Background of the Invention

This invention relates generally to Hepatitis C Virus (HCV), and more particularly, relates to mammalian expression systems capable of generating HCV proteins and uses of these proteins.

Descriptions of Hepatitis diseases causing jaundice and icterus have been known to man since antiquity. Viral hepatitis is now known to include a group of viral agents with distinctive viral organization protein structure and mode of replication, causing hepatitis with different degrees of severity of hepatic damage through different routes of transmission. Acute viral hepatitis is clinically diagnosed by well-defined patient symptoms including jaundice, hepatic tenderness and an elevated level of liver transaminases such as Aspartate Transaminase and Alanine Transaminase. Serologicai assays currently are employed to further distinguish between

Hepatitis-A and Hepatitis-B. Non-A Non-B Hepatitis (NANBH) is a term first used in 1975 that described cases of post-transfusion hepatitis not caused by either Hepatitis A Virus or Hepatitis B Virus. Feinstone et al., New Enαl. J. Med.

292:454-457 (1975). The diagnosis of NANBH has been made primarily by means of exclusion on the basis of serologicai analysis for the presence of Hepatitis

A and Hepatitis B. NANBH is responsible for about 90% of the cases of post- transfusion hepatitis. Hollinger et al. in N. R. Rose et al., eds., Manual of Clinical Immunology. American Society for Microbiology, Washington, D. C, 558-572 ( 1 986) . Attempts to identify the NANBH virus by virtue of genomic similarity to one of the known hepatitis viruses have failed thus far, suggesting that NANBH has a distinctive genomic organization and structure. Fowler et al., J. Med. Virol. 12:205-213 (1983), and Weiner et al., J. Med. Virol. 21 :239-247 (1987). Progress in developing assays to detect antibodies specific for NANBH has been hampered by difficulties encountered in identifying antigens associated with the virus. Wards et al., U. S. Patent No. 4,870,076; Wards et al., Proc. Natl. Acad. Sci. 83:6608-6612 (1986); Ohori et al., J. Med. Virol. 12:161-178 (1983); Bradly et al., Proc. Natl. Acad. Sci. 84:6277-6281 (1987); Akatsuka et al., J\ Med. Virol. 20:43-56 (1986). In May of 1988, a collaborative effort of Chiron Corporation with the

Centers for Disease Control resulted in the identification of a putative NANB agent, Hepatitis C Virus (HCV). M. Houghton et al. cloned and expressed in E. coli a NANB

agent obtained from the infectious plasma of a chimp. Cuo et al., Science 244:359- 361 (1989); Choo et al., Science 244:362-364 (1989). CDNA sequences from HCV were identified which encode antigens that react immunologicaily with antibodies present in a majority of the patients clinically diagnosed with NANBH. Based on the information available and on the molecular structure of HCV, the genetic makeup of the virus consists of single stranded linear RNA (positive strand) of molecular weight approximately 9.5 kb, and possessing one continuous translational open reading frame. J. A. Cuthbert, Amer. J. Med. Sci. 299:346-355 (1990). It is a small enveloped virus resembling the Flaviviruses. Investigators have made attempts to'identify the NANB agent by ultrastructural changes in hepatocytes in infected individuals. H, Gupta, Liver 8:111-115 (1988); D.W. Bradly J. Virol. Methods 10:307-319 (1985). Similar ultrastructural changes in hepatocytes as well as PCR amplified HCV RNA sequences have been detected in NANBH patients as well as in chimps experimentally infected with infectious HCV plasma. T. Shimizu et al., Proc. Natl. Acad. Sci. 87:6441-6444 (1990).

Considerable serologicai evidence has been found to implicate HCV as the etiological agent for post-transfusion NANBH. H. Alter et al., N. Eno. J. Med. 321:1494-1500 (1989); Estaben et a!., The Lancet: Aug. 5:294-296 (1989); C. Van Der Poel et al., The Lancet Aug. 5:297-298 (1989); G. Sbolli, J. Med. Virol. 30:230-232 (1990); M. Makris et al., The Lancet 335:1117-1119 (1990). Although the detection of HCV antibodies eliminates 70 to 80% of NANBH infected blood from the blood supply system, the antibodies apparently are readily detected during the chronic state of the disease, while only 60% of the samples from the acute NANBH stage are HCV antibody positive. H. Alter et al., New Eno. J. Med. 321:1994-1500 (1989). The prolonged interval between exposure to HCV and antibody detection, and the lack of adequate information regarding the profile of immune response to various structural and non-structural proteins raises questions regarding the infectious state of the patient in the latent and antibody negative phase during NANBH infection. Since discovery of the putative HCV etiological agent as discussed supra, investigators have attempted to express the putative HCV proteins in human expression systems and also to isolate the virus. To date, no report has been published in which HCV has been expressed efficiently in mammalian expression systems, and the virus has not been propagated in tissue culture systems. Therefore, there is a need for the development of assay reagents and assay systems to identify acute infection and viremia which may be present, and not currently detected by commercially-available assays. These tools are needed to

help distinguish between acute and persistent, on-going and/or chronic infection from those likely to be resolved, and to define the prognostic course of NANBH infection, in order to develop preventive and/or therapeutic strategies. Also, the expression systems that allow for secretion of these glycosylated antigens would be helpful to purify and manufacture diagnostic and therapeutic reagents.

Summary Of The Invention

This invention provides novel mammalian expression systems that are capable of generating high levels of expressed proteins of HCV. In particular, full- length structural fragments of HCV are expressed as a fusion with the Amyloid

Precursor Protein (APP) or Human Growth Hormone (HGH) secretion signal. These unique expression systems allow for the production of high levels of HCV proteins, contributing to the proper processing, gycolsylation and folding of the viral protein(s) in the system. In particular, the present invention provides the plasmids pHCV-162, pHCV-167, pHCV-168, pHCV-169 and pHCV-170. The

APP-HCV-E2 fusion proteins expressed by mammalian expression vectors pHCV- 162 and pHCV-167 also are included. Further, HGH-HCV-E2 fusion proteins expressed by a mammalian expression vectors pHCV-168, pHCV-169 and pHCV- 170 are provided. The present invention also provides a method for detecting HCV antigen or antibody in a test sample suspected of containg HCV antigen or antibody, wherein the improvement comprises contacting the test sample with a glycosylated HCV antigen produced in a mammalian expression system. Also provided is a method for detecting HCV antigen or antibody in a test sample suspected of containg HCV antigen or antibody, wherein the improvement comprises contacting the test sample with aan antibody produced by using a glycosylated HCV antigen produced in a mammalian expression system. The antibody can be monoclonal or polyclonal.

The present invention further provides a test kit for detecting the presence of HCV antigen or HCV antigen in a test sample suspected of containing said HCV antigen or antibody, comprising a container containing a glycosylated HCV antigen produced in a mammalian expression system. The test kit also can include an antibody produced by using a glycosylated HCV antigen produced in a mammalian expression system. Another test kit provided by the present invention comprises a container containing an antibody produced by using a glycosylated HCV antigen produced in a mammalian expression system. The antibody provided by the test kits can be monoclonal or polyclonal.

Brief Description of the Drawings

Figure . presents a schematic representation of the strategy employed to generate and assemble HCV genomic clones.

Figure 2 presents a schematic representation of the location and amino acid composition of the APP-HCV-E2 fusion proteins expressed by the mammalian expression vectors pHCV-162 and pHCV-167.

Figure 3 presents a schematic representation of the mammalian expression vector pRC/CMV.

Figure 4 presents the RIPA results obtained for the APP-HCV-E2 fusion protein expressed by pHCV-162 in HEK-293 cells using HCV antibody positive human sera.

Figure 5 presents the RIPA results obtained for the APP-HCV-E2 fusion protein expressed by pHCV-162 in HEK-293 cells using rabbit polyclonal sera directed against synthetic peptides. Figure 6 presents the RIPA results obtained for the APP-HCV-E2 fusion protein expressed by pHCV-167 in HEK-293 cells using HCV antibody positive human sera.

Figure 7 presents the Endoglycosidase-H digestion of the immunoprecipitated APP-HCV-E2 fusion proteins expressed by pHCV-162 and pHCV-167 in HEK-293 cells.

Figure 8 presents the RIPA results obtained when American HCV antibody positive sera were screened against the APP-HCV-E2 fusion protein expressed by pHCV-162 in HEK-293 cells.

Figure 9 presents the RIPA results obtained when the sera from Japenese volunteer blood donors were screened against the APP-HCV-E2 fusion protein expressed by pHCV-162 in HEK-293 cells.

Figure 10 presents the RIPA results obtained when the sera from Japanese volunteer blood donors were screened against the APP-HCV-E2 fusion protein expressed by pHCV-162 in HEK-293 ceils. Figure 11 presents a schematic representation of the mammalian expression vector pCDNA-l.

Figure 12 presents a schematic representation of the location and amino acid composition of the HGH-HCV-E1 fusion protein expressed by the mammalian expression vector pHCV-168. Figure 13 presents a schematic representation of the location and amino acid composition of the HGH-HCV-E2 fusion proteins expressed by the mammalian expression vectors pHCV-169 and pHCV-170.

Figure 14 presents the RIPA results obtained when HCV E2 antibody positive sera were screened against the HGH-HCV-E1 fusion protein expressed by pHCV-

168 in HEK-293 cells.

Figure 15 presents the RIPA results obtained when HCV E2 antibody positive sera were screened against the HGH-HCV-E2 fusion proteins expressed by pHCV-

169 and pHCV-170 in HEK-293 cells.

Detailed Description of the Invention

The present invention provides full-length genomic clones useful in a variety of aspects. Such full-length genomic clones can allow culture of the HCV virus which in turn is useful for a variety of purposes. Successful culture of the HCV virus can allow for the development of viral replication inhibitors, viral proteins for diagnostic applications, viral proteins for therapeutics, and specifically structural viral antigens, including, for example, HCV putative envelope, HCV putative E1 and HCV putative E2 fragments.

Cell lines which can be used for viral replication are numerous, and include (but are not limited to), for example, primary hepatocytes, permanent or semi¬ permanent hepatocytes, cultures transfected with transforming viruses or transforming genes. Especially useful cell lines could include, for example, permanent hepatocyte cultures that continuously express any of several heterologous RNA polymerase genes to amplify HCV RNA sequences under the control of these specific RNA polymerase sequences.

Sources of HCV viral sequences encoding structural antigens include putative core, putative E1 and putative E2 fragments. Expression can be performed in both prokaryotic and eukaryotic systems. The expression of HCV proteins in mammalian expression systems allows for glycosylated proteins such as the E1 and E2 proteins, to be produced. These glycosylated proteins have diagnostic utility in a variety of aspects, including, for example, assay systems for screening and prognostic applications. The mammalian expression of HCV viral proteins allows for inhibitor studies including elucidation of specific viral attachment sites or sequences and/or viral receptors on susceptible cell types, for example, liver cells and the like.

The procurement of specific expression clones developed as described herein in mammalian expression systems provides antigens for diagnostic assays which can determine the stage of HCV infection, such as, for example, acute versus on-going or persistent infections, and/or recent infection versus past exposure. These specific expression clones also provide prognostic markers for resolution of disease such as to distinguish resolution of disease from chronic hepatitis caused by HCV. It is

contemplated that earlier seroconversion to glycosylated structural antigens possibly may be detected by using proteins produced in these mammalian expression systems. Antibodies, both monoclonal and polyclonal, also may be produced from the proteins derived from these mammalian expression systems which then in turn may be used for diagnostic, prognostic and therapeutic applications. Also, reagents produced from these novel expression systems described herein may be useful in the characterization and or isolation of other infectious agents.

Proteins produced from these mammalian expression systems, as well as reagents produced from these proteins, can be placed into appropriate container and packaged as test kits for convenience in performing assays. Other aspects of the present invention include a polypeptide comprising an HCV epitope attached to a solid phase and an antibody to an HCV epitope attached to a solid phase. Also included are methods for producing a polypeptide containing an HCV epitope comprising incubating host cells transformed with a mammalian expression vector containing a sequence encoding a polypeptide containing an HCV epitope under conditions which allow expression of the polypeptide, and a polypeptide containing an HCV epitope produced by this method.

The present invention provides assays which utilize the recombinant or synthetic polypeptides provided by the invention, as well as the antibodies described herein in various formats, any of which may employ a signal generating compound in the assay. Assays which do not utilize signal generating compounds to provide a means of detection also are provided. All of the assays described generally detect either antigen or antibody, or both, and include contacting a test sample with at least one reagent provided herein to form at least one antigen/antibody complex and detecting the presence of the complex. These assays are described in detail herein.

Vaccines for treatment of HCV infection comprising an immunogenic peptide obtained from a mammalian expression system containing an HCV epitope, or an inactivated preparation of HCV, or an attenuated preparation of HCV also are included in the present invention. Also included in the present invention is a method for producing antibodies to HCV comprising administering to an individual an isolated immunogenic polypeptide containing an HCV epitope in an amount sufficient to produce an immune response in the inoculated individual.

Also provided by the present invention is a tissue culture grown cell infected with HCV. The term "antibody containing body component"(or test sample) refers to a component of an individual's body which is the source of the antibodies of interest. These components are well known in the art. These samples include biological

samples which can be tested by the methods of the present invention described herein and include human and animal body fluids such as whole blood, serum, plasma, cerebrospinal fluid, urine, lymph fluids, and various external sections of the respiratory, intestinal and genitourinary tracts, tears, saliva, milk, white blood cells, myelomas and the like, biological fluids such as cell culture supernatants, fixed tissue specimens and fixed cell specimens.

After preparing recombinant proteins, as described by the present invention, the recombinant proteins can be used to develop unique assays as described herein to detect either the presence of antigen or antibody to HCV. These compositions also can be used to develop monoclonal and or polyclonal antibodies with a specific recombinant protein which specifically binds to the immunoiogical epitope of HCV which is desired by the routineer. Also, it is contemplated that at least one recombinant protein of the invention can be used to develop vaccines by following methods known in the art. It is contemplated that the reagent employed for the assay can be provided in the form of a kit with one or more containers such as vials or bottles, with each container containing a separate reagent such as a monoclonal antibody, or a cocktail of monoclonal antibodies, or a polypeptide (either recombinant or synthetic) employed in the assay. "Solid phases" ("solid supports") are known to those in the art and include the walls of wells of a reaction tray, test tubes, polystyrene beads, magnetic beads, nitrocellulose strips, membranes, microparticles such as latex particles, and others. The "solid phase" is not critical and can be selected by one skilled in the art. Thus, latex particles, microparticles, magnetic or non-magnetic beads, membranes, plastic tubes, walls of microtiter wells, glass or silicon chips and sheep red blood cells are all suitable examples. Suitable methods for immobilizing peptides on solid phases include ionic, hydrophobic, covalent interactions and the like. A "solid phase", as used herein, refers to any material which is insoluble, or can be made insoluble by a subsequent reaction. The solid phase can be chosen for its intrinsic ability to attract and immobilize the capture reagent. Alternatively, the solid phase can retain an additional receptor which has the ability to attract and immobilize the capture reagent. The additional receptor can include a charged substance that is oppositely charged with respect to the capture reagent itself or to a charged substance conjugated to the capture reagent. As yet another alternative, the receptor molecule can be any specific binding member which is immobilized upon (attached to) the solid phase and which has the ability to immobilize the capture reagent through a specific binding reaction. The receptor molecule enables

the indirect binding of the capture reagent to a solid phase material before the performance of the assay or during the performance of the assay. The solid phase thus can be a plastic, derivatized plastic, magnetic or non-magnetic metal, glass or silicon surface of a test tube, microtiter well, sheet, bead, microparticle, chip, and other configurations known to those of ordinary skill in the art.

It is contemplated and within the scope of the invention that the solid phase also can comprise any suitable porous material with sufficient porosity to allow access by detection antibodies and a suitable surface affinity to bind antigens. Microporous structures are generally preferred, but materials with gel structure in the hydrated state may be used as well. Such useful solid supports include: natural polymeric carbohydrates and their synthetically modified, cross- linked or substituted derivatives, such as agar, agarose, cross-linked alginic acid, substituted and cross-linked guar gums, cellulose esters, especially with nitric acid and carboxylic acids, mixed cellulose esters, and cellulose ethers; natural polymers containing nitrogen, such as proteins and derivatives, including cross- linked or modified gelatins; natural hydrocarbon polymers, such as latex and rubber; synthetic polymers which may be prepared with suitably porous structures, such as vinyl polymers, including polyethylene, polypropylene, polystyrene, polyvinylchloride, polyvinylacetate and its partially hydrolyzed derivatives, polyacrylamides, polymethacrylates, copolymers and terpolymers of the above polycondensates, such as polyesters, poiyamides, and other polymers, such as polyurethanes or polyepoxides; porous inorganic materials such as sulfates or carbonates of alkaline earth metals and magnesium, including barium sulfate, calcium sulfate, calcium carbonate, silicates of alkali and alkaline earth metals, aluminum and magnesium; and aluminum or silicon oxides or hydrates, such as clays, alumina, talc, kaolin, zeolite, silica gel, or glass (these materials may be used as filters with the above polymeric materials); and mixtures or copolymers of the above classes, such as graft copolymers obtained by initializing polymerization of synthetic polymers on a pre-existing natural polymer. All of these materials may be used in suitable shapes, such as films, sheets, or plates, or they may be coated onto or bonded or laminated to appropriate inert carriers, such as paper, glass, plastic films, or fabrics.

The porous structure of nitrocellulose has excellent absorption and adsorption qualities for a wide variety of reagents including monoclonal antibodies. Nylon also possesses similar characteristics and also is suitable. It is contemplated that such porous solid supports described hereinabove are preferably in the form of sheets of thickness from about 0.01 to 0.5 mm, preferably about 0J mm. The pore

size may vary within wide limits, and is preferably from about 0.025 to 15 microns, especially from about 0J5 to 15 microns. The surfaces of such supports may be activated by chemical processes which cause covalent linkage of the antigen or antibody to the support. The irreversible binding of the antigen or antibody is obtained, however, in general, by adsorption on the porous material by poorly understood hydrophobic forces. Suitable solid supports also are described in U.S. Patent Application Serial No. 227,272.

The "indicator reagent "comprises a "signal generating compound" (label) which is capable of generating a measurable signal detectable by external means conjugated (attached) to a specific binding member for HCV. "Specific binding member" as used herein means a member of a specific binding pair. That is, two different molecules where one of the molecules through chemical or physical means specifically binds to the second molecule. In addition to being an antibody member of a specific binding pair for HCV, the indicator reagent also can be a member of any specific binding pair, including either hapten-anti-hapten systems such as biotin or anti-biotin, avidin or biotin, a carbohydrate or a lectin, a complementary nucleotide sequence, an effector or a receptor molecule, an enzyme cofactor and an enzyme, an enzyme inhibitor or an enzyme, and the like. An immunoreactive specific binding member can be an antibody, an antigen, or an antibody/antigen complex that is capable of binding either to HCV as in a sandwich assay, to the capture reagent as in a competitive assay, or to the ancillary specific binding member as in an indirect assay.

The various "signal generating compounds" (labels) contemplated include chromogens, catalysts such as enzymes, luminescent compounds such as fluorescein and rhodamine, chemiluminescent compounds, radioactive elements, and direct visual labels. Examples of enzymes include alkaline phosphatase, horseradish peroxidase, beta-galactosidase, and the like. The selection of a particular label is not critical, but it will be capable of producing a signal either by itself or in conjunction with one or more additional substances. The various "signal generating compounds" (labels) contemplated include chromogens, catalysts such as enzymes, luminescent compounds such as fluorescein and rhodamine, chemiluminescent compounds such as acridinium, phenanthridinium and dioxetane compounds, radioactive elements, and direct visual labels. Examples of enzymes include alkaline phosphatase, horseradish peroxidase, beta-galactosidase, and the like. The selection of a particular label is not critical, but it will be capable of producing a signal either by itself or in conjunction with one or more additional substances.

