, unknown 470081 470650 Transport of small molecules 1 476333 4777904 Hypothetical, unclassified, unknown 484404 4848381, Hypothetical, unclassified, unknown i 496478 496362J Transport of small molecules 496871 498361 ; Related to phage ; transposon, or plasmid 501120 5001041 Hypothetical, unclassified, unknown 502599 501376 Transcriptional regulators.'~ 504121 505029, H othetical, unclassified, unknown 510499 509825 t Adaptation, protection 514775WiX Hypothetical, unclassified, unknown {526877 527179 J Hypothetical, unclassified, unknown 535085 535489 ! Transcriptional requlators 535539 5361081 -4 Transcriptional regulators 539143 538217i Transcriptional regulators ! 540735 539785 Hypothetical, unclassified, unknown ! 549614 549294 Putative enzymes 5503811 Hypothetical, unclassified, unknown 550813 550520 Putative enzymes, 552746 5529941 Hypothetical, unclassified, unknown 558361 557354 Biosynthesis of cofactors, prosthetic grou 560808 562013 Biosynthesis of cofactors, prosthetic grou 562006 562728 ! Biosynthesis of cofactors, prosthetic grou 562721 563545 Biosynthesis of cofactors, prosthetic grou 563549 564235 Hypothetical unclassified, unknown 564344 564574 Biosynthesis of cofactors, prosthetic grou 576040 575516 Transcriptional regulators ! 586663 585980 Putativeenzvmes {590105 590821 S 0 w Transcriptional regulators 594580 595134 Hypothetical, unclassified, unknown 598608 598994i Hypothetical, unclassified, unknown 600176 599757i Hypothetical, unclassified, unknown J 600426 601394 Hypothetical, unclassified, unknown Central intermediary metabolism 604896 603706i Hypothetical, unclassified, unknown 609999 609202 Energy metabolism 611281 612444i jpothetica), unciassified, unknown J61M Carbon compound cataboiism ! 613338 614402 j Hypotheticat. unciassified, unknown J 617549 616371J j Hypothetical, unclassified, unknown" ! 620488T 620135t Hypothetical, unclassified, unknown 621695 622033i Hypothetical, unclassified, unknown J 622726 622884J Hypothetical, unclassified, unknown f, 624199', 6238524 w f Hypothetical ! unclassified, unknown S 624803 624189 l Hypothetical, unclassified, unknown 1 629884 628763i , Hypothetical, unclassified, unknown 638830 6383811 Translation, post-trånslational modificatiogl 639115J 638900' Translation, post-translational modificatioh 639316'640341 i Biosynthesis of cofactors, prosthetic grou ! 641073 641426i Hypothetical, unclassified, unknown 643714 643208J Hypothetical, unclassified, unknown 649263 648931 l Hypothetical, unclassified, unknown 650538 650158 Biosynthesis of cofactors, prosthetic grou 652483 651497 I w- Chaperones & heat shock proteins 653772 652480 Adaptation, protection 656527 653753 Energy metabolism 669415 670089 Transcriptional regulators l 673091 672777} Transcriptional regulators 673961 673191 Hypothetical, unclassified, unknown 674667 675026r' Hypothetical, unclassified, unknown 675390 675839 Related to phage, transposon, or plasmid 677083 677409 Hypothetical, unclassified, unknown 685846 6867. 183 Hypothetical, unclassified, unknown 686693 686899 Related to phage, transposon, or plasmid 688605 668967' s Related to phage, transposon, or plasmid 689236 689466 Related to phage, transposon, or plasmid 690420 690674f Hypothetical unclassified, unknown 693596 6943661 Hypothetical, unclassified, unknown 1 698932 699720i Hypothetical, unclassified, unknown 1 699744 7008351 ! h Related to phage, transposon, or plasmid 700835 701170i Hypothetical, unclassified, unknown 701477 7025291 Related to phage, transposon, or plasmid ; 702529 702831 ; Related to phage, transposon, or plasmid 702828 7030588; Transcriptiona (regu (ators 706672 706028 ; 0 0 Hypothetical, unclassified, unknown 706944 707366 Hypothetical unclassified unknown 1 709182 708535 Hypothetical, unclassified, unknown 714247 713279J P l Hypothetical, unclassified, unknown 714686 714264 j Hypothetical, unclassified, unknown 1 717231 175'81 Proteinsecretion/export apparatus 737530 737081 Hypothetical unclassified unknown 737677 738108= Protein secretion/export apparatus 738485 738111 j Protein secretion/export apparatus 741335 741928 Protein secretion/export apparatus* 744333 745742 Protein secretion/export apparatus 745742 746956 Hypothetical, unclassified, unknown 748662 749774 Hypothetical, unclassified, unknown 770156 770818 Hypothetical, unclassified, unknown"770847 771326 ! Hypothetical, unclassified, unknown 772275 772700 j 'Hypothetical, unciassified, unknown 775321 774416J . ; , Putative enzymes 1 778181 1 7767871 Putative 779208s... _... _ 778309 ; 'Transcriptionaf regutators 782113 781259j X, unclassified, unknown w 782525 ; Central intermediary metabolism'S 782570g 782965i Hvpothetical, unclassified, unknown l 7838331 783576 ! Hypothetical, unclassified, unknown i 784698j 785174J Hypothetical, unclassified, unknown 785969) 786925! Hypothetical, unclassified, unknown f 786928 788253J Related to phage, transposon, or ptasmidj 7j39144 789356J Related to phage, transposon, or plasmicf, 7901661 9060011 Related to phage, transposon, or plasmid'795793 796776} Hypothetical, unclassified, unknown 797251 797598' Putative enzymes"")'"798827 797925 Transcription, RNA processing and decirF 801967 8012751 Hypothetical, unclassified, unknown ! 802239 801967J Hypothetical, unclassified, unknown 805228 805473l Hypothetical, unclassified, unknown 1 809882 8095743 Hypothetical unclassified, unknown 828344 827400 Secreted Factors (toxins, enzymes, algin 831914 832498 Protein secretionlexport apparatus 835523 837322 8 3 Protein secretion/export apparatus 837328 838182 Translation, post-translational modificatio 839407 840324 j Hypothetical, unclassified, unknown ! 844295 843723 Hypothetical, unclassified, unknown ! 845682 845278 Transport of small molecules 858646 858951 Hypothetical, unclassified, unknown 859007 8601701 Hypothetical, unclassified, unknown (866451 865636 Hypothetical, unclassified, unknown 881077 881400 Hypothetical, unclassified, unknown 883216 882989 4 Hypothetical, unclassified, unknown 885635 886108 Transcriptional regulators 893041 893994 Hypothetical, unclassified, unknown J 895668, 8953961 Hypothetical, unclassified, unknown 896416, 897228i Hypothetical, unclassified, unknown 898886i 8984400 e wl Hypothetical, unclassified, unknown 900165 8998301 Hypothetical, unclassified, unknown 901046-9004081 Putative enzymes ; 903692 904633i Chaperones & heat shock proteins 913086 9135fit Hypothetical, unclassified, unknown 929084 929503 Hypothetical, unclassified, unknown Putative enzymes 932725 932102 Ce) t division935989 936294 Hypothetical, unclassified, unknown, 9426481 9434301 , Hypothetical, unclassified, unknown 948776 9491591 Hypothetical, unclassified, unknown 949280 949693 Cell wall/LPS/capsule 1 9506481 9497161 Amino acid biosynthesis and metabolism 952514 952158 Hypothetical, unclassified, unknown 955722 955456 Putative enzymes 9625451 962925 __... __. Hypothetical, unclassified, unknown 9777431 9774201 ! , Hypothetical, unclassified, unknown ì 9842451 984535, Translation, post-translational modification-986818 989442J Amino acid biosynthesis and metabolism, 989590.'990828 gTranscriptional requlators 991013 : 991198 jTranscriptional regulators', _ 992543= 991830 : {Hypotheticat, undassified, unknown F'993409993783j 993409-993783 Hypothetical, unclassified, unknown. 993776j 994051. Transport of small molecules'996038r 9974861 Hypothetical, unclassified, unknown J1007548 {10072341 Carbon compound catabolism ! 1012972i 1011983 : Amino acid biosynthesis and metabollsm ; 1020708'j 1021607'- Hypothetical, unclassified, unknown i 1027445i 1027984. j l 2 Hypothetical, unclassified, unknown 1030151 1029825i Nucleotide biosynthesis and metabolism 1032763j'1032095' Nucleotide biosynthesis and metabolism 1033824 10327631 jHypothetical, unclassified, unknown 1 1035277'1035981 i Putative enzymes 1039968 1040432i Putative enzymes 1040432 1040707 ! Translation, post-translational modificatio 1 1043404 1041689. Hypothetical, unclassified, unknown 1046462 104667 Adaptation, protection"1048019"10475491 S sl Transport of small molecules 1053848 10545431 Transport of small molecules 1054566 1 05S006 j Transport of small molecules 1055009 1056052i Transport of small molecules 1056049 1 0573474j Transport of small molecules 1 1057400 10579061 i Hypothetical, unclassified, unknown 1059622 1060296i Hypothetical, unclassified, unknown 1062034 1061207i Hypothetical, unclassified, unknown 1062369 106206 Hypothetical, : unclassified, unknown 1062601 10628851 . w.... ______ Hypothetical unclassified unknown'1062921 10635441 1062921 1063544 ! Hypothetical, unclassified, unknown 1065138 10654251 Secreted Factors (toxins, enzymes, algina 1067817 1066321 -l r fo Hypothetical, unclassified unknown ! 1068193 1068456, Hypothetical, unclassified, unknown, 1071877 10712391 Hypothetical unclassified, unknown i 1072462 1072839, Chaperones & heat shock proteins 1073960 10746731. Hypothetical, unclassified unknown 0 1082949 1083854, Hypothetical, unclassified, unknown 1090606 109085741 Adaptation, protection 1092498 Amino acid biosynthesis and metabolism 10932511 1094129' Hypothetical, unclassified, unknown t 10952761 10960341 Nucleotide biosynthesis and metabotism J 1096063 1096773j t Putative enzymes 1 110700_0 1107761 Hypothetical, unclassified, unknown _i 1113050 1112574 ; i Hypothetical, unclassified, unknown 1123850 11233'561 Hypothetical unclassified, unknown 1125865 1125548J Hypothetical, unclassified, unknown 1126338 1125865 j Biosynthesis of cofactors, prosthetic grou 1136388 1137035j Hypothetical, unclassified. unknown 1144862 11452001 Hypothetical, unclassified, unknown 1150470 1150721 ; Chaperones & heat shock proteins 1153637 1155547) Hypothetical, unclassified, unknown 1 1163660 1164022 ; Hypothetical, unclassified, unknown, 1175614t 1176375tt ! Hypothetical, unclassified, unknown 1176380, 1176982 Hypothetical, unclassified, unknown 1176958 1177620' Hypothetical, unclassified, unknown Two-component regutatory systems 1*189172 1190380 j Motility&Attachment i, 1194207'1 1195223i Motili & Attachment 11973901 1197833'. Hypothetical, unclassified, un 11985511 1197838i Hypothetical, unclassified, unknown 1 1199946 1198750 HYPothetiCal, unclassified, unknown 1 1207875 1207540 Hypothetical, unclassified, unknown 1212571 12118881 l-i Hypothetical, unclassified, unknown 1214502 1213195if Translation post-translational modification 1215284 1215727i t-P Biosynthesis of cofactors, prosthetic group 1218181 1218933J Antibiotic resistance and susceptibility 1 1221691 | 1222098, Hypothetical, unclassified, unknown 1226278 1225757 Hypothetical, unclassified ; unknown 1227302 1226427' Transcrintional requlators 1229344 12302 1 9, Transcriptional regulators 1 1236644 1237546i Hypothetical, unclassified, unknown 1243118 1242750 Adaptation, protection 1245665 1245928 Hypothetical, unclassified, unknown 1249552 1249019 Nucleotide biosynthesis and metabolism 1251154 1249907 Nucleotide biosynthesis and metabolism 1254309 1251418t Two-component regulatory systems'1255042 1255752' Adaptation, protection 1257772| 12579811 Hypothetica !, unciassified, unknown 1258470 1258087 ; Amino acid biosynthesis and metabolism. 1260442 92 Hypothetical, unclassified, unknown 1263378 1264190 Hypothetical, unclassified, unknown 1264919 12641911 Hypothetical, unclassified, unknown 1266111 1266782 Hypothetical unclassified unknown 1267282 12676021 l l C Energy metabolism ! 1272796 12722001 Energy metabolism 12728071, Energy metabolism 1276608 w Energy metabolism 1276784 Transport of small molecules 1284513 12858231 Hypothetical, unclassified, 1294138 1294809, Hypothetical unclassified, unknown 1303457 1303864 Hypothetical, unclassified, unknown 1303892 1304449 ; Hypothetical, unclassified, unknown 1305583 1306056i Hypothetical, unclassified, unknown-1 Hypothetical, unclassified, unknown 1317202 1315916 Hypothetical, unclassified, unknown 1 1318150 1317404l Amino acid biosynthesis and metabolism, 1319514 1318147i Hypothetical, unclassified, unknown 1321000 13203861, Transcriptionai regulators 1 1326939 326046 Putative enzymes 1327024 1327803 ,'Hypothetical, unclassified, unknown 1 1330467 1330117 . Hypothetical, unclassified, unknown 1331517 1331714 HHpothetical, unclassified, unknown _ 1334671 1334928i Antibiotic resistance and susceptibiiity f 1339082 1337931J , ISecreted Factors (toxins, enzymes algina 1'573'7 1357712, in, a 13573171 1357712 ! i I Transcriptional reg Transcriptional regulators __i_ 1370092j 1369418i tTranscrlptiotulators i3 1379168 1378500' Hypothetical, unclassified, unknown 1 1391277 ! 1391861 Transcriptional_gulators i 1396731 1397180 Hypotheticai, undassified, unknown j OS4S9'1406752] .. I_. _1406752 Hypothetical, unclassified, unknown j 1408999 1409274J Transcriptional regulators 1 14099491 1410476 ; -a f Putative enzymes 1417500 j 1417961) 3Hypothetical, unclassified, unknown í 14179651 1418738'; Transcriptional regulators 14257545411425140 Energy metabolism 14320373 14329271 Hypothetical, unclassified, unknown 1435493 1435825 Transcriptional reguiators1441547 1440639 Hypothetical, unclassified, unknown 1444451 1442904 ! Transport of small molecules 1456723 14558151 Hypothetical, unclassified, unknown 1462696 14634031 Hypothetical, unclassified, unknown 1463586 1463936 Hypothetical, unclassified, unknown 1464111 1464860 Hypothetical, unclassified, unknown 1467320 1466109 Hypothetical, unclassified, unknown 1467901 1467488 Hypothetical, unclassified, unknown 1468510 1468890 Hypothetical, unclassified, unknown ! 1470978 1470580 Hypothetical, unclassified, unknown 1474391 1474714i Transcriptional regulators 147546 Hypothetical, unclassified, unknown 1479791 14790211 Hypothetical, unclassified unknown 1483123 1483875 Hypothetical, unclassified, unknown Hypothetical, unclassified, unknown 1486967 1486266 Hypothetical, unclassified, unknown 148909 Biosynthesis of cofactors, prosthetic groul 1491913 149305543 i Hypothetical, unclassified, unknown 1494959 14954921 : Hypothetical, unclassified, unknown 1495635 1495997 t l Putative enzymes _ ! 1496920 14960871 Hypothetical, unclassified, 1516433 1516687i s Two-component regulatory systems 1 1518914 15195463 Hypothetical unclassified, unknown i 1519627 1519968 Hypothetical, unclassified, unknown 1526657 1526430 f f Carbon compound catabolism 1533238 1534278in Hypothetical, unclassified, unknown) 1552641 1552997 ! Hypothetical, unclassified, unknown 1553112 1553675 ! Transcriptional reaulators ß 1559122 1558880i Adaptation, protection 1559254 1559859 ! t Hypothetical unclassified, unknown 1572023 1572544' Motility & Attachment 1575290 15755591 E lWo l 1575559 i Motility & Attachment 1583956 15847981 J w Two-component regulatory systems 1585640 1586014 Chemotaxis 1591286 15921761 Cell division i 1592271 1593059i Chemotaxis i 15940871 1594566' Hypothetical, unclassified, unknown 1 15945971 1595004 Hypothetical, unclassified, unknown 15968891 15973051 Hypothetical, unclassified, unknown 1 1599982 1599428' Transport of small molecules Transport of small molecules 16028771 ! 1603548 Transport of small molecules, 16036711 16044292 Hypothetical, unclassified, unknown i. 1604426i 1604602' « E gy metabolism l 1605088i 1607061' Energ metabolism 1607065i 1607607'i Energy metabolism. 1607604 ! 1608071, u Ls lHypothetical, unclassified, unknown, 1615908. 1614670i Hypothetical, unclassified, unknown''1617085 ! 1615895 F Hypothetical, unclassified, unknown1619907) 1620263 Transport of small molecules 1 1624715', 1623864l Transcriptiona ! regutators j"'1634492"1633842 j Hypothetical, unclassified, unknown i 1638639t 1638379 ! Hypothetical, unclassified, unknown 1639794 1638652 Hypothetical, unclassified, unknown j 1647046 1646537 j Hypothetical, unclassified, unknown 1649555 1648629 e Hypothetical, unclassified, unknown 1649928 1650308 Transcriptional regulators 16607271 1661386 s m Cell division 1665065 1665934 DNA replication, recombination, modifica 1666025 1668409 DNA replication, recombination, modifica 1669989 1672034 Hypothetical, unclassified unknown 1672080 1672406 Putative enzymes 1673204 1674352 Hypothetical, unclassified, unknown 1677559 1678407 Hypothetical, unclassified, unknown 1678590 1678261 Transport of small molecules 1678952 1678584 Hypothetical, unclassified, unknown 1684962 1684753 Energy metabolism 1689339 1687924 w 0 Energy metabolism 16940451 16931191 Hypothetical, unclassified, unknown 1696467 1697099 Hypothetical, unclassified, unknown 1697188 1697919 Hypothetical, unclassified, unknown _ 1697916 1698344 Hypothetical, unclassified, unknown 1704522 1704761t Hypothetical unclassified, unknown i 1709765 1709412i S m Hypothetical, unclassified, unknown 1 1712698 17125191 mo w m Energy metabolism 1 1720744 17211300 w _ w Energy metabolism'1721124"1721492 Energy metabolism 1721496 17232, 68 Energy metabolism 1723280 1723987 Energy metabolism 1728416, 1729852 Energy metabolism 17301811 1731347 Energy metabolism 1 31347 1732234 Hypothetical, unclassified, unknown 1734004 1734768 w Hypothetical, unclassified, unknown 1735063 1734827i Hypotheticai, unclassified, unknown j 173S236, 17357091 Hypothetical, unclassified, unknown 1735706 1736152) Hypothetical, unclassified, unknown 1736189 17374181 Fatty acid and phospholipid metabolism 1753418 1752903i Hypothetical, unclassified, unknown 1762433 17628701 iTranscriptional regulators i 1762928i 1763752t Putative enzymes 17653451 1766205i (pothetical,, unclassified, unknown""-.. ,.,.-,-. 1766285i. 1788947 ; IHYpotheticall unclassified, unknown 17669561'1767762 Transcriptiona ! regujatorsJ 1773682 1774548 j Transport of small molecules. p ; _1775945 ; 177 0 Transport of small molecules i 1779877 ! 