Other embodiments which utilize various other solid phases also are contemplated and are within the scope of this invention. For example, ion capture procedures for immobilizing an immobiiizabie reaction complex with a negatively charged polymer, described in co-pending U. S. Patent Application Serial No. 150,278 corresponding to EP publication 0326100, and U. S. Patent Application Serial No. 375,029 (EP publication no. 0406473) both of which enjoy common ownership and are incorporated herein by reference, can be employed according to the present invention to effect a fast solution-phase immunochemical reaction. An immobiiizabie immune complex is separated from the rest of the reaction mixture by ionic interactions between the negatively charged poly-anion/immune complex and the previously treated, positively charged porous matrix and detected by using various signal generating systems previously described, including those described in chemiluminescent signal measurements as described in co-pending U.S. Patent Application Serial No.921 ,979 corresponding to EPO Publication No. 0 273J 15, which enjoys common ownership and which is incorporated herein by reference.

Also, the methods of the present invention can be adapted for use in systems which utilize microparticle technology including in automated and semi-automated systems wherein the solid phase comprises a microparticle. Such systems include those described in pending U. S. Patent Applications 425,651 and 425,643, which correspond to published EPO applications Nos. EP 0 425 633 and EP 0 424 634, respectively, which are incorporated herein by reference.

The use of scanning probe microscopy (SPM) for immunoassays also is a technology to which the monoclonal antibodies of the present invention are easily adaptable. In scanning probe microscopy, in particular in atomic force microscopy, the capture phase, for example, at least one of the monoclonal antibodies of the invention, is adhered to a ^' solid phase and a scanning probe microscope is utilized to detect antigen/antibody complexes which may be present on the surface of the solid phase. The use of scanning tunnelling microscopy eliminates the need for labels which normally must be utilized in many immunoassay systems to detect antigen/antibody complexes. Such a system is described in pending U. S. patent application Serial No. 662,147, which enjoys common ownership and is incorporated herein by reference.

The use of SPM to monitor specific binding reactions can occur in many ways. In one embodiment, one member of a specific binding partner (analyte specific substance which is the monoclonal antibody of the invention) is attached to a surface suitable for scanning. The attachment of the analyte specific substance may be by adsorption to a test piece which comprises a solid phase of a plastic or

metal surface, following methods known to those of ordinary skill in the art. Or, covalent attachment of a specific binding partner (analyte specific substance) to a test piece which test piece comprises a solid phase of derivatized plastic, metal, silicon, or glass may be utilized. Covalent attachment methods are known to those skilled in the art and include a variety of means to irreversibly link specific binding partners to the test piece. If the test piece is silicon or glass, the surface must be activated prior to attaching the specific binding partner. Activated silane compounds such as triethoxy amino propyl silane (available from Sigma Chemical Co., St. Louis, MO), triethoxy vinyl silane (Aldrich Chemical Co., Milwaukee, Wl), and (3-mercapto-propyl)-trimethoxy silane (Sigma Chemical Co., St. Louis, MO) can be used to introduce reactive groups such as amino-, vinyl, and thiol, respectively. Such activated surfaces can be used to link the binding partner directly (in the cases of amino or thiol) or the activated surface can be further reacted with linkers such as glutaraldehyde, bis (succinimidyl) suberate, SPPD 9 succinimidyl 3-[2-pyridyldithio] propionate), SMCC (succinimidyl-4-[N- maleimidomethyl] cyclohexane-1 -carboxylate), SIAB (succinimidyl [4- iodoacetyl] aminobenzoate), and SMPB (succinimidyl 4-[1-maleimidophenyl] butyrate) to separate the binding partner from the surface. The vinyl group can be oxidized to provide a means for covalent attachment. It also can be used as an anchor for the polymerization of various polymers such as poly acrylic acid, which can provide multiple attachment points for specific binding partners. The amino surface can be reacted with oxidized dextrans of various molecular weights to provide hydrophilic linkers of different size and capacity. Examples of oxidizable dextrans include Dextran T-40 (molecular weight 40,000 daltons), Dextran T- 110 (molecular weight 110,000 daltons), Dextran T-500 (molecular weight

500,000 daltons), Dextran T-2M (molecular weight 2,000,000 daltons) (all of which are available from Pharmacia, LOCATION), or Ficoll (molecular weight 70,000 daltons (available from Sigma Chemical Co., St. Louis, MO). Also, polyelectrolyte interactions may be used to immobilize a specific binding partner on a surface of a test piece by using techniques and chemistries described by pending

U. S. Patent applications Serial No. 150,278, filed January 29, 1988, and Serial No. 375,029, filed July 7, 1989, each of which enjoys common ownership and each of which is incorporated herein by reference. The preferred method of attachment is by covalent means. Following attachment of a specific binding member, the surface may be further treated with materials such as serum, proteins, or other blocking agents to minimize non-specific binding. The surface also may be scanned either at the site of manufacture or point of use to verify its suitability for assay

purposes. The scanning process is not anticipated to alter the specific binding properties of the test piece.

Various other assay formats may be used, including "sandwich" immunoassays and competitive probe assays. For example, the monoclonal antibodies produced from the proteins of the present invention can be employed in various assay systems to determine the presence, if any, of HCV proteins in a test sample. Fragments of these monoclonal antibodies provided also may be used. For example, in a first assay format, a polyclonal or monoclonal anti-HCV antibody or fragment thereof, or a combination of these antibodies, which has been coated on a solid phase, is contacted with a test sample which may contain HCV proteins, to form a mixture. This mixture is incubated for a time and under conditions sufficient to form antigen/antibody complexes. Then, an indicator reagent comprising a monoclonal or a polyclonal antibody or a fragment thereof, which specifically binds to the HCV fragment, or a combination of these antibodies, to which a signal generating compound has been attached, is contacted with the antigen/antibody complexes to form a second mixture. This second mixture then is incubated for a time and under conditions sufficient to form antibody/antigen/antibody complexes. The presence of HCV antigen present in the test sample and captured on the solid phase, if any, is determined by detecting the measurable signal generated by the signal generating compound. The amount of HCV antigen present in the test sample is proportional to the signal generated.

Alternatively, a polyclonal or monoclonal anti-HCV antibody or fragment thereof, or a combination of these antibodies which is bound to a solid support, the test sample and an indicator reagent comprising a monoclonal or polyclonal antibody or fragments thereof, which specifically binds to HCV antigen, or a combination of these antibodies to which a signal generating compound is attached, are contacted to form a mixture. This mixture is incubated for a time and under conditions sufficient to form antibody/antigen/antibody complexes. The presence, if any, of HCV proteins present in the test sample and captured on the solid phase is determined by detecting the measurable signal generated by the signal generating compound. The amount of HCV proteins present in the test sample is proportional to the signal generated.

In another alternate assay format, one or a combination of one or more monoclonal antibodies of the invention can be employed as a competitive probe for the detection of antibodies to HCV protein. For example, HCV proteins, either alone or in combination, can be coated on a solid phase. A test sample suspected of containing antibody to HCV antigen then is incubated with an indicator reagent

comprising a signal generating compound and at least one monoclonal antibody of the invention for a time and under conditions sufficient to form antigen/antibody complexes of either the test sample and indicator reagent to the solid phase or the indicator reagent to the solid phase. The reduction in binding of the monoclonal antibody to the solid phase can be quantitatively measured. A measurable reduction in the signal compared to the signal generated from a confirmed negative NANB hepatitis test sample indicates the presence of anti-HCV antibody in the test sample.

In yet another detection method, each of the monoclonal antibodies of the present invention can be employed in the detection of HCV antigens in fixed tissue sections, as well as fixed cells by immunohistochemical analysis.

In addition, these monoclonal antibodies can be bound to matrices similar to CNBr-activated Sepharose and used for the affinity purification of specific HCV proteins from cell cultures, or biological tissues such as blood and liver.

The monoclonal antibodies of the invention can also be used for the generation of chimeric antibodies for therapeutic use, or other similar applications.

The monoclonal antibodies or fragments thereof can be provided individually to detect HCV antigens. Combinations of the monoclonal antibodies (and fragments thereof) provided herein also may be used together as components in a mixture or "cocktail" of at least one anti-HCV antibody of the invention with antibodies to other

HCV regions, each having different binding specificities. Thus, this cocktail can include the monoclonal antibodies of the invention which are directed to HCV proteins and other monoclonal antibodies to other antigenic determinants of the HCV genome. The polyclonal antibody or fragment thereof which can be used in the assay formats should specifically bind to a specific HCV region or other HCV proteins used in the assay. The polyclonal antibody used preferably is of mammalian origin; human, goat, rabbit or sheep anti-HCV polyclonal antibody can be used. Most preferably, the polyclonal antibody is rabbit polyclonal anti-HCV antibody. The polyclonal antibodies used in the assays can be used either alone or as a cocktail of polyclonal antibodies. Since the cocktails used in the assay formats are comprised of either monoclonal antibodies or polyclonal antibodies having different HCV specificity, they would be useful for diagnosis, evaluation and prognosis of HCV infection, as well as for studying HCV protein differentiation and specificity. In another assay format, the presence of antibody and/or antigen to HCV can be detected in a simultaneous assay, as follows. A test sample is simultaneously contacted with a capture reagent of a first analyte, wherein said capture reagent

comprises a first binding member specific for a first analyte attached to a solid phase and a capture reagent for a second analyte, wherein said capture reagent comprises a first binding member for a second analyte attached to a second solid phase, to thereby form a mixture. This mixture is incubated for a time and under conditions sufficient to form capture reagent/first analyte and capture reagent/second analyte complexes. These so-formed complexes then are contacted with an indicator reagent comprising a member of a binding pair specific for the first analyte labelled with a signal generating compound and an Indicator reagent comprising a member of a binding pair specific for the second analyte labelled with a signal generating compound to form a second mixture. This second mixture is incubated for a time and under conditions sufficient to form capture reagent/first analyte/indicator reagent complexes and capture reagent/second analyte/indicator reagent complexes. The presence of one or more analytes is determined by detecting a signal generated in connection with the complexes formed on either or both solid phases as an indication of the presence of one or more analytes in the test sample.

In this assay format, proteins derived from human expression systems may be utilized as well as monoclonal antibodies produced from the proteins derived from the mammalian expression systems as disclosed herein. Such assay systems are described in greater detail in pending U.S. Patent Application Serial No. 07/574,821 entitled Simultaneous Assay for Detecting One Or More Analytes, filed

August 29, 1990, which enjoys common ownership and is incorporated herein by reference.

In yet other assay formats, recombinant proteins may be utilized to detect the presence of anti-HCV in test samples. For example, a test sample is incubated with a solid phase to which at least one recombinant protein has been attached.

These are reacted for a time and under conditions sufficient to form antigen/antibody complexes. Following incubation, the antigen/antibody complex is detected. Indicator reagents may be used to facilitate detection, depending upon the assay system chosen. In another assay format, a test sample is contacted with a solid phase to which a recombinant protein produced as described herein is attached and also is contacted with a monoclonal or polyclonal antibody specific for the protein, which preferably has been labelled with an indicator reagent. After incubation for a time and under conditions sufficient for antibody/antigen complexes to form, the solid phase is separated from the free phase, and the label is detected in either the solid or free phase as an indication of the presence of HCV antibody. Other assay formats utilizing the proteins of the present invention are contemplated. These include contacting a test sample with a solid phase to which at

least one recombinant protein produced in the mammalian expression system has been attached, incubating the solid phase and test sample for a time and under conditions sufficient to form antigen/antibody complexes, and then contacting the solid phase with a labelled recombinant antigen. Assays such as this and others are described in pending U.S. Patent Application Serial No. 07/787,710, which enjoys common ownership and is incorporated herein by reference.

While the present invention discloses the preference for the use of solid phases, it is contemplated that the proteins of the present invention can be utilized in non-solid phase assay systems. These assay systems are known to those skilled in the art, and are considered to be within the scope of the present invention.

The present invention will now be described by way of examples, which are meant to illustrate, but not to limit, the spirit and scope of the invention.

EXAMPLES Example 1 : Generation of HCV Genomic Clones

RNA isolated from the serum or plasma of a chimpanzee (designated as "CO") experimentally infected with HCV, or an HCV seropositive human patient (designated as "LG") was transcribed to cDNA using reverse transcriptase employing either random hexamer primers or specific anti-sense primers derived from the prototype HCV-1 sequence. The sequence has been reported by Choo et al.

(Choo et al., Proc. Natl Acad. Sci. USA 88:2451-2455 [1991], and is available through GenBank data base, Accession No. M62321). This cDNA then was amplified using PCR and AmpliTaq® DNA polymerase (available in the Gene Amp Kit® from Perkin Elmer Cetus, Norwalk, Conneticut 06859) employing either a second sense primer located approximately 1000-2000 nucleotides upstream of the specific antisense primer or a pair of sense and antisense primers flanking a 1000-2000 nucleotide fragment of HCV. After 25 to 35 cycles of amplification following standard procedures known in the art, an aliquot of this reaction mixture was subjected to nested PCR (or "PCR-2"), wherein a pair of sense and antisense primers located internal to the original pair of PCR primers was employed to further amplify HCV gene segments in quantities sufficient for analysis and subcloning, utilizing endonuclease recognition sequences present in the second set of PCR primers. In this manner, seven adjacent HCV DNA fragments were generated which then could be assembled using the generic cloning strategy presented and described in FIGURE 1. The location of the specific primers used in this manner are presented in Table 1 and are numbered according to the HCV-1 sequence reported by Choo et al (GenBank data base, Accession No. M62321). Prior to

assembly, the DNA sequence of each of the individual fragments was determined and translated into the genomic amino acid sequences presented in SEQUENCE ID. NO. 1 and 2, respectively, for CO and LG, respectively. Comparison of the genomic polypeptide of CO with that of HCV-1 demonstrated 98 amino acid differences. Comparison of the genomic polypeptide of CO with that of LG. demonstrated 150 amino acid differences. Comparison of the genomic polypeptide of LG with that of HCV-1 demonstrated 134 amino acid differences.

Example 2. Expression of the HCV E2 Protein As A Fusion With The Amyloid Precursor Protein (APP)

The HCV E2 protein from CO developed as described in Example 1 was expressed as a fusion with the Amyloid Precursor Protein (APP). APP has been described by Kang et al., Nature 325:733-736 (1987). Briefly, HCV amino acids 384-749 of the CO isolate were used to replace the majority of the APP coding sequence as demonstrated in FIGURE 2. A Hindlll-Styl DNA fragment representing the amino-terminal 66 amino acids and a Bglll-Xbal fragment representing the carboxyl-terminal 105 amino acids of APP were ligated to a PCR derived HCV fragment from CO representing HCV amino acids 384-749 containing Styl and Bglll restriction sites on its 5' and 3' ends, respectively. This APP-HCV-E2 fusion gene cassette then was cloned into the commercially available mammalian expression vector pRC/CMV shown in FIGURE 3, (available from Invitrogen, San Diego, CA) at the unique Hindlll and Xbal sites. After transformation into E. coli DH5a, a clone designated pHCV-162 was isolated, which placed the expression of the APP-HCV-E2 fusion gene cassette under control of the strong CMV promotor. The complete nucleotide sequence of the mammalian expression vector pHCV-162 is presented in

SQUENCE ID. NO. 3. Translation of nucleotides 922 through 2535 results in the complete amino acid sequence of the APP-HCV-E2 fusion protein expressed by pHCV-162 as presented in SEQUENCE ID. NO. 4.

A primary Human Embryonic Kidney (HEK) cell line transformed with human adenovirus type 5, designated as HEK-293, was used for all transfections and expression analyses. HEK-293 cells were maintained in Minimum Essential Medium (MEM) which was supplemented with 10% fetal calf serum (FCS), penicillin and streptomycin.

Approximately 20 μg of purified DNA from pHCV-162 was transfected into HEK-293 cells using the modified calcium phosphate protocol as reported by Chen et al., Molecular and Cellular Biology 7(8):2745-2752 (1987). The calcium- phosphate-DNA solution was incubated on the HEK-293 cells for about 15 to 24

hours. The solution was removed, the cells were washed twice with MEM media, and then the cells were incubated in MEM media for an additional 24 to 48 hours. In order to analyze protein expression, the transfected cells were metabolically labelled with 100 μCi/ml S-35 methionine and cysteine for 12 to 18 hours. The culture media was removed and stored, and the cells were washed in MEM media and then lysed in phosphate buffered saline (PBS) containing 1% Triton X-100® (available from Sigma Chemical Co., St. Louis, MO), 0.1 % sodium dodecyl sulfate (SDS), and 0.5% deoxychloate, designated as PBS-TDS. This cell lysate then was frozen at -70°C for 2 to 24 hours, thawed on ice and then clarified by centrifugation at 50,000 x g force for one hour at 4°C. Standard radio- immunoprecipitation assays (RIPAs) then were conducted on those labelled cell lysates and/or culture medias. Briefly, labelled cell lysates and/or culture medias were incubated with 2 to 5 μl of specific sera at 4°C for one hour. Protein-A sepharose then was added and the samples were further incubated for one hour at 4°C with agitation. The samples were then centrifuged and the pellets washed several times with PBS-TDS buffer. Proteins recovered by immunoprecipitation were eluted by heating in an electrophoresis sample buffer (50 mM Tris-HCI, pH 6.8, 100 mM dithiothreitol [DTT], 2% SDS, 0.1% bromophenol blue, and 10% glycerol) for five minutes at 95°C. The eluted proteins then were separated by SDS polyacrylamide gels which were subsequently treated with a fluorographic reagent such as Enlightening® (available from NEN [DuPont], Boston, MA), dried under vacuum and exposed to x-ray film at -70°C with intensifying screens. FIGURE 4 presents a RIPA analysis of pHCV-162 transfected HEK cell lysate precipitated with normal human sera (NHS), a monoclonal antibody directed against APP sequences which were replaced in this construct (MAB), and an HCV antibody positive human sera (#25). Also presented in FIGURE 4 is the culture media (supernatant) precipitated with the same HCV antibody positive human sera (#25). From FIGURE 4, it can be discerned that while only low levels of an HCV specific protein of approximately 75K daltons is detected in the culture media of HEK-293 cells transfected with pHCV-162, high levels of intracellular protein expression of the

APP-HCV-E2 fusion protein of approximately 70K datons is evident.

In order to further characterize this APP-HCV-E2 fusion protein, rabbit polyclonal antibody raised against synthetic peptides were used in a similar RIPA, the results of which are illustrated in FIGURE 5. As can be discerned from this Figure, normal rabbit serum (NRS) does not precipitate the 70K dalton protein while rabbit sera raised against HCV amino acids 509-551 (6512), HCV amino

acids 380-436 (6521), and APP amino acids 45-62 (anti- N-terminus) are highly specific for the 70K dalton APP-HCV-E2 fusion protein.

In order to enhance secretion of this APP-HCV-E2 fusion protein, another clone was generated which fused only the amino-terminal 66 amino acids of APP, which contain the putative secretion signal sequences to the HCV-E2 sequences. In addition, a strongly hydrophobic sequence at the carboxyl-terminal end of the HCV- E2 sequence which was identified as a potential transmembrane spanning region was deleted. The resulting clone was designated as pHCV-167 and is schematically illustrated in FIGURE 2. The complete nucleotide sequence of the mammalian expression vector pHCV-167 is presented inSEQUENCE ID. NO. 5 Translation of nucleotides 922 through 2025 results in the complete amino acid sequence of the APP-HCV-E2 fusion protein expressed by pHCV-167 as presented in SEQUENCE ID. NO. 6. Purified DNA of pHCV-167 was transfected into HEK-293 cells and analyzed by RIPA and polyacrylamide SDS gels as described previously herein. FIGURE 6 presents the results in which a normal human serum sample (NHS) failed to recognize the APP-HCV-E2 fusion protein present in either the cell lysate or the cell supernatant of HEK-293 cells transfected with pHCV-167. The positive control HCV serum sample (#25), however, precipitated an approximately 65K dalton APP-HCV-E2 fusion protein present in the cell lysate of HEK-293 cells transfected with pHCV-167. In addition, substantial quantities of secreted APP- HCV-E2 protein of approximately 70K daltons was precipitated from the culture media by serum #25.

Digestion with Endoglycosidase-H (Endo-H) was conducted to ascertain the extent and composition of N-linked glycosylation in the APP-HCV E2 fusion proteins expressed by pHCV-167and pHCV-162 in HEK-293 cells. Briefly, multiple aliquots of labelled cell lysates from pHCV-162 and pHCV-167 transfected HEK- 293 cells were precipitated with human serum #50 which contained antibody to HCV E2 as previously described. The Proteϊn-A sepharose pellet containing the immunoprecϊpitated protein-antibody complex was then resuspended in buffer (75mM sodium acetate, 0.05% SDS) containing or not containing 0.05 units per ml of Endo-H (Sigma). Digestions were performed at 37°C for 12 to 18 hours and all samples were analyzed by polyacrylamide SDS gels as previously described. FIGURE 7 presents the results of Endo-H digestion. Carbon-14 labelled molecular weight standards (MW) (obtained from Amersham, Arlington Heights, IL) are common on ail gels and represent 200K, 92.5K, 69K, 46K, 30K and 14. 3K daltons, respectively. Normal human serum (NHS) does not immunoprecϊpitate the APP-HCV-E2 fusion protein expressed by either pHCV-162 or pHCV-167, while

human serum positive for HCV E2 antibody (#50) readily detects the 72K dalton APP-HCV-E2 fusion protein in pHCV-162 and the 65K dalton APP-HCV E2 fusion protein in pHCV-167. Incubation of these immunoprecipitated proteins in the absence of Endo-H (#50 -Endo-H) does not significantly affect the quantity or mobility of either pHCV-162 or pHCV-167 expressed proteins. Incubation in the presence of Endo-H (#50 +Endo-H), however, drastically reduces the mobility of the proteins expressed by pHCV-162 and pHCV-167, producing a heterogenous size distribution. The predicted molecular weight of the non-glycosylated polypeptide backbone of pHCV-162 is approximately 59K daltons. Endo-H treatment of pHCV- 162 lowers the mobility to a minimum of approximately 44K daltons, indicating that the APP-HCV-E2 fusion protein produced by pHCV-162 is proteolytically cleaved at the carboxyl-terminal end. A size of approximately 44K daltons is consistent with cleavage at or near HCV amino acid 720. Similarly, Endo-H treatment of pHCV-167 lowers the mobility to a minimum of approximately 41 K daltons, which compares favorably with the predicted molecular weight of approximately 40K daltons for the intact APP-HCV-E2 fusion protein expressed by pHCV-167.

Example 3 Detection of HCV E2 Antibodies Radio-immunoprecipitation assay (RIPA) and polyacrylamide SDS gel analysis previously described was used to screen numerous serum samples for the presence of antibody directed against HCV E2 epitopes. HEK-293 cells transfected with pHCV-162 were metabolically labelled and cell lysates prepared as previously described. In addition to RIPA analysis, all serum samples were screened for the presence of antibodies directed against specific HCV recombinant antigens representing distinct areas of the HCV genome using the Abbott Matrix® System, (available from Abbott Laboratories, Abbott Park, IL 60064, U.S. No. Patent 5,075,077). In the Matrix data presented in Tables 2 through 7, C100 yeast represents the NS4 region containing HCV amino acids 1569-1930, C100 E.coli represents HCV amino acids 1676-1930, NS3 represents HCV amino acids 1192-

1457, and CORE represents HCV amino acids 1-150.

FIGURE 8 presents a representative RIPA result obtained using pHCV-162 cell lysate to screen HCV antibody positive American blood donors and transfusion recipients. Table 2 summarizes the antibody profile of these various American blood samples, with seven of seventeen (41%) samples demonstrating HCV E2 antibody. Genomic variability in the E2 region has been demonstrated between different HCV isolates, particularly in geographically distinct isolates which may

lead to differences in antibody respones. We therefore screened twenty-six Japanese volunteer blood donors and twenty Spanish hemodiaiysis patients previously shown to contain HCV antibody for the presence of specific antibody to the APP-HCV E2 fusion protein expressed by pHCV-162. Figures 9 and 10 present the RIPA analysis on twenty-six Japanese volunteer blood donors. Positive control human sera (#50) and molecular weight standards (MW) appear in both figures in which the specific immunoprecipitation of the approximately 72K dalton APP- HCV-E2 fusion protein is demonstrated for several of the serum samples tested. Table 3 presents both the APP-HCV-E2 RIPA and Abbott Matrix® results summarizing the antibody profiles of each of the twenty-six Japanese samples tested. Table 4 presents similar data for the twenty Spanish hemodiaiysis patients tested. Table 5 summarizes the RIPA results obtained using pHCV-162 to detect HCV E2 specific antibody in these various samples. Eighteen of twenty-six (69%) Japanese volunteers blood donors, fourteen of twenty (70%) Spanish hemodiaiysis patients, and seven of seventeen (41%) American blood donors or transfusion recipients demonstrated a specific antibody response against the HCV E2 fusion protein. The broad immunoreactivity demonstrated by the APP-HCV-E2 fusion protein expressed by pHCV-162 suggests the recognition of conserved epitopes within HCV E2. Serial bleeds from five transfusion recipients which seroconverted to HCV antibody were also screened using the APP-HCV-E2 fusion protein expressed by pHCV-162. This analysis was conducted to ascertain the time interval after exposure to HCV at which E2 specific antibodies can be detected. Table 6 presents one such patient (AN) who seroconverted to NS3 at 154 days post transfusion (DPT). Antibodies to HCV E2 were not detected by RIPA until 271 DPT. Table 7 presents another such patient (WA), who seroconverted to CORE somewhere before 76 DPT and was positive for HCV E2 antibodies on the next available bleed date (103 DPT). Table 8 summarizes the serologicai results obtained from these five transfusion recipients indicating (a) some general antibody profile at seroconversion (AB Status); (b) the days post transfusion at which an ELISA test would most likely detect HCV antibody (2.0 GEN); (c) the samples in which HCV E2 antibody was detected by RIPA (E2 AB Status); and (d) the time interval covered by the bleed dates tested (Samples Tested). The results indicate that antibody to HCV E2, as detected in the RIPA procedure described here, appears after seroconversion to at least one other HCV marker (CORE, NS3, C100, etc.) and is persistent in nature once it appears. In addition, the absence of antibody to the structural gene CORE appears highly correlated with the absence of detectable antibody to E2,

another putative structural antigen. Further work is ongoing to correlate the presence or absence of HCV gene specific antibodies with progression of disease and/or time interval since exposure to HCV viral antigens.

Example 4 Expression of HCV E1 and E2 Using

Human Growth Hormone Secretion Signal HCV DNA fragments representing HCV E1 ( HCV amino acids 192 to 384) and HCV E2 ( HCV amino acids 384-750 and 384-684) were generated from the CO isolate using PCR as described in Example 2. An Eco Rl restriction site was used to attach a synthetic oligonucleotide encoding the Human Growth Hormone (HGH) secretion signal (Blak et al, Oncooene. 3 129-136, 1988) at the 5' end of these HCV sequence. The resulting fragment was then cloned into the commercially available mammalian expression vector pCDNA-l, (available from Invitrogen, San Diego, California) illustrated in FIGURE 11. Upon transformation into E. coli MC1061/P3, the resulting clones place the expression of the cloned sequence under control of the strong CMV promoter. Following the above outlined methods, a clone capable of expressing HCV-E1 ( HCV amino acids 192-384) employing the HGH secretion signal at the extreme amino-terminal end was isolated. The clone was designated pHCV-168 and is schematically illustrated in FIGURE 12. Similarly, clones capable of expressing HCV E2 ( HCV amino acids 384-750 or 384-684) exmploying the HGH secretion signal were isolated, designated pHCV-169 and pHVC-170 respectively and illustrated in FIGURE 13. The complete nucleotide sequence of the mammalian expression vectors pHCV-168, pHCV-169, and pHCV- 170 are presented in Sequence ID. NO. 7, 9, and 11 respectively. Translation of nucleotides 2227 through 2913 results in the complete amino acic sequence of the

HGH-HCV-E1 fusion protein expressed by pHCV-168 as presented in Sequence ID. NO. 8. Translation of nucleotides 2227 through 3426 results in the complete amino acic sequence of the HGH-HCV-E2 fusion protein expressed by pHCV-169 as presented in Sequence ID. NO. 10. Translation of nucleotides 2227 through 3228 results in the complete amino acic sequence of the HGH-HCV-E2 fusion protein expressed by pHCV-170 as presented in Sequence ID. NO. 12. Purified DNA from pHCV-168, pHCV-169, and pHCV-170 was transfected into HEK-293 cells which were then metabolically labelled, cell lysates prepared, and RIPA analysis performed as described previously herein. Seven sera samples previously shown to contain antibodies to the APP-HCV-E2 fusion protein expressed by pHCV-162 were screened against the labelled cell lysates of pHCV-168, pHCV-169, and pHCV-170. Figure 14 presents the RIPA analysis for pHCV-168 and demonstrated that five

sera containing HCV E2 antibodies also contain HCV E1 antibodies directed against as approximately 33K dalton HGH-HCV-E1 fusion protein ( #25, #50, 121, 503, and 728 ), while two other sera do not contain those antibodies ( 476 and 505 ). Figure 15 presents the RIPA results obtained when the same sera indicated above were screened against the labelled cell lysates of either pHCV-169 or pHCV-170. All seven HCV E1 antibody positive sera detected two protein species of approximately 70K and 75K daltons in cells transfected with pHCV-168. These two different HGH-HCV-E2 protein species could result from incomplete proteolytic cleavage of the HCV E2 sequence at the carboxyl-terminal end (at or near HCV amino acid 720) or from differences in carbohydrate processing between the two species.

All seven HCV E2 antibody positive sera detected a single protein species of approximately 62K daltons for the HGH-HCV-E2 fusion protein expressed by pHCV-170. Table 9 summarizes the serologicai profile of six of the seven HCV E2 antibody positive sera screened against the HGH-HCV-E1 fusion protein expressed by pHCV-170. Further work is ongoing to correlate the presence or absence of HCV gene specific antibodies with progression of disease and/or time interval since exposure to HCV viral antigens.

Clones pHCV-167 and pHCV-162 have been deposited at the American Type Culture Collection, 12301 Parklawn Drive, Rockville, Maryland, 20852, as of

January 17, 1992 under the terms of the Budapest Treaty, and accorded the following ATCC Designation Numbers: Clone pHCV-167 was accorded ATCC deposit number 68893 and clone pHCV-162 was accorded ATCC deposit number 68894. Clones pHCV-168, pHCV-169 and pHCV-170 have been deposited at the American Type Culture Collection, 12301 Parklawn Drive, Rockville, Maryland, 20852, as of January 26, 1993 under the terms of the Budapest Treaty, and accorded the following ATCC Designation Numbers: Clone pHCV-168 was accorded ATCC deposit number 69228, clone pHCV-169 was accorded ATCC deposit number 69229 and clone pHCV-170 was accorded ATCC deposit number 69230. The designated deposits will be maintained for a period of thirty (30) years from the date of deposit, or for five (5) years after the last request for the deposit; or for the enforceable life of the U.S. patent, whichever is longer. These deposits and other deposited materials mentioned herein are intended for convenience only, and are not required to practice the invention in view of the descriptions herein. The HCV cDNA sequences in all of the deposited materials are incorporated herein by reference.

Other variations of applications of the use of the proteins and mammalian expression systems provided herein will be apparent to those skilled in the art.

Accordingly, the invention is intended to be limited only in accordance with the appended claims.

TABLE 1

TABLE 2 AMERICAN HCV POSITIVE SERA

TABLE 3 JAPANESE HCV POSITIVE POSITIVE BLOOD DONORS

TABLE 4 SPANISH HEMODIALYSIS PATIENTS

SPANISH

HEMODIALYSIS 16/20 16/20 19/20 17/20 14/20

PATIENTS

JAPANESE

BLOOD 12/26 14/26 20/26 26/26 18/26

DONORS

TABLE 6 HUMAN TRANSFUSION RECIPIENT (AN)

TABLE 7 HUMAN TRANSFUSION RECIPIENT (WA)

TABLE 9 SELECTED HCV E2 ANTIBODY POSITIVE SAMPLES

SEQUENCE LISTING

(1) GENERAL INFORMATION:

(i) APPLICANT: CASEY, JAMES M. BODE, SUZANNE L. ZECK, BILLY J. YAMAGUCHI, JULIE FRAIL, DONALD E. DESAI, SURESH M. DEVARE, SUSHIL G.

(ii) TITLE OF INVENTION: MAMMALIAN EXPRESSION SYSTEMS FOR HCV PROTEINS

(iii) NUMBER OF SEQUENCES: 12

(iv) CORRESPONDENCE ADDRESS:

(A) ADDRESSEE: ABBOTT LABORATORIES D377/AP6D

(B) STREET: ONE ABBOTT PARK ROAD

(C) CITY: ABBOTT PARK

(D) STATE: IL

(E) COUNTRY: USA

(F) ZIP: 60064-3500

(v) COMPUTER READABLE FORM:

(A) MEDIUM TYPE: Floppy disk

(B) COMPUTER: IBM PC compatible

(C) OPERATING SYSTEM: PC-DOS/MS-DOS

(D) SOFTWARE: Patentin Release #1.0, Version #1.25

(vi) CURRENT APPLICATION DATA:

(A) APPLICATION NUMBER:

(B) FILING DATE:

(C) CLASSIFICATION:

(viii) ATTORNEY/AGENT INFORMATION:

(A) NAME: POREMBSKI, PRISCILLA E.

(B) REGISTRATION NUMBER: 33,207

(C) REFERENCE/DOCKET NUMBER: 5131.PC.01

(ix) TELECOMMUNICATION INFORMATION:

(A) TELEPHONE: 708-937-6365

(B) TELEFAX: 708-937-9556

(2) INFORMATION FOR SEQ ID NO:l:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 3011 amino acids

(B) TYPE: amino acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: protein

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:l:

Met Ser Thr Asn Pro Lys Pro Gin Arg Lys Thr Lys Arg Asn Thr Asn 1 5 10 15

Arg Arg Pro Gin Asp Val Lys Phe Pro Gly Gly Gly Gin lie Val Gly 20 25 30

Gly Val Tyr Leu Leu Pro Arg Arg Gly Pro Arg Leu Gly Val Arg Ala 35 40 45

Thr Arg Lys Thr Ser Glu Arg Ser Gin Pro Arg Gly Arg Arg Gin Pro 50 55 60 lie Pro Lys Ala Arg Arg Pro Glu Gly Arg Thr Trp Ala Gin Pro Gly 65 70 75 80