1780428 ! Hypothetical, unclassified, unknown, 1784108§ 1785016. Hypothetical, unclassified, unknown i 17868881 17871661 Hypothetical undassified, unknowni 1790879 j 1790472 Hypothetical, unclassified, unknown 1805218 1805724 Hypothetical, unclassified, unknown i 1814995 1815135, Hypothetical, unclassified, unknown 1816352 1816858 Hypothetical, unclassified, unknown'1824969 1825430 Biosynthesis of cofactors, prosthetic grou 1826040 1825495' Hypothetical, unclassified, unknown 1826675 1826118i Hypothetical, unclassified, unknown 1827052 18267325 # l 5 Amino acid biosynthesis and metabolism 1836367 1837227 Protein secretion/export apparatus 18415171 18404681* w Protein secretion/export apparatus 1842302 1841514 J - Protein secretion/export apparatus i-1846717 18452411 Protein secretion/export apparatus 1 1847227 1848093 Hypothetical, unclassified, unknown 184. 8707 18490720 Hypothetical, unclassified, unknown 1849077 1849406 Protein secretion/export apparatus 1851982 1852278 Protein secretion/export apparatus 1855862 1856299 Hypothetical, unclassified, unknown 1856308 1856553 Protein secretion/export apparatus 1862558 1862761. Protein secretion/export apparatus 1 1862764 1863021 Protein secretion/export apparatus 1 1863024 1863371 Protein secretion/export apparatus 1863799 1864137 Hypothetical unclassified, unknown 1874967 1875767 Hypothetical, unclassified, unknown 1875849 1876580 Hypothetical, unclassified, unknown 1887297 1887058 Hypothetical, unclassified, unknown 1888186 1887698 Hypothetical, unclassified, unknown 1889173 1888985 Amino acid biosynthesis and metabolisml 1891815 1890739 Amino acid biosynthesis and metabolism 1896630 1897247 Hypothetical, Unclassified, unknown 1912756 1912217 s w Biosynthesis of cofactors, prosthetic grou 1917599 1918087 Transport of small molecules 1921174 1922226 Central intermediary metabolism 1926123 1925797 Transport of small molecules ! 1931775 1930564 Hypothetical, unclassified, unknown i 1933632 1933054 Energy metabolism 1937644 1935035 t Hypothetical unclassified, unknown 1939583| 1940206 s Hvpothetical, unclassified, unknown 19424421 1941720 Translation, post-transtationa ! modification 1943067 1944737 Transition, post-transtationat modification 1944747 1946129 ! Biosynthesis of cofactors, prosthetic grou 1947041 1946187 i Translation, post-translational modificatioh 19562271 1958623i DNA replication, recombination, modifica 1972959 1973405 ; Nucleotide biosynthesis and metabolism 1 1973470 1974210 ; E f unclassified, unknown 1974627 9742 small molecules 1979941, 1978439 rs r Hypothetical, 1984389 19850331 tHypotheticalz unclassified, unknown s 19894233 1989109 Hypothetical, unclassified, unknown 1993898 1993461i Hypothetical, unclassified, unknown 1 1995164, 1994667' Hypothetical, unclassified, unknown X 1998587 ! 1998949 Hypothetical, unclassified, unknown Y 199 1999512 j Hypothetical, undassified, unknown ! 2004892J 2004374 ! Hypothetical, unclassified, unknown _ j 2007665i 2008249 i Hypothetical, unclassified, unknown rv-Lrv2012530 y2012255 Hypothetical, unclassified, unknown j 2015136i 2014915i Transcriptional regulators 2019690 20188031 Transport of small molecules 2022398i 2021712' Hypothetical, unclassified, unknown 2028454 2028981 ! -|s s Fatty acid and phospholipid metabolism 2031466 2031705 Hypothetical, unclassified, unknown 2034857 2034066 Transport of small molecules 2052941 2053264i En Energy metabolism 2053277 205367 Transcriptional regulators 2054223 2053672 Hypothetical, unclassified, unknown 2054309 2054842 Hypothetical unclassified, unknown 2062401 2061664 Hypothetical, unclassified, unknown 2065545 2064853 Hypothetical unclassified, unknown 2066767 2065493 Hypothetical, unclassified, unknown 2067955 2066786 Hypothetical, unclassified, unknown 2068728 2067961 Secreted Factors (toxins, enzymes, algma 2q70685 2071 3 Secreted Factors (toxins, enzymes, algn 2071209 2071697j Secreted Factors (toxins, enzymes, algin : 2076311 207 5 Transcriptional regulators 1 2085426 2084476 Transcriptional regulators t 2085929 20854B1 Hypothetical, unclassified, unknown 2088034 2086808 Hypothetical, unclassified, unknown 209183 209149 Hypothetical unclassified, unknown 2103294 2103770 ! HYDothetical unclassified unknown 2103770 2104096 j Translation, post-translational modificatio i 2109511 2108942 n 2109511 Hypothetical, unclassified, unknown 2109854 2109558i l Hypothetical, unclassified, unknown 2117897 211809 ! Hypothetical, unclassified, unknown 2118585 211889 ! Hypothetical, unclassified, unknown 2118926 2119747 Hypothetical, unclassified, unknown 2122222 2120225 Hypothetical, unclassified, unknown,'I 2137785 2136520t Hypothetical, unclassified, unknown 2138598 2137846j Hypothetical, unclassified, unknown 2141002 2140433 a Hypothetical, unclassified, unknown 2141487 21409991 Hypothetical, unclassified, unknown 2145894 21465021 Hypothetical, unclassified, unknown 2146609 2146875gt Hypothetical, unclassified, unknown 2148854 2149189 P w Hypothetical, unclassified, unknown 2150364 2149864 Hypothetical, unclassified, unknown 2150524 2150781 i Hypothetical, unclassified, unknown 2157968 21'59167 ; Transcriptional regu tators"2164548 2163883J Two-component regulatory systems 2165876"2166553 Biosynthesis of cofactors, prosthetic grou 2171865 _, 2171936= Biosynthesis of cofactors, prosthetic grouj"2171989 ! 2172903i ; Biosynthesis of cofactors, prosthetic grouj'"217362j 2173940 fHypothetical, unclassified, unknown í 2181744'2181181' iHypothetical, unclassified, unknown'. 21820971 2181741 Chaperones&heatshockproteins'_21823941 21821168 'fatty acid and phospholipid metabolism 1 21870651 2188246 ; Hypothetical, unclassified, unknown 2188459 21898831 Carbon compound catabq ! jsm ! 2196132 2195494 : Transcriptiona ! regutatorsY 2198891 2199694 j Putative enzymes i 2203446 2202649i Ho Fatty acid and phospholipid metabolism 2206353 1 2205190 t # o- Transcriptional regulators 2206806 1 2206402 i Hpothetical, unclassified, unknown 2206999 2207928 v z w Hypothetical, unclassified, unknown 2213539 2213315 Hypothetical, unclassified, unknown 2219099 2218098J Hypothetical, unclassified, unknown 2220275 2220574J Hypothetical, unclassified, unknown 2221157 2220903 ; Hypothetical, unclassified, unknown 2223804 2224478 Hypothetical, unclassified, unknown 2227541 2229001 Transport of small molecules. 2234080 2235309f Transcriptional regulators 2244995 2245948i Central intermediary metabolism 2246456 2245986 Transport of small molecules 2256704 2257720 Putative enzymes 2259478 2260659i Hypothetical, unclassified, unknown 2265764 22651261 Translation, post-translational modificatio 2272460 2274568 Transport of small molecules 2277552 2278982 S s Hypothetical, unclassified, unknown 2278982. 2279794 Hypothetical, unclassified, unknown 2281578 2279917 Hypothetical, unclassified, unknown 2290335 2289085 Hypothetical, unclassified, unknown 2297072 2297920 Hypothetical, unclassified, unknown 1 2300676 2301755f Transport of small molecules 2303022 2304215 HvDothetical unclassified, unknown 1 23066271 2305782i t v Carbon compound catabolism 2307957 2309432 Hypothetical, unclassified, unknown 2312899 2313789 Biosynthesis of cofactors, prosthetic grou 2314512 2315690 Putative enzymes2316708 2317403 Hypothetical, unclassified, unknown 2318624 2318226 0 Hypothetical, unclassified, unknown 2322071 2321130 DNA replication, recombination, modificai, 2329348 2330424 Putative enzymes2332059 2330959J iHypothetical, unclassified, unknown 2332820 2332392, Transcriptional regulators 1 2335172f 23361041 Hypothetical, unclassified unknown""2339987 2339352 ! Hypothetical, unclassified, unknown 2346477 2347838 Hypothetical, unclassified, unknown 1 2352430i 2351906 Putative enzymes 1 23567131 23575731 HYpotheticat, unciassified, unknown2358024 2358311) 'Hypothetical, unclassified, unknown I 2361706j 2361873i L ! Hypothetical, unclassified, unknown 2364816 2365058 ! IHym hetical, unclassified, unknown j 237747 2376538i Hypothetical, unclassified, unknown 23817781 23814732 Hypothetical, unclassified, unknqwn 2390255 2390620j Hypothetical, unclassified, unknown"2390949 2392046J Hypothetical unclassified, unknown 2393424 ! 2393633a Hypothetical, unclassified, unknown 23937081 23941781 Hypothetical, unclassified, unknown i 23962521 2395944' ------------1------- Hypothetical, unclassified, unknown Hypothetical, unclassified, unknown"12404655 ! 2404386 j Hvpothetical unclassified unknown, 2405233. 24049495 Hypothetical, unclassified, unknown 2405739 2405230 ; Hypothetical, unclassified, unknown i 24. 05993 2406877' Hypothetical, unclassified, unknown 2406961 24071311, Hypothetical, unclassified, unknown 2407236 24076611, Hypothetical, unclassified, unknown 2409837 2410181, Hypothetical, unclassified unknown 2411709 2412122 l Transcriptional regulators 24 Hypothetical, unclassified unknown 2416376 241741 31 Hypothetical, unclassified, unknown 2424400 2423924j Hypothetical, unclassified, unknown 2427017 2425497 j Hypothetical, unclassified, unknown 2430170 2431129 Transport of small molecules. 2433748 2435070 Transport of small molecules 2441555 2440347 Transcriptional regulators 2441771 2442691 X -ffi Hypothetical, unclassified, unknown 2443161 2444366 Hypothetical, unclassified, unknown 2445533 2444886 b r Hypothetical, unclassified, unknown 2446564 2445545 Hypothetical, unclassified, unknown 2447325 24465971 Hypothetical, unclassified, unknown 2447989 2447573 Hypothetical, unclassified, unknown 2448533 2448533, 2448033 Transcriptional regutatorsj 2449545) 2448568 Hypothetical, unclassified, unknown 2450765 24495541 Hypothetical, unclassified, unknown 2451707 2452426i Transport of small molecules | 24575101 2458280, HYPothetiCal unclassified unknown 1 2468032|. 24690991 Hypothetical, unclassified, unknown 2472104 2472409 Hypothetical, unclassified, unknown 2479130 2478312 Amino acid biosynthesis and metabolism 2480844 2481830 Secreted Factors (toxins, enzymes, aigina 2485471 2486118 I Transcriptional regulators 1 2487293 24863551 Hypothetical, unclassified, unknown 1 2488950 2489732 Hypothetical, unclassified, unknown 2508775 2509467 Hypothetical unclassified, unknown 2510622 2511050 Hypothetical, unclassified, unknown 2512511"2513182J t i Y Hypothetical, unclassified, unknown 2523239 2522958 Hypothetical, unclassified, unknown 2524201 2523236 Transport of small molecules 1 25250521 2524198' Transport of small molecules 2525846 2525049i Energy metabolism i 25276571 2527412t Putative enzymes 2529467 25277431 IHypothetical, unclassified, unknown 1 2540082'2539063ri r J ~vt Hypothetical, unclassified unknown ! 2549748| 2549906. Transcript onal regulators 25539651 25548581 itransport of small molecules 1 2571691 2570855 ; Hypothetical'unclassified, unknown 25733651 2572805) , Hypothetical, unclassified, unknown, 25795421 2580882 {Transport of small molecules ~. ~ 2582097'2583407 3Carbon compound catabolism 1 2587906 2589414i [iJiypothetical, unciassified, unknöwn f 25933301 25945471 i Hypothetical, unclassified, unknown s 2595985i 25967795 Transport of small molecules i 2597869 i 2598522 f Hypothetical unclassified, unknown 1 2614893 2615438 Hypothetical, unclassified, unknown 2617019 2617516 i Hypothetical unclassified, unknown (2617529 31 Hypothetical, unclassified, unknown 2619695-26207-1-11 Hypothetical, unclassified, unknown 2623284 26238561 Hypothetical, unclassified, unknown 2627175 2626780 s t l Transcriptional regulators 2635971 2635051 ? Transport of small molecules 2645303 2646727 j Hypothetical, unclassified, unknown, 2689240 2689569} Hypothetical ; unclassified, unknown 268956895690126 0 w E Putative enzymes 2694544 2693780 Hypothetical, unclassified, unknown 2694763 2694545 Hypothetical, unclassified, unknown 2701205 2702065 Hypothetical unclassified unknown. 2705772 27060861 Hypothetical, unclassified, unknown 2723221 2722754J Hypothetical, unclassified, unknown 2724222 2723308 Hypothetical, unclassified, unknown _ 2724484 2724729 Hypothetical, unclassified, unknown l 2730520j 27299750 Hypothetical, unclassified, unknown 2732965 2732531 Hypothetical, unclassified, unknown 2738840 2739715 Amino acid biosynthesis and metabolism 2740882 2739761ii Amino acid biosynthesis and metabolism 2747072 2746689 ; Hypothetical, unclassified, unknown 2753464 2752865J Hypothetical, unclassified, unknown 275. 4601 2754822 j Hypothetical, unclassified, unknown, 2755724 27562511 Hypothetical, unclassified, unknown 2756308 2756649' Hypothetical, unclassified, unknown 2760086 2759481J Hypothetical, unclassified unknown 3 2760618 2760322 Hypothetical, unclassified, unknown 2761350 27608711. Hypothetical, unclassified, unknown 2781450 2780926 Tfanscriptionaireguiatorsi 2786355 2785369 Transcriptional regulators 2787885 2786971 Central intermediarg metabolism i 2791219} 2791863 Hypothetical, unclassified, unknown j 2791905 2792816 Putative enzymes 1 2794132} 27927981 o Hypothetical, unclassified, unknown t 2803344j 2803622i Hypothetical unclassified, unknown} 28041311 2803859' Hypothetical, unclassified, unknown j 2805916 280629Q ! Putative enzymes I 2807368 28063491 Transcriptional regulators i 28074681 2808511 Hypothetical, unclassified, 2814766i 2815281 Transport of small moiecu ! es 2817448 2818674J Hypothetical, unclassified, unknown''2818885 2818718 ! Hypothetical, unclassified, unknown'. 2822321L 2821704 ! Carbon cornpound catabolism 1 28255903 28246589 Carbon compound cataboiism 2833129 ; 2832368 Carbon compound catabolism, 2H Transport of small molecules _ __ 1 28419651 2840511, Fatty acid and phospholipid metabolism 2865104 28641691 ; Hypothetical, unclassified, unknown Hypothetical, unclassified, unknown 2867505 2866192 Hypothetical, unclassified unknown | 2876409} 2875696' Hvpothetical unclassified unknown i 2880519i 28815621 Transcriptional regulators Putative enzymes 2885332 2884205 Putative enzymes Putative enzymes,'2888551 2885361 Putative enzvmes 2887336 2886569l _.,.. _Y _-. »-- Transcriptional requlators Fatty acid and phospholipid metabolism 2923926 2923366i Transcriptional regulators'2934387 2933581 Hypothetical, unclassified, unknown 2945263 2945868 Hypothetical unclassified, unknown 2948581 2948976 Hypothetical, unclassified, unknown 2948973 2949332 Hypothetical, unclassified, unknown 1 2949332 2949637 HYPothetiCal unclassified, unknown 2949634 2949969 z Translation, post-translational modificatio 2954695 2953415 Chaperones & heat shock proteins 2956778 2956152 S-m > Cell division 2959239 2956804 Translation, post-translational modificatio 1 2û6û455 2961135i Translation, post-translational modificatioh 2962002 2962220 Hypothetical, unclassified, unknown ! 2964842 2964606 w Transcription, RNA processing and degra 2969986 2971113 Nucleotide biosynthesis and metabolism j 2972698 2974068 Energy metabolism2983204 2983881 Z E Energy metabolism 2986242 29875881, Energy metabolism 1 29920010 2992501' Energy metaboiism2992547 2992855' Hypothetical, unclassified, unknown 1 3008323 3008009 j Hypothetical, unclassified, unknown 3012536 3012279 Biosynthesis ofcofactors, prosthetic grou 3015581 3015937 Hypothetical, unclassified, unknown 3016245 3016598 Hypothetical, unclassified, unknown 3016883 3016674 Protein secretion/export apparatus 3020928 3020503 < Protein secretion/export apparatus 3021338 30209 8 S-l Protein secretion/export apparatus 3021741 3021307 Transport of small molecules 3025507 3024704 Transcriptional regulators ^ t 30280741 30290031 Amino acid biosynthesis and metabolism 3031397 3030435} Hypothetical, unclassified, unknown 3042501 3043415 Related to phage, transposon, or ptasmic 3044765 3043749 Energy metabolism 3047969 3047643 Hypothetical, unclassified, unknown 3050611 30 Hypothetical, unclassified, unknown 3057375 3057, 608. ypo e ica, unclassified, unknown 3060659"060264 Energy metaboiismj 3070365 3070703 3070365 ypothetical, unclassified, unknown 1 30737311 3074417'. rHypothetical, unclassified, unknown ! 3075217 ! 3074579 j Hypothetical, unclassified, unknown 30754101. 30758892 ! ; Hypothetical, unclassified, unknown 3075921 i 3076313' : 'Hypothetical, unclassified, unknown 1 3076343i 30766211 : Central intermediarv metabolism I 3079196 3079834'. 'Hypothetical, unclassified, unknown 1 3089612'3088659 ; Hypothetical, unclassified, unknown , Hypothetical, unclassified, unknown 3094184f 3093657 Hypothetical, unclassified, unknown 3096051 j 3094756 Hypothetical, unclassified unknown 1 30989781 3098625 Hypothetical, unclassified, unknown 30997291 3099472i DNA replication, recombination, modifica 310011111 3099809 i Translation, post-trans) at ! qna) modificatiop 3102493 3100115 : Translation, post-translational modificatio 1 3103544 3102528, Translation, post-translational modificatio 3103998 3103642 Transtation, post-transiationatmodificatiol 3104216 3104022 Translation, post-translational modificatio 3104829 3104278 Translation post-translational modificatio 3106751 3104829 DNA replication, recombination, mod ! fica 3110900 3111613 Hypothetical, unclassified, unknown 3114818 3115192' Hypothetical, unclassified, unknown 3117610 3117176@, Hypothetical, unclassified, unknown 3119707 3119393 j Hypothetical, unclassified, unknown 3122264 3121920J Hypothetical, unclassified, unknown 31. 22585 3122376'. Putative enzymes 3126251 31272191 0 t 9 Hypothetical, unclassified, unknown 3127225 3127707 r Hypothetical, unclassified unknown 3128269 3127859. Hypothetical, unclassified, unknown _ 3132815 313. 2228 Hypothetical, unclassified, unknown 31332571 3132820t Hypothetical, unclassified, unknown 1 31378491 3138193j Hypothetical, unclassified, unknown 3138190 3138531 Hypothetical, unclassified, unknown 3139010 3139669) Hypothetical, unclassified, unknown 3141718 3142275i Hypothetical, unclassified, unknown 3142284 3142502' z Hypothetical, unclassified, unknown 3142617 314308 ?, Hypothetical, unclassified, unknown 3149318 3148722j Hypothetical unclassified, unknown 3152201 3150885 i Hypothetical, unclassified, unknown 3155074 3154592 Hypothetical, unclassified, unknown 3156502 3156801 Hypothetical, unclassified, unknown 1 31575631 3156859 ! Hypothetical, unclassified unknown 3158925 3159668 Hypothetical, unclassified, unknown 1 3160320 31605831 Hypothetical, unclassified, unknown 3162215 31615981 Hypothetical, unclassified, unknown 3162578 Transport of small molecules 3166641 3164762 ; Hypothetical, unclassified, unknown 3173174 3171597J Hypothetical, unclassified, unknown 3173243 3173722 Hypothetical, unclassified, unknown 3180951 31805531 Hypothetical, unclassified, unknown 3182399 3182851 ! Hypothetical, unclassified, unknown 3184001 3185128 ; Adaptation, protection 3185816 3185160 ! Hypothetical, unclassified, runknown 3192748 3193518 ; tative 3197641 a . 