Tyr Pre Trp Pro Leu Tyr Gly Asn Glu Gly Cys Gly Trp Ala Gly Trp 85 90 95

Leu L<?-i Ser Pro Arg Gly Ser Arg Pro Ser Trp Gly Pro Thr Asp Pro 100 105 110

Arg Arg Arg Ser Arg Asn Leu Gly Lys Val lie Asp Thr Leu Thr Cys 115 120 125

Gly Phe Ala Asp Leu Met Gly Tyr lie Pro Leu Val Gly Ala Pro Leu 130 135 140

Gly Gly Ala Ala Arg Ala Leu Ala His Gly Val Arg Val Leu Glu Asp 145 150 155 160

Gly Val Asn Tyr Ala Thr Gly Asn Leu Pro Gly Cys Ser Phe Ser lie 165 170 175

Phe Leu Leu Ala Leu Leu Ser Cys Leu Thr Val Pro Ala Ser Ala Tyr 180 185 190

Gin Val Arg Asn Ser Ser Gly Leu Tyr His Val Thr Asn Asp Cys Pro 195 200 205

Asn Ser Ser lie Val Tyr Glu Ala Ala Asp Ala lie Leu His Thr Pro 210 215 220

Gly Cys Val Pro Cys Val Arg Glu Gly Asn Ala Ser Arg Cys Trp Val 225 230 235 240

Ala Val Thr Pro Thr Val Ala Thr Arg Asp Gly Lys Leu Pro Thr Thr 245 250 255

Gin Leu Arg Arg His lie Asp Leu Leu Val Gly Ser Ala Thr Leu Cys

260 265 270

Ser Ala Leu Tyr Val Gly Asp Leu Cys Gly Ser Val Phe Leu Val Gly 275 280 285

Gin Leu Phe Thr Phe Ser Pro Arg Arg His Trp Thr Thr Gin Asp Cys 290 295 300

Asn Cys Ser lie Tyr Pro Gly His lie Thr Gly His Arg Met Ala Trp 305 310 315 320

Asp Met Met Met Asn Trp Ser Pro Thr Ala Ala Leu Val Val Ala Gin 325 330 335

Leu Leu Arg lie Pro Gin Ala lie Leu Asp Met lie Ala Gly Ala His 340 345 350

Trp Gly Val Leu Ala Gly lie Ala Tyr Phe Ser Met Val Gly Asn Trp 355 360 365

Ala Lys Val Leu Val Val Leu Leu Leu Phe Ala Gly Val Asp Ala Glu 370 375 380

Thr His Val Thr Gly Gly Ser Ala Gly His Thr Thr Ala Gly Leu Val 385 390 395 400

Arg Leu Leu Ser Pro Gly Ala Lys Gin Asn lie Gin Leu lie Asn Thr 405 410 415

Asn Gly Ser Trp His lie Asn Ser Thr Ala Leu Asn Cys Asn Glu Ser 420 425 430

Leu Asn Thr Gly Trp Leu Ala Gly Leu Phe Tyr His His Lys Phe Asn 435 440 445

Ser Ser Gly Cys Pro Glu Arg Leu Ala Ser Cys Arg Arg Leu Thr Asp 450 455 460

Phe Ala Gin Gly Gly Gly Pro lie Ser Tyr Ala Asn Gly Ser Gly Leu 465 470 475 480

Asp Glu Arg Pro Tyr Cys Trp His Tyr Pro Pro Arg Pro Cys Gly lie 485 490 495

Val Pro Ala Lys Ser Val Cys Gly Pro Val Tyr Cys Phe Thr Pro Ser 500 505 510

Pro Val Val Val Gly Thr Thr Asp Arg Ser Gly Ala Pro Thr Tyr Ser 515 520 525

Trp Gly Ala Asn Asp Thr Asp Val Phe Val Leu Asn Asn Thr Arg Pro 530 535 540

Pro Leu Gly Asn Trp Phe Gly Cys Thr Trp Met Asn Ser Thr Gly Phe 545 550 555 560

Thr Lys Val Cys Gly Ala Pro Pro Cys Val He Gly Gly Val Gly Asn 565 570 575

Asn Thr Leu Leu Cys Pro Thr Asp Cys Phe Arg Lys His Pro Glu Ala 580 585 590

Thr Tyr Ser Arg Cys Gly Ser Gly Pro Trp He Thr Pro Arg Cys Met 595 600 605

Val Asp Tyr Pro Tyr Arg Leu Trp His Tyr Pro Cys Thr He Asn Tyr 610 615 620

Thr He Phe Lys Val Arg Met Tyr Val Gly Gly Val Glu His Arg Leu 625 630 635 640

Glu Ala Ala Cys Asn Trp Thr Arg Gly Glu Arg Cys Asp Leu Glu Asp 645 650 655

Arg Asp Arg Ser Glu Leu Ser Pro Leu Leu Leu Ser Thr Thr Gin Trp 660 665 670

Gin Val Leu Pro Cys Ser Phe Thr Thr Leu Pro Ala Leu Ser Thr Gly 675 680 685

Leu He His Leu His Gin Asn He Val Asp Val Gin Tyr Leu Tyr Gly 690 695 700

Val Gly Ser Ser He Ala Ser Trp Ala He Lys Trp Glu Tyr Val Val 705 710 715 720

Leu Leu Phe Leu Leu Leu Ala Asp Ala Arg Val Cys Ser Cys Leu Trp 725 730 735

Met Met Leu Leu He Ser Gin Ala Glu Ala Ala Leu Glu Asn Leu Val 740 745 750

He Leu Asn Ala Ala Ser Leu Ala Gly Thr His Gly Phe Val Ser Phe 755 760 765

Leu Val Phe Phe Cys Phe Ala Trp Tyr Leu Lys Gly Arg Trp Val Pro 770 775 780

Gly Ala Ala Tyr Ala Leu Tyr Gly He Trp Pro Leu Leu Leu Leu Leu 785 790 795 800

Leu Ala Leu Pro Gin Arg Ala Tyr Ala Leu Asp Thr Glu Val Ala Ala 805 810 815

Ser Cys Gly Gly Val Val Leu Val Gly Leu Met Ala Leu Thr Leu Ser 820 825 830

Pro Tyr Tyr Lys Arg Tyr He Ser Trp Cys Met Trp Trp Leu Gin Tyr 835 840 845

Phe Leu Thr Arg Val Glu Ala Gin Leu His Val Trp Val Pro Pro Leu 850 855 8S0

Asn Val Arg Gly Gly Arg Asp Ala Val He Leu Leu Met Cys Ala Val 865 870 875 880

His Pro Thr Leu Val Phe Asp He Thr Lys Leu Leu Leu Ala He Phe 885 890 895

Gly Pro Leu Trp He Leu Gin Ala Ser Leu Leu Lys Val Pro Tyr Phe 900 905 910

Val Arg Val Gin Gly Leu Leu Arg He Cys Ala Leu Ala Arg Lys He 915 920 925

Ala Gly Gly His Tyr Val Gin Met He Phe He Lys Leu Gly Ala Leu 930 935 940

Thr Gly Thr Tyr Val Tyr Asn His Leu Thr Pro Leu Arg Asp Trp Ala 945 950 955 960

His Asn Gly Leu Arg Asp Leu Ala Val Ala Val Glu Pro Val Val Phe 965 970 975

Ser Arg Met Glu Thr Lys Leu He Thr Trp Gly Ala Asp Thr Ala Ala 980 985 990

Cys Gly Asp He He Asn Gly Leu Pro Val Ser Ala Arg Arg Gly Gin 995 1000 1005

Glu He Leu Leu Gly Pro Ala Asp Gly Met Val Ser Lys Gly Trp Arg 1010 1015 1020

Leu Leu Ala Pro He Thr Ala Tyr Ala Gin Gin Thr Arg Gly Leu Leu 1025 1030 1035 1040

Gly Cys He He Thr Ser Leu Thr Gly Arg Asp Lys Asn Gin Val Glu 1045 1050 1055

Gly Glu Val Gin He Val Ser Thr Ala Thr Gin Thr Phe Leu Ala Thr 1060 1065 1070

Cys He Asn Gly Val Cys Trp Thr Val Tyr His Gly Ala Gly Thr Arg 1075 1080 1085

Thr He Ala Ser Pro Lys Gly Pro Val He Gin Met Tyr Thr Asn Val 1090 1095 1100

Asp Gin Asp Leu Val Gly Trp Pro Ala Pro Gin Gly Ser Arg Ser Leu 1105 1110 1115 1120

Thr Pro Cys Thr Cys Gly Ser Ser Asp Leu Tyr Leu Val Thr Arg His 1125 1130 1135

Ala Asp Val He Pro Val Arg Arg Gin Gly Asp Ser Arg Gly Ser Leu

1140 1145 1150

Leu Ser Pro Arg Pro He Ser Tyr Leu Lys Gly Ser Ser Gly Gly Pro 1155 1160 1165

Leu Leu Cys Pro Ala Gly His Ala Val Gly Leu Phe Arg Ala Ala Val 1170 1175 1180

Cys Thr Arg Gly Val Ala Lys Ala Val Asp Phe He Pro Val Glu Asn

1185 1190 1195 1200

Leu Glu Thr Thr Met Arg Ser Pro Val Phe Thr Asp Asn Ser Ser Pro 1205 1210 1215

Pro Ala Val Pro Gin Ser Phe Gin Val Ala His Leu His Ala Pro Thr 1220 1225 1230

Gly Ser Gly Lys Ser Thr Lys Val Pro Ala Ala Tyr Ala Ala Gin Gly 1235 1240 1245

Tyr Lys Val Leu Val Leu Asn Pro Ser Val Ala Ala Thr Leu Gly Phe 1250 1255 1260

Gly Ala Tyr Met Ser Lys Ala His Gly Val Asp Pro Asn He Arg Thr

1265 1270 1275 1280

Gly Val Arg Thr He Thr Thr Gly Ser Pro He Thr Tyr Ser Thr Tyr 1285 1290 1295

Gly Lys Phe Leu Ala Asp Gly Gly Cys Ser Gly Gly Ala Tyr Asp He 1300 1305 1310

He He Cys Asp Glu Cys His Ser Thr Asp Ala Thr Ser He Leu Gly 1315 1320 1325

He Gly Thr Val Leu Asp Gin Ala Glu Thr Ala Gly Ala Arg Leu Val 1330 1335 1340

Val Leu Ala Thr Ala Thr Pro Pro Gly Ser Val Thr Val Pro His Pro

1345 1350 1355 1360

Asn He Glu Glu Val Ala Leu Ser Thr Thr Gly Glu He Pro Phe Tyr 1365 1370 1375

Gly Lys Ala He Pro Leu Glu Val He Lys Gly Gly Arg His Leu He 1380 1385 1390

Phe Cys His Ser Lys Lys Lys Cys Asp Glu Leu Ala Ala Lys Leu Val 1395 1400 1405

Ala Leu Gly He Asn Ala Val Ala Tyr Tyr Arg Gly Leu Asp Val Ser 1410 1415 1420

Val He Pro Ala Ser Gly Asp Val Val Val Val Ser Thr Asp Ala Leu

1425 1430 1435 1440

Met Thr Gly Phe Thr Gly Asp Phe Asp Pro Val He Asp Cys Asn Thr 1445 1450 1455

Cys Val Thr- Gin Thr Val Asp Phe Ser Leu Asp Pro Thr Phe Thr He 1460 1465 1470

Glu Thr Thr Thr Leu Pro Gin Asp Ala Val Ser Arg Thr Gin Arg Arg 1475 1480 1485

Gly Arg Thr Gly Arg Gly Lys Pro Gly He Tyr Arg Phe Val Ala Pro 1490 1495 1500

Gly Glu Arg Pro Ser Gly Met Phe Asp Ser Ser Val Leu Cys Glu Cys 1505 1510 1515 1520

Tyr Asp Ala Gly Cys Ala Trp Tyr Glu Leu Thr Pro Ala Glu Thr Thr 1525 1530 1535

Val Arg Leu Arg Ala Tyr Met Asn Thr Pro Gly Leu Pro Val Cys Gin 1540 1545 1550

Asp His Leu Glu Phe Trp Glu Gly Val Phe Thr Gly Leu Thr His He 1555 1560 1565

Asp Ala His Phe Leu Ser Gin Thr Lys Gin Ser Gly Glu Asn Phe Pro 1570 1575 1580

Tyr Leu Val Ala Tyr Gin Ala Thr Val Cys Ala Arg Ala Gin Ala Pro 1585 1590 1595 1600

Pro Pro Ser Trp Asp Gin Met Trp Lys Cys Leu He Arg Leu Lys Pro 1605 1610 1615

Thr Leu His Gly Pro Thr Pro Leu Leu Tyr Arg Leu Gly Ala Val Gin 1620 1625 1630

Asn Glu He Thr Leu Thr His Pro Val Thr Lys Tyr He Met Thr Cys 1635 1640 1645

Met Ser Ala Asn Pro Glu Val Val Thr Ser Thr Trp Val Leu Val Gly 1650 1655 1660

Gly Val Leu Ala Ala Leu Ala Ala Tyr Cys Leu Ser Thr Gly Cys Val 1665 1670 1675 1680

Val He Val Gly Arg He Val Leu Ser Gly Lys Pro Ala He He Pro 1685 1690 1695

Asp Arg Glu Val Leu Tyr Gin Glu Phe Asp Glu Met Glu Glu Cys Ser 1700 1705 1710

Gin His Leu Pro Tyr He Glu Gin Gly Met Met Leu Ala Glu Gin Phe 1715 1720 1725

Lys Gin Glu Ala Leu Gly Leu Leu Gin Thr Ala Ser Arg Gin Ala Glu

1730 1735 1740

Val He Thr Pro Ala Val Gin Thr Asn Trp Gin Lys Leu Glu Ala Phe 1745 1750 1755 1760

Trp Ala Lys His Met Trp Asn Phe He Ser Gly Thr Gin Tyr Leu Ala 1765 1770 1775

Gly Leu Ser Thr Leu Pro Gly Asn Pro Ala He Ala Ser Leu Met Ala 1780 1785 1790

Phe Thr Ala Ala Val Thr Ser Pro Leu Thr Thr Ser Gin Thr Leu Leu 1795 1800 1805

Phe Asn He Leu Gly Gly Trp Val Ala Ala Gin Leu Ala Ala Pro Gly

1810 1815 1820

Ala Ala Thr Ala Phe Val Gly Ala Gly Leu Ala Gly Ala Ala He Gly 1825 1830 1835 1840

Ser Val Gly Leu Gly Lys Val Leu Val Asp He Leu Ala Gly Tyr Gly 1845 1850 1855

Ala Gly Val Ala Gly Ala Leu Val Ala Phe Lys He Met Ser Gly Glu 1860 1865 1870

Val Pro Ser Thr Glu Asp Leu Val Asn Leu Leu Pro Ala He Leu Ser 1875 1880 1885

Pro Gly Ala Leu Val Val Gly Val Val Cys Ala Ala He Leu Arg Arg

1890 1895 1900

His Val Gly Pro Gly Glu Gly Ala Val Gin Trp Met Asn Arg Leu He 1905 1910 1915 1920

Ala Phe Ala Ser Arg Gly Asn His Val Ser Pro Thr His Tyr Val Pro 1925 1930 1935

Glu Ser Asp Ala Ala Ala Arg Val Thr Ala He Leu Ser Asn Leu Thr 1940 1945 1950

Val Thr Gin Leu Leu Arg Arg Leu His Gin Trp He Gly Ser Glu Cys 1955 1960 1965

Thr Thr Pro Cys Ser Gly Ser Trp Leu Arg Asp He Trp Asp Trp He

1970 1975 1980

Cys Glu Val Leu Ser Asp Phe Lys Thr Trp Leu Lys Ala Lys Leu Met 1985 1990 1995 2000

Pro Gin Leu Pro Gly He Pro Phe Val Ser Cys Gin Arg Gly Tyr Arg 2005 2010 2015

Gly Val Trp Arg Gly Asp Gly He Met His Thr Arg Cys His Cys Gly

2020 2025 2030

Ala Glu He Thr Gly His Val Lys Asn Gly Thr Met Arg He Val Gly 2035 2040 2045

Pro Arg Thr Cys Arg Asn Met Trp Ser Gly Thr Phe Pro He Asn Ala 2050 2055 2060

Tyr Thr Thr Gly Pro Cys Thr Pro Leu Pro Ala Pro Asn Tyr Lys Phe 2065 2070 2075 2080

Ala Leu Trp Arg Val Ser Ala Glu Glu Tyr Val Glu He Arg Arg Val 2085 2090 2095

Gly Asp Phe His Tyr Val Ser Gly Met Thr Thr Asp Asn Leu Lys Cys 2100 2105 2110

Pro Cys Gin He Pro Ser Pro Glu Phe Phe Thr Glu Leu Asp Gly Val 2115 2120 2125

Arg Leu His Arg Phe Ala Pro Pro Cys Lys Pro Leu Leu Arg Glu Glu 2130 2135 2140

Val Ser Phe Arg Val Gly Leu His Glu Tyr Pro Val Gly Ser Gin Leu 2145 2150 2155 2160

Pro Cys Glu Pro Glu Pro Asp Val Ala Val Leu Thr Ser Met Leu Thr 2165 2170 2175

Asp Pro Ser His He Thr Ala Glu Ala Ala Gly Arg Arg Leu Ala Arg 2180 2185 2190

Gly Ser Pro Pro Ser Met Ala Ser Ser Ser Ala Ser Gin Leu Ser Ala 2195 2200 2205

Pro Ser Leu Lys Ala Thr Cys Thr Thr Asn His Asp Ser Pro Asp Ala 2210 2215 2220

Glu Leu He Glu Ala Asn Leu Leu Trp Arg Gin Glu Met Gly Gly Asn 2225 2230 2235 2240

He Thr Arg Val Glu Ser Glu Asn Lys Val Val He Leu Asp Ser Phe 2245 2250 2255

Asp Pro Leu Val Ala Glu Glu Asp Glu Arg Glu Val Ser Val Pro Ala 2260 2265 2270

Glu He Leu Arg Lys Ser Gin Arg Phe Ala Arg Ala Leu Pro Val Trp 2275 2280 2285

Ala Arg Pro Asp Tyr Asn Pro Pro Leu He Glu Thr Trp Lys Glu Pro 2290 2295 2300

Asp Tyr Glu Pro Pro Val Val His Gly Cys Pro Leu Pro Pro Pro Arg 2305 2310 2315 2320

Ser Pro Pro Val Pro Pro Pro Arg Lys Lys Arg Thr Val Val Leu Thr 2325 2330 2335

Glu Ser Thr Leu Ser Thr Ala Leu Ala Glu Leu Ala Thr Lys Ser Phe 2340 2345 2350

Gly Ser Ser Ser Thr Ser Gly He Thr Gly Asp Asn Thr Thr Thr Ser 2355 2360 2365

Ser Glu Pro Ala Pro Ser Gly Cys Pro Pro Asp Ser Asp Val Glu Ser 2370 2375 2380

Tyr Ser Ser Met Pro Pro Leu Glu Gly Glu Pro Gly Asp Pro Asp Phe 2385 2390 2395 2400

Ser Asp Gly Ser Trp Ser Thr Val Ser Ser Gly Ala Asp Thr Glu Asp 2405 2410 2415

Val Val Cys Cys Ser Met Ser Tyr Ser Trp Thr Gly Ala Leu Val Thr 2420 2425 2430

Pro Cys Ala Ala Glu Glu Gin Lys Leu Pro He Asn Ala Leu Ser Asn 2435 2440 2445

Ser Leu Leu Arg His His Asn Leu Val Tyr Ser Thr Thr Ser Arg Ser 2450 2455 2460

Ala Cys Gin Arg Gin Lys Lys Val Thr Phe Asp Arg Leu Gin Val Leu 2465 2470 2475 2480

Asp Ser His Tyr Gin Asp Val Leu Lys Glu Val Lys Ala Ala Ala Ser 2485 2490 2495

Arg Val Lys Ala Asn Leu Leu Ser Val Glu Glu Ala Cys Ser Leu Thr 2500 2505 2510

Pro Pro His Ser Ala Lys Ser Lys Phe Gly Tyr Gly Ala Lys Asp Val 2515 2520 2525

Arg Cys His Ala Arg Lys Ala Val Ala His He Asn Ser Val Trp Lys 2530 2535 2540

Asp Leu Leu Glu Asp Ser Val Thr Pro He Asp Thr Thr He Met Ala 2545 2550 2555 2560

Lys Asn Glu Val Phe Cys Val Gin Pro Glu Lys Gly Gly Arg Lys Pro 2565 2570 2575

Ala Arg Leu He Val Phe Pro Asp Leu Gly Val Arg Val Cys Glu Lys 2580 2585 2590

Met Ala Leu Tyr Asp Val Val Ser Lys Leu Pro Leu Ala Val Met Gly 2595 2600 2605

Ser Ser Tyr Gly Phe Gin Tyr Ser Pro Gly Gin Arg Val Glu Phe Leu 2610 2615 2620

Val Gin Ala Trp Lys Ser Lys Lys Thr Pro Met Gly Phe Ser Tyr Asp 2625 2630 2635 2640

Thr Arg Cys Phe Asp Ser Thr Val Thr Glu Ser Asp He Arg Thr Glu 2645 2650 2655

Glu Ala He Tyr Gin Cys Cys Asp Leu Asp Pro Gin Ala Arg Val Ala 2660 2665 2670

He Lys Ser Leu Thr Glu Arg Leu Tyr Val Gly Gly Pro Leu Thr Asn 2675 2680 2685

Ser Arg Gly Glu Asn Cys Gly Tyr Arg Arg Cys Arg Ala Ser Gly Val 2690 2695 2700

Leu Thr Thr Ser Cys Gly Asn Thr Leu Thr Cys Tyr He Lys Ala Arg 2705 2710 2715 2720

Ala Ala Cys Arg Ala Ala Gly Leu Gin Asp Arg Thr Met Leu Val Cys 2725 2730 2735

Gly Asp Asp Leu Val Val He Cys Glu Ser Ala Gly Val Gin Glu Asp 2740 2745 2750

Ala Ala Ser Leu Arg Ala Phe Thr Glu Ala Met Thr Arg Tyr Ser Ala 2755 2760 2765

Pro Pro Gly Asp Pro Pro Gin Pro Glu Tyr Asp Leu Glu Leu He Thr 2770 2775 2780

Ser Cys Ser Ser Asn Val Ser Val Ala His Asp Gly Ala Gly Lys Arg 2785 2790 2795 2800

Val Tyr Tyr Leu Thr Arg Asp Pro Thr Thr Pro Leu Ala Arg Ala Ala 2805 2810 2815

Trp Glu Thr Ala Arg His Thr Pro Val Asn Ser Trp Leu Gly Asn He 2820 2825 2830

He Met Phe Ala Pro Thr Leu Trp Ala Arg Met He Leu Met Thr His 2835 2840 2845

Phe Phe Ser Val Leu He Ala Arg Asp Gin Phe Glu Gin Ala Leu Asn 2850 2855 2860

Cys Glu He Tyr Gly Ala Cys Tyr Ser He Glu Pro Leu Asp Leu Pro 2865 2870 2875 2880

Pro He He Gin Arg Leu His Gly Leu Ser Ala Phe Ser Leu His Ser 2885 2890 2895

Tyr Ser Pro Gly Glu He Asn Arg Val Ala Ala Cys Leu Arg Lys Leu

2900 2905 2910

Gly Val Pro Pro Leu Arg Ala Trp Lys His Arg Ala Arg Ser Val Arg 2915 2920 2925

Ala Arg Leu Leu Ser Arg Gly Gly Arg Ala Ala He Cys Gly Lys Tyr 2930 2935 2940

Leu Phe Asn Trp Ala Val Arg Thr Lys Pro Lys Leu Thr Pro He Ala 2945 2950 2955 2960

Ala Ala Gly Arg Leu Asp Leu Ser Gly Trp Phe Thr Ala Gly Tyr Ser 2965 2970 2975

Gly Gly Asp He Tyr His Ser Val Ser His Ala Arg Pro Arg Trp Ser 2980 2985 2990

Trp Phe Cys Leu Leu Leu Leu Ala Ala Gly Val Gly He Tyr Leu Leu 2995 3000 3005

Pro Asn Arg 3010

(2) INFORMATION FOR SEQ ID NO:2:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 3011 amino acids