3198987 ì Hr S 'Hypothetical, unclassified, unknown 32005521 3200319 ! iTranslation, post-translational modification 3205079i 3204513i Hypothetical, unclassified, unknown 32062521 3205122 ; Membrane proteins 3207165 ! Hypothetical, _un_classified, unknown- _ _ 3208260 3207289 =Transcription, RNA processing and degrã 3212524. 32130301 Secreted Factors (toxi ns, enzymes, a ! g ina 3215422T 3216288 Hypothetical, unclassified, unknown 32212051 3221564i s HYpotheticäl unciassified unknöwn A227383 t Nucleotide biosynthesis and metabolism 32301811 3229483 1 Transcriptiona ! reguiatorsJ 3231171 3230278' Transcriptional regulators Hypothetical, unclassified, unknown 3235793 32359600 Hypothetical, unclassified, unknown 3249630 3249208 . Hypothetical, unclassified, unknown 3253679, 32533111 Hypothetical, unclassified, unknown 3255765 3255406 Hypothetical unclassified, unknown'32652101 3264641 v-S Hypothetical, unclassified, unknown 3271692 3270826 Hypothetical unclassified, unknown 32724051 3271812s Putative enzymes 3277408 3278577 Hypothetical, unclassified, unknown 3284597 3283374 Hypothetical, unclassified, unknown 3291172 3290693 Hypothetical, unclassified, unknown 3291282 3291863 Hypothetical, unclassified, unknown 3291959 3292267 Putative enzymes 32971117 3295978 Putative enzymes 3308390 3309337 Energy metabolism 33117201. 3310791 Enegy metabolism 3312469 3311720 Energy metabolism 3312790 3314445 Motility & Attachment 3320029 3319673 r DNA replication, recombination, modificat 3321049 3320063 Nucleotide biosynthesis and metabolism ! ism3321674 3321042 Hypothetical, unclassified, unknown ß 3322752 3321703 Fatty acid and phospholipid metabolism 3325182 3324946 Fatty acid and phosphoiipid metabotism 3326121 33253781 ; Fatty acid and phosphoiipidmetabotism 3327082 3326144 Translation, post-translational modificatio 3328384 3328202 Hypothetical, unclassified, unknown | 3328934 3328398 r s Transcription, RNA processing and degr'3332303 3331347 Cell wall/LPS/capsule 1 3337230 {333621 1 Translation, post-transtationa ! modificatioh 3337691 3337227 Cell wa ! !/LPS/capsule.''J 3338455 3337691 I Hypothetical, unclassified, unknown _. L 33386401 3338455 Cell wall/LPS/capsule 3339676 3338678 Hypotheticat, undassified, unknown 3340116 3339676 Transport of small molecules 3340748 3340113 Hypothetical, unclassified, unknown 3343177 3343710 Hypothetical, unclassified, unknown 33450991 33437981 Transport of. small molecules _"3345795fi 33451121 Hypotiietica), undassified, unknown j 3347038 3345788 ! H) Lpothefical, unclassified, unknown 334 ___. __________. __. _ J _. ~__ ucieotide biosynthesis and metabolism,'3350231, 3348B37 ; Hypothetical, unclassified, unknown,'33508371, 3350410' Energy metabotismJ''3354171 j 3353497J Putative enzymes ______. 33606531 3359268, INucleotide biosynthesis and metabotism 3365743 336500 33657431 3365006'i (Hypothetical, unclassified, unknown f 3369268 3369035 lDNA repiication, rec. ombination, modificai 3372705i 333370099i Hypothetical, unclassified, unknown 33731701 3372796'' Hypothetical, unclassified, unknown 3378511t 3378948 { Hypothetical, unclassified, unknown'3383947 3384333J Hypothetical, unclassified, unknown, 3385183 3384377 ; Putative enzymes 33878 3386269i 28 Biosynthesis of cofactors, prosthetic grou 3394087 3394683j Hypothetical, unclassified, unknown 3397373 3397095, Hypothetical, unclassified, unknown 3398716 3399648' m--W 3404i40i Hypothetical, unclassified, unknown 3403811 3404140 ! Hypothetical, unclassified, unknown 3404144 34045241 Hypothetical, unclassified, unknown 3404538 3404861 Hypothetical, unclassified, unknown 3409952 3409608 Transtation, post-tfansfationat modification 3414400 34146121 Hypothetical, un_classified, unknown '3416066 3415785. Transcriptional regutators3436429 3435986J Hypothetical, unclassified, unknown 3457909 34566421 Hypothetical, unclassified, unknown 3464151 3463888, Hypothetical, unclassified, unknown 3466959 3466072 Hypothetical, unclassified, unknown 3467084 34680491 Hypothetical, unclassified, unknown 3473017 3474135 z Protein secretion/export apparatus j 3476479 3475955 Protein secretion/export apparatus I Protein secretion/export apparatus, 34802381 34797201 Protein secretion/export apparatus {3483421 3481913 j Hypothetical, unclassified, unknown _ 3491410 3490751'. Fatty acid and phospholipid metabolism 3493572 34927001 H H Amino acid biosynthesis and metabolism 3500597 3499485i Hypothetical, unclassified, unknown 3506241 3505864i Hypothetical, unclassified, unknown 3524489 3524160} Hypothetical, unclassified, unknown 1 3527733 35274281 Hypothetical, unclassified, unknown 3528349 3528230 Cell wall/LPS/capsule 3529446 3528427 Cell wall/LPS/capsule 3530457 3529507 Cell wall/LPS/capsule,, 1 3531707 3530466 Ce) f wat)/LPS/capsu) e) 3532814 3531750 Cell wall/LPS/capsule ! 3533932 3532811 í _ < ve Cell wall/LPS/capsule 3535080 35339471- Amino acid biosynthesis and metabolism-3535970 3535215l Amino acid biosynthesis and metabolism 3536578 3535970 Cell wall/LPS/capsule 3537810 3536575 Cell wall/LPS/capsuie 3539123 35378071 'Cell wall I LPS/capsule ___ _ _ 3540206 35391271 jCe) twa !)/LPS/capsu) e i 3540784 3540209 J cell wall/LPS/capsule 35426701'35407811 1Cell wail/LPS/capsule 3543689 3542739 Ce) i wa ! i/LPS'/capsule"""""""""'"""" ?"3545073 3543763 ! Cell wall/LPS/capsule t 3546926 0 3545880 í DNA replication, recombination, modificaj ; 3547972 i 3547688J Translation, post-translational modificatioh 3549788 3548109 Nucleotide biosynthesis and metabolism 3550745 3550056 Biosynthesis of cofactors, prosthetic grou 3556338 3555253J DNA replication, recombination, modificai 3559197 556426= Biosynthesis of cofactors, prosthetic grou 35621M 3562803 j Hypothetical, unclassified, unknown 3569012 3568635j Carbon compound catabolism 35716021, 3570940 Hypothetical, unclassified, unknown 3575696.-35748451, Carbon compound cataboJism j 3587432r'3588436' ! Hypothetical, unclassified, unknown 3593732 35940311 Hypothetical, unclassified, unknown {35942070 3594569 Hypothetical, unclassified, unknown --... I. 35973181 35977974 : Transport of small molecules f i 36004531'3601598 Transcriptional regu ! ators J"3609075 3609839, Chaperones & heat shock proteins 3614303 614866 Hypothetical, unclassified, unknown. 3618466 3617342 Putative enzymes 3619618 3618992 Hypothetical, unclassified, unknown 3624836} 3625057 Cell wall/LPS/capsule 3630604 3629666 Cell division3632429 3632683J Transcription, RNA processing and degree 3632787 3633422J Transcriptionatregujators 3636256 3635540J Hypothetical, unclassified, unknown ; 3641232 3641810 Transcriptional reguiators'3648065 3647754 Adaptation, protection 3653666 3653875 j 3653666 Hypothetical, unclassified, unknown _ 36685_2 3667923 Hypothetical, unclassified, unknown 3668473 3668760 Hypothetical, unclassified, unknown 3669168 3668839 Hypothetical, unclassified, unknown 36710571 36707584 Transport of small molecules 3674323 3673007t Hypothetical, unclassified, unknown 3675159 3674569I v-S Hypothetical, unclassified, unknown 3679945 3680460 Hypothetical, unclassified, unknown 3680967 3680464 Hypothetical, unclassified, unknown 1 3681047 3681460 Hypothetical, unclassified, unknown 3684716 3684162 Hypothetical, unclassified, unknown 3685761 3684904 Hypothetical, unclassified, unknown 3695480 3695178 Hypothetical, unclassified, unknown 3700315 3700785 . Hypothetical, unclassified, unknown g 37102241 3710679' x Putative enzymes} 3715804 3714914J Transport of small molecules 3716807 3717610' Transport of small molecules 3717604 , Hypothetical, unclassified, unknown 1 3719328, 3720056, Hypothetical, unclassified, unknown 3720122 3720622 Hypothetical, unclassified, unknown 3723444 3722989 Translation, post-transtationaimodificatiol 3730075 3729470 j Putative enzymes 3740104 3741018J Hypothetical, unclassified, unknown j 3742264j 3742689. Fatty acid and phospholipid metabolism j 3743699 3743938 : Hypotheticaf, undassified, unknown f 3747455 ? 3747165 Transcriptional regulators 37520431 ! Hypothetical, unclassified, unknown 3 59680 37593 5 Chemotaxis 3760819 3759995 HYpöthëticalX unclassified, unknown T 3762803 3763126 j Hypothetical, undassified, unknown 3763680 3764471 Hypothetical, unclassified, unknown ; 765086 ; 3764475' Secreted Factors (toxins, enzymes, a) gin 3772560 3771502 Hypothetical, unclassified, unknown 3778265J 3778603 . Putative enzymes 37787031 37793681 Hypotheticai, undassified. unknown j 3779929 3780075J Hypothetical, unclassified, unknown 3780127 _3780312i Hypothetical, unclassified, unknown 3787478 3787020 Transport of small molecules 3 1790985 37901491 Hypothetical, unclassified, unknown 3794116 3793814 Energy metabolism 3801103 3801930 Energy metabolism 3801947 3802483 ! i ffi w i- Energy metabolism 3803342 3802566, Jr. hs 9S _ _ Hypothetical, unclassified, unknown 1 3808802 3808317 Transport of small molecules 3814574 3813957 Hypothetical, unclassified, unknown 3820005 3820271 Hypothetical, unclassified, unknown 3820307 38208911 Energy metabolism 3823515 38225141, Hypothetical, unclassified, unknown 3840747 3840358 Transcriptiona) regutators 3840843 3841736J Related phage, transposon, or plasmid 3843289 1 w l f Hypothetical, unclassified, unknown 3844104 3843652J Biosynthesis of cofactors, prosthetic grou, _ 3845774 3846334_ Biosynthesis of cofactors, prosthetic grou 3846336 3846707 ; Transport of small molecules 3849582 3848794i Hypothetical, unclassified, unknown 3850751 3849603 -l X Hypothetical, unclassified, unknown 3851800 38508_291 Hypothetical, unclassified, unknown 3852512 3851919 ww Adaptation, prptectiqn, 3856130 3855492 j Hypothetical, unclassified, unknown 3856336 w Putative enzymes _ _ _ 3867706 38694631 Hypothetical, unclassified unknown 3882979 3883437 s Hypothetical, unclassified, unknown 3885710 3886306 Transcriptional regulators 3890649 3889924 ! Translation, post-translational modificatio 3895323 38973564 Hypothetical, unclassified, unknown 3906390 3907088 Hypothetical unclassified, unknown 7851 t Hypothetical, unclassified, unknown 39107381 39117721 Hypothetical, unclassified, unknown 3912409 3913131 DNA replication, recombination, modifica 3913128 39137661 Hyp. othetical, unclassified, unknown 3913875 39140541 Hypothetical, unclassified, unknown 3918468 3918250 Hypothetical, unclassified, unknown 3918787 3918470 Hypothetical, unclassified, unknown 3922072 3921269 Transport of small molecules 3928333 3927557 Hypothetical, unclassified, unknown 3936836 3935799 Hypothetical, unclassified, unknown 1 39374311 3937237i ; Transport of small molecules 3943806 39426491 Transcription, RNA processing and degr"'"3948137 !'3948811J Hypothetical, unclassified, unknown 55 39498521 3950073 3040852 3960073 ; Hypothetical, unclassified, unknown 3952386 3952060s Secreted Factors (toxins, enzymes, alt 3965841 3967010' Secreted Factors (toxins, enzymes, aigina 3977183 3977833J Hypothetical, unclassified, unknown 3986879, i 3987292' Hypothetical, unclassified, unknown ! 3997821 3998114 Carbon compound cataboiism'''4003600"4002107 Hypothetical, unclassified, unknown ; 4004782 4004958 Hypothetical, unclassified, unknown ! 4007506 4008036 Hypothetical unclassified, unknown 1 4010326 40095411 Carbon compound catabolism 4023176 4021 9715 _ t7 t Hypothetical, unclassified, unknown 40357557 4035605, Hypothetical, unclassified, unknown 4036020 40357577 Hypothetical, unclassified, unknown 4039173 4040099 Hypothetical, unclassified, unknown 4040819 4040103 t | Transport of small molecules 4043060 404383041 Hypothetical, unclassified unknown 4045159 4045569 5. S Hypothetical, unclassified unknown 1 4045588 4045809 H othetical, unclassified, unknown 4051557 4051096j. DNA replication, recombination, modifiât 4052603 4051563 w < Hypothetical, unclassified, unknown 4062898 4062425 Hypothetical, unclassified, unknown 40672941 4067542 Hypothetical, unclassified, unknown-,- z f 7 Hypothetical, unclassified, unknown 4068611 40683271 Carbon compound catabolism 4069965 4068676i Cell wall/LPS/capsule 4070856 407001 1 Nucleotide biosynthesis and metabolism 4072487 4070859= Hypothetical, unclassified, unknown 4073986 4072658 s Fatty acid and phospholipid metabolism 40750061 4074056 DNA replication recombination, modificat 4078677 4075156a Cell wall/LPS/capsule 4082180 4081044 Cell wall/LPS/capsule 1 40829605 40821841 Fatty acid and phospholipid metabolism 4083397 4082957 ; Cell wat)/LPS/capsutej 4084504 4083443 Transport of small molecules 40845041 4085010 Transport of small molecules 4087454 40850611 ? Biosynthesis ofcofactors, prosthetic grou ! 4090093 4088903J 40889031 Fatty acid and phospholipid metabolism 1 4090905 40900901 Cell waft/LPS/capsufe'4091654 4090899) Translation, post-translational modificatio 4092227 4091670 Nucleotide biosynthesis and metabolism 1 4092967 ITranslation, post-translational modificatioh 40940351 40931661 Trans ! ation, post-transtationa) modificatiop 4094906 4094166 Translation, post-translational modificatioh 4095171 4095956 Hypothetical, unclassified, unknown 4102767 4103060 Hypothetical, unclassified, unknown 4103731 4104078 Cell wall/LPS/capsule 4104744 4105778 Transport o small molecules 41110801 4110346 ( Hypothetical, unclassified, unknown 411493 1 41153301 Transcriptional regulators H pothetical, unclassified, unknown i 41239551 4123260 pothetical, unclassified, unknown Hypotheticat, undassified, unknown ! 4126843} 4126163j lhypothetical, unclassified, unknown 4130598'41309425Z Hypothetical, unclassified. unknown ! 4135972 f 4135451', Translation, post-translational modification 4143664j 4142569 i Chemotaxis I 4148251 ; 4145942 Hypothetical, unclassified, unknown 4165879i 4165718 ; DNA replication recombination modifica 4172484 4170769 ! Hvpothetica, unclassified, unknown 4173067 172528,,' lhypothetical, unclassified, unknown 4182018 4181377'' lhypothetical, unclassified, unknown 4182769 4182074 Hypothetical, unclassified, unknown 4183709 4184938f Translation, post-translational modificatio i 4195357 4195007j Transcription, RNA processing. and degr 4196157 41953994 Transcription, RNA processing and degr 4196691 41961641 Trans ! ation, post-transtationaf modificatio i 4196958 4196707 j Protein secretion/export apparatus 4198541 4197168 D < Hypothetical, unclassified, unknown 4204736 42045421 m Hypothetical unclassified, unknown 4205906 4205295 4205906 Hypothetical, unclassified, unknown 4206664 4207164i. Putative enzymes4209255 4210277 Hypothetical, unclassified, unknown 4221796 4221212 Hypothetical, unclassified, unknown 4224115 4223567 w e Nucleotide biosynthesis and metabolism 42272361 4225659 ! Hypothetical, unclassified, unknown 4231070 4232227 Transcriptional regulators 4234274 4235182 1 DNA replication, recombination_odifica _ 4235219 4236598 Transcriptionai regulators 4242294 4241341 Hypothetical, unclassified, unknown 4243651 Hypothetical, unclassified, unknown 4244184 4243708 Hypothetical, unclassified, unknown 4244875 4245723 Hypothetical, unclassified, unknown i 42458091 42462041 Hypothetical unclassified unknown 4255323 4254736. Hypothetical, unclassified, unknown 4260396 4259254 Hypothetical, unclassified, unknown 4263492 4262377 Motili ity& Attachment4265287 4264529 Hypothetical, unclassified, unknown 4266444 4265305 Nucleotide biosynthesis and metabolism L 4266900 4266469 Hypothetical, undassified, unknown 4267344 4267144 Energy metabolism 1 42677090 4267371' l Chaperones & heat shock proteins 4269575 4267716 Chaperones & heat shock proteins4270139 4269618 ; Biosynthesis of cofactors, prosthetic grou 4270470 4270147 Biosynthesis of cofactors, prosthetic grou 4270884 1 Hypothetical, unclassified, unknown 4272655 4272164 Protein secretion/export apparatus 4278946 4277084 Hypothetical, unclassified, unknown 4279344 4279006 Hypothetical, unclassified, unknown 4285483 42844161 Hypothetical, unclassified, unknown 4286594 4285476 ; unclassified, unknown 4287709 4286786 ; tHypothetaf. undassifiedu j 4290866r4291234J Translation, post-translational modificatio 4291355i4294207' al, unclassified, unknown j 604 4294254 ! lhypothetical, unclassified, unknown ! 4302040 4303050il w Hypothetical, unclassified, unknown"j4305063 4305425 j Hypothetical, unclassified, unknown T 4311861 4310950'. Hypothetical, unclassified, unknown 4312702 43119501. Hypothetical, unclassified, unknown 4314794 4314480) Hypothetical, unclassified, unknown 4316037 4315555 l j 4 Hypothetical, unclassified, unknown 4316824 4316108 Putative enzymes 4318258 4318905 DNA replication, recombination, modifical4330320 4330895; Hypothetical, unclassified, unknown 4331451 4332461j f Hypothetical, unclassified, unknown 43326521 43330351 Transport of small molecules 4343527 Hypothetical, unclassified, unknown 4350352 43507381 Hypothetical, unclassified, unknown 4351594 4352493 Transport of small molecules 4354543 43552651 Transport of small molecules 4356200 4356862 Transport of small molecules 4356875 4358038 Hypothetical, unclassified, unknown 4359074 4358166r Hypothetical, unclassified, unknown 4373937 4374332 Hypothetical, unclassified, unknown 4374329 4374856 Hypothetical, unclassified, unknown 4374849 43752321 Hypothetical, unclassified, unknown 4382015 4381500i Biosynthesis of cofactors, prosthetic grou 4386351 4385899 Biosynthesis of cofactors, prosthetic grou 4386607 4386356 Biosynthesis of cofactors, prosthetic grou 4387086 4386604 Transport of small molecules 4414949 44141311 DNA replication, recombination, modifica 4418302 4418583t Hypothetical, unclassified, unknown 4439376 44397831 Hypothetical, unclassified, unknown 4442452 44428681 Transcriptional regulators 4444977 4445486 ; Hypothetical, unclassified, unknown 4445998 4446390 Hypotheticat, undassified, unknown 4447132 4448217 Transcriptional regulators 4453182 4452535 Biosynthesis of cofactors, prosthetic grou 4457361 4458644 Hypothetical, unclassified, unknown 4459749 4459417 Hypothetical, unclassified, unknown 4461387 4462409 Hypothetical, unclassified, unknown 4462399 4462881 j Translation, post-trans ! ationa ! modification 4463903 4465438 Hypothetical, unclassified, unknown L 4466753-4466322 Translation, post-transtationa ! modification 4466924 4469545 ! Hypothetical, unclassified, unknown 4469612 4470235 DNA replication, recombination, modificat 4470273} 44713108 Hypothetical, unclassified, unknown 4471402 4471554J Related to phage, transposon, or plasmic 4473622 4474638 Biosynthesis of cofactors, prosthetic grou 4477976 4476993 Hypothetical, unclassified, unknown 4478907 4478626 Cell wall/LPS/capsule 4483363 4482260 Hypothetical, unclassified, unknown j 4486179 4485823 Hypothetical, unclassified, unknown 4486846 4486202 Putative enzymes 4488409 4489632 jSecreted Factors (toxins, enzymes, algina3 4713795i 4714283, ! Secreted Factors (toxins, enzymes'aigin », 4714313'4714801 ! t oP secreted Factors (toxins, enzymes, algin-47148-4716042 : Secreted Factors (toxins, enzymes'a Zna< 4719sQZ Secreted Factors (toxins, enzymes'algin, 9 4719418 4720062i Hypothetical, unclassified, unknown j 4724034 4722850 : Transport of small molecules 47448181 4745123 ; DNA repficatior), recombination, modificat 4747136 4746639J Translation, post-translational m. odificatioh 47543781 4753989T Transcription, RNA processing and degr ; +j Translation, post-translational modification 4756066, 1 4755446" T nslation, post-translational modificatio 4756472 47560831, |Translation, post-translational modificatioh 4756847 4756491 i Translation, ost-translationaf modificatioi 4757094 47569781 Protein secretion/export apparatus 4758451 47571231 Translation, post-translational modificatiol _4758888 04758452, Translation, post-translational modificatio 4759066 4758890 { Translation, post-translational modificatio 4759569 47590691 Translation, post-translational modificatio 4759923 4759573T Translation, post-translational modificatio 1 4760467 4759934 Translation, post-translational modificatio 4760871 4760479 Translation, post-translational modificatio 4761366 4761061 Translation, post-translational modificatio 4761919 4761380 Translation, post-translational modificatio 4762253 4761939 Translation, post-translational modificatio 4762634 4762266 ; , 4762634 4762266 ! Translation, post-translational modificatio 4762924, 47626581 Translation, post-translational modificatio 4763118 4762927 ; Transtation, post-trans) ationaimodifjcatioi 4763531 4763118 Translation, post-trans) ationat modificatio i 4764229 4763543 w w-z Translation, post-translational modificatio i 4764574 4764242 Translation, post-translational modificatio 4764862 4764587 Translation, post-translational modificatio i 4765700 4764879 Translation, post-transtationa ! modificatio i 4766011 4765712 Translation, post-transtationa ! modificatfoi 4766610 4766008 Translation, post-translational modificatio 4767259 47666241 Translation, post-translational modificatio i 4767653 4767342 Translation, post-transtationa) modificatio 1 4771655 4771185 l w- Translation, post-translational modification 4772126 4771755 Transcription, RNA processing and degree 4776477 4772278 Transcription, RNA processing and degr 4780616 4776543 j Translation, post-translational modificatio 1 Xtwl Translation, post-translational modificatio 4781785 4781285 ! Translation, post-translational modificatio 1 4782679 4781984i J Translation, post-translational modificatioi 4783110 4782679 ! Transcription, RNA processing and de 4783760 4783227 Protein secretion/export apparatus 4784138 4783770) Hypothetical, unclassified, unknown 4787479 47867331 Hypothetical, unclassified, unknown 4819927 4820409 Two-component regulatory systems 4 4820531 4821358i Hypothetical, unclassified, unknown _s _4823363 48230794 Hypothetical, unclassified, unknown S 4824123 4823386 I Hypothetical, unclassified, unknown L 4830553 Lv642 ! Hypothetical unclassified, unknown 4830963 4831181j Nudeotide biosynthesis and metabolism 4843551 48427001 ; Hypothetical, unclassified, unknown 4851309 4852316j Hypothetical, unclassified, unknown 4854008 48536491, Carbon compound catabolism 4856959 4858410i Putative enzymes4859262 4858489j g Transcriptiona) regujators) 4870327 ! 4869557 j Hypothetical, unclassified, unknown")"'4875244 4874654 j Hypothetical, unclassified, unknown 4877605 48768201 H othetical, unclassified, unknown 4878586 487769_ 487- Hypothetical, unclassified, unknown 4878789 9544l Hypothetical, unclassified, unknown 4882052 4882354j Hypothetical, unclassified, unknown 4884960 4884718 Hypothetical, unclassified, unknown 4887505 4887278J Adaptation, protection4893696 4894277J Hypothetical, unclassified, unknown 4902201 4903295 Hypothetical, unclassified, unknown 4909403 4909161 Hypothetical, unclassified, unknown 4914509 4914126 Chaperones & heat shock proteins 4917123 4915480 0 Chaperones & heat shock proteins 4917467 4917174 Hypothetical, unclassified, unknown 4918932 4918198 Putative enzymes 4919041 4919799 Hypothetical, unclassified, unknown 4921980 4922363 Hypothetical, unclassified unknown 4925794 4925315 : l v Protein secretion/export apparatus 1 4936615 4933865 Hypothetical, unclassified, unknown 4937819 4938214 Cell wall/LPS/capsule 4939186 4938275 Cell division 4940483 4939299 Cell division 4941787 4940534t Celt division 4942672 4941809 Cell wall/LPS/capsule 4945074 4943632i Cell wall/LPS/capsule _ 4946140 4945067 w Cell division 4946130 L Cell wall/LPS/caPsule I 4948675 49473291 Cell wall/LPS/capsule 4949771 4§4868911 v Cell _,, +__ _9J Cell wall/LPS/caDsule I 49526031 4951140ii Cell wall/LPS/capsule 4954342 4952C) 0311 Cell division 4954632 4954339 Hypothetical, unclassified, unknown ! 495557J 4954629 j Hypothetical, unclassified, unknown 4956028 4955573J ne Hypothetical, unclassified, unknown 4959519 4959896 Putative enzymes 4959925 4960518 Adaptation, protection 4961557 4961150 Adaptation, protection 4962186 4961569 Energy metabolism 4964265 Translation, post-translational modificatio 4965500 4965108 Translation, post-translational modificatio 4965943 4966515 -jv Transcriptiona) regutators 4968711 4969610 Hypothetical, unclassified, unknown 4971890 49707961 Translation, post-translational modification 49733311 49719851 tHypothetical, unclassified, unknown 4974, 028 49733991 Cell wall/LPS/capsule 4985469, 1 49842, 04i FHypothetìcal, unclassified, unknown | 49861541 4985846' Hypothetical, unclassified, unknown i 49867981 4986151 Hypothetical, unclassified, unknown 0 49872831 4986810 Transport of small molecules 1 4988081'4987284} Hypothetical, unclassified, unknown 4989304 4990284- Hypothetical, unclassified unknown i'4990832 4991404- Hypothetical,-un. clas'sified, unknown 4991391 4991918 Transport of small molecules 4991918 49926431 Transcriptional regulators 4992869 49943621 Hypothetical, unclassified, unknown 4994440 4994748 Transport of small molecules _ 4994762 4995226i Transport of sma)) mofecutes4996118"996390J Hypothetical, unclassified, unknown 5000303 4999908 Cell. division 5012313 50113211 Cell division 5013430 5012393 Translation, post-translational modificatio 5013670 5013960 Translation, post-transtationai modificatjoi 5013973 50154271 Translation, post-translational modificatio 5015534 5016979 Hypothetical, unclassified, unknown 5017039 5017416 Hypothetical, unclassified, unknown 5028230 5027421 Transcriptions) regulators5036244 5036807 Hypothetical, unclassified, unknown 5046663 50460311 Biosynthesis of cofactors, prosthetic grou 5068031 5068879 Motili & Attachment5069530 5069081 Motility & Attachment 5069762 5071462 M S x Motility & Attachment5071566 5072690 Hypothetical, unclassified, unknown 5073563 5074174 5074171 0-74 Hypothetica, unclassified, unknown 5074171 50743710 . Hypothetical, unclassified, unknown 5077535 5077705 ; Transcription, RNA processing and deers 5091814 5090852 Two-component regulatory systems 5094984 5096321 Motility & Attachment 5099262 5100086 Motility & Attachment 5100083 51006703 Adaptation, protection 51059290 5104985' Translation, post-translationa_I modificatio 5 5106957 5106448s Translation, post-translation-1 5109781 51069501 Biosynthesis of cofactors, prosthetic grou 5110743 5109805 Translation, post-translational modificatio 1 5112662 61129371 Hypothetical, unclassified, unknown 5113468 5113004 Amino acid biosynthesis and metabolism 5114598 5113480 Adaptation, protection 5115889 5114669 Translation, post-translational modification 5116288 5116031 Tra nsiation, post-translational modification 5116623 5116312 Biosynthesis of cofactors, prosthetic grou 5116864 5117832 Hypothetical, unclassified, unknown 5122386 5121898 Hypothetical, unclassified, unknown 5122901"5122560 Hypothetical, unclassified, unknown 5125930 5125604 Hypothetical, unclassified, unknown 5135815 5136192 Transcriptiona ! regutators 5155560 5156123 ! Hypothetical unclassified, unknown 5162448 5162068 H pothetical, unclassified, unknown ! 5168754 5169188 ; j Hypothetica). undassified, unknown 5169504T 5169250 2thet Hypothetical, unclassified, unknown 51749791 5176103 Hypothetical, unclassified, unknown 5197880 5198323 Hypöthetical, unc, assified, unknown g 52049271 5206087. Hypothetical, unclassified, unknown 1 5206486'S 5206208. Hypothetical, unclassified, unknown !'52070061'5206719 Hypothetica), undassified, unknown J 5210995) 5210705 Hypothetical, unclassified, unknown) 5212104j 5211631 Nucleotide biosynthesis and metabolism 52128321 5213470 ; Hypothetical, unclassified, unknown 5215669 52162021 f Chaperones & heat shock proteins 5216763 5217551 ---223-561 5222539i Biosynthesis of cofactors, prosthetic grou Get ! wa))/LPS/capsu ! e5230910" 5230113 ! Biosynthesis of cofactors, prosthetic grou 5231658 5230900 Translation, post-translational modificatio 5233566 5232484 ! Translation, post-translational modificatioi 5234852 52335841 l Cell wall I LPS/capsule 5236773 52373901 9 Biosynthesis of cofactors, prosthetic grou 5237392 5238240 e Nucleotide biosynthesis and metabolism 5238407 5239348 S Transtation, post-transfaiiona) modificatiol 5239465 5240079 Translation, post-translational modificatiofi 5240121 5240705 Hypothetical, unclassified, unknown 5242558 5242253 Putative enzymes 5246122 52454751 Hypothetical, unclassified, unknown 5248771 52480701 Hypothetical, unclassified, unknown 5250617 52495998 Fatty acid and phospholipid metabolism 5272299 5271484i Amino acid biosynthesis and metabolism 5275731 5274007 Hypothetical, unclassified, unknown 5276282 5276734J Hypothetical, unclassified, unknown"5277171 5276842 Hypothetical, unclassified, unknown 5277950 5277171 i Hypothetical, unclassified, unknown 5282158 52825051 Transport of small molecules 5286034 5285267 j Hypothetical, unclassified, unknown 5291613, 5291960 Hypothetical, unclassified, unknown 5297482 52970061 Biosynthesis of cofactors, prosthetic grou 5310725 5311213 Biosynthesis of cofactors, prosthetic grou 5311463 53122631 Biosynthesis of cofactors, prosthetic grou 5313205 5313585 Energy metabolism 5313675 5315339i Hypothetical, unclassified, unknown 5322234 5322449 Hypothetical, unclassified, unknown j 5322706 5322509 | Hypothetical, unclassified unknown 5323100 5322756 Transcription, RNA processing and degri 5325478 53233731 i < Transtation. post-transfationat modificatiol 5325921 5325652 Transtation, post-transiationa) modificat ! oi 5329948 5327426 Transcription, RNA processing and deara 53314571 5329976t t Hypothetical, unclassified, unknown 5331960 53315021 Protein secretion/export apparatus5332742 5332353) Energy metabolism-5333500 5332745 Cell wa))/LPS/capsu) e 5334903 5333566 Biosynthesis of cofactors, prosthetic grou 5335771 5334920 j Ce) t division5338522 5337899 Hypothetical unclassified, unknown 5338617 5338931 pothetical, unclassified, unknown'ì 5343754 i 5343104 i Amino acid biosYnthesis and metabofism S 5345891} 53450851 'Chaperones & heat shock proteins ; 5349760 j 53492001 Transcriptiona, _gu_tors_ 1 5352078i 5351674. jTransportofsmall molecu, es T 5352176'5352706t Hypothetical unclassified, unknown 53535061 5353072 ! Hypothetical, unclassified, unknown T"5361585 5362067 Two-component regulatory systems 5364070 5364735.' Transcriptional ret _62561 5366654 Hypothetical, unclassified, unknown 53707201 5370475 Hypothetical, unclassified, unknown 53768511 5377708' Hypothetical, unclassified, unknown 5377790 5378095. _ Hypothetical, unclassified, unknown 53780924 53788411 Hypothetical, unclassified, unknown 53804471 5379512 w Related to phage, transposon, or ptasmic 5383811 5382795 Hypothetical, unclassified, unknown 5386999 5387721 Energy metabolism 5396858 53959291 Fatty acid and phospholiPid metabolism 54029441 54020153 Hypothetical, unclassified, unknown 5414487 5414278l s Hypothetical, unclassified, unknown 5418568 5418350 Hypothetical, unclassified,. unknown-. 5419862 5420317 Transcriptional regulators 1 5422505 54230651 Hypothetical, unclassified, unknown 5434651 5434115j Fatty acid and phospholipid metabolism 5442303 5442773 Fatty acid and phospholipid metabolism-5442791 5444140c, Translation, post-transfationa ! modificatto i 5445198 5446082 i Transcription, RNA processing and deers 5448642 5448965 w o Transport of small molecules 5460525 5461382 Central intermediary metabolism _ _ 5462762 5463604 Hypothetical, unclassified, unknown 5463917 5464435 Central intermediary metabolism 5464820 5466520 Hypothetical, unclassified, unknown 5468282 5468016 . Hypothetical, unclassified, unknown 5468408 5469022 Hypothetical, unclassified, unknown 5472041 5471625 Hypothetical, unclassified, unknown 5472438 5472734 w=S--5472438 H Transcriptional regulators 5473765 5474577 Two-component regutatory systems"5480401 5481090 Transport of small molecules 1 54837671 54824511 Hypothetical, unclassified, unknown 5486355 5486984 Biosynthesis of cofactors, prosthetic grou 5487714 5488385 Hypothetical, unclassified, unknown 5489040 5489612 Transcriptional regulators 5490651 548962911 Transcriptional regulators 1 55057831 55050704 Hypothetical, undassified, unknown 5507897 5506965J Hypothetica), unctassified, unknown""""5517093 5516398) Biosynthesis of cofactors, prosthetic grou 5519710 55205371 Hypothetical, unclassified, unknown 5522385 5522972 S Hypothetical, unclassified, unknown 5524718 5523867 H othetical, unclassified, unknown 5525904 5524969 ; DNA replication, recombination, modificat 5535555 553 iTrans, ation, post-translational modificatioh 5537288aj 5537058t {Translation, post-trans. ationa. modificatioh ç 5537318 Nucleotide biosynthesis and metabolism 55433641 55420729* Hypothetical, unclassified, unknown Hypothetical, unclassified, unknown 5548644 5548396 i m X 5549721 5548750 0 Hypothetical, unclassified, unknown j"5553583) 5553116) Hypothetical, unclassified, unknown 55569341 5557953 Central intermediary metabolism jHypotheticai, undassified, unknown 5569089 5570627J Hypothetical, unclassified, unknown, 5570624j 55711602 DNA replication, recombination, modificat 5574485 Hypothetica"unclassified, unknown X 55750171 55744931 Hypothetica), undassified, unknown j 5576027 5575014, 11 DNA replication, recombination, modificat 5577916 5576027 ; Hypothetical, unclassified, unknown _L_ 6579498 5578680 Hypothetical, unclassified, unknown j 55809611 5581707 Putative enzymes 5593423 5592632 Cell wall/LPS/capsule 303820 Antibiotic resistance and susceptibility 5606102 5606434 Hypothetical, unclassified, unknown 5606495 5607670i Hypothetical, unclassified, unknown'S607667 5608479 Transport of small molecules 5615543 5613732 Hypothetical, unclassified, unknown j 561565T 5616301 Hypothetical, unclassified, unknown 5626378 5624900 Hypothetical, unclassified, unknown 5627133 5626375 Hypothetical, unclassified, unknown 5627864 5627130 Cell wa/LPS/capsule 5628670 5627864 j Ce wall/LPS/capsule 5629788 56286671 Cell wall/LPS/5630852 5629785 Cell wall/LPS/capsule 5631886 5630849 Hypothetical, unclassified, unknown 5659184 5658417 Transcriptional regulators 56627411 5663814 Hypothetical, unclassified, unknown 5663888 ! 5664868 j Biosynthesis of cofactors, prosthetic gro 5666056 56649891 Amino acid biosynthesis and metabolism 5675701 567-51831 Motility & Attachment 5680712 5679648 DNA rep. ication, recombination, modificat 56874Wi 56897i41 Translation, post-translational modification 5689963 5691726i Hypothetical, unclassified, unknown 5691761 Hypothetical, unclassified, unknown 5702113 5701697 Biosynthesis of cofactors, prosthetic groul 57026681 57034381 Hypothetical, unclassified, unknown 5703453 5704079 Hypothetical, unclassified, unknown 5704076 5705677 Amino acid biosynthesis and metabolism 5706190 5706525 Protein secretion/export apparatus") 5706551 5706799 . Hypothetical, unclassified, unknown 5708035 5708742, Chemotaxis 5708954 5710897 Hypothetical, unclassified, unknown 5720468 5720944 Transcriptional regulators 57235811 57245371 Carbon compound catabolism 5752463 Central intermediary metabolism 5754143 5753613 fTranscriptiona. requ. ators-r mbWi Amino acid biosynthesis and metabolism 57, 66483 57678921 , Protein secretion/exDort apDaratus i 5777096i 5776605 ! Nucleotide biosynthesis and metabolism 57773871 5777133 Hypothetical unclassified, unknown _ ; _ t _ ___ p, 5777808j 5777389 ; Carbon compound catabotismj 5778133 5779680 Hypothetical, unclassified, unknown 57799941 57808121 Amino acid biosynthesis and metabolism 5791085'5790444s Hypothetical, unclassified, unknown's 5791836 5792234J Hypothetical, unclassified, unknown !'5797064J 5797336 ! Transport of small molecule 128-. 