(B) TYPE: amino acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: re ■-.-:n

(Xi) SEQUENCE DESCRIPTION _-EQ ID NO:2:

Met Ser Thr Asn Pro Lys _ ro Gin Arg Lys Thr Lys Arg Asn Thr Asn 1 5 10 15

Arg Arg Pro Gin Asp Val Lys Phe Pro Gly Gly Gly Gin He Val Gly 20 25 30

Gly Val Tyr Leu Leu Pro Arg Arg Gly Pro Arg Leu Gly Val Arg Ala 35 40 45

Thr Arg Lys Thr Ser Glu Arg Ser Gin Pro Arg Gly Arg Arg Gin Pro 50 55 60

He Pro Lys Ala Arg Arg Pro Glu Gly Arg Thr Trp Ala Gin Pro Gly 65 70 75 80

Tyr Pro Trp Pro Leu Tyr Gly Asn Glu Gly Cys Gly Trp Ala Gly Trp 85 90 95

Leu Leu Ser Pro Arg Gly Ser Arg Pro Ser Trp Gly Pro Thr Asp Pro 100 105 110

Arg Arg Arg Ser Arg Asn Leu Gly Lys Val He Asp Thr Leu Thr Cys 115 120 125

Gly Phe Ala Asp Leu Met Gly Tyr He Pro Leu Val Gly Ala Pro Leu 130 135 140

Gly Gly Ala Ala Arg Ala Leu Ala His Gly Val Arg Val Leu Glu Asp 145 150 155 160

Gly Val Asn Tyr Ala Thr Gly Asn Leu Pro Gly Cys Ser Phe Ser He 165 170 175

Phe Leu Leu Ala Leu Leu Ser Cys Leu Thr Val Pro Ala Ser Ala Tyr 180 185 190

Gin Val Arg Asn Ser Ser Gly Leu Tyr His Val Thr Asn Asp Cys Pro 195 200 205

Asn Ser Ser He Val Tyr Glu Thr Ala Asp Thr He Leu His Ser Pro 210 215 220

Gly Cys Val Pro Cys Val Arg Glu Gly Asn Thr Ser Lys Cys Trp Val 225 230 235 240

Ala Val Ala Pro Thr Val Thr Thr Arg Asp Gly Lys Leu Pro Ser Thr 245 250 255

Gin Leu Arg Arg His He Asp Leu Leu Val Gly Ser Ala Thr Leu Cys 260 265 270

Ser Ala Leu Tyr Val Gly Asp Leu Cys Gly Ser Val Phe Leu Val Ser 275 280 285

Gin Leu Phe Thr Phe Ser Pro Arg Arg His Trp Thr Thr Gin Asp Cys 290 295 300

Asn Cys Ser He Tyr Pro Gly His He Thr Gly His Arg Met Ala Trp 305 310 315 320

Asp Met Met Met Asn Trp Ser Pro Thr Thr Ala Leu Val Val Ala Gin 325 330 335

Leu Leu Arg He Pro Gin Ala He Leu Asp Met He Ala Gly Ala His 340 345 350

Trp Gly Val Leu Ala Gly He Ala Tyr Phe Ser Met Val Gly Asn Trp 355 360 365

Ala Lys Val Leu Val Val Leu Leu Leu Phe Ser Gly Val Asp Ala Ala 370 375 380

Thr Tyr Thr Thr Gly Gly Ser Val Ala Arg Thr Thr His Gly Leu Ser

385 390 395 400

Ser Leu Phe Ser Gin Gly Ala Lys Gin Asn He Gin Leu He Asn Thr 405 410 415

Asn Gly Ser Trp His He Asn Arg Thr Ala Leu Asn Cys Asn Ala Ser 420 425 430

Leu Asp Thr Gly Trp Val Ala Gly Leu Phe Tyr Tyr His Lys Phe Asn 435 440 445

Ser Ser Gly Cys Pro Glu Arg Met Ala Ser Cys Arg Pro Leu Ala Asp 450 455 460

Phe Asp.Gin Gly Trp Gly Pro He Ser Tyr Thr Asn Gly Ser Gly Pro 465 470 475 480

Glu His Arg Pro Tyr Cys Trp His Tyr Pro Pro Lys Pro Cys Gly He 485 490 495

Val Pro Ala Gin Ser Val Cys Gly Pro Val Tyr Cys Phe Thr Pro Ser 500 505 510

Pro Val Val Val Gly Thr Thr Asp Lys Ser Gly Ala Pro Thr Tyr Thr 515 520 525

Trp Gly Ser Asn Asp Thr Asp Val Phe Val Leu Asn Asn Thr Arg Pro 530 535 540

Pro Pro Gly Asn Trp Phe Gly Cys Thr Trp Met Asn Ser Ser Gly Phe 545 550 555 560

Thr Lys Val Cys Gly Ala Pro Pro Cys Val He Gly Gly Ala Gly Asn 565 570 575

Asn Thr Leu His Cys Pro Thr Asp Cys Phe Arg Lys His Pro Glu Ala 580 585 590

Thr Tyr Ser Arg Cys Gly Ser Gly Pro Trp He Thr Pro Arg Cys Leu 595 600 605

Val His Tyr Pro Tyr Arg Leu Trp His Tyr Pro Cys Thr He Asn Tyr 610 615 620

Thr Leu Phe Lys Val Arg Met Tyr Val Gly Gly Val Glu His Arg Leu 625 630 635 640

Glu Val Ala Cys Asn Trp Thr Arg Gly Glu Arg Cys Asp Leu Asp Asp 645 650 655

Arg Asp Arg Ser Glu Leu Ser Pro Leu Leu Leu Ser Thr Thr Gin Trp 660 665 670

Gin Val Leu Pro Cys Ser Phe Thr Thr Leu Pro Ala Leu Thr Thr Gly 675 680 685

Leu He His Leu His Gin Asn He Val Asp Val Gin Tyr Leu Tyr Gly 690 695 700

Val Gly Ser Ser He Val Ser Trp Ala He Lys Trp Glu Tyr Val He 705 710 715 720

Leu Leu Phe Leu Leu Leu Ala Asp Ala Arg He Cys Ser Cys Leu Trp 725 730 735

Met Met Leu Leu He Ser Gin Ala Glu Ala Ala Leu Glu Asn Leu Val 740 745 750

Leu Leu Asn Ala Ala Ser Leu Ala Gly Thr His Gly Leu Val Ser Phe 755 760 765

Leu Val Phe Phe Cys Phe Ala Trp Tyr Leu Lys Gly Lys Trp Val Pro 770 775 780

Gly Val Ala Tyr Ala Phe Tyr Gly Met Trp Pro Phe Leu Leu Leu Leu 785 790 795 800

Leu Ala Leu Pro Gin Arg Ala Tyr Ala Leu Asp Thr Glu Met Ala Ala 805 810 815

Ser Cys Gly Gly Val Val Leu Val Gly Leu Met Ala Leu Thr Leu Ser 820 825 830

Pro His Tyr Lys Arg Tyr He Cys Trp Cys Val Trp Trp Leu Gin Tyr 835 840 845

Phe Leu Thr Arg Ala Glu Ala Leu Leu His Gly Trp Val Pro Pro Leu 850 855 860

Asn Val Arg Gly Gly Arg Asp Ala Val He Leu Leu Met Cys Val Val 865 870 875 880

His Pro Ala Leu Val Phe Asp He Thr Lys Leu Leu Leu Ala Val Leu 885 890 895

Gly Pro Leu Trp He Leu Gin Thr Ser Leu Leu Lys Val Pro Tyr Phe 900 905 910

Val Arg Val Gin Gly Leu Leu Arg He Cys Ala Leu Ala Arg Lys Met 915 920 925

Ala Gly Gly His Tyr Val Gin Met Val Thr He Lys Met Gly Ala Leu 930 935 940

Ala Gly Thr Tyr Val Tyr Asn His Leu Thr Pro Leu Arg Asp Trp Ala 945 950 955 960

His Asn Gly Leu Arg Asp Leu Ala Val Ala Val Glu Pro Val Val Phe 965 970 975

Ser Gin Met Glu Thr Lys Leu He Thr Trp Gly Ala Asp Thr Ala Ala 980 985 990

Cys Gly Asp He He Asn Gly Leu Pro Val Ser Ala Arg Arg Gly Arg 995 1000 1005

Glu He Leu Leu Gly Pro Ala Asp Gly Met Val Ser Lys Gly Trp Arg 1010 1015 1020

Leu Leu Ala Pro He Thr Ala Tyr Ala Gin Gin Thr Arg Gly Leu Leu 1025 1030 1035 1040

Gly Cys He He Thr Ser Leu Thr Gly Arg Asp Lys Asn Gin Val Glu 1045 1050 1055

Gly Glu Val Gin He Val Ser Thr Ala Ala Gin Thr Phe Leu Ala Thr 1060 1065 1070

Cys He Asn Gly Val Cys Trp Thr Val Tyr His Gly Ala Gly Thr Arg 1075 1080 1085

Thr He Ala Ser Pro Lys Gly Pro Val He Gin Met Tyr Thr Asn Val 1090 1095 1100

Asp Arg Asp Leu Val Gly Trp Pro Ala Pro Gin Gly Ala Arg Ser Leu 1105 1110 1115 1120

Thr Pro Cys Thr Cys Gly Ser Ser Asp Leu Tyr Leu Val Thr Arg His 1125 1130 1135

Ala Asp Val He Pro Val Arg Arg Arg Gly Asp Ser Arg Gly Ser Leu 1140 1145 1150

Leu Ser Pro Arg Pro He Ser Tyr Leu Lys Gly Ser Ser Gly Gly Pro 1155 1160 1165

Leu Leu Cys Pro Ala Gly His Ala Val Gly He Phe Arg Ala Ala Val 1170 1175 1180

Cys Thr Arg Gly Val Ala Lys Ala Val Asp Phe He Pro Val Glu Ser 1185 1190 1195 1200

Leu Glu Thr Thr Met Arg Ser Pro Val Phe Thr Asp Asn Ser Ser Pro 1205 1210 1215

Pro Ala Val Pro Gin Ser Phe Gin Val Ala His Leu His Ala Pro Thr 1220 1225 1230

Gly Ser Gly Lys Ser Thr Lys Val Pro Ala Ala Tyr Ala Ala Gin Gly 1235 1240 1245

Tyr Lys Val Leu Val Leu Asn Pro Ser Val Ala Ala Thr Leu Gly Phe 1250 1255 1260

Gly Ala Tyr Met Ser Lys Ala His Gly He Asp Pro Asn He Arg Thr

1265 1270 1275 1280

Gly Val Arg Thr He Thr Thr Gly Ser Pro He Thr Tyr Ser Thr Tyr 1285 1290 1295

Gly Lys Phe Leu Ala Asp Gly Gly Cys Ser Gly Gly Ala Tyr Asp He 1300 1305 1310

He He Cys Asp Glu Cys His Ser Thr Asp Ala Thr Ser He Leu Gly 1315 1320 1325

He Gly Thr Val Leu Asp Gin Ala Glu Thr Ala Gly Ala Arg Leu Val 1330 1335 1340

Val Leu Ala Thr Ala Thr Pro Pro Gly Ser Val Thr Val Pro His Pro 1345 1350 1355 1360

Asn He Glu Glu Val Ala Leu Ser Thr Thr Gly Glu He Pro Phe Tyr 1365 1370 1375

Gly Lys Ala He Pro Leu Glu Ala He Lys Gly Gly Arg His Leu He 1380 1385 1390

Phe Cys His Ser Lys Lys Lys Cys Asp Glu Leu Ala Ala Lys Leu Val 1395 1400 1405

Thr Leu Gly He Asn Ala Val Ala Tyr Tyr Arg Gly Leu Asp Val Ser 1410 1415 1420

Val He Pro Thr Ser Gly Asp Val Val Val Val Ala Thr Asp Ala Leu 1425 1430 1435 1440

Met Thr Gly Phe Thr Gly Asp Phe Asp Ser Val He Asp Cys Asn Thr 1445 1450 1455

Cys Val Thr Gin Ala Val Asp Phe Ser Leu Asp Pro Thr Phe Thr He 1460 1465 1470

Glu Thr Thr Thr Leu Pro Gin Asp Ala Val Ser Arg Thr Gin Arg Arg 1475 1480 1485

Gly Arg Thr Gly Arg Gly Lys Pro Gly He Tyr Arg Phe Val Ala Pro 1490 1495 1500

Gly Glu Arg Pro Ser Gly Met Phe Asp Ser Ser Val Leu Cys Glu Cys 1505 1510 1515 1520

Tyr Asp Ala Gly Cys Ala Trp Tyr Glu Leu Thr Pro Ala Glu Thr Thr 1525 1530 1535

Val Arg Leu Arg Ala Tyr Met Asn Thr Pro Gly Leu Pro Val Cys Gin 1540 1545 1550

Asp His Leu Glu Phe Trp Glu Gly Val Phe Thr Gly Leu Thr His He 1555 1560 1565

Asp Ala His Phe Leu Ser Gin Thr Lys Gin Ser Gly Glu Asn Leu Pro 1570 1575 1580

Tyr Leu Val Ala Tyr Gin Ala Thr Val Cys Ala Arg Ala Gin Ala Pro 1585 1590 1595 1600

Pro Pro Ser Trp Asp Gin Met Trp Lys Cys Leu He Arg Leu Lys Pro 1605 1610 1615

Thr Leu His Gly Pro Thr Pro Leu Leu Tyr Arg Leu Gly Ala Val Gin 1620 1625 1630

Asn Glu Val Thr Leu Thr His Pro He Thr Lys Tyr He Met Thr Cys 1635 1640 1645

Met Ser Ala Asp Leu Glu Val Val Thr Ser Thr Trp Val Leu Val Gly 1650 1655 1660

Gly Val Leu Ala Ala Leu Ala Ala Tyr Cys Leu Ser Thr Gly Cys Val 1665 1670 1675 1680

Val He Val Gly Arg He Val Leu Ser Gly Lys Pro Ala He He Pro 1685 1690 1695

Asp Arg Glu Val Leu Tyr Arg Glu Phe Asp Glu Met Glu Glu Cys Ser 1700 1705 1710

Gin His Leu Pro Tyr He Glu Gin Gly Met Met Leu Ala Glu Gin Phe 1715 1720 1725

Lys Gin Lys Ala Leu Gly Leu Leu Gin Thr Ala Ser His Gin Ala Glu 1730 1735 1740

Val He Ala Pro Ala Val Gin Thr Asn Trp Gin Arg Leu Glu Thr Phe 1745 1750 1755 1760

Trp Ala Lys His Met Trp Asn Phe He Ser Gly He Gin Tyr Leu Ala 1765 1770 1775

Gly Leu Ser Thr Leu Pro Gly Asn Pro Ala He Ala Ser Leu Met Ala 1780 1785 ^■ 1790

Phe Thr Ala Ala Val Thr Ser Pro Leu Thr Thr Ser Gin Thr Leu Leu 1795 1800 1805

Phe Asn He Leu Gly Gly Trp Val Ala Ala Gin Leu Ala Ala Pro Ser 1810 1815 1820

Ala Ala Thr Ala Phe Val Gly Ala Gly Leu Ala Gly Ala Ala He Gly 1825 1830 1835 1840

Ser Val Gly Leu Gly Lys Val Leu Val Asp He Leu Ala Gly Tyr Gly 1845 1850 1855

Ala Gly Val Ala Gly Ala Leu Val Ala Phe Lys He Met Ser Gly Glu 1860 1865 1870

Val Pro Ser Thr Glu Asp Leu Val Asn Leu Leu Pro Ala He Leu Ser 1875 1880 1885

Pro Gly Ala Leu Val Val Gly Val Val Cys Ala Ala He Leu Arg Arg 1890 1895 1900

His Val Gly Pro Gly Glu Gly Ala Val Gin Trp Met Asn Arg Leu He 1905 1910 1915 1920

Ala Phe Ala Ser Arg Gly Asn His Val Ser Pro Thr His Tyr Val Pro 1925 1930 1935

Gly Ser Asp Ala Ala Ala Arg Val Thr Ala He Leu Ser Ser Leu Thr 1940 1945 1950

Val Thr Gin Leu Leu Arg Arg Leu His Gin Trp Val Ser Ser Glu Cys 1955 1960 1965

Thr Thr Pro Cys Ser Gly Ser Trp Leu Arg Asp He Trp Asp Trp He 1970 1975 1980

Cys Glu Val Leu Ser Asp Phe Lys Thr Trp Leu Lys Ala Lys Leu Met 1985 1990 1995 2000

Pro Gin Leu Pro Gly He Pro Phe Val Ser Cys Gin Arg Gly Tyr Lys 2005 2010 2015

Gly Val Trp Arg Gly Asp Gly He Met His Thr Arg Cys His Cys Gly 2020 2025 2030

Ala Glu He Ala Gly His Val Lys Asn Gly Thr Met Arg He Val Gly 2035 2040 2045

Pro Lys Thr Cys Arg Asn Met Trp Ser Gly Thr Phe Pro He Asn Ala 2050 2055 2060

Tyr Thr Thr Gly Pro Cys Thr Pro Leu Pro Ala Pro Asn Tyr Lys Phe 2065 2070 2075 2080

Ala Leu Trp Arg Val Ser Ala Glu Glu Tyr Val Glu He Arg Gin Val 2085 2090 2095

Gly Asp Phe His Tyr Val Thr Gly Met Thr Ala Asp Asn Leu Lys Cys 2100 2105 2110

Pro Cys Gin Val Pro Ser Pro Glu Phe Phe Thr Glu Leu Asp Gly Val 2115 2120 2125

Arg Leu His Arg Phe Ala Pro Pro Cys Lys Pro Leu Leu Arg Asp Glu 2130 2135 2140

Val Ser Phe Arg Val Gly Leu His Asp Tyr Pro Val Gly Ser Gin Leu

2145 2150 2155 2160

Pro Cys Glu Pro Glu Pro Asp Val Ala Val Leu Thr Ser Met Leu Thr 2165 2170 2175

Asp Pro Ser His He Thr Ala Glu Thr Ala Gly Arg Arg Leu Ala Arg 2180 2185 2190

Gly Ser Pro Pro Ser Met Ala Ser Ser Ser Ala Ser Gin Leu Ser Ala 2195 2200 2205

Pro Ser Leu Lys Ala Thr Cys Thr Thr Asn His Asp Ser Pro Asp Ala 2210 2215 2220

Glu Leu Leu Glu Ala Asn Leu Leu Trp Arg Gin Glu Met Gly Gly Asn 2225 2230 2235 2240

He Thr Arg Val Glu Ser Glu Asn Lys Val Val Val Leu Asp Ser Phe 2245 2250 2255

Asp Pro Leu Val Ala Glu Glu Asp Glu Arg Glu Val Ser Val Pro Ala 2260 2265 2270

Glu He Leu Arg Lys Ser Arg Arg Phe Ala Gin Ala Leu Pro Ser Trp 2275 2280 2285

Ala Arg Pro Asp Tyr Asn Pro Pro Leu Leu Glu Thr Trp Lys Lys Pro 2290 2295 2300

Asp Tyr Glu Pro Pro Val Val His Gly Cys Pro Leu Pro Pro Pro Gin 2305 2310 2315 2320

Ser Pro Pro Val Pro Pro Pro Arg Lys Lys Arg Thr Val Val Leu Thr 2325 2330 2335

Glu Ser Thr Val Ser Ser Ala Leu Ala Glu Leu Ala Thr Lys Ser Phe 2340 2345 2350

Gly Ser Ser Ser Thr Ser Gly He Thr Gly Asp Asn Thr Thr Thr Ser 2355 2360 2365

Ser Glu Pro Ala Pro Ser Val Cys Pro Pro Asp Ser Asp Ala Glu Ser 2370 2375 2380

Tyr Ser Ser Met Pro Pro Leu Glu Gly Glu Pro Gly Asp Pro Asp Leu 2385 2390 2395 2400