5802823 Cell wall/LPS/capsule 5810280 5581131338; . m em ! Cell wa) t/LPS/capsule5811335 5812243} Cell wall/LPS/5812240 58131211 Cell wall I capsule 5813121 5813666 Amino acid biosynthesis and metabolism 5824786 5825718 Hypothetical, unclassified, unknown 58294691 5828903 Hypothetical, unclassified unknown 5830749 5830312 Hypothetical, unclassified, unknown 5835481 5835894 Putative enzymes 5840894 5839104 Putative enzymes 5843799 584 9 Chaperones & heat shock proteins 5848366 5847971' Putative enzymes 5 5879970 5878753 Hypothetical, unclassified, unknown 58804811 58799991 Trans) ation, post-trans) ationa) mod) ficatioh 5883012 5881678 Hypothetical, unclassified, unknown 1 58835760 58830225 Hypothetical, unclassified, unknown j 5883971 5884285 m v Hypothetical, unclassified, unknown 5885485 5885943 Hypothetical, unclassified, unknown 5896726 589526011 Transcription, RNA processing and degri 5900123 5898864 Energy metabolism 1 5900694 5900368 Hypothetical, unclassified, unknown 5907389J 5907862 Hypothetical, unclassified, unknown 5908329 5907847 Biosynthesis of cofactors, prosthetic grou 5921496 5920741 s Biosynthesis of cofactors, prosthetic grou 5922434 5921493 e H othetical, unclassified, unknown 5941081 5940746 Cell wa/LPS/capsule 5941335 5941475t Amino acid biosynthesis and metabolism 5942744 5943574 Putative enzymes5945234 5945932 Central intermedia metabolism 5952820 5952482 DNA replication, recombination, modifica 5962715 5964724 Energy metabolism 1 5969764 59693541 Hypothetical, unclassified, unknown. r-5972202 5971849 Transtation, post-transiationatmodificatiol 5986118 5985882 DNAreplication, recombination modificat 5989319 5988645 DNA replication, recombination, modifica 5989459 5990667 4- Nucleotide biosynthesis and metabolism 5990675 5991130t Hypothetical, unclassified, unknown 5996035 5996985 Energy metabolism 6000142 5999753 Hypothetical, unclassified, unknown 6001377 6000760 Nucleotide biosynthesis and metabolism 1 6002039 6001398 Hypothetical, unclassified, unknown 6003329 6002958J Transcription, RNA processing and degree 6004095 60033761 unclassified, unkno i Hypothetical, unclassified, _known J _8004276 60051 39ì Nucleotide biosynthesis and metabolism 6005198 6005809i ! Hypothetical, unclassified, unknown 6008397 6008777 Hypothetical, unclassified, unknown 6017013 601662_1 Carbon compound catabolism t 60189961 6018829' Carbon compound catabotism 6019347"6019180 Biosynthesis of cofactors, prosthetic groul 6025304 ! 6026194 Two-component regutatory systems 6032267 6031365 Hypothetical, unclassified, unknown Hypothetical, unclassified, unknown'"'6063831 ! 6063352 j Putative enzymes, 6069098 6067944i #s Hypothetical, unclassified, unknown'6074229 6075206 Transcriptional regulators. ~ 6082866 6083072 i Hypothetical, unclassified, unknown 60831041 6083508 f P. l . Hypothetical, unclassified, unknown 6083752 6084084 Hypothetical, unclassified, unknown 60845441 60843681 Carbon compound catabolism 6096706 6097026 Hypothetical, unclassified, unknown 6148318 6149181 j Hypothetical, unclassified, unknown 6151313 6151525 Hypothetical, unclassified, unknown l 6155202 61547831 Hypothetical, unclassified, unknown 6158178 6158942 Translation, post-translational modificatio 6159562 6158948 v 6171732 1 Hypothetical, unclassified, unknown 6171732 6171926 Hypothetical, unclassified, unknown 6172872 6172711 1 Hypothetical, unclassified, unknown 6188854 6188165 Transport of small molecules 6195308-6196315 Hypothetical, unclassified, unknown 6219068 6218856 Transport of small molecules 6221100 Transport of small molecules 6225924 6224896 . Hypothetical, unclassified, unknown 6227081 62274491 Hypothetical, unclassified unknown 6228243 6227602 Hypothetical, unclassified, unknown 6236228 62358301 Central intermediary metabolism 6244945 6243110 Cell wa))/LPS/capsuje !' 6247689 6246325 Energy metabolism. I 6248235 6247810 Energy metabolism 6249653 62482771 Energy metabolism 6250544 6249684 Energy metabolism 6252139 62505951 Energy metabolism 6252694 6252158 Energy metabolism 6253176 62527061 Energy metabolism 6253491 6253234 _. _-.. Energy metabolism 62544101 6253541 Energy metabolism 6254807 6254427 Cell division 6255843 6254971 Hypothetical, unclassified, unknown ___ __ 6260053 6259670 Hypothetical, unclassified, unknown 6263563 6261827 Transtation, post-transtationaf modificatio i 6264211 6263804 Transiation, post-transiationai modificatio i 6264360 6264226 4269 4200 0. 9999719 . 4270'4074 i 0. 9999719, rpoB 3640 3522 0. 9999058 : _11_56 _ 2892 995D54 ; nrdA 3834 2853 0. 9994519 vals 4560 2832 0. 999420 osta 595 2775 0. 999327 3168 2772 0. 9993217 i gyrA 1t__sqq_qq_ 4403 2751 0. 9992831 I secA a 0. 9990012 alas 3987 2622 s 0 3987 i 2622 5 0. 9989933 5 leuS i 1787 T"2610'0. 998961 i acnB. I toA 3011 i 2607 | 0. 9989528 5 4744 B 2523 0. 9986937 2436 0. 9983575 ftsk o__q qqe a_s 4"2421 0. 9982914"gyrB'j L- ! 3 ! 2397"0. 99818 | lon _j 3648 2394 0. 9981655 1529 1 2385 0. 9981216"Jig ! s 2739 2379 0. 9980917 pheT 5 3704 1 2310 0. 9977116 . 4964 2265 0. 9974239 parC 5050'2220 0. 9971 priA 260 ! 2151 0. 9965225 21-36 0. 9963824 1372 5 2136 0. 9963824 5 2071 1 2109 0. 996116 fusA2 0. 9960852 n 4740 2106 p P--i 8 2055 0. 9955228 giyS -3 qq--q 2046 0. 9954155 5 dnaX 1532 3482 j 2034 0. 9952684 5 metG 55 5296 2010 0. 9949599 ! rep 1939 1998 0. 9947981 1480 1974 0. 9944589 ccmF 5072 i 1944 0. 9940037 2744 1923 0. 9936629 thrS 1068 1911 0. 9934596 1596 1905 0. 9933555 htpG 3157 1890 0. 9930879 4967 !"1890'0. 9930879 parE 4044 i 1884 0 9929779 i dxs 5 1 1370 1 1866 0. 9926372 3821 i 1863 5 0. 9925788 secD 5 al-PM 3810"1860 0. 99252 j hscA 3821 1810 1848 099228 5549 1 1836 0. 992032------qlm-s 4gg7 _ 1812 0. 9915127 msbAV _ 5 4997 1812 0. 9915127 I msbA 767 i 1800 0. 9912404 1. epA 5187 1791 0. 9910304 1583 i 1773 t 0. 9905952 i sdhA r S _ __. _ L _ __. _. _,, _,, _, 1764 0, 9903698 arqs 0. 9902165 740,. . M 0. 98974B1 B. M.. _.. _ m. _. _.. ftsl.. ^. y . 1758 0. 9902165 ! 4418 1740 : 0. 9897418 ; i fts. . 5568. 1737. 0. 9896605 2298 i 1725 0. 9893287 ; j _4696 _. _ L_ t__ _____ ___ i ; 3725'1716 0. 9893287 956 1716 0. 9890729 4868 , 170'1 ; 0. 9886329 l ureC !""4526""'i'7oi'""""0. 9886329" """p ! ! B"""" 3162 1 1680 1 0. 9879869 ; rpsA 3162 1680 0. 9879869 r-psa ! t 1794 1671 0. 987699 gins 2075 i 1662 0. 9874041 2953 1656 : 0. 9872036 in i 4385 1644 ! 0. 986793 i groEL 2953 1 1656 0. 9872036 3637 1629 i 0. 9862612'i pyrG 1251 1 1626 0. 9861522 ; j 1629 0. 9862612 ; ; 183 ; 1611 i 0. 9855946 1 atsA 183 1611 0. 9855946 atsa 5065 1602 0. 9852493 3769 1578 0. 9842875 jguaA ! 2818'""1578"1"0. 9842875"'j'"i i 3024 1560 i 0. 9835251 f 5131 ! 1548 10. 9829965pgm M 1331 i 1548 0. 9829965 r 555611545) 0. 9828617)"'atpA 5556 1545 0. 9828617 atpa 3984 r'1536 j 0. 9825889 _ 1536 _ t Int 3984 i 1536 ! 0. 9824509 l Int 2207 Si 1521 i 0. 9817442 fj _. __ i mtlY 2343 ! 1509 0. 9811584 3103 ii 1509 1 0. 9811584 I xcpR 3103 1509 0. 9811584 xcp I b2u 1503 0. 9808585 ! 985 1497'j"0. 9805538T"'"'j r 3570 1494 0. 9803997 ; mmsA 4462 1494 ; 0. 9803997' rpoN ( l s 443 ii 1491 0. 9802443 "'4745'"" ! 1482'" !'0. 9797707 T'nusA"""" 5006 1 1479 i 0. 9796104 2097 ! 1476 ! 0. 9794487 ! ! 5237 j 1467 ! 0. 9789561 0. 9787893 4417 1464 iure 2037 1461 A 0. 9786211 2037 1461 0. 9786211 202 8 1458 ! 0. 9784516 { om 202 14bb 0. 9784516 1 1458 1 0. 9784516 --1455 1 0. 9782808 CZCB 4483 j 1455 v 0. 97. 82808 gatA 4329 1452) 0. 9781087 pykA 913 1449 j 0. 9779351 nTgtE 4484'1446 J 0, 9777602'gatB 1443 0, 9775839 murc 1587 1 1437 0. 9772271 1 lpdg LZ3.- !"11"''. 0. 9768646' T""" ! z 2073 i 1431', 0. 9768646 1 ! 1425 0. 9764964 2391 1425 0. 9764964 1416 0. 975933 .'"""'5119" :"1410"""09755499"f'g) nA ..... _., _ __. __ _ _. ; _ _. _ _ ___ __. _ _ __. ___ _4__ _ _ _ _-__-_ 4 686, 1410 _ 3876 1407 0. 9753561 i narK2 704,, 5 _... . ... __ 139 974565 E I 1395 , i 704 1395 0. 9745653 1395-0. 9745653...... .'""4931r""1395 ;"" 0. 9745653""IdnaB 1386 0. 9739556 1795 1383 0. 9737491 cys 3-8-0 0, 973541 xsea S 4416 1377 0. 9733313 | murF 1377 0. 9733313 3746 1 1374'0. 9731199 f ffh * * ; 9 2629 1371 ¢ 0. 9729068 f ; purB f 3746 374 0. 9731199 ffh !'1217 j1368,'0. 972692 1j 3081 1368 0. 972692 I ß 3081 1368 t 0. 972692 lu 2131 1362 0. 9722573 0. 9713671 l- 119 1350'. 0. 9713671 0. 9713671 accc z e z '. 4439 j 1347 0. 9711402 1 trpS 4414 1347lIV 0. 9711402_W murD I 2641 _ 1347, 0. 9711402 i nuoF f l >o 2843 1347 ! 0. 9711402 f 2336 1341 ! 0. 9706808 0. 9704484 ----l 4-9 1338 0. 9704484 gimm '5224) 1335' ! 0. 9702141 pep ?) 2475 1335 0. 9702141 r'3638 ! 1329""""' :"0. 96974"" J 4243 1329 v 0. 96974 J secY w oM-M-^ 716 1326 0. 9695001 2214 1323 0. 9692583 2794 0. 968769 1 | 4887 1317 ! 0. 968769 ! 3280 1317 1 0. 968769 opro e yyw y 3154 1317'. 0. 968769 wzy r--53-9 1314 0. 968521 3159 1311 0. 9682719 ___ybpA 2338 1-311 0. 9682719.......... 1183 ft 1311. 0. 9682719 I dctA f _........ _... _.. _........ 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DNA polymerase III, alpha chain ribonucleoside s, large chanM ; valyl-tRNA synthetase ; organic solvent_t_oIe_rance protein OstA, precursor, , m... _. _. _.,. _. _..... _. _..... _ __. : DNA yrase subunit A_ _,-. _. _.. _.. _ _. __,. _____... _. _, _. _. __ secretion protein Sec_A DN rase sub _^ ___ _ _ __ _ _. _ ; tfe'ucyl-tRNA synthetase . _-.. _.. _, ... _. ___.., . _. _. _.. . _ < cA___ ~__-_ g e nv'i NA 511'bdi <-_ _ wow wwR~_w_. w__w _w ___tw_ £aconitate h dratase 2 {DNA topoisomerase I translation initiation factor IF-2 'cell division protein FtsK _ _. _. T_. _,. _ _ : .. iLonprotease ; a . robable outerelembrane protein F-P J DNA ligase phenylalanyl-tRNA synthetase, beta subunit probable chemotaxis sensorleffector fusion protein topoisomerase IV subunit A primosomal protein N' _. __.. _ __ __ _ __ _w_ _ _ v. ____ __w ___ __ ___ _ hypotheticaf protein hypothetical protein , elongation factor G lelongation factor G pojyrtbonucieotide nucieotidyitransferase . . . . .-.- ! glycyl-tRNA synthetase beta chain DNA polymerase subunits gamma and tau methionyt-tRNA synthetase J ATP-dependent DNA helicase Rep hypothetical protein jcytochromeC-type biogenesis prpteinC ! tmwS_rOtein CcmF probabtechemqtaxist ! besut tHNA sEthetcse probable heat shock protein (hsp90 family)... _.. ! heat shock Drotein HtpG _ E iprobable acetyltransferase itopoisomerase) V subunit B j t a r _w. _w _} % _ ______w_____fl_ P__ __9_ww~__~_ _ _ _ hypothetica) prqte) n j secretion protein SecD heat shock protein HscA robable binding rotein compon_ent of ABC transporter __ rhate aminotransferase *__ transport protein MsbA _ ' GTP-binding protein LepA probable acyl-CoA dehydrogena_s_e _ _ _ _ arginyt-tRNA synthetase.'- ! _arginyl-tRNA_synthetase _ probable acetyltransferase ; p_enic''_.. g. ! conserved hypothetical protein ! probable oxidoreductase , _. iacetolactate synthase larqe subunit isingte-stranded-DNA-specific exonuciease RecJ . prolyl-tRNA synthetase iprolyl-tRNA synthetase urease alpha subunit ttyEria mesis pro ñ i il ~~~ 30S ribosomal protein S1 ! 130S ribosomal protein Sl Igluco_se_-6-phospha_te. isomerase--.. _... ____. _.. _.... _.. _. _. __.. _-__... _... ___. __.. _. __.. _.... _... _. _.... , __. _... _. _. __.. _....-_.... w__.. __. __. ___. ___... __. __.... __... _.. _.... _.. __. _.. _..... ______... __.. ___.. ... _. _.. __ glucose-6-phosphate isomerase hypothetical protein GroËL protein CTP synthas_e _ __ _ _ Cbabsynchemotaxie arylsulfatase conservedjhypothet) cajjp j i . gap hypothetical protein probable carbohydrate kinase phosphoglycerate mutase conserved hypothetical protein chromosoma ! repfication initiator protein DnaA J ATP synthase alpha chain hypothetical protein H g. _ _ S H apolipoprotein N-acyltransferase - hypothetical protein xytuiose kinase ! general secretion pathway protein E ^ M v_ __ _*w__ sodium/proton antiporter NhaB _ _ _ _ _ _ i probable colicin-like toxin probabtecojicin-iike toxin') methyinDaJonate-semiaidehyde dehydrogenase. 1""""""" wmerase sigma-54 factor probable transporter N utilization substance protein A hypothetical protein probable flavin-binding monooxygenase conserved hypotheticål protein hypothetical protein h pothetical rotein probabie amidase"] outer membrane protein OprM precursor probable amidase RND divalent metal cation efflux membrane fusion protein precursor _ § Glu-tRNA (Gln) amidotransferase subunit A pyruvate kinase 11 probable Mg transporter MgtE Glu-tRNA (Gln) amidotransferase subunit B UDP-N-acet ly muramate_--a_lan_ine ligase w -. ^-.. __ I lli, ooamide dehydroqenase-glc probable transporter (membrane subunit) conserved hypothetical protein robable outer ; probab) e ferredoxin -.-....-.--....-.-.........-..... EglutamT y ; ine senthetase ___... ___ =nitrile extrusion protein 2 ! Probable amidase '9 Pi"-I ! i° !.' ?"shydrpgenase *""' soluble pyridine nucleotide transhydrogenase probabfegjycerafdehyde-S-phospha"'.--- cysteinyf-tRNAsYnthetase "'-- jexodeoxyribonuctease VJ ! targe subunit'"----.-..-.... UDP-N-acety) muramoy) atanyD-gfutamy2, 6-djam'""""" ATP synthase beta chais signal recognition particle rotem Ffh adenylosuccinate lyase _ _ _ signat recognitiqn partide rec" ! {probabte 2-is 'probable 2-isopropylmaiate s nthase , . _, conse_rved hypothetical protein''"""''-- protein . h othetical rotein probable dicarboxylate transporter biotin carboxylase"" ; tryptophanyl-tRNA synthetase UDP-N-acety) muramoy ! atanine-D-g ! utamate ! igase , NADH dehydrogenase I chain F" probable aldolase I hypothetica ! protejn''"""" two-component response reg_ulator PiIR _ phosphoglucosamine mutas aminopeptidase P probable cytochrome P450 secretion protein SecY hypothetical protein hypothetica) ! probable MFS transporter hypothetical protein jprobabte MFS transporter porin 0 precursor 3 B-bandO-antigenpoiymerase"j conserved hypothetical protein probabte UDP-gjucose/GDP-mannose dehydrogenase WbpA *'" probabte binding protein component of ABC maltose/mannitol transporter C4-dicarboxylate transport protein 'conserved hypothetical protein conserved hypothetical protein A, TflIB roteW _ _ _ _ _ _ conserved hypothetical protein jTotB protein"''' (hypotheticat protein -----j ; adenytosuccinate synthetase '""""""*'''"'' ! peptidyi-prolyi cis-trans isomerase SurA ., _. ___. _.... _... _........ __. _........ _.. _. _......... _.. _... _.. _. _. _.. _. __.... _.... __... __.. __........ hvpothetical Drotein Y _ glutamate-1-semialdeh de 2 1 Vamnom. utase........ _.. _, ____. _.... _... hypothetical protein glutamate-l-semialdehvde 2, 1-aminomutase h otheticalrotein -... _.. _. _.. _..... _.-_... _.. _. __... __... _.. _.. _... ___.. _. _.. _........-. _.... _.... _...-... _........ _.. _...... . _. __.... e......... __. __. _. _.. _.. _... _. _... _... ____r.. __.... ; -.. Yp p : 3-oxoacyl-ac i cy _arrier protein synthase I I ''------------------_---___.... _ hypothetical protein ; glutamyl-tRNA reductase "" ""°°--------. __. __.. _........ _. _.... ___., , 3-oxoacvl-acvl carrier protein synthase 11 me 1 carbox vin Itransferase, glutamyl-tRNA reductase 'UDP-N-acetylglucosamine 1-carboxyvinyltransferase hypothetical protein 'hypqtheticai protein-.---...-.-....- ; transcription termination factor Rho Ecell division protein FtsA ihypothetical protein __ _ __. _.. __.-_. ___....... _. _. __.. _. _. ___...... _.. _....... _... WH. Hy- i_conserved hypotheticai protein _ _-- ---3 ''conserved hypotheticaf protein II. L. 1I'.. I-"'"""'' ! nbonucieoside reductase, small chain probable glycosy ! transferase WbpJ, hypothetical protein laspartate kinase alpha and beta chain O-antiaen translocåse 0-anti en translocase hypothetical protein i . hypothetical protein probable transporter (membrane subunit) probable transporter Cr-jr-i : Er-n-e's-ubunit)""'*"""-'--"'--'"",--"-"",-,--"-"-",",""-,-* "",---"""----,--", *----,--,------"---"-, fconserved hypothetica) protein j probable MFS transporter conserved hypothetical protein ! probabie hyd ro ! ase ! hypothetical protein GTP-binding protein Obg . probable FAD-dependent monooxygenase J - p. p p--.. __ -M _.. _ hypothetical prOteln _ _rwrwrrrr~~~~'~rr__________ _S hypothetical protein phenazine biosynthesis protein PhzC probable type 11 secretion system protein probable MFS transporter conserved hvpothetical Drotein ~ _ ____"________ _ _> _r_ __^____ ___~__ probable cytochrome b probable cytochrome b hypal protein--- probable cytochrome b hypothetical protein DNA/pantothenate meta bolis membrane protein OpdE membrane protein OpdE two-component sensor probable acyl-CoA thiolase - M 7thase amno 7 oxononanoate s nthase lconserved hvpothetical Drotein ! conserved hypothetical protein cei) division protein FtsW hypothetjca) protein ! probabte FAD-dependent monooxygenase conserved hypothetical protein probable MFS transporter probable acyl-CoA thiolase _ypothetical ypothetical protein 11-deox--lulose 5-phosphate reductoisomerase methionine adenosyltransferase t-cell division protein FtsZ __. ¢_ypothetical protein _ prpbabje pyndoxai-phqsp enzyme"""'"' acetyl-CoA acetyltransferase ) probable mdybdqpterin biosynthesis protein MoeB------ hypothetical protein (hypothetica) protein----. ! hypothetical protein Yp p : _. _ 'h othetical rotein __. __... __.. ___. _... __.. _.... . v. _^. .. _. _. _. ^......... _.. ___. ___. _.......... _..... _.... _... _. __.. _. _... _. __....... _. . _. _. _... __.. _..... __.. ! fsuccmy-Asyntheta""'""'" Y Y _ , probable hydrolase lhypothetical protein probable al-CoA dehydrogenase .......... phosphoglycerate kinase _ hypothetica ! protein hypothetica ! protein"" hypothetical hypothetical protein probable petidic bond hydrolase__y phosphoglycerate kinase hypothetical protein probable precursor probable peptidic bond hydrolase probable multidrug resistanc conserved hypothetical protein probable acyl-CoA dehydrogenase x general secretion pathway protein L probable permease of ABC transporter erythronate-4-phosphate dehyÇse rV w P Y 9 conserved hypothetical hypothetical protein probable acyl-CoA thiolase conserved hypothetica) protein'"""J lipid A-disaccharide synthase , LPS biosynthesis protein WbpG hypothetical protein probable methyltransferase 2robable acyl-CoA dh conserved hypothetical protein conserved hypotheticai protein conserved hypqtheticai prqtei n still tochrome c oxidase, subunit II v cytochrome c oxidase, subunit 11 conserved hypothetical protein riboflavin-specific deaminase/reductase probable giycosyttransferaseWbpH-- . gtycinecjeavageystem protein T2 muconate cycloisomerase I , glycine cleavage system protein T2 .. ----. _ ___ hypothetical protein ___ _ glutamate 5-kinase conserved hypothetical protein hypothetical protein 'conserved hypothetical protein , hypothetical protein I'hypothetical protein conserved hypothetical protein ,--. .. _. __. rod sha e determmm rote n g , _co oi dehy ge ase Zn-d pe dent) _ _ _ _ __ _ conserved hypothetica ! protein-.