Ser Asp Gly Ser Trp Ser Thr Val Ser Ser Gly Ala Asp Thr Glu Asp 2405 2410 2415

Val Val Cys Cys Ser Met Ser Tyr Ser Trp Thr Gly Ala Leu He Thr 2420 2425 2430

Pro Cys Ala Ala Glu Glu Gin Lys Leu Pro He Asn Ala Leu Ser Asn 2435 2440 2445

Ser Leu Leu Arg His His Asn Leu Val Tyr Ser Thr Thr Ser Arg Asn 2450 2455 2460

Ala Cys Leu Arg Gin Lys Lys Val Thr Phe Asp Arg Leu Gin Val Leu 2465 2470 2475 2480

Asp Asn His Tyr Gin Asp Val Leu Lys Glu Val Lys Ala Ala Ala Ser 2485 2490 2495

Lys Val Lys Ala Asn Leu Leu Ser Val Glu Glu Ala Cys Ser Leu Thr 2500 2505 2510

Pro Pro His Ser Ala Arg Ser Lys Phe Gly Tyr Gly Ala Lys Asp Val 2515 2520 2525

Arg Cys His Ala Arg Lys Ala Val Ser His He Asn Ser Val Trp Lys 2530 2535 2540

Asp Leu Leu Glu Asp Ser Val Thr Pro He Asp Thr Thr He Met Ala 2545 2550 2555 2560

Lys Asn Glu Val Phe Cys Val Gin Pro Glu Lys Gly Gly Arg Lys Pro 2565 2570 2575

Ala Arg Leu He Val Phe Pro Asp Leu Gly Val Arg Val Cys Glu Lys 2580 2585 2590

Met Ala Leu Tyr Asp Val Val Ser Lys Leu Pro Leu Ala Val Met Gly 2595 2600 2605

Ser Ser Tyr Gly Phe Gin Tyr Ser Pro Gly Gin Arg Val Glu Phe Leu 2610 2615 2620

Val Gin Ala Trp Lys Ser Lys Lys Thr Pro Met Gly Phe Ser Tyr Asp 2625 2630 2635 2640

Thr Arg Cys Phe Asp Ser Thr Val Thr Glu Ser Asp He Arg Thr Glu 2645 2650 2655

Glu Ala He Tyr Gin Cys Cys Asp Leu Asp Pro Gin Ala Arg Val Ala 2660 2665 2670

He Lys Ser Leu Thr Glu Arg Leu Tyr Val Gly Gly Pro Leu Thr Asn 2675 2680 2685

Ser Arg Gly Glu Asn Cys Gly Tyr Arg Arg Cys Arg Ala Ser Gly Val 2690 2695 2700

Leu Thr Thr Ser Cys Gly Asn Thr Leu Thr Cys Tyr He Lys Ala Arg 2705 2710 2715 2720

Ala Ala Cys Arg Ala Ala Gly Leu Gin Asp Cys Thr Met Leu Val Cys 2725 2730 2735

Gly Asp Asp Leu Val Val He Cys Glu Ser Gin Gly Val Gin Glu Asp 2740 2745 2750

Ala Ala Ser Leu Arg Ala Phe Thr Glu Ala Met Thr Arg Tyr Ser Ala 2755 2760 2765

Pro Pro Gly Asp Pro Pro Gin Pro Glu Tyr Asp Leu Glu Leu He Thr 2770 2775 2780

Pro Cys Ser Ser Asn Val Ser Val Ala His Asp Gly Ala Gly Lys Arg 2785 2790 2795 2800

Val Tyr Tyr Leu Thr Arg Asp Pro Thr Thr Pro Leu Ala Arg Ala Ala 2805 2810 2815

Trp Glu Thr Ala Arg His Thr Pro Val Asn Ser Trp Leu Gly Asn He 2820 2825 2830

He Met Phe Ala Pro Thr Leu Trp Ala Arg Met He Leu Met Thr His 2835 2840 2845

Phe Phe Ser Val Leu He Ala Arg Asp Gin Leu Glu Gin Ala Leu Asp 2850 2855 2860

Cys Glu He Tyr Gly Ala Cys Tyr Ser He Glu Pro Leu Asp Leu Pro 2865 2870 2875 2880

Pro He He Gin Arg Leu His Gly Leu Ser Ala Phe Ser Leu His Ser 2885 2890 2895

Tyr Ser Pro Gly Glu He Asn Arg Val Ala Ala Cys Leu Arg Lys Leu 2900 2905 2910

Gly Val Pro Pro Leu Arg Ala Trp Arg His Arg Ala Arg Ser Val Arg 2915 2920 2925

Ala Arg Leu Leu Ser Arg Gly Gly Arg Ala Ala He Cys Gly Lys Tyr 2930 2935 2940

Leu Phe Asn Trp Ala Val Arg Thr Lys Leu Lys Leu Thr Pro He Ala 2945 2950 2955 2960

Ala Ala Gly Gin Leu Asp Leu Ser Gly Trp Phe Thr Ala Gly Tyr Gly 2965 2970 2975

Gly Gly Asp He Tyr His Ser Val Ser Arg Ala Arg Pro Arg Trp Phe 2980 2985 2990

Trp Phe Cys Leu Leu Leu Leu Ala Ala Gly Val Gly He Tyr Leu Leu 2995 3000 3005

Pro Asn Arg 3010

(2) INFORMATION FOR SEQ ID NO:3 :

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 7298 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: circular

(ii) MOLECULE TYPE: DNA (genomic)

(ix) FEATURE:

(A) NAME/KEY: CDS

(B) LOCATION: 922..2532

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:3:

GACGGATCGG GAGATCTCCC GATCCCCTAT GGTCGACTCT CAGTACAATC TGCTCTGATG 60

CCGCATAGTT AAGCCAGTAT CTGCTCCCTG CTTGTGTGTT GGAGGTCGCT GAGTAGTGCG 120

CGAGCAAAAT TTAAGCTACA ACAAGGCAAG GCTTGACCGA CAATTGCATG AAGAATCTGC 180

TTAGGGTTAG GCGTTTTGCG CTGCTTCGCG ATGTACGGGC CAGATATACG CGTTGACATT 2 0

GATTATTGAC TAGTTATTAA TAGTAATCAA TTACGGGGTC ATTAGTTCAT AGCCCATATA 300

TGGAGTTCCG CGTTACATAA CTTACGGTAA ATGGCCCGCC TGGCTGACCG CCCAACGACC 360

CCCGCCCATT GACGTCAATA ATGACGTATG TTCCCATAGT AACGCCAATA GGGACTTTCC 420

ATTGACGTCA ATGGGTGGAC TATTTACGGT AAACTGCCCA CTTGGCAGTA CATCAAGTGT 480

ATCATATGCC AAGTACGCCC CCTATTGACG TCAATGACGG TAAATGGCCC GCCTGGCATT 540

ATGCCCAGTA CATGACCTTA TGGGACTTTC CTACTTGGCA GTACATCTAC GTATTAGTCA 600

TCGCTATTAC CATGGTGATG CGGTTTTGGC AGTACATCAA TGGGCGTGGA TAGCGGTTTG 660

ACTCACGGGG ATTTCCAAGT CTCCACCCCA TTGACGTCAA TGGGAGTTTG TTTTGGCACC 720

AAAATCAACG GGACTTTCCA AAATGTCGTA ACAACTCCGC CCCATTGACG CAAATGGGCG 780

GTAGGCGTGT ACGGTGGGAG GTCTATATAA GCAGAGCTCT CTGGCTAACT AGAGAACCCA 840

CTGCTTAACT GGCTTATCGA AATTAATACG ACTCACTATA GGGAGACCGG AAGCTTTGCT 900

CTAGACTGGA ATTCGGGCGC G ATG CTG CCC GGT TTG GCA CTG CTC CTG CTG 951

Met Leu Pro Gly Leu Ala Leu Leu Leu Leu 1 5 10

GCC GCC TGG ACG GCT CGG GCG CTG GAG GTA CCC ACT GAT GGT AAT GCT 999 Ala Ala Trp Thr Ala Arg Ala Leu Glu Val Pro Thr Asp Gly Asn Ala 15 20 25

GGC CTG CTG GCT GAA CCC CAG ATT GCC ATG TTC TGT GGC AGA CTG AAC 1047 Gly Leu Leu Ala Glu Pro Gin He Ala Met Phe Cys Gly Arg Leu Asn 30 35 40

ATG CAC ATG AAT GTC CAG AAT GGG AAG TGG GAT TCA GAT CCA TCA GGG 1095 Met His Met Asn Val Gin Asn Gly Lys Trp Asp Ser Asp Pro Ser Gly 45 50 55

ACC AAA ACC TGC ATT GAT ACC AAG GAA ACC CAC GTC ACC GGG GGA AGT 1143 Thr Lys Thr Cys He Asp Thr Lys Glu Thr His Val Thr Gly Gly Ser 60 65 70

GCC GGC CAC ACC ACG GCT GGG CTT GTT CGT CTC CTT TCA CCA GGC GCC 1191 Ala Gly His Thr Thr Ala Gly Leu Val Arg Leu Leu Ser Pro Gly Ala 75 80 85 90

AAG CAG AAC ATC CAA CTG ATC AAC ACC AAC GGC AGT TGG CAC ATC AAT 1239 Lys Gin Asn He Gin Leu He Asn Thr Asn Gly Ser Trp His He Asn 95 100 105

AGC ACG GCC TTG AAC TGC AAT GAA AGC CTT AAC ACC GGC TGG TTA GCA 1287 Ser Thr Ala Leu Asn Cys Asn Glu Ser Leu Asn Thr Gly Trp Leu Ala 110 115 120

GGG CTC TTC TAT CAC CAC AAA TTC AAC TCT TCA GGT TGT CCT GAG AGG 1335 Gly Leu Phe Tyr His His Lys Phe Asn Ser Ser Gly Cys Pro Glu Arg 125 130 135

TTG GCC AGC TGC CGA CGC CTT ACC GAT TTT GCC CAG GGC GGG GGT CCT 1383 Leu Ala Ser Cys Arg Arg Leu Thr Asp Phe Ala Gin Gly Gly Gly Pro 140 145 150

ATC AGT TAC GCC AAC GGA AGC GGC CTC GAT GAA CGC CCC TAC TGC TGG 1431 He Ser Tyr Ala Asn Gly Ser Gly Leu Asp Glu Arg Pro Tyr Cys Trp 155 160 165 170

CAC TAC CCT CCA AGA CCT TGT GGC ATT GTG CCC GCA AAG AGC GTG TGT 1479 His Tyr Pro Pro Arg Pro Cys Gly He Val Pro Ala Lys Ser Val Cys 175 180 185

GGC CCG GTA TAT TGC TTC ACT CCC AGC CCC GTG GTG GTG GGA ACG ACC 1527 Gly Pro Val Tyr Cys Phe Thr Pro Ser Pro Val Val Val Gly Thr Thr 190 195 200

GAC AGG TCG GGC GCG CCT ACC TAC AGC TGG GGT GCA AAT GAT ACG GAT 1575 Asp Arg Ser Gly Ala Pro Thr Tyr Ser Trp Gly Ala Asn Asp Thr Asp 205 210 215

GTC TTT GTC CTT AAC AAC ACC AGG CCA CCG CTG GGC AAT TGG TTC GGT 1623 Val Phe Val Leu Asn Asn Thr Arg Pro Pro Leu Gly Asn Trp Phe Gly 220 225 230

TGC ACC TGG ATG AAC TCA ACT GGA TTC ACC AAA GTG TGC GGA GCG CCC 1671 Cys Thr Trp Met Asn Ser Thr Gly Phe Thr Lys Val Cys Gly Ala Pro 235 240 245 250

CCT TGT GTC ATC GGA GGG GTG GGC AAC AAC ACC TTG CTC TGC CCC ACT 1719 Pro Cys Val He Gly Gly Val Gly Asn Asn Thr Leu Leu Cys Pro Thr 255 260 265

GAT TGC TTC CGC AAG CAT CCG GAA GCC ACA TAC TCT CGG TGC GGC TCC 1767 Asp Cys Phe Arg Lys His Pro Glu Ala Thr Tyr Ser Arg Cys Gly Ser 270 275 280

GGT CCC TGG ATT ACA CCC AGG TGC ATG GTC GAC TAC CCG TAT AGG CTT 1815 Gly Pro Trp He Thr Pro Arg Cys Met Val Asp Tyr Pro Tyr Arg Leu 285 290 295

TGG CAC TAT CCT TGT ACC ATC AAT TAC ACC ATA TTC AAA GTC AGG ATG 1863 Trp His Tyr Pro Cys Thr He Asn Tyr Thr He Phe Lys Val Arg Met 300 305 310

TAC GTG GGA GGG GTC GAG CAC AGG CTG GAA GCG GCC TGC AAC TGG ACG 1911 Tyr Val Gly Gly Val Glu His Arg Leu Glu Ala Ala Cys Asn Trp Thr 315 320 325 330

CGG GGC GAA CGC TGT GAT CTG GAA GAC AGG GAC AGG TCC GAG CTC AGC 1959 Arg Gly Glu Arg Cys Asp Leu Glu Asp Arg Asp Arg Ser Glu Leu Ser 335 340 345

CCG TTA CTG CTG TCC ACC ACG CAG TGG CAG GTC CTT CCG TGT TCT TTC 2007 Pro Leu Leu Leu Ser Thr Thr Gin Trp Gin Val Leu Pro Cys Ser Phe 350 355 360

ACG ACC CTG CCA GCC TTG TCC ACC GGC CTC ATC CAC CTC CAC CAG AAC 2055 Thr Thr Leu Pro Ala Leu Ser Thr Gly Leu He His Leu His Gin Asn 365 370 375

ATT GTG GAC GTG CAG TAC TTG TAC GGG GTA GGG TCA AGC ATC GCG TCC 2103 He Val Asp Val Gin Tyr Leu Tyr Gly Val Gly Ser Ser He Ala Ser 380 385 390

TGG GCT ATT AAG TGG GAG TAC GAC GTT CTC CTG TTC CTT CTG CTT GCA 2151 Trp Ala He Lys Trp Glu Tyr Asp Val Leu Leu Phe Leu Leu Leu Ala 395 400 405 410

GAC GCG CGC GTT TGC TCC TGC TTG TGG ATG ATG TTA CTC ATA TCC CAA 2199 Asp Ala Arg Val Cys Ser Cys Leu Trp Met Met Leu Leu He Ser Gin 415 420 425

GCG GAG GCG GCT TTG GAG ATC TCT GAA GTG AAG ATG GAT GCA GAA TTC 2247 Ala Glu Ala Ala Leu Glu He Ser Glu Val Lys Met Asp Ala Glu Phe 430 435 440

CGA CAT GAC TCA GGA TAT GAA GTT CAT CAT CAA AAA TTG GTG TTC TTT 22 5 Arg His Asp Ser Gly Tyr Glu Val His His Gin Lys Leu Val Phe Phe 445 450 455

GCA GAA GAT GTG GGT TCA AAC AAA GGT GCA ATC ATT GGA CTC ATG GTG 2343 Ala Glu Asp Val Gly Ser Asn Lys Gly Ala He He Gly Leu Met Val

460 465 470

GGC GGT GTT GTC ATA GCG ACA GTG ATC GTC ATC ACC TTG GTG ATG CTG 2391 Gly Gly Val Val He Ala Thr Val He Val He Thr Leu Val Met Leu 475 • 480 485 490

AAG AAG AAA CAG TAC ACA TCC ATT CAT CAT GGT GTG GTG GAG GTT GAC 2439 Lys Lys Lys Gin Tyr Thr Ser He His His Gly Val Val Glu Val Asp 495 500 505

GCC GCT GTC ACC CCA GAG GAG CGC CAC CTG TCC AAG ATG CAG CAG AAC 2487 Ala Ala Val Thr Pro Glu Glu Arg His Leu Ser Lys Met Gin Gin Asn 510 515 520

GGC TAC GAA AAT CCA ACC TAC AAG TTC TTT GAG CAG ATG CAG AAC 2532 Gly Tyr Glu Asn Pro Thr Tyr Lys Phe Phe Glu Gin Met Gin Asn 525 530 535

TAGACCCCCG CCACAGCAGC CTCTGAAGTT GGACAGCAAA ACCATTGCTT CACTACCCAT 2592

CGGTGTCCAT TTATAGAATA ATGTGGGAAG AAACAAACCC GTTTTATGAT TTACTCATTA 2652

TCGCCTTTTG ACAGCTGTGC TGTAACACAA GTAGATGCCT GAACTTGAAT TAATCCACAC 2712

ATCAGTATTG TATTCTATCT CTCTTTACAT TTTGGTCTCT ATACTACATT ATTAATGGGT 2772

TTTGTGTACT GTAAAGAATT TAGCTGTATC AAACTAGTGC ATGAATAGGC CGCTCGAGCA 2832

TGCATCTAGA GGGCCCTATT CTATAGTGTC ACCTAAATGC TCGCTGATCA GCCTCGACTG 2892

TGCCTTCTAG TTGCCAGCCA TCTGTTGTTT GCCCCTCCCC CGTGCCTTCC TTGACCCTGG 2952

AAGGTGCCAC TCCCACTGTC CTTTCCTAAT AAAATGAGGA AATTGCATCG CATTGTCTGA 3012

GTAGGTGTCA TTCTATTCTG GGGGGTGGGG TGGGGCAGGA CAGCAAGGGG GAGGATTGGG 3072

AAGACAATAG CAGGCATGCT GGGGATGCGG TGGGCTCTAT GGAACCAGCT GGGGCTCGAG 3132

GGGGGATCCC CACGCGCCCT GTAGCGGCGC ATTAAGCGCG GCGGGTGTGG TGGTTACGCG 3192

CAGCGTGACC GCTACACTTG CCAGCGCCCT AGCGCCCGCT CCTTTCGCTT TCTTCCCTTC 3252

CTTTCTCGCC ACGTTCGCCG GCTTTCCCCG TCAAGCTCTA AATCGGGGCA TCCCTTTAGG 3312

GTTCCGATTT AGTGCTTTAC GGCACCTCGA CCCCAAAAAA CTTGATTAGG GTGATGGTTC 3372

ACGTAGTGGG CCATCGCCCT GATAGACGGT TTTTCGCCTT TACTGAGCAC TCTTTAATAG 3432

TGGACTCTTG TTCCAAACTG GAACAACACT CAACCCTATC TCGGTCTATT CTTTTGATTT 3492

ATAAGATTTC CATCGCCATG TAAAAGTGTT ACAATTAGCA TTAAATTACT TCTTTATATG 3552

CTACTATTCT TTTGGCTTCG TTCACGGGGT GGGTACCGAG CTCGAATTCT GTGGAATGTG 3612

TGTCAGTTAG GGTGTGGAAA GTCCCCAGGC TCCCCAGGCA GGCAGAAGTA TGCAAAGCAT 3672

GCATCTCAAT TAGTCAGCAA CCAGGTGTGG AAAGTCCCCA GGCTCCCCAG CAGGCAGAAG 3732

TATGCAAAGC ATGCATCTCA ATTAGTCAGC AACCATAGTC CCGCCCCTAA CTCCGCCCAT 3792

CCCGCCCCTA ACTCCGCCCA GTTCCGCCCA TTCTCCGCCC CATGGCTGAC TAATTTTTTT 3852

TATTTATGCA GAGGCCGAGG CCGCCTCGGC CTCTGAGCTA TTCCAGAAGT AGTGAGGAGG 3912

CTTTTTTGGA GGCCTAGGCT TTTGCAAAAA GCTCCCGGGA GCTTGGATAT CCATTTTCGG 3972

ATCTGATCAA GAGACAGGAT GAGGATCGTT TCGCATGATT GAACAAGATG GATTGCACGC 4032

ACCTGTCGTG CCAGCTGCAT TAATGAATCG GCCAACGCGC GGGGAGAGGC GGTTTGCGTA 5352

TTGGGCGCTC TTCCGCTTCC TCGCTCACTG ACTCGCTGCG CTCGGTCGTT CGGCTGCGGC 5412

GAGCGGTATC AGCTCACTCA AAGGCGGTAA TACGGTTATC CACAGAATCA GGGGATAACG 5472

CAGGAAAGAA CATGTGAGCA AAAGGCCAGC AAAAGGCCAG GAACCGTAAA AAGGCCGCGT 5532

TGCTGGCGTT TTTCCATAGG CTCCGCCCCC CTGACGAGCA TCACAAAAAT CGACGCTCAA 5592

GTCAGAGGTG GCGAAACCCG ACAGGACTAT AAAGATACCA GGCGTTTCCC CCTGGAAGCT 5652

CCCTCGTGCG CTCTCCTGTT CCGACCCTGC CGCTTACCGG ATACCTGTCC GCCTTTCTCC 5712

CTTCGGGAAG CGTGGCGCTT TCTCAATGCT CACGCTGTAG GTATCTCAGT TCGGTGTAGG 5772

TCGTTCGCTC CAAGCTGGGC TGTGTGCACG AACCCCCCGT TCAGCCCGAC CGCTGCGCCT 5832

TATCCGGTAA CTATCGTCTT GAGTCCAACC CGGTAAGACA CGACTTATCG CCACTGGCAG 5892

CAGCCACTGG TAACAGGATT AGCAGAGCGA GGTATGTAGG CGGTGCTACA GAGTTCTTGA 5952

AGTGGTGGCC TAACTACGGC TACACTAGAA GGACAGTATT TGGTATCTGC GCTCTGCTGA 6012

AGCCAGTTAC CTTCGGAAAA AGAGTTGGTA GCTCTTGATC CGGCAAACAA ACCACCGCTG 6072

GTAGCGGTGG TTTTTTTGTT TGCAAGCAGC AGATTACGCG CAGAAAAAAA GGATCTCAAG 6132

AAGATCCTTT GATCTTTTCT ACGGGGTCTG ACGCTCAGTG GAACGAAAAC TCACGTTAAG 6192

GGATTTTGGT CATGAGATTA TCAAAAAGGA TCTTCACCTA GATCCTTTTA AATTAAAAAT 6252

GAAGTTTTAA ATCAATCTAA AGTATATATG AGTAAACTTG GTCTGACAGT TACCAATGCT 6312

TAATCAGTGA GGCACCTATC TCAGCGATCT GTCTATTTCG TTCATCCATA GTTGCCTGAC 6372

TCCCCGTCGT GTAGATAACT ACGATACGGG AGGGCTTACC ATCTGGCCCC AGTGCTGCAA 6432

TGATACCGCG AGACCCACGC TCACCGGCTC CAGATTTATC AGCAATAAAC CAGCCAGCCG 6492

GAAGGGCCGA GCGCAGAAGT GGTCCTGCAA CTTTATCCGC CTCCATCCAG TCTATTAATT 6552

GTTGCCGGGA AGCTAGAGTA AGTAGTTCGC CAGTTAATAG TTTGCGCAAC GTTGTTGCCA 6612

TTGCTACAGG CATCGTGGTG TCACGCTCGT CGTTTGGTAT GGCTTCATTC AGCTCCGGTT 6672

CCCAACGATC AAGGCGAGTT ACATGATCCC CCATGTTGTG CAAAAAAGCG GTTAGCTCCT 6732

TCGGTCCTCC GATCGTTGTC AGAAGTAAGT TGGCCGCAGT GTTATCACTC ATGGTTATGG 6792

CAGCACTGCA TAATTCTCTT ACTGTCATGC CATCCGTAAG ATGCTTTTCT GTGACTGGTG 6852

AGTACTCAAC CAAGTCATTC TGAGAATAGT GTATGCGGCG ACCGAGTTGC TCTTGCCCGG 6912

CGTCAATACG GGATAATACC GCGCCACATA GCAGAACTTT AAAAGTGCTC ATCATTGGAA 6972

AACGTTCTTC GGGGCGAAAA CTCTCAAGGA TCTTACCGCT GTTGAGATCC AGTTCGATGT 7032

AACCCACTCG TGCACCCAAC TGATCTTCAG CATCTTTTAC TTTCACCAGC GTTTCTGGGT 7092

GAGCAAAAAC AGGAAGGCAA AATGCCGCAA AAAAGGGAAT AAGGGCGACA CGGAAATGTT 7152

GAATACTCAT ACTCTTCCTT TTTCAATATT ATTGAAGCAT TTATCAGGGT TATTGTCTCA 7212

TGAGCGGATA CATATTTGAA TGTATTTAGA AAAATAAACA AATAGGGGTT CCGCGCACAT 7272

TTCCCCGAAA AGTGCCACCT GACGTC 7298

(2) INFORMATION FOR SEQ ID NO:4:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 537 amino acids

(B) TYPE: amino acid (D) TOPOLOGY: linear

(ii) MOLECULE TYPE: protein

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:4:

Met Leu Pro Gly Leu Ala Leu Leu Leu Leu Ala Ala Trp Thr Ala Arg 1 5 10 15

Ala Leu Glu Val Pro Thr Asp Gly Asn Ala Gly Leu Leu Ala Glu Pro 20 25 30

Gin He Ala Met Phe Cys Gly Arg Leu Asn Met His Met Asn Val Gin 35 40 45

Asn Gly Lys Trp Asp Ser Asp Pro Ser Gly Thr Lys Thr Cys He Asp 50 55 60

Thr Lys Glu Thr His Val Thr Gly Gly Ser Ala Gly His Thr Thr Ala 65 70 75 80

Gly Leu Val Arg Leu Leu Ser Pro Gly Ala Lys Gin Asn He Gin Leu 85 90 95

He Asn Thr Asn Gly Ser Trp His He Asn Ser Thr Ala Leu Asn Cys 100 105 110

Asn Glu Ser Leu Asn Thr Gly Trp Leu Ala Gly Leu Phe Tyr His His 115 120 125

Lys Phe Asn Ser Ser Gly Cys Pro Glu Arg Leu Ala Ser Cys Arg Arg 130 135 140

Leu Thr Asp Phe Ala Gin Gly Gly Gly Pro He Ser Tyr Ala Asn Gly 145 150 155 160

Ser Gly Leu Asp Glu Arg Pro Tyr Cys Trp His Tyr Pro Pro Arg Pro 165 170 175

Cys Gly He Val Pro Ala Lys Ser Val Cys Gly Pro Val Tyr Cys Phe 180 185 190

Thr Pro Ser Pro Val Val Val Gly Thr Thr Asp Arg Ser Gly Ala Pro 195 200 205

Thr Tyr Ser Trp Gly Ala Asn Asp Thr Asp Val Phe Val Leu Asn Asn 210 215 220

Thr Arg Pro Pro Leu Gly Asn Trp Phe Gly Cys Thr Trp Met Asn Ser 225 230 235 240

Thr Gly Phe Thr Lys Val Cys Gly Ala Pro Pro Cys Val He Gly Gly 245 250 255

Val Gly Asn Asn Thr Leu Leu Cys Pro Thr Asp Cys Phe Arg Lys His 260 265 270

Pro Glu Ala Thr Tyr Ser Arg Cys Gly Ser Gly Pro Trp He Thr Pro 275 280 285

Arg Cys Met Val Asp Tyr Pro Tyr Arg Leu Trp His Tyr Pro Cys Thr 290 295 300

He Asn Tyr Thr He Phe Lys Val Arg Met Tyr Val Gly Gly Val Glu 305 310 315 320

His Arg Leu Glu Ala Ala Cys Asn Trp Thr Arg Gly Glu Arg Cys Asp 325 330 335

Leu Glu Asp Arg Asp Arg Ser Glu Leu Ser Pro Leu Leu Leu Ser Thr 340 345 350

Thr Gin Trp Gin Val Leu Pro Cys Ser Phe Thr Thr Leu Pro Ala Leu 355 . 360 365

Ser Thr Gly Leu He His Leu His Gin Asn He Val Asp Val Gin Tyr 370 375 380

Leu Tyr Gly Val Gly Ser Ser He Ala Ser Trp Ala He Lys Trp Glu 385 390 395 400

Tyr Asp Val Leu Leu Phe Leu Leu Leu Ala Asp Ala Arg Val Cys Ser 405 410 415

Cys Leu Trp Met Met Leu Leu He Ser Gin Ala Glu Ala Ala Leu Glu 420 425 430

He Ser Glu Val Lys Met Asp Ala Glu Phe Arg His Asp Ser Gly Tyr 435 440 445

Glu Val His His Gin Lys Leu Val Phe Phe Ala Glu Asp Val Gly Ser

450 455 460

Asn Lys Gly Ala He He Gly Leu Met Val Gly Gly Val Val He Ala 465 470 475 480

Thr Val He Val He Thr Leu Val Met Leu Lys Lys Lys Gin Tyr Thr 485 490 495

Ser He His His Gly Val Val Glu Val Asp Ala Ala Val Thr Pro Glu 500 505 510

Glu Arg His Leu Ser Lys Met Gin Gin Asn Gly Tyr Glu Asn Pro Thr 515 520 525

Tyr Lys Phe Phe Glu Gin Met Gin Asn 530 535

(2) INFORMATION FOR SEQ ID NO:5:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 7106 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: circular

(ii) MOLECULE TYPE: DNA (genomic)

(ix) FEATURE:

(A) NAME/KEY: CDS

(B) LOCATION: 922..2022

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:5:

GACGGATCGG GAGATCTCCC GATCCCCTAT GGTCGACTCT CAGTACAATC TGCTCTGATG 60

CCGCATAGTT AAGCCAGTAT CTGCTCCCTG CTTGTGTGTT GGAGGTCGCT GAGTAGTGCG 120

CGAGCAAAAT TTAAGCTACA ACAAGGCAAG GCTTGACCGA CAATTGCATG AAGAATCTGC 180

TTAGGGTTAG GCGTTTTGCG CTGCTTCGCG ATGTACGGGC CAGATATACG CGTTGACATT 240

GATTATTGAC TAGTTATTAA TAGTAATCAA TTACGGGGTC ATTAGTTCAT AGCCCATATA 300

TGGAGTTCCG CGTTACATAA CTTACGGTAA ATGGCCCGCC TGGCTGACCG CCCAACGACC 360

CCCGCCCATT GACGTCAATA ATGACGTATG TTCCCATAGT AACGCCAATA GGGACTTTCC 420

ATTGACGTC ATGGGTGGAC TATTTACGGT AAACTGCCCA CTTGGCAGTA CATCAAGTGT 480

ATCATATGCC AAGTACGCCC CCTATTGACG TCAATGACGG TAAATGGCCC GCCTGGCATT 540

ATGCCCAGTA CATGACCTTA TGGGACTTTC CTACTTGGCA GTACATCTAC GTATTAGTCA 600

TCGCTATTAC CATGGTGATG CGGTTTTGGC AGTACATCAA TGGGCGTGGA TAGCGGTTTG 660

ACTCACGGGG ATTTCCAAGT CTCCACCCCA TTGACGTCAA TGGGAGTTTG TTTTGGCACC 720

AAAATCAACG GGACTTTCCA AAATGTCGTA ACAACTCCGC CCCATTGACG CAAATGGGCG 780

GTAGGCGTGT ACGGTGGGAG GTCTATATAA GCAGAGCTCT CTGGCTAACT AGAGAACCCA 840

CTGCTTAACT GGCTTATCGA AATTAATACG ACTCACTATA GGGAGACCGG AAGCTTTGCT 900

CTAGACTGGA ATTCGGGCGC G ATG CTG CCC GGT TTG GCA CTG CTC CTG CTG 951

Met Leu Pro Gly Leu Ala Leu Leu Leu Leu 1 5 10

GCC GCC TGG ACG GCT CGG GCG CTG GAG GTA CCC ACT GAT GGT AAT GCT 999 Ala Ala Trp Thr Ala Arg Ala Leu Glu Val Pro Thr Asp Gly Asn Ala 15 20 25

GGC CTG CTG GCT GAA CCC CAG ATT GCC ATG TTC TGT GGC AGA CTG AAC 1047 Gly Leu Leu Ala Glu Pro Gin He Ala Met Phe Cys Gly Arg Leu Asn 30 35 40

ATG CAC ATG AAT GTC CAG AAT GGG AAG TGG GAT TCA GAT CCA TCA GGG 1095 Met His Met Asn Val Gin Asn Gly Lys Trp Asp Ser Asp Pro Ser Gly 45 50 55

ACC AAA ACC TGC ATT GAT ACC AAG GAA ACC CAC GTC ACC GGG GGA AGT 1143 Thr Lys Thr Cys He Asp Thr Lys Glu Thr His Val Thr Gly Gly Ser 60 65 70

GCC GGC CAC ACC ACG GCT GGG CTT GTT CGT CTC CTT TCA CCA GGC GCC 1191 Ala Gly His Thr Thr Ala Gly Leu Val Arg Leu Leu Ser Pro Gly Ala 75 80 85 90

AAG CAG AAC ATC CAA CTG ATC AAC ACC AAC GGC AGT TGG CAC ATC AAT 1239 Lys Gin Asn He Gin Leu He Asn Thr Asn Gly Ser Trp His He Asn 95 100 105

AGC ACG GCC TTG AAC TGC AAT GAA AGC CTT AAC ACC GGC TGG TTA GCA 1287 Ser Thr Ala Leu Asn Cys Asn Glu Ser Leu Asn Thr Gly Trp Leu Ala 110 115 120

GGG CTC TTC TAT CAC CAC AAA TTC AAC TCT TCA GGT TGT CCT GAG AGG 1335 Gly Leu Phe Tyr His His Lys Phe Asn Ser Ser Gly Cys Pro Glu Arg 125 130 135

TTG GCC AGC TGC CGA CGC CTT ACC GAT TTT GCC CAG GGC GGG GGT CCT 1383 Leu Ala Ser Cys Arg Arg Leu Thr Asp Phe Ala Gin Gly Gly Gly Pro 140 145 150

ATC AGT TAC GCC AAC GGA AGC GGC CTC GAT GAA CGC CCC TAC TGC TGG 1431 He Ser Tyr Ala Asn Gly Ser Gly Leu Asp Glu Arg Pro Tyr Cys Trp 155 160 165 170

CAC TAC CCT CCA AGA CCT TGT GGC ATT GTG CCC GCA AAG AGC GTG TGT 1479

His Tyr Pro Pro Arg Pro Cys Gly He Val Pro Ala Lys Ser Val Cys 175 180 185

GGC CCG GTA TAT TGC TTC ACT CCC AGC CCC GTG GTG GTG GGA ACG ACC 1527 Gly Pro Val Tyr Cys Phe Thr Pro Ser Pro Val Val Val Gly Thr Thr 190 195 200

GAC AGG TCG GGC GCG CCT ACC TAC AGC TGG GGT GCA AAT GAT ACG GAT 1575 Asp Arg Ser Gly Ala Pro Thr Tyr Ser Trp Gly Ala Asn Asp Thr Asp 205 210 215

GTC TTT GTC CTT AAC AAC ACC AGG CCA CCG CTG GGC AAT TGG TTC GGT 1623 Val Phe Val Leu Asn Asn Thr Arg Pro Pro Leu Gly Asn Trp Phe Gly 220 225 230

TGC ACC TGG ATG AAC TCA ACT GGA TTC ACC AAA GTG TGC GGA GCG CCC 1671 Cys Thr Trp Met Asn Ser Thr Gly Phe Thr Lys Val Cys Gly Ala Pro 235 240 245 250

CCT TGT GTC ATC GGA GGG GTG GGC AAC AAC ACC TTG CTC TGC CCC ACT 1719 Pro Cys Val He Gly Gly Val Gly Asn Asn Thr Leu Leu Cys Pro Thr 255 260 265

GAT TGC TTC CGC AAG CAT CCG GAA GCC ACA TAC TCT CGG TGC GGC TCC 1767 Asp Cys Phe Arg Lys His Pro Glu Ala Thr Tyr Ser Arg Cys Gly Ser 270 275 280

GGT CCC TGG ATT ACA CCC AGG TGC ATG GTC GAC TAC CCG TAT AGG CTT 1815 Gly Pro Trp He Thr Pro Arg Cys Met Val Asp Tyr Pro Tyr Arg Leu 285 290 295

TGG CAC TAT CCT TGT ACC ATC AAT TAC ACC ATA TTC AAA GTC AGG ATG 1863 Trp His Tyr Pro Cys Thr He Asn Tyr Thr He Phe Lys Val Arg Met 300 305 310

TAC GTG GGA GGG GTC GAG CAC AGG CTG GAA GCG GCC TGC AAC TGG ACG 1911 Tyr Val Gly Gly Val Glu His Arg Leu Glu Ala Ala Cys Asn Trp Thr 315 320 325 330

CGG GGC GAA CGC TGT GAT CTG GAA GAC AGG GAC AGG TCC GAG CTC AGC 1959 Arg Gly Glu Arg Cys Asp Leu Glu Asp Arg Asp Arg Ser Glu Leu Ser 335 340 345

CCG TTA CTG CTG TCC ACC ACG CAG TGG CAG GTC CTT CCG TGT TCT TTC 2007 Pro Leu Leu Leu Ser Thr Thr Gin Trp Gin Val Leu Pro Cys Ser Phe 350 355 360