--.-.-..----.-...... -.. . . , hypothetica) protein"'''""'"--"--""--""-"- : _hypoth_etical protein _.... _... _... _ _. __ 'chain release factor 2 r., hypothetical protein conserved hypothetical protein thypotheticai protein _ _ _ ____ __ conserved r) ypotheHcaj protein--...-.-. -.... hypothetical protein conserved hypothetical protein 3 pho õserin an iñotran fera é ~ ~ ~ ~ ~ ~ ~-~~~~~~~ ~6 , peptide chain release factor 1 phospho-N-acetylmuramoyl-pentapeptide-transferase i thypothetical protein iprobable aminotransferase WbpE ~~~~ ~ ~ ~ ~~ ~~ ~ -~ ~~~~~~ hypothetical protein phospho-2-dehydro-3-deoxyheptonate alll o ase O methyiguanine-DNA methyltransferase .' probable transcriptional regulator ! phospho-2-dehydro-3-deoxyheptonate aidoiase ; f UDP-N-acety glucosamine--N-acetylmuramvl-(pentapeptide) pyrophosphoryl-undecaprenol N-ac_y giucosamine tr were , uroporphyrinogen decarboxylase ; hypothetical protein _ jhypothetK ; a) protein' he tos rltransferase I conserved hypothetical protein fructose-1, 6-bisphosphate aldolase type 4 fimbrial biogenesis protein PilM probable UDP-N-acetylglucosamine 2-epimerase Wbpl- phosphoribosylaminoimidazole synthetase UDP-3-0-r3 ~ 1ucosamine N-acvitransferase yL iquinolinate synthetase A probable alcohol dehydrogenase (Zn.. dependent) . _ conserved hypothetical protein. conserved hypothetical protein _ g dTDP-D-glucose 4, 6-dehydratase out-er membrane protein OprF precurso hypothetical protein lconserved hypothetical protein jtransjocation protein jntype"'"j T_ _. __ To) __. _.. _.. _... rTolA protein __~ p __ conserved hypothetical protein acetyipodyamine ãmiiiohydroïase conserved hypothetical protein conserved hypothetical protein t ; RecA _. ..... _-_... _.. _..... __. _ theptosyltransferase 11 ____ ls P=--- conserved hypothetical protein t ng protein MreB _ __ _ g rod shape-determining protein MreB hypothetical protein hypothetical protein Ste ydrodipicolinate succinyiase _ ____ __ ___ +_. _ w... __. _. __ _ tMat omponent of ABC transpo ter _ ___ tetrahyd rod ipico linate succin) Lase., __. jDNA potymerase i)), deita subunit'"II'IITIIZ'" hetrahydrpdipiconate su"""'"*'" iprobab ! e binding protein cqmppnent of ABC transporter ! hypotheticat protein'j Sconserved hypothetical protein lprobable binding protein component of ABC transporter _ n p tei _ _ _ _ _ _ 'O-sialolycorotein endopeptidase y. T.... ... . _ .,..,. ^.. _ _. _. ... w_,-_ ... ...... ,, , ferrochelatase-.. ; conserved hypothetical , iconserve hypotheMcaprotejn"""" ; probable aminotransferase tprobable transmembrane sensor jUDP-N-acetylpyruvoylgiucosamine reductase ,'hypothetical protein' iacetoin catabolism protein AcoB conserved hypothetical protein probable oxidoreductase Nw, ,., , W "''''-"'' hypothetical (giycosyltransferase WbpL _ hypothetical protein probable transposase flagella motor switch protein Fui6 hypothetical protein probable transposase P P probable transposase probably robable trans osase phenylalanyl-tRNA synthetase, alpha-subunit _ _ probable permease of ABC transporter hypothetical protein hypothetical protein fructose-1, 6-bisphosphatase conserved hypothetical protein ___ probable ATP-binding component of ABC transporter conserved hypothetical protein hypotheticat protein""''"''"'''""j probable 0-methyltransferase partate carbamoyltransferase jg) ycera) dehyde 3-phosphate dehydrogenase'" jconserved hypothetical protein D directed RNA polymerase alpha chain _"rv. probable pyruvate dehydrogenase E1 component, beta chain tetraacyldisaccharide 4-kinase rod shape-determining protein MreC D-lactate dehydrogenase (fermentative) sulfate transport protein CysA ' " pyridoxal phosphate biosynthetic protein PdxA probable transmembrane sensor =DNA polymerase lil, deita prime subunit' L-aspa iginase I hypothetical protein probable bacteriophage integrate tipoate synthase J hypothetical protein (y othetical protein _ p ion nscriptional conserved .. __. _.-otheticaj...... protei_...... _.... ___.. __...... _.. _...... _....... __. _. _........ _... ___.. _... _.. ____.. _..... _........ __... _.. _. _... ___. ___.. _. __........ __. __... _. _. _...... _. _. _......... __. _.. _. _.. __... ___.. __... _... __ . hyp p t n _ . conserved hvpothetical protein 'pctaprenyt-diphosphatesynthase *"" j hypotheticat protein ; hypothetical protein' hypothet'. thase Shpothetical proten-__ hypothetical protein . conserved hypothetical protein hypothetical protein __ ___ probable enoyl-CoA hydratase/is_erase_ ____ _ _ _ 'thiamine monophosphate kinase hypothetica) protein" ical roten ; , hypothetical rotein hypothetical protein hypothetical protein , hvpothetical protein 'p probable serine/threonine dehydratase, degradative 1 seudouridine synthase conserved hypothetical protein hypothetical protein MLee . ribosomal larae subunit pseudouridine synthase C ,,. _, « ___ _____ ____ ~ . h pothetical rotein Ty -, hvpothetical Drotein probable transmembrane sensor probable transmembrane sensor hypothetical protein probabietransmembrane sensor'" p = probable transcriptional regulator nypuUI=cH pTto n _ _ _ _ _ _ _ glutathione synthetase hypothetical protein probabie adhesion protein acetyi-coenzyme A carboxytase carboxyt transferase (a) pha subunit) probabie transcriptiona) regutator probable transcriptional regulator probable NAD-dependent epimerase/dehydratase WbpK probable NAD-dependentepimerase/dehydrataseW" ! probable oxidoreductase WpbB probable transmembrane sensor hypothetical protein hypothetical protein gtycyl-tRNA synthetase alpha chain,, ro ; p bable h ase l-Lyt_-B protein [conserved hypothetical protein probable hydrolase _ - p Y p _. _.. _. _. _. __. ribose-phosphate pyrophosphokinase hypothetical protein conserved hypothetica) protein porphobiiinogen deaminase ! probabte transcriptiona) regutator" ! probable transcriptional reaulator probable lauroyl acyltransferase transcriptional regulator PtxR probable transcriptional regulator probable transcriptional regulator hypothetical protein ____ __ __. _ _---> malonyl-CoA- [acyl-carrier-protein] transacylase maionyt-CoA- [acyi-carrier-protejn] transacyiase"j riboflavin kinase/FAD synthase, conserved hypothetica) protein j conserved hypothetical protein ; probable phosphatidate cytiltransferase '''..-. _. _. _______... _. ___-- hypothetical protein carbamate kinase hypotheticai protein ____ probable transcriptional regulator catechol 1, 2-dioxygenase D-alanyl-D-alanine-endopeptidase ! probable transcriptional regulator l hypothetical protein probabie epimerase.""""""" ; _lipase Lip_C.. w' probable epimerase ranscnptional reguiator __ electron transfer flavoprotein alpha-subunit FdhE protein jFdhE protein hy othetical rotem __TT hyEthetical protein conserved hypothetical protein probable ATP-binding component of ABC transporter .. tprobable cytochrome c _ __ __ zg u I ato r _ _ __ _ _ probable permease of ABC transporter j probable cytochrome c probable transcriptional regulator hypotheticat protein' probabte transcriptiona) reguiator' probable permease of ABC transporter hypothetical protein p obable transcriptional regulator probable transcriptional regulator probabte transcriptiona ! regutator GTP-bindinprotein Era_"Vw---.-_. _. ... . . . ..-_. __. __... _. _. ___. ... ____.. _ hvpothetical protein GTP-binding protein Era pyrroloquinoline quinone biosynthesis protein B probab ! e cytochrome c oxidaseassembjy factor' hypotheticat protein"""'"''"""""'----"------.-..--. hypothetical protein probable short chain dehydrogenase hypothetical protein hypothetical protein hypothetical protein hypothetical protein UDP-3-O-acyt-N-acetyfgtucosamine deacetyiase probabte probable transcriptional regulator _ probable hypothetical protein conserved hypothetical protein probable binding protein component of ABC transporter' Mv dTDP-4-dehydrorhamnose reductase"'--------.---. ... _ _ __ _....... J conserved hypothetical protein probable transcriptional regulator hypothetical protein""" hypothetical protein probable transcriptional regulator probable transferase i _ S probable two-component response regulator p bable transcriptional regulator i _. } probable transcriptional reoulator L hvpothetical Drotein ? cys eine syn hase 8.. _'... .. _. _.., _ ,.... ...... ...... _.-... _.. .. M,. _,.. __. _., _. __ cystine synthase B probable glycosyl transferase fh. __thetica_r - ; prokable_transcriptional regulator, -. 'W----.. __. _. _. _. _ _. ... _... __.. _. . ____.. _. __... _.. _. ...... pra_bable transcrip_tional reg_ulato_r ---------.--m-. __-_., pro iprcbabte transcriptionai reguiator"' : '. probabte transcriptionai regutator"' yprobabie transcriptional regulator _ _ : cytochrome o ubiqui. nqioxidase protein CyoE'"' probable transcriptional regulator ; probable transcriptional regulator 4-hydroxybenzoate-octapreny ! transferase *' p _s_ rotem. _. _, _. . _. ........ , . . _.. _ lprobable ch motaxis protein cons__. _..... _ probable 2-OH-lauroyltransferase probable transcriptional regulator succnyl CoA synthetase alpha chaW conserved hypothetical protein geranyitranstransferase __w____... _... .. __ _. __ . _.. __ hypothetical protein ribosome protein L11 methyltransferase' r glucose-1-phosphate thymidylyltransferase conserved hypothetical rotem _ hypothetical protein dihydrodipicolinate synthase ; meth Itransferase PiIK , _."" M, _ methyttransferase PitK . "'''" ! probable transcriptional regulator conserved hypothetical protein hypothetical protein chromosome partitioning protein P p g rotein S o0J _ _ _ ; hypothetical protein acety !-CoA carboxylase beta subunit élongation factor Ts cell dmsion roten zi A ATP s nthase A chain outer membrane protein Pope lipase modulator protein hypothetical protein probable transcriptional regulator ; conserved hypothetical protein cell division protein Fis mafonate decarboxyfase beta subunit _ ATP synthase gamm . LJI'IHIIHI ! probabie hydrolase h pothetical rotein probabie short-chain dehydrogenase' ! probable ATP-binding component of ABC transporter spermidine synthase h pothetical protein __ _ ' . 2robable ATP-binding component of ABC transporter pro_'I rotein _ _. othetical protein probabie transcriptionat reguiator' !' IIL---"---'- 5, 10-methyiene-tetrahydrofo) ate dehydrogenase/cyc ! ohydro ! ase-...-.--- _qrolase s signal peptidase)---- isignal eptidase I ! hypothetical protein Idihydropteroate synthase conserved hypothet) capoteir) *""t probable potassium channel isopentenyt monophosphate kinase""'I hypothetica) protein"'"' hypothetical protein tncotmate-nucleotide pyrophosphorylase, w nicotinate-nucleotide p rophosphorylase hypotheticai protein--- 2-dehydro-3-deoxyphosphooctonate aldolase-_,. _,-" , Y. w u. _. _.. _. __. _________. __. .. probable transmembrane censor, urease accessory protejn flagellar synthesis regulator FIeN _ l probable ATP-binding component of ABC transporter"""" probable phenazine biosynthesis protein ATP-binding component of ABC phosphonate transporter probable ATP-binding component of ABC transporter probable permease of ABC transporter probable short-chain dehydrogenase diaminopimelate epimerase _ , . .e _. I conserved hypothetica) protein JinI NH3-dependent NAD synthase probable biotin synthesis protein BioC probab ! e transcriptionai reguiator j type4 fimbria) biogenesis prqteinPiJW"""j probable chemotaxis protein methyltransferase conserved hypothetical protein 50S ribosomal protein *" probable oxidoreductase _ j probable permease of ABC taurine transporter __ conserved hypothetical protein hypothetical protein _ hypothetical protein ; phosphatidate cytidylyltrans_ferase bhosphatidyiserine synthase""--.---.--j _. _ h pothetical protem . _. ____. _. i thiosulfate suifurtransferase hypothetical protein conserved hypothetical protein protein protein hYpothetical proteW---_.. _. _. ____.. _.... __ ____.. _... _ .. _. j hypothetica) protein P ! ? ! 'P3i''egu) atqr"*-------. . _ robable".. _.. mese of ABC p pe a tr n porter : conserved hypothetical _ ___ sprobable transcriptional regulator_. .. _. _. ___. , _v.... . v_. _.. _w... _. Wprobable transcriptionai reguiator i hypothetical protein. ___.. _. . ___. _ Lqypo ! e al tein 5, tydrodipicolinate reductase j dihydrodipicoiinate reductase'"""""""""'""" ) hypothetical erotein ______ _ lhypothetical protein ; tryptophan synthas_e a_Ipha chain __-T-- __. _... _. _. __ _. _.. _. _. ______. __. ______. _. _. _.. ___. ____.. . __ 3tryptophan synthase alpha chain probable ATP-bindinq component of ABC transporter çprotein hypothetical protein_ probable permease of ABC-2 transporter iprobable transcriptional regulator conserved hypothetical protein acylglyceryl transferase | 3-methyl-2-oxobutanoate hydroxymethy, transferase glutamate racemase igiutamate racemase probable hypothetical protein probable permease of ABC transporter _ probabieperrnease of ABC transporter probable enoyl-CoA hydratase/isomerase probable transcriptional regulator conserved hypothetical protein thymidylate synthase hypothetica) protein'j l, hypothetical protein probable enoyl-CoA hydratase/isomerase translocation protein in type III secretion M _ hypothetical protein probable pili assembly chaperone robable ili ass _, lprobable transcriptional regulator probable plasmid partitioning protein probable permease of ABC transporter conserYed hypothetica ! protei'""'' conse vo tas t conserved hyppthetica) protein ihypothetical protein hypothetical protein probable transcriptional regulator ! probable permease of ABC-2 transporter tprobable NAD (P) H dehydrogenase lconservedpothetical ihypotheticat protein j fUDPacetytgiucpsam"J . ife_rredoxin--NAD_P+ reductas_e __ _ conserved hypothetical protein fflo. heI z j probabie enoyi-CoA hydratase/isomerase""' lconserved hypothetical protein Iferredoxin--NADP+ reductase potyamine transport protein PotC ubiquinone biosynthesis methyttransferase UbiE--- _.-- h. ypothetical protein __ __ v... _. ,.... .. _.. ____. M__. _. _ _r... _ llubiquinone biosynthesis methyitra_sfe ase UbiE _ ____ __ ___<3 tXyÇp otein _ __ transcriptional regulator PrtR_ ! transcriptional r ator PRLR 0 ein j probable transcriptiona ! regulator''"""------ p na eg ator __ , conserved hypothetical protein kprobable exopolysaccharide transporter_-. -'"- Y ". .-. _.. _.. _ _ PI°J '"9--29TP°I ! f' °f. BC transporter' jhypothetical protein probable short-chain dehydrogenase conserved hypothetical protein [3-deoxymanno-octu) osonate cytidytyttransferase"'"" jprobabfe transporter H. JI-1"""""" conserved hypo__ p prothetical rotein _ probable thioesterase" , hypothetical rotein hypotheticat protein ; hypothetical protein cis-1, 2-dihydroxycyclohexa-3, 4-diene carbox late deh dro enase p robab ! e enoyt-CoA hyd ratase/iso m erase"-.-.....--j molybdopterin biosynthesis MoeB protein jprobabfe short-chain dehydrpg'j conserved hypothetica) protein" heme exporter protein CcmC hypothetical protein tRNA (guanine-N1)-methyltransferase yw, 3 type 4 fimbrial biogenesis protein Pilaf imidazoleglycerol-phosphate synthase, cyclase subunit conserved hypothetical protein triosephosphate isomerase j uroporphyrinogen-))) synthetase i i undeca renyrophosphanthetase hypotheticat protein j ) probab) e cobaiamin biosynthetic protein _ _ YP P.. _ hypothetical protein hypothetical protein probable short-chain dehydrogenase" h pothetical protein électron transfer flavoprotein beta-subunit conserved hypothetica ! protein hypothetical protein _ hypothetical protein hypothetical protein hvpothetical protein hypothetical protein 3^oxoacyl- [acyl-carrier-protein] reductase,. _. __.. _.. ___.. _. ___...... _.. 3 hypothetical proteW __ _ hypothetical protein hypothetical protein i DNA polymerase III, epsilon chain ' ! DNA polymerase III, epsilon chain , h othetcal roteW ! hypothetica) protein, probab) ernjdeoseosphoryfase j 'uridylate kinase _ _ __ __ j hypothetica ! p rote ! n"-.-...-.-- , h pothetical rotein ..... i hypothetica ! prptejn''"'""""""""""'" j hypothetical protein---.--......--......... : hypo_theticai pro_tein kinase hell conserved hypothet_p '- __.. _.. _ r. ^ Y.'______.. ____. _. __. . _.. _v. _..... .. _. _. _. ical roten ! conserved h pothetical protein hypothetical protein _ypothetical protein _ _ _ _ ____ _ _ _ m __. ____ _. *. _. ~. _ _<_w~ _ ~ m_ n _ _ _ _-- i transcriptional regulator RHIR £prob_ab_le p_hosphoribosyl transfer_a_se_ __ _ 'fprobab ! e ATPndingcomppnentpf ABC transporter'"-------- Shypothetical protein rconserve^ d hyipotheticai protem w. -'. _.... _.. ___ _... _. _ _ _.. _......... M. . ^. ^T-m V. _. _... . __,. _. _ ____. ? _ypothetical Protein LIansCLD 0 K h R ^ ^ iconserved hypothetical protein prpbabiebotin biosynthesis protein bioH probabje permease of ABC transporter' hypotheticat protein 7conserved hypothetical protein ribonudeasePH,"" probabfe gfutamine amidotransferase-..-..--.-... Nhypothetical protein probable b » <_teín bioH = ! conserved hypothetical protein (robabie transcri tional regulator . . V. __-_.... __.,.. . ^. .. _. _ ___ f bdb e g utam ne ar amrsferase _ _ __ __ _ _______< j DNA-specific endonuc ! ease t *'"*"""" E obable transcriW conserved hypothetical protein (hypothetical prote_in. V. ___.. __ _ . _.. __. _. _.. __.. _. _ ___. ___.. _____.... ______. _. ; hypothetical protein thypothetical protein probable pili assembly chaperone , probable two-component response regulator ! phosphonbosy) aminoimidazoie-succinocarboxamide synthase""" h pothet_ical isuccinate hypothetical protein hypothetical protein conserYed hyp'*"""*"*' ! hyppthetjcat protein conserved hypothetical protein"' conserved hypothetical protein ypothetical protein su serveN e : Ch theme exporter component of ABC fan-sporter heme exporter protein CcmA hypothetical protein hypothetical protein hypothetical protein t5Y _. __. _ _. w_ _ ___. _. _,. v, + We__ {~ Xo R w v----___s__E^___ __________v_e lprobable hydrolase -.----___. __ V nsne-9 w methyltrans se _ _ __ __ = , orotidine 5'-phosphate decarboxylase '" ! probable hydrolase hypothetical protein hypotheticaj protein'----. ; 50S ribosomai protein L1 __ __o _ = w"~. _ M..,,,, «,,, _, w, _~~~_o_~__ _ 'hypothetical protein , probable permease of ABC transporter hypothetica) protein'-.--.--............................................. ......................... protein : hypothetical prole-i : ToiQ-protein __..... .. w_. _. w-__, _. .... _. ____, _w.. _-.. _-__. ___.... _____ __________. _. : TdQ pjrotein"'""""""""'' YP P _ _ _ _ hypothetical protein. p _'"""wm-.. _. _. .. __ . . __w. _ Y.. ,. . __. ___... _. ____ _.. _ conserved hypothetical protein d late s nthase c tid late knase _ '... . _... _.o__ _..a... _ 'hypothetica ! protein----- jprobabie pseudouridyfate synthase'"' jcytidyfatekinase '''"''" ihypothetica ! protein""'" : [. 9°ni6° ! UE° [ ! -. '"'i SOSribosomai protein S3......'."*""i hypothetical protein jprobabie transcriptionat regutator" dethiobiotin synthase'"'" molybdenum transport protein Mod_B y ' transcriptional regulator Dnr hypothetical protein fprobabietp-bindin probable ATP-binding component of ABC transporter conserved hypothetica) protein'' NADH dehydrogenase) chain B probabie two-component response regujator P P P 9 hypothetical protein proba_ble transcriptional regulator Na+-translocating NADH : uniquinone oxidoreductase subunit Nqr4 Iribonuclease T ''. ' hypothetical protein conserved hypothetical protein DNA repair protein Rads ribulose-phosphate 3-epimerase hypothetical protein. lhypothetical protein ; heme expg j ihYpothetical protein urease accesso p (ribose 5-hos ha e isoemerase P P _... _ phosphoribosylaminoimidazole synthetase probable transcriptional regu_lato_r probabtetranscriptionaf regulator'""i probable transcriptional regulator hypotheticai protein hypothetical protein iconserved hypo probabfe transcriptiona) reguiator"'" lprobable two-component response regulator 'conserved hypothetica) proteinIIIL---"I'''"'"" , conserved hypothetical protein hypothetical protein -, hypothetica) protein"* i hypothetical protein ... _____w. _. ______w ____.l__. _____ yp cal p e ypothet ! caiproteHT"''""-"""-- conserved hypothetical. protein S conserved hypothetica) protein" f_.-.. _. _.... _. ___ : thi permease of ABC transporte _ _ ; atginate o-acetyitransferase A) gF'' conserved hypothetica) protein""""'"''" ! conserved hypothetical protein probable transcriptional regulator ! pyridoxine 5'-phosphate oxidase Yp p h othetical rotein h othetical rotem probable carboxylesterase pyoverdine biosynthesis protin Pvc-D probable transcriptional ; robabve carbonic anhydraseW ; conserved hypgtheticafpr conserved hypothetical protein probable pyridoxamine 5'phosphate oxidase----------------------- probable glutathione S-transferase, hypo hetical protein- h pothetical protein probable glutathione S-transferase transcriptionat regutator Vfr------- s transcriptional regulator Vfr giutamineami dptra n sfe ra s e"" ! .......... b pothetical protein maleylacetoacetate isomerase probabfe radicaf activating enzyme Ih) Lpothetical erot (Lin uracil phosphoribosyltransferase hypothetical protein endonuclease III . probable trantcn tional re u a. hypothetical protein conserved hypothetical protein hypothetical protein probable tolQ-type transport protein pseudouridine synthase R) uA'' conserved hypothetical protein 50S ribosoma_ .rotein _. conserved hypothetical protein_ hypothetical protein _ hypothetical protein probable two-component response regulator thyMIq conserved hypothetica) ? 1'*. II""""""III] Econserved hypotheticalprotein ' L ^' If , conserved hypothetical protein Iprobabie aromatic acid decat-boxylase --'""''. _"w"'-=.. _.. _.. ... _... ___. ru ! probabfe nuctease . ."""'"" iperiplasmic chaperone LOIA 'ihypothetical protein L__. _____ iprobable oxidoreductase jhypothetical protein ; hypothetical protein hypothetical protein fhypothetica ! protein '"'""""''""" ! hypothetica) protein"""" [hypotheticat protejr) ! j30S ribosomai protein S4 *''"' homoserine hypothetical protein "robable li o rotem localization rotem LoiB P P P P hypothetica t protein"""'"'j GTP c cloh drolase II hypothetical protein probable transcriptional regulator _ probable peptide chain release factor probable ribosomal protein L25 hypothetical protein hypothetical protein hypothetical protein hypothetical protein conserved hypothetical protem guanylate kinase glutamine amidotransferase conserved hvpothetical protein hypothetical protein protocatechuate 3, 4-diox genase, alpha subunit hypothetical protein probable Clp-family ATP-dependent protease _ _ lhypothetical protein hypothetica) protein 503 ribosomai protein L4' conserved hypothetical protein , hypothetical protein hypothetical protein hypothetical protein hypptheticai protei n .) probable malybdopterin-, guanine dinucleotide biosynttesi ! prgtei MobA hypotheticai protein _''-"" hypothetical protein , conserved h othetical proten _. _.... _ conserved hypothetical protein ihypothetical protein I hypothetical protein iprobable hosphoheptose isomerase E ; p type I e etion sys#em protein ; hypothetical protein.,............... ...""' ; hypothetical protein _ __. _ ____. _. _.. _.... _ Fin hypothetical protein i'hypothetical protein type 4 fimbrial biogenesis "YJ2ZEi9'r-. ' antStgmafatqrjuc "' prpbabietranscriptionat regu ! ator''"'""" ; , I hypothetical protein conserved hypotheticai protem" conserved h ypothetical rotein hypothetical protein hypotheticat protein superoxide dismutase, hypothetica) protein ! hypothetical protein' hypothetical protein hypotheticai protein''hypotheticai protein probabie DNA invertase. . . i probable DNA invertase probable. acetyltransferase _. _. _. hypothetical protein hypothetical protein hypothetical protein conserved hypothetica) protem J hypothetical protein conserved hypothetical protein ri osomal protein alanine acetyltransferase ; probable transcriptional regulator jhpothetical protein------... _...... onserve. j conserve hypotheticai protein j conserved hypothetical protein conserved hypothetical protein ; hypothetical protein. _. __ [conserved hypothetical protein probabie protease robable protease g_. x... ____. _.. _-. . tt __.. __. ___... _... _.. _ aikyi hydrpperpxide reductase subu ! probable protease fhYpothetical protein . _... ". __. _. .. _ hypotheticai protein'"" Lhothetical protein hypothetical protein P -probable transcriptional regulator , heat shock rotein Gr E.. _ CDP-diacylgl cerol--gcerol-3-phosph, ate 3-hosphatidyltransferase, __4W iCDP-diacyJgtycero1'IIIIIT conserved hypotheticai prptein nserved hypothetical protein ribosomerecyding factor ome ecyc ng facto i hypothetical protein"'""''""""""'"' : hypothetical protein : __.. ; hypothetical protein. ___.. ... _. _.. . _.... . . _. _..... _. _. -. .... . _.,. ,..... _. _...... . _, _... ...... _,... __... __. ___.,.. _. _. . ; h pothetical protein : ribosome recyclinq factor ! conserved hypothetical protein ! _hypothetical protein rra I t o mrtration factor iF 3 _ _ ein itranslation initiation factor IF-3 rconseryedjiypothettcapro"'""' , probable transcriptional regulator {conserved hypothetica ! protein *---- iadenine phosphonbosyttransferase"''"--- conserved hypothetica) protein"'"'* dTDP-4-dehydrorhamnose 3, 5-epimej'ase""'' tprobable_si ma 70 factor, ECF subfamii probabte sigma-70 factprE * ! GTP cyclohydrolase I precursor 2othetical protein . qu 50S ribosomal protein L5 hypothetical protein conserved hypothetical protein conserved hypothetical protein ATP synthase delta chais NosL protein conserved hypothetical protein conserved hypotheticai protein. j conserved hypothetica_I protein ; 50S ribosomap _rotein L6 _. conserved hypothetrcal rotem, : conserved hypothet) ca) prptejn conseryed "" ! conserved hypothetical protein hypptheticat prptetn hypptheticaf protein""" conserved hypothetica) protein' jtactoyigtutathione tyase J hypothetical protein r Ypothe_. __... _. h othetrcal protem -----..... ____. _.. _. _. __. _. _. ___. __. _ ____. ___. inorganic pyrophosphatase ! probable sigma-70 factor, ECF_subfamily probable sigma-70 factor, ECF subfamily___, conserved h pothetical protein - othetical conserved hyp p hypothetica) prptetn-----j roteW^M hypothetica ! protein l hypothetrcal protem __ h pothetical protein _., __. _ hyothetical protein _.----__.. _.... _. ____. __. ____ __. _. ___. _,-. _. _ ____.. __. _. _ r_... ___.. _, _.... _.. 'genera ! secretion pathway protein M.-..-..-.....-...-.......-...-............-.. hypothetica) prptein"""" P __ w__, _ ihypothetical e ; conserved hypothetica ! protein"'..---.-.............. ,.. _____.. _... .......... __... __. _. _. _..... _. _. _.. _..... _.. __.... __...... _....... __...... _.. _. _..... _....... _.... _. __.......... _........ _..... _...... _. __..... __..... _. _... _... _. _.. _. _. _....... __... __... _. _. __... _....... __.. _..-..... _. _... __..... _..... ; conserved hypothetical protein _ _ ; conserved hypothetical protein heat shock protein HscB conserve_d_hypothetical protein ' -'"'". hypothetical protein' genera ! secretion pathway prpteinH"'" !. __. ... r__.. _ shikimatekinase *"-.--, h pothetical protein _ : conserved h othetical rotein phosphatidylglycerophosphatase A beta-hydroxydecanoyl-ACP dehydrase conserved hypothetical protein transcriptional re conserved hypotheticaf protein """""< conserved hypothetical protein spermidine acetyltransferase prolipoprotein signal eptidase hypothetical protein conserved hypothetical protein probable transcriptional regulator _ _ - conserved hypothetical protein probable sima-70 factor, ECF subfamy"j conserved hypothetical protein hypothetical protein hypothetical protein probable outer membrane protein precursor probabie outer membrane protein hypothetical protein transcription elo, g dih drofolate reductase hypothetical protein hypothetical protein 50S nbosomal protein L_10 30S ribosomal protein S5 hypothetical protein hypothetical protein hypothetical protein ADH dehydrogenase I chain J hypothetica) protein hypothetical protein hypotheticat protein"''"''""""'"""""'"""""'"""""j lhypothetical protein hypothetical protein thypothetical protein hypothetical protein conserved h p j^ ypothetical rotein _ __ _ ferredoxm protein NapF _. _........ , ... ...., .. .. _. W _ p phenazine bios nthesis proten _ ' : probabie phenazine bipsynt-..-..-*""'-- : probable phenazine biosynthesis protein ---..-.-..-...-.-.....--.-..-......... ° YP p i_.. __ ; hypothetical proten _ _ j probable phenazine biosynthesis protein 'conserved hypothetical protein 'tofuate 1, 2-dioxygenase beta subunit"' 2-amino-4-hydroxy-6-hydrox methyldihydropteridine p rophos hokinase hypotheticat protein""'"""' _.. _. _. Y.... Y P lhypothetical protein lpeptidyi-prolyl cis-trans iso mrase hypothetica ! protein'" h pothetical protein , hypothetical protein hypothetical protein molybdopterYpbiosynthetic protein C conserved hypothetical protein hypothetical protein conserved hypothetical protein hypothetical protein F NusB protein _______ _ phosphopantetheine adenylyltransferase hypothetica) protein : phosphopantetheine adenylyltransferase hypothetical protein hypothetical protein_ hypothetical protein conserved hypothetical protein conserved hypothetical protein hypothetical protein roba mdi jprobabte purine-binding chemotaxis protein hypothetical protein hypothetical protein jjiypothettcajproteLJL-1111111111111111"] 6, 7-dimethyl-8-ribityllumazine synthase.""_ hypotheticai protein. ."""'"---'-----...---j co"nserved h othetical protein translocation protein in type III secretion M, _ w translocation protein in type III secretion hypothetical protein hypothetical protein lhypothetical protein YP'_p Jconserved hypotheticai protein"""j conserved hypothetical protein bacteno {bacterioferrittncomigratory protein''"'} conserved hypothetical protein hypothetical protein . Yp ical rotein conserved hypothet p sconserved h pothetical protem yhase 8 h i , 30S ribosoml_grgt ir S cyanate Jyase""""""''""" biotin carboxyl carrier protein (BCCP) probable dna-binding stress protein . conserved hypothetical protein - ghypothetical protein . h othetical rotein _ _ _ ihypothetical-protein' ,, cytochrome C-pe biiogenesis protein CcmH hypothetical protein hypothetica ! protein ! lhypothetical protein t lprobable thioredoxin hypothetical protein _ _-- ; nitrogen regulatory IIA protein conserved hypothetical protein phosphotyrosine protein phosphatase probable HIT famil protein conserved hypothetical protein lhypothetical protein hypothetical protein ___, conserved hypothetical protein conserved hypothetical protein _ _ _ _'--" conserved hypothetical protein hypothetical protem hypothetical proteW_. _ . . conserved hypothetical protein conserved hypothetica) protein j psmoticaj) yir) ducib) e protein smC' deox uridine 5'-triphosphate conserved hypothetical protein hypothetical protein hypothetical protein moiybdopterir) converting facto '"''"' conserved hypothetical protein conserved hypothetical protein_ conserved hypothetical protein_ probable type)) secretion system protein jtype 4 fimbria) precursor PijA ! lprobable transcript_ional regulator __ "' hypothetica) protein J II. I 11'"""'".,"'"i hypothetical protein 3-dehydroquinate dehydratase ribonuclease H ____ ____... . _. _.. _ hypothetical protein j probabte peptide defprmyiase'j probable peptide deformylase ; ToIR pfo etem .. _.. __. _ _.... _. _ _. __,.. __.... _ _.. , w -_. __.. __... _. _. __... _. __ __. _. __. _. ____. ___. : conserved hypothetical protein _ ; ToiR protein"''""""""" iconserved hypotheticai protein"""'"""""' ' (3R)-hydroxymyristqy [M protein] dehydratase""""" ; _hrpothetical protein ihyppthetica) protein" conserved hypothetical protein i conserved hypotheticai protein---- ' : probabtetranscripM. "-111''' 9 f {hypothettcai protein" I_exoenzyme S synthesis protein C precursor iprobable transcriptional regulator _. hypothetical protein hypothetical protein -50S ribosomal protein Let hypothetical protein conserved hypothetical protein hypotheticai protein hypothetical protein conserved hypothetical protein hypotheticat protein""" ! probable type 11 secretion system protein helix destabilizing_ protein of bacteriopag, PL hypothetical protein nucleoside diphosphate kinase 50S ribosomal protein Lu I hypothetical protein hypotheticaf protein-.--..- 50S ribosomal rotem L13 n , hypothetical protein hypothetical protein conserved hypothetical protein probabfe transcriptional reguiator _ hypothetical protein probable. tT e ! I secrefion system __... _... ___. _.. _ hypothetical protein ATP synthase epsilon chain hypothetical protein conserved hypothetical proten ; hypothetical protein hypothetical protein conserved hypothetical protein conserved hypothetical protein conserved hypothetical protein conserved hypothetical protein hypothetical protein J i 30S ribosomal protein S6 hypothetical protein , hypothetical protein jconsejyedjhypotheticap""'""f _erved hypothetical protem -o-tei-n h_.. othetical ppotem, p.. ! hypothetical protein hypothetical protein 'hypotheticat protein.---.-..-......-....-..-..,-...-. jhypothecaj protein""" f"hYpothetical protein. .. _. _. __. __.. __. _. w. j hypotheticai protein"" conserved hypothetical protein h othetical protein ; p' p wy o p ihypotheticai protein ! hypothetical protein hypothet cal protein '"''-w° . ihypothetical protein hyhypothetical protein _. V... _. ___.. ___... ___ »,.... _ ribonucjeasep protein component.' stringent starvation protein B probable fosfom cin resistance protein hypothetical hypothetical rotem hypothetical protein hypothetical protein ferric uptake regulation protein hypothetical protein hypothetical protein hypothetical protein probable transcription l reLulator jprobabte transcriptiona) reguiator probabteNADH-ubiquinone/p ! ) hypothetica) protein) nserved hypothetica) protein""" ! hypothetical rotem hypothetirotein ; E hypothetica) protein"'' proba bfe transcription at regulator yp theti p .. _....... _......... __....... _..... _.......... _..... _......... _. _. _... _. _. _,.. _. _......... _..... _..... _... _... _. _....... __... _....... _. __.. _. _. __.. ____.. _.. _.. _... _____.. ____. _. l aikaiine proteinase inhibitqr Aprt iactoyigiutathione Jyase . . jhypotheticai protein... I-.. l. . .. 11""' '""''"'''--'-"'''-"'''"-j h o'cal rotem hypothetical protein lalka. line proteinase inhibitor Ap-rl lacto I lutathione lyase hypothetical prote _ !, hypothetical protein hypothetical protein , conserved hypothetical protein hypotheticat protein j hypothetical protein 130S ribosomal protein S8 t ; hpothet p ; ical rotein . N ! 5-carboxymethyt-2-hydroxym) jcqftejsornerase'"" : -30S ribosomal protein Su 'hypothetical protein _ __ __ __ __ _ _ 'hypotheticai protem_ ! hypothetical protein t 30S ribosoma p_tein S 1 pr bable cytochrome c (mono-heme type) 30S ribosomal protein Sl l hl2_ _-_-_ ___ _ « _ _ _ __ _ _ _ __ _ _ _ __< j_yEhe ribosimal rotei lsecretion protein SecG 'hypothetical p, rotein hypothetical hypothetica ! protein'"''"" probabiejron-binding protem iscU'"""") IhEot rot in.............................................. _ _ Lypothetical protein ___ al protein conserved hypothetical, protein conserved hypothetical protein glycine cleavage system protein H2 ___ _ __ __ _ _ _ _ypothetical protein _ _ __ _ YP __ P - my hypothetical protein hypothetical protein hypotheticat protein--- YP P hypothetical protein _ __ _ _ t hypothetical prote, in.. aspartate 1-decarboxylase precursor hypothetical protein .. _ f hypothetical protein hypothetical. protein....... pr bable rinq-cleaving xygenase w... ___ __ _s hypothetical protein ATP synthase protein I conserved hypothetical protein conserved hypothetical protein hypothetical protein,,. conserved hypothetical protein conserved hypotheticai protein"*'"'" hypothetical protein, YP P __, _ _.--- p hypothetical protein, hypothetical protein hypothetical protein hypothetical protein conserved hypothetica) protein probabie type)) secretipn system protein j 1 consenred hypothetical protein _ . , _"" .. ", _, _....- , probable type II secretion system protenw .. . ",. . , ___Mw", "_, yothetical protein- _ _ : two-component response regulator CheY _ __ _ _ _ _ _ ß hypothetical protein _ _ _ hypothetical protein transcriptional regulator MvaT, P16 subunit d-ervthro-7, 8-dihvdroneopterin triphosphate epimeras_ _ _ _ __ _ __ 30S ribosomal protein Sl. 2 in :. _.. _. _. _ _ pr__' Yt r hY. potheticalp ____. _.. ___........ ___. _.... ___. _.. _. __... _.... _.... _. _..,.. _. _ ____ __.. ________. . ____. _. _. ! probable drug efflux trans orter _50S ribosomal prot_ein L14 hypothetica } hypotheticai protein'" o ribosomal protein L7 Ll 2 s secretion protein Secte ! succinate dehydeo e inate dehydrogenase (D, hypothetical protein., hyppthetica ! proteinL-. J. IL.'' conserved hypothetical protein in type III secretion hy_pothetical. proten__ hypothetical protein hypothetical. protein hypothetical rotein hypothetical protem YP hypothetical protein hypothetical protein _ a hypothetical protein hypothetical rotem. _ _PP conserved hypothetical protein , _ m _ hypothetical protein. hypothetica ! protein. i hypothetical protein conserved hypothetica) protein *" 50S ribosomal protein L20 probable 6-pyruvoyl tetrahydrobiopterin synthase , , m, ""_', hypothetical protein_^ __^w__ 30S ribosomal protein S13 t_. _ type 4 fimbrial biogenesis protein Pilez pterin-4-alpha-carbinolamine dehydratase hypothetical protein conserved hypothetical rotein- Yp P ihydroneopterin aldolase , conserved hypothetical protein YP P. _ ical rotein conserved hypothetical protein conserved hypothetical rotein conserved,. hypothetical protein 50S ribosomal protein L1_8 hypothetical protein hypothetical hypothetical protein hypothetical.. protein hypothetical protein, _ I [hypothe'p tem __ _ Yp tical rorotein pothetical protein, ____... _....,. .... ".-... .. _. hypothetical protein zu =,hypothetical protein h thetical protein 'conserved hypotheticat protein *'"''" ;, conserved hypothetical protein iconserved hypqthetica) qtein ."''""' ; hypothet ; conserved hypothetical protein. conserved hypothetical protein ; hypothetical protem . h pothetical protein nserved hypothet ! cajprotejn"------ hypotheticat protein hypothetical protein hYpothetical protein _.-. . _. _.. _.... __........ __. _.... __. __.. __. _. _.. __.. _.. _.. ,.. ...., _. _,. _. . : hypothetical protein probable ferredoxin ! nitrogen regulatory protein P-II 2 - _ ferredoxin [2Fe-2S] conserved hypothetical protein conserved h pothetical protein conserved h pothetical protein - conserved_h pothetical protein hypothetical protein _ _ _ __"__ hypothetical protein hypothetical protein h pothetical protein conserved hypothetical protein ___ _'""- phosphoribosyf-ATP pyrophosphohydrotase I""I11 conserved hypo_t_he_tic_al protein conserved hypothetical protein iSMR multidrug efflux transporter ; , hypothetical protein' h othetical protein hypothetical protein '-"' , hypothetical protein (hypothetica) protein t fhypotheticat protein j ; hypothetical protein, iconserved hypothetical protein ,, hypothetical protein h potheticalrotei_n _ "-II''"'4'"-' ) hypothetica) protein ! conserved hypothetical protein ssimiiatory nitrite reductase smat) subunit ! hypothetical protein Jconserved hypothetical protein, '"*'*'] probabte thioredoxin J thioredqxtn. ! ! prqbabie bacteriophage protein t 'conserved hypotheticat protein . . ! s hypotheticai protein j Iconserved hypothetical protein 'conserved hypothetical protein ; £hYpothetical protein [hypothetical protein __ _ __ hypothetical protein .. _.. _..... __ _. _. _... ___, __ . --. -y ^s W. __... -mr W.. __.. ihypothetical protem _, oh hypotheti p ^_. __... _-. _.. _.. _.. . _.... _. _ __ _.. ___. ______. __.. ____... Jprobable iron-bndmg protem IscA. ! hypothetical protein =DNA-binding protein _,.. ., _. __ __.. ___... ___... _.. ___ jhypotheticaj protein----. fDNA-binding protein Fis'"""*"" ., hypothetical protein' conserved h pothetical protein hypothetical protein hypotheticai protein."""" : h othetical protein sarcosine qxidase deita subunit"'""''""---, hypotheticai protein------ hypothetical protein hypothetical protein hypothetical protein h, pothetical protein transcriptional regulator PrtN -OS ribosomalprtein Lq4 hypothetical protein hypothetical protein conserved hypothetical protein hypothetical protein hypothetical protein conserved hypothetic"" 30S ribosomaiprpte) nS1p"------------- probable transcriptional regulator 50S ribosoma_I pro_tem L21 hypothetical protein hypothetical protein NADH dehydrogenase I chain K conserved hypothetical protein conserved h pothetical pr_o_te_in conserved hypothetical protein hypothetical protein ' '""""-'"'_"- conserved hypothetical protein , probable transporter i ! hypothetica) protein""-" hypothetical protein conserved hypothetical protein hypothetical protein ; salicylate biosynthesis protein PchB"""""" , hypothetical protein conserved hypothetical protein -os ribosom conserved hypothetical protein 'morphogene protein BoIA _ _ _ _ _ ; hypothetica ! protein'"'' hypothetical protein ypothetical protem ! hypothetical protein __. _ _.... f Snit_ __ _ ______ __ _ _ _ Iconserved hypoíhetical protein_ rc_al rotem. r50S ribosomal protein thypothetical protein 3 hypothetical protein lhpothetical protein fhypothetica) protein"""'" hypothetical _ . _ein hy hy_p_othetical protein ""°""' hypothetical protein . __g. _. : __y hypothetica) protejn" ! regutator inype) ! i secretion,. . ."'""'"j hypothetical protein 3hypothetical protein ___w_-__ _____o _ _ j3 ; cell division protein FtsL (GroES protein _ _ conserved hypothetical conserved hvDothetical protein conserved hypothetical tconserved hypotheticalerot in hypothetical protein hypothetical protein 3 G ! u-tRNA (Gln) amidotransferase subunit C . hypothetical protein hypothetical protein hypothetica) protein hypothetical protein , _., Y. ...... w. ^..... _.. w. ___..... __. w__.. _... _. _.. _.. __. __. ___. __y_... _ hypothetical protein hypothetical protein conserved hypothetical protein hypthetrcal rtern hypothetical protein hypothetical protein __~_w ww _ E conserved hypothetical protein j hypothetica) protein""J h othetical protein integration host factor beta subunit hypothetical protein conserved hypothetica ! protem' h othettcai roten @t rotein_ __ _ _ .. I hypothetical protein _ _ _ _ _ __ __ hypothetical Drotein probabie DNA binding protein j probable DNA binding protein peptidyl-prolyl cis-trans isomerase C2 hypothetical protein hypothetical protein hypothetical protein _pyrroloquinoli_ne quinone biosy_nthesis protein D e fid I rol I''e C1L hypothetica) protein h othetical rotem = jhypotheticaf protein j h othetical rotern _ jhypotheticaprotein j sS.... _ _............. t Lhypothetical pro ein _ __ __ _ _ _ _ _ ___ ___4 hypothetical protein } 30S ribogttK S20 , 30S ribosomal protein S19 mal rotem S19 ... _.. _____.... _. _. __... ____. __.... _ _ _ __ probable acylphosphatase : hYpothet p hypothetical protein conserved hypothetical proteln yp.. _. __. _... _. _.. __. _ . ____... _... _.. ____ _. _... _ conserved hypothetical protein ; Conserved hypothetical protein ihypothetical proteln . conserved hypothetical protein : conserved hypothetical protein ! _ f W hypothetica-otein ' ; hypothetical protein ".. _. __..... _. _.. __....... _... _.. _... _.. __ _... __.. ___.. _.. _ _ Eflagellar biosynthetic protein FliQ hypothetical protein ; ffagellar biosynthetic protein FiiQ W. ypthetical protein ww wwwwwww____ _.... _....... _. 30S ribosomal conserved hypothetical protein Iconserved hypothetical protein S ribosomal protein S17 ihypotheticai protean ! 'hvDothetical protein v _. __w_wwww_ww_ __ w___ww _ _ s __wwwwmwww_ __* w_ wwww__ l hypothetical protein lhypothetical protein _. conserved hypothetical protein hypothetical protein conserved hypothetical protein pyocin S2 immunity protein pyocin S2 immunity protein___ _ _ _ __ _ _ _ _ _í hypotheticai protein _ _.. __ _. _. _..... ..,., _, _ ~ _,. t..... __. __. _,. .. _ _. _. _. _. _ _ ___ __ « j hypothetical protein hypothetical protein p.. _. _ hypothetical proteln type III export protein PscF atp synthase C chain j hypothetical protein hypothetical protein 50S ribosomal protein L27 hypothetical protein o_ hypothetical protein 50S ribosomal protein L27 hypothetical protein klutaredoxin. hypothetical hypothetical proteln h othetical rotem -----.---.. __. _ ___.. .. .. glutaredoxin thypothetical protein thvpothetical protein 'hypothetical protein tein hypothetical protein cetf division topobgica J motybdopterin converting factor, smaf) subunit t molybdopterin converting factor, small subunit molybdoDterin convertinq factor, small subunit major outer membrane lipoprotein precursor hypothetical protein 4 Yp P _ 30S ribosomal protein ferredoxin [4Fe-4S] , j _. ____. __. _. ___.. _. __.. _. ___. __. _. __.. _. ... _. _. _. __ _ ferredox_m _4Fe 4S conserved hypothetical protein conserved hypothetical protein _} probable biotin-requiring enzyme translocation protein TatA conserved hypothetical protein . .. .. _ . _.. _.. ___. __.. w. ____ _.. _ _. 'hypothetical protein p p. _. _..... _ ; hypothetical, Eein _ __ _, _o : rp cal rot m wY., : __. __ i probable ferredoxin'-'--"-""--- ; _pr_obable ferredoxin _.. ____.,. _.. __, __.... 'nypothetica) protein --.-.-...-......... .......... ihYpothetical protein hypothetical {conserved hypothetica ! protein" regulatory prot_n_saL _ __ _ _ _ __ \ conserved hypothe_tical protein "'"''""'"'w° . conserved hypothetical protein lhypothetical protein probable acyl carrier protein __ ____ _ _ lprobable acyl carrier protein # conserved hypothetical protein probable acyl carrier protein . hypothetical protein conserved hypothetical protein hypothetical protein _ 60S nbosoma ! protein L28ZIIII'*'""J 50S ribosomal protein L28 acyl carrier protein hypothetical rotein hypothet ! ca) protein' hyDpjthecai protein j conserved hypothetical protein hypothetical protein hypothetical protein 30S ribosomal protein S18 hypotheticat protein l. i.-J -.-.-Lj hypothetical protein hypothetical protein hypothetical protein yap p hypothetical protein hypothetica ! protein t hypothetica) protein) conserved hypothetica ! protein' hypothetical protein ! conserved_hypothetical protein hypothetical protein , conserved hypothetical protein , conserved hypothetical protein ypothetical protein) h"ypothetical protein hypothetical protein conserved hypothetica) protein'"'' hypotheticat protein hypothetical protein , conserved hypothetical protein wshe p initiation factor Yp'p h othetical rotem . __ _ p.. __, hypothetical protein ihypothetical protein _ hypothetical protein p __.. _. _. __. _ _ _ __ _.. 30S ribosomal protein S21 '! tofbacteriop age Pf1 _ _ _ _ _ __ _ r bnssme o1 : acbr ribosome modulation factor_ W ribosome modulation factor .......... protein O hypothetical protein i hypothetica) prgtein . "" ! hypothetical protein conserved hypothetical protein 3hypothetical protein 'probable cold-shock protein j hypotheticat protein."*"" 3, hypothetical protein 3hypothetical protein ih othetical protein ypothetical protein hypothetical protein probable tra_ns_criptionai regulator probable transcriptional regulator cold acclimation protein B rt protein PscE conserved hypothetical protein'' wgulator probable transcriptional regulator leonserved hypothetical protein hypothetical protein conserved hypothetical protein hypothetical protein conserved hypothetical protein hypothetical protein hypothetical protein hwgMgn_ hvDothetical Drotein hypothetical protein hypothetical protein hypotheticai protein : 50S ribosomal protein L35 hypothetical proteln 50S ribosomal protein L29 hypothetical protein carbon storage regulator hypothetical Protein hypothetical protein h othetical Iprotein __" conserved hypothetical protein hvpothetical protein wiz hypothetical s hypothetical protein hypothetical protein hypotheticai protein hypothetical protein hvpothetical protein hypothetical protein hypothetical protein 50S nbosomal protein L30 50S ribosomal protein L30 hypothetical protein hypothetical protein hypothetical protein hypothetical protein periplasmic nitrate reductase 3 rubredoxin f-conserved hypothetical hypotheticai protein . , h othet 'hypothetical protein i conserved hypotheticaj protein..,.,...,..,.........,. hypotheticai protein iconserved h othetical rotein _., . _. __. _. _. __ P ___. _. __p _.. _. ____.. _. __... __...... _.. __..... _. _.. _. __... __. _.. _........ _... _... _......... __ qat hypothetical. ,, conserved hypothetical protein : hypothetical protein----------- hypothetica, protein................... ih othetical rotein _, _.. __ jiipopeptide precursor S hypothetical protein_, _____- 50S ribosomal protein L34 hypothetical. protein......... hypotheticai protein 50S ribosomal protein L36 k KdpF protein pyrrobquirolnj . i Y-& 'F,'4... f Y S, Vr fV'1 h : 'M ta tylar'e g rltserr, i, r, . kPnt, "ho'tt !'i ., 001 A01 1 3254036 ; PAK 001A03 1 42145261 ; PAK 001A04 800208ji PAK'. OOlAO4 PAK 001A05 3627210. ; PAK 001A06 | 4931913j PAK 001A07. 1034469 ; i PAK 001A08 ij 6144940 PAK 001AO9 ì 4102307í ; PAK 001A11 1515737ji PAK 1244688 l l 001B02 5924618 001B03 3267350 PAK 001B04 3302119"PAK" 001 B05 3108121 PAK _ _j 001 B06 j 3987545 PAK 001 B07 5585518 PAK ; i e 6 001B09 4682141 PAK 001B10"5041902 PAK _.... _. __ 001B11 3187871 PAK ! 001 C01 5924618 PAK 001 C02 2871514 PAK 001C03 001 C03 4746191 PAK 001 C04 515693 PAK r 001C04 5156931 PAK Si 001 C05 61557441 PAK 001C06 _ 5741543j PAK 001C08 5945561 PAK i 001 CO9 23707631 PAK 001 C10 4186878 PAK . 001 C12'5610441) PAK t PAK s 001D01 6193619 PAK 001 D03 60780341 PAK 001 D04 61398781 PAK _ 1 5225311 PAK ) 001 D06 i 3567982 PAK j 001 D08 3998726ij PAK 001DO9 21402511 PAK 13011691 PAK OOlDlO i ' 001D11 34235191 PAK 6701 001D12 3472 PAK 8009361 PAK I f"001E03'4246525""PAK1 o w i j 001 E04477959 PAK ; 001 E05 li 4779591 PAK ! 001E06 5939777 PAK ; 001 E07 5471284 PAK __ !'001 E08"5582311 PAK 001 EO9 1 442285i PAK ; i 001E10 ; 1 45148791 PAK r 001 E11'4514879 ! PAK'"' i001 E12 5064081T PAK J i. _. __.-. 001 F01.-.. _______. _... _.. _... _.. 903376 ! pat 001 F02-'PAK 001 F02 i 18895811 PAK i ; 001 F03 i 123081 3i PAK '''''Lf J1230813 !"PAK'" : 001F04 j 5755640 ! PAK' ) 001 F05" !'" 5355403)""PAK" 001 F06 ; ; 4493238i PAK ; 001F07'850016 _ PAK_ j'j 001F08'j 5286479ii PAK 001 FO9 3035834 PAK 001 F10 isi 270414 PAK 001F11 4300290 PAK 00IF12 3471740 PAK 001 G01 5431778 PAK 001 G06 5064081 PAK j 001 G07 353196 PAK 001 G08, 4881356 PAK 001 G094684466 PAK i 001G10 1340542 PAK -e 001G11 2659761 PAK 001 G 12 4553600 PAK _ ; 001 G12 4553600 PAK ji 001G01 5431778 PAK 001 G02 627197 PAK i 001G03 30056161 PAK 001GO5 2300131 PAK 5064081 001 G07 353195 PAK m a 001G08 48813561 PAK 'j 001 G08 4881356 PAK 001GO9 4684465 PAK 001 H01 ii 4600925jj PAK 001H02 4465691 PAK 001 H03435295 PAK 001 H04 i 558396T PAK 001 H06 5823734 PAK 001 H08 3880466 PAK 001 H09 271720 PAK j 001H10 433381 PAK 001H11 395319 PAK 002A01 3652524 PAK 002A02 3987593 PAK I 002A03 530429 PAK g 002A04 6232877 PAK 002A05 1543845 PAK 002A06 619210i PAK 002A07 4795351 PAK 002AO9 1152202 PAK 002A09 1 11522021 PAK 002A11 449470i PAK 002B01 5643797 PAK 002B02 3423910 PAK" 002B03 1 36169161 PAK 002B04 1511758 PAK 002B05 3809953 PAK ! 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PAK j Medline Ul Gene Organism PA_ID 94049123 metK Escherichia coli 546 97075927 rpoD Rhodobacter capsulatus 576 98414051 ygjD Escherichia coli 580 90330537 surA Escherichia coli 594 88262245 era Escherichia coli 771 97295239 era Salmonella typhimurium 771 97113525 tolQ Pseudomonas aeruginosa 969 97113525 tolR Pseudomonas aeruginosa 970 97113525 tolA Pseudomonas aeruginosa 971 92355498 dapE Escherichia coli 1162 95095976 flhA Paracoccus denitrificans 1452 98414051 ycfB Escherichia coli 1678 98429489 clpP Caulobacter crescentus 1801 98429489 clpX Caulobacter crescentus 1802 93224448 Ion Myxococcus xanthus 1803 99065127 lolA Escherichia coli 2614 94110226 infA Escherichia coli 2619 98241618 IpxK Escherichia coli 2981 91348525 metZ Escherichia colis,. 3107 92193258 folC Escherichia cöli 3111 96405645 htrB Escherichia coli 3242 95014035 surE Escherichia coli 3625 94240115 frr Escherichia coli 3653 93077430 smbA Escherichia coli 3654 96228708 ispA Shigella flexneri 4043 88163790 tufA Escherichia coli 4265 90264268 rpoB Escherichia coli 4270 96107188 IpxC Escherichia coli 4406 84236117 ftsZ Escherichia coli 4407 84236117 ftsA Escherichia coli 4408 93003529 murG Bacillus subtilis 4412 97361813 ftsW Escherichia coli 4413 99047598 mraY Escherichia coli 4415 99029898 rluD Escherichia coli 4544 99000128 obg Streptomyces coelicolor 4566 97284515 ispB Escherichia coli 4569 93259941 murl Escherichia coli 4662 91311678 infB Escherichia coli 4744 92355498 dnaj Escherichia coli 4760 92355498 dnak Escherichia coli 4761 98414051 yjeQ Escherichia coli 4952 96070892 kdtA Escherichia coli 4988 96405645 msbA Escherichia coli 4997 96347399 trxA Synechocytis 5240 97177775 trxA Rhodobacter sphaeroides 5240 93125123 polA Streptococcus pneumoniae 5493 94012475 glmU Escherichia coli 5552 REFERENCES Hardalo, C. , Edberg, S. 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