ACG ACC CTG CCA GCC TAGATCTCTG AAGTGAAGAT GGATGCAGAA TTCCGACATG 2062 Thr Thr Leu Pro Ala 365

ACTCAGGATA TGAAGTTCAT CATCAAAAAT TGGTGTTCTT TGCAGAAGAT GTGGGTTCAA 2122

ACAAAGGTGC AATCATTGGA CTCATGGTGG GCGGTGTTGT CATAGCGACA GTGATCGTCA 2182

TCACCTTGGT GATGCTGAAG AAGAAACAGT ACACATCCAT TCATCATGGT GTGGTGGAGG 2242

TTGACGCCGC TGTCACCCCA GAGGAGCGCC ACCTGTCCAA GATGCAGCAG AACGGCTACG 2302

AAAATCCAAC CTACAAGTTC TTTGAGCAGA TGCAGAACTA GACCCCCGCC ACAGCAGCCT 2362

CTGAAGTTGG ACAGCAAAAC CATTGCTTCA CTACCCATCG GTGTCCATTT ATAGAATAAT 2422

GTGGGAAGAA ACAAACCCGT TTTATGATTT ACTCATTATC GCCTTTTGAC AGCTGTGCTG 2482

TAACACAAGT AGATGCCTGA ACTTGAATTA ATCCACACAT CAGTAATGTA TTCTATCTCT 2542

CTTTACATTT TGGTCTCTAT ACTACATTAT TAATGGGTTT TGTGTACTGT AAAGAATTTA 2602

GCTGTATCAA ACTAGTGCAT GAATAGGCCG CTCGAGCATG CATCTAGAGG GCCCTATTCT 2662

ATAGTGTCAC CTAAATGCTC GCTGATCAGC CTCGACTGTG CCTTCTAGTT GCCAGCCATC 2722

TGTTGTTTGC CCCTCCCCCG TGCCTTCCTT GACCCTGGAA GGTGCCACTC CCACTGTCCT 2782

TTCCTAATAA AATGAGGAAA TTGCATCGCA TTGTCTGAGT AGGTGTCATT CTATTCTGGG 2842

GGGTGGGGTG GGGCAGGACA GCAAGGGGGA GGATTGGGAA GACAATAGCA GGCATGCTGG 2902

GGATGCGGTG GGCTCTATGG AACCAGCTGG GGCTCGAGGG GGGATCCCCA CGCGCCCTGT 2962

AGCGGCGCAT TAAGCGCGGC GGGTGTGGTG GTTACGCGCA GCGTGACCGC TACACTTGCC 3022

AGCGCCCTAG CGCCCGCTCC TTTCGCTTTC TTCCCTTCCT TTCTCGCCAC GTTCGCCGGC 3082

TTTCCCCGTC AAGCTCTAAA TCGGGGCATC CCTTTAGGGT TCCGATTTAG TGCTTTACGG 3142

CACCTCGACC CCAAAAAACT TGATTAGGGT GATGGTTCAC GTAGTGGGCC ATCGCCCTGA 3202

TAGACGGTTT TTCGCCTTTA CTGAGCACTC TTTAATAGTG GACTCTTGTT CCAAACTGGA 3262

ACAACACTCA ACCCTATCTC GGTCTATTCT TTTGATTTAT AAGATTTCCA TCGCCATGTA 3322

AAAGTGTTAC AATTAGCATT AAATTACTTC TTTATATGCT ACTATTCTTT TGGCTTCGTT 3382

CACGGGGTGG GTACCGAGCT CGAATTCTGT GGAATGTGTG TCAGTTAGGG TGTGGAAAGT 3442

CCCCAGGCTC CCCAGGCAGG CAGAAGTATG CAAAGCATGC ATCTCAATTA GTCAGCAACC 3502

AGGTGTGGAA AGTCCCCAGG CTCCCCAGCA GGCAGAAGTA TGCAAAGCAT GCATCTCAAT 3562

TAGTCAGCAA CCATAGTCCC GCCCCTAACT CCGCCCATCC CGCCCCTAAC TCCGCCCAGT 3622

TCCGCCCATT CTCCGCCCCA TGGCTGACTA ATTTTTTTTA TTTATGCAGA GGCCGAGGCC 3682

GCCTCGGCCT CTGAGCTATT CCAGAAGTAG TGAGGAGGCT TTTTTGGAGG CCTAGGCTTT 3742

TGCAAAAAGC TCCCGGGAGC TTGGATATCC ATTTTCGGAT CTGATCAAGA GACAGGATGA 3802

GGATCGTTTC GCATGATTGA ACAAGATGGA TTGCACGCAG GTTCTCCGGC CGCTTGGGTG 3862

GAGAGGCTAT TCGGCTATGA CTGGGCACAA CAGACAATCG GCTGCTCTGA TGCCGCCGTG 3922 TTCCGGCTGT CAGCGCAGGG GCGCCCGGTT CTTTTTGTCA AGACCGACCT GTCCGGTGCC 3982 CTGAATGAAC TGCAGGACGA GGCAGCGCGG CTATCGTGGC TGGCCACGAC GGGCGTTCCT 4042 TGCGCAGCTG TGCTCGACGT TGTCACTGAA GCGGGAAGGG ACTGGCTGCT ATTGGGCGAA 4102 GTGCCGGGGC AGGATCTCCT GTCATCTCAC CTTGCTCCTG CCGAGAAAGT ATCCATCATG 4162 GCTGATGCAA TGCGGCGGCT GCATACGCTT GATCCGGCTA CCTGCCCATT CGACCACCAA 4222 GCGAAACATC GCATCGAGCG AGCACGTACT CGGATGGAAG CCGGTCTTGT CGATCAGGAT 4282 GATCTGGACG AAGAGCATCA GGGGCTCGCG CCAGCCGAAC TGTTCGCCAG GCTCAAGGCG 4342 CGCATGCCCG ACGGCGAGGA TCTCGTCGTG ACCCATGGCG ATGCCTGCTT GCCGAATATC 4402 ATGGTGGAAA ATGGCCGCTT TTCTGGATTC ATCGACTGTG GCCGGCTGGG TGTGGCGGAC 4462 CGCTATCAGG ACATAGCGTT GGCTACCCGT GATATTGCTG AAGAGCTTGG CGGCGAATGG 4522 GCTGACCGCT TCCTCGTGCT TTACGGTATC GCCGCTCCCG ATTCGCAGCG CATCGCCTTC 4582 TATCGCCTTC TTGACGAGTT CTTCTGAGCG GGACTCTGGG GTTCGAAATG ACCGACCAAG 4642 CGACGCCCAA CCTGCCATCA CGAGATTTCG ATTCCACCGC CGCCTTCTAT GAAAGGTTGG 4702 GCTTCGGAAT CGTTTTCCGG GACGCCGGCT GGATGATCCT CCAGCGCGGG GATCTCATGC 4762 TGGAGTTCTT CGCCCACCCC AACTTGTTTA TTGCAGCTTA TAATGGTTAC AAATAAAGCA 4822 ATAGCATCAC AAATTTCACA AATAAAGCAT TTTTTTCACT GCATTCTAGT TGTGGTTTGT 4882

CCAAACTCAT CAATGTATCT TATCATGTCT GGATCCCGTC GACCTCGAGA GCTTGGCGTA 4942 ATCATGGTCA TAGCTGTTTC CTGTGTGAAA TTGTTATCCG CTCACAATTC CACACAACAT 5002 ACGAGCCGGA AGCATAAAGT GTAAAGCCTG GGGTGCCTAA TGAGTGAGCT AACTCACATT 5062 AATTGCGTTG CGCTCACTGC CCGCTTTCCA GTCGGGAAAC CTGTCGTGCC AGCTGCATTA 5122 ATGAATCGGC CAACGCGCGG GGAGAGGCGG TTTGCGTATT GGGCGCTCTT CCGCTTCCTC 5182 GCTCACTGAC TCGCTGCGCT CGGTCGTTCG GCTGCGGCGA GCGGTATCAG CTCACTCAAA 5242

GGCGGTAATA CGGTTATCCA CAGAATCAGG GGATAACGCA GGAAAGAACA TGTGAGCAAA 5302 AGGCCAGCAA AAGGCCAGGA ACCGTAAAAA GGCCGCGTTG CTGGCGTTTT TCCATAGGCT 5362 CCGCCCCCCT GACGAGCATC ACAAAAATCG ACGCTCAAGT CAGAGGTGGC GAAACCCGAC 5422 AGGACTATAA AGATACCAGG CGTTTCCCCC TGGAAGCTCC CTCGTGCGCT CTCCTGTTCC 5482

GACCCTGCCG CTTACCGGAT ACCTGTCCGC CTTTCTCCCT TCGGGAAGCG TGGCGCTTTC 5542

TCAATGCTCA CGCTGTAGGT ATCTCAGTTC GGTGTAGGTC GTTCGCTCCA AGCTGGGCTG 5602

TGTGCACGAA CCCCCCGTTC AGCCCGACCG CTGCGCCTTA TCCGGTAACT ATCGTCTTGA 5662

GTCCAACCCG GTAAGACACG ACTTATCGCC ACTGGCAGCA GCCACTGGTA ACAGGATTAG 5722

CAGAGCGAGG TATGTAGGCG GTGCTACAGA GTTCTTGAAG TGGTGGCCTA ACTACGGCTA 5782

CACTAGAAGG ACAGTATTTG GTATCTGCGC TCTGCTGAAG CCAGTTACCT TCGGAAAAAG 5842

AGTTGGTAGC TCTTGATCCG GCAAACAAAC CACCGCTGGT AGCGGTGGTT TTTTTGTTTG 5902

CAAGCAGCAG ATTACGCGCA GAAAAAAAGG ATCTCAAGAA GATCCTTTGA TCTTTTCTAC 5962

GGGGTCTGAC GCTCAGTGGA ACGAAAACTC ACGTTAAGGG ATTTTGGTCA TGAGATTATC 6022

AAAAAGGATC TTCACCTAGA TCCTTTTAAA TTAAAAATGA AGTTTTAAAT CAATCTAAAG 6082

TATATATGAG TAAACTTGGT CTGACAGTTA CCAATGCTTA ATCAGTGAGG CACCTATCTC 6142

AGCGATCTGT CTATTTCGTT CATCCATAGT TGCCTGACTC CCCGTCGTGT AGATAACTAC 6202

GATACGGGAG GGCTTACCAT CTGGCCCCAG TGCTGCAATG ATACCGCGAG ACCCACGCTC 6262

ACCGGCTCCA GATTTATCAG CAATAAACCA GCCAGCCGGA AGGGCCGAGC GCAGAAGTGG 6322

TCCTGCAACT TTATCCGCCT CCATCCAGTC TATTAATTGT TGCCGGGAAG CTAGAGTAAG 6382

TAGTTCGCCA GTTAATAGTT TGCGCAACGT TGTTGCCATT GCTACAGGCA TCGTGGTGTC 6442

ACGCTCGTCG TTTGGTATGG CTTCATTCAG CTCCGGTTCC CAACGATCAA GGCGAGTTAC 6502

ATGATCCCCC ATGTTGTGCA AAAAAGCGGT TAGCTCCTTC GGTCCTCCGA TCGTTGTCAG 6562

AAGTAAGTTG GCCGCAGTGT TATCACTCAT GGTTATGGCA GCACTGCATA ATTCTCTTAC 6622

TGTCATGCCA TCCGTAAGAT GCTTTTCTGT GACTGGTGAG TACTCAACCA AGTCATTCTG 6682

AGAATAGTGT ATGCGGCGAC CGAGTTGCTC TTGCCCGGCG TCAATACGGG ATAATACCGC 6742

GCCACATAGC AGAACTTTAA AAGTGCTCAT CATTGGAAAA CGTTCTTCGG GGCGAAAACT 6802

CTCAAGGATC TTACCGCTGT TGAGATCCAG TTCGATGTAA CCCACTCGTG CACCCAACTG 6862

ATCTTCAGCA TCTTTTACTT TCACCAGCGT TTCTGGGTGA GCAAAAACAG GAAGGCAAAA _. 6922

TGCCGCAAAA AAGGGAATAA GGGCGACACG GAAATGTTGA ATACTCATAC TCTTCCTTTT 6982

TCAATATTAT TGAAGCATTT ATCAGGGTTA TTGTCTCATG AGCGGATACA TATTTGAATG 7042

TATTTAGAAA AATAAACAAA TAGGGGTTCC GCGCACATTT CCCCGAAAAG TGCCACCTGA 7102

CGTC 7106

(2) INFORMATION FOR SEQ ID NO:6:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 367 arr.ino acids

(B) TYPE: amino acid (D) TOPOLOGY: linear

(ii) MOLECULE TYPE: protein

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:6:

Met Leu Pro Gly Leu Ala Leu Leu Leu Leu Ala Ala Trp Thr Ala Arg 1 5 10 15

Ala Leu Glu Val Pro Thr Asp Gly Asn Ala Gly Leu Leu Ala Glu Pro 20 25 30

Gin He Ala Met Phe Cys Gly Arg Leu Asn Met His Met Asn Val Gin 35 40 45

Asn Gly Lys Trp Asp Ser Asp Pro Ser Gly Thr Lys Thr Cys He Asp 50 55 60

Thr Lys Glu Thr His Val Thr Gly Gly Ser Ala Gly His Thr Thr Ala 65 70 75 80

Gly Leu Val Arg Leu Leu Ser Pro Gly Ala Lys Gin Asn He Gin Leu 85 90 95

He Asn Thr Asn Gly Ser Trp His He Asn Ser Thr Ala Leu Asn Cys 100 105 110

Asn Glu Ser Leu Asn Thr Gly Trp Leu Ala Gly Leu Phe Tyr His His 115 120 125

Lys Phe Asn Ser Ser Gly Cys Pre Glu Arg Leu Ala Ser Cys Arg Arg 130 135 140

Leu Thr Asp Phe Ala Gin Gly Gly Gly Pro He Ser Tyr Ala Asn Gly 145 150 155 160

Ser Gly Leu A.sp Glu Arg Pro Tyr Cys Trp His Tyr Pro Pro Arg Pro 165 170 175

Cys Gly He Val Pro Ala Lys Ser Val Cys Gly Pro Val Tyr Cys Phe 180 185 190

Thr Pro Ser Pro Val Val Val Gly Thr Thr Asp Arg Ser Gly Ala Pro 195 200 205

Thr Tyr Ser Trp Gly Ala Asn Asp Thr Asp Val Phe Val Leu Asn Asn 210 215 220

Thr Arg Pro Pro Leu Gly Asn Trp Phe Gly Cys Thr Trp Met Asn Ser 225 230 235 240

Thr Gly Phe Thr Lys Val Cys Gly Ala Pro Pro Cys Val He Gly Gly 245 250 255

Val Gly Asn Asn Thr Leu Leu Cys Pro Thr Asp Cys Phe Arg Lys His 260 265 270

Pro Glu Ala Thr Tyr Ser Arg Cys Gly Ser Gly Pro Trp He Thr Pro 275 280 285

Arg Cys Met Val Asp Tyr Pro Tyr Arg Leu Trp His Tyr Pro Cys Thr 290 295 300

He Asn Tyr Thr He Phe Lys Val Arg Met Tyr Val Gly Gly Val Glu 305 310 315 320

His Arg Leu Glu Ala Ala Cys Asn Trp Thr Arg Gly Glu Arg Cys Asp 325 330 335

Leu Glu Asp Arg Asp Arg Ser Glu Leu Ser Pro Leu Leu Leu Ser Thr 340 345 350

Thr Gin Trp Gin Val Leu Pro Cys Ser Phe Thr Thr Leu Pro Ala 355 360 365

(2) INFORMATION FOR SEQ ID NO:7 :

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 4810 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: circular

(ii) MOLECULE TYPE: DNA (genomic)

Jx) FEATURE:

(A) NAME/KEY: CDS

(B) LOCATION: 2227..2910

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:7 :

GCGTAATCTG CTGCTTGCAA ACAAAAAAAC CACCGCTACC AGCGGTGGTT TGTTTGCCGG 60

ATCAAGAGCT ACCAACTCTT TTTCCGAAGG TAACTGGCTT CAGCAGAGCG CAGATACCAA 120

ATACTGTCCT TCTAGTGTAG CCGTAGTTAG GCCACCACTT CAAGAACTCT GTAGCACCGC 180

CTACATACCT CGCTCTGCTA ATCCTGTTAC CAGTGGCTGC TGCCAGTGGC GATAAGTCGT 240

GTCTTACCGG GTTGGACTCA AGACGATAGT TACCGGATAA GGCGCAGCGG TCGGGCTGAA 300

CCAAGTCTCC ACCCCATTGA CGTCAATGGG AGTTTGTTTT GGCACCAAAA TCAACGGGAC 2040

TTTCCAAAAT GTCGTAACAA CTCCGCCCCA TTGACGCAAA TGGGCGGTAG GCGTGTACGG 2100

TGGGAGGTCT ATATAAGCAG AGCTCTCTGG CTAACTAGAG AACCCACTGC TTAACTGGCT 2160

TATCGAAATT AATACGACTC ACTATAGGGA GACCGGAAGC TTGGTACCGA GCTCGGATCT 2220

GCCACC ATG GCA ACA GGA TCA AGA ACA TCA CTG CTG CTG GCA TTT GGA 2268 Met Ala Thr Gly Ser Arg Thr Ser Leu Leu Leu Ala Phe Gly 1 5 10

CTG CTG TGT CTG CCA TGG CTG CAA GAA GGA TCA GCA GCA GCA GCA GCG 2316

Leu Leu Cys Leu Pro Trp Leu Gin Glu Gly Ser Ala Ala Ala Ala Ala

15 20 25 30

AAT TCG GAT CCC TAC CAA GTG CGC AAT TCC TCG GGG CTT TAC CAT GTC 2364

Asn Ser Asp Pro Tyr Gin Val Arg Asn Ser Ser Gly Leu Tyr His Val

35 40 45

ACC AAT GAT TGC CCT AAT TCG AGT ATT GTG TAC GAG GCG GCC GAT GCC 2412

Thr Asn Asp Cys Pro Asn Ser Ser He Val Tyr Glu Ala Ala Asp Ala

50 55 60

ATC CTA CAC ACT CCG GGG TGT GTC CCT TGC GTT CGC GAG GGT AAC GCC 2460

He Leu His Thr Pro Gly Cys Val Pro Cys Val Arg Glu Gly Asn Ala

65 70 75

TCG AGG TGT TGG GTG GCG GTG ACC CCC ACG GTG GCC ACC AGG GAC GGC 2508

Ser Arg Cys Trp Val Ala Val Thr Pro Thr Val Ala Thr Arg Asp Gly 80 85 90

AAA CTC CCC ACA ACG CAG CTT CGA CGT CAT ATC GAT CTG CTC GTC GGG 2556

Lys Leu Pro Thr Thr Gin Leu Arg Arg His He Asp Leu Leu Val Gly

95 100 105 110

AGC GCC ACC CTC TGC TCG GCC CTC TAC GTG GGG GAC CTG TGC GGG TCT 2604

Ser Ala Thr Leu Cys Ser Ala Leu Tyr Val Gly Asp Leu Cys Gly Ser

115 120 125

GTC TTT CTT GTT GGT CAA CTG TTT ACC TTC TCT CCC AGG CGC CAC TGG 2652

Val Phe Leu Val Gly Gin Leu Phe Thr Phe Ser Pro Arg Arg His Trp

130 135 140

ACG ACG CAA GAC TGC AAT TGT TCT ATC TAT CCC GG" CAT ATA ACG GGT 2700

Thr Thr Gin Asp Cys Asn Cys Ser He Tyr Pro G.. His He Thr Gly

145 150 155

CAT CGT ATG GCA TGG GAT ATG ATG ATG AAC TGG TCC CCT ACG GCA GCG 2748

His Arg Met Ala Trp Asp Met Met Met Asn Trp Ser Pro Thr Ala Ala 160 165 170

TTG GTG GTA GCT CAG CTG CTC CGG ATC CCA CAA GCC ATC TTG GAC ATG 2796

Leu Val Val Ala Gin Leu Leu Arg He Pro Gin Ala He Leu Asp Met

175 180 185 190

ATC GCT GGT GCC CAC TGG GGA GTC CTG GCG GGC ATA GCG TAT TTC TCC 2844 He Ala Gly Ala His Trp Gly Val Leu Ala Gly He Ala Tyr Phe Ser 195 200 205

ATG GTG GGG AAC TGG GCG AAG GTC CTG GTA GTG CTG CTG CTA TTT GCC 2892 Met Val Gly Asn Trp Ala Lys Val Leu Val Val Leu Leu Leu Phe Ala 210 215 220

GGC GTT GAC GCG GAG ATC TAATCTAGAG GGCCCTATTC TATAGTGTCA 2940 Gly Val Asp Ala Glu He 225

CCTAAATGCT AGAGGATCTT TGTGAAGGAA CCTTACTTCT GTGGTGTGAC ATAATTGGAC 3000

AAACTACCTA CAGAGATTTA AAGCTCTAAG GTAAATATAA AATTTTTAAG TGTATAATGT 3060

GTTAAACTAC TGATTCTAAT TGTTTGTGTA TTTTAGATTC CAACCTATGG AACTGATGAA 3120

TGGGAGCAGT GGTGGAATGC CTTTAATGAG GAAAACCTGT TTTGCTCAGA AGAAATGCCA 3180

TCTAGTGATG ATGAGGCTAC TGCTGACTCT CAACATTCTA CTCCTCCAAA AAAGAAGAGA 3240

AAGGTAGAAG ACCCCAAGGA CTTTCCTTCA GAATTGCTAA GTTTTTTGAG TCATGCTGTG 3300

TTTAGTAATA GAACTCTTGC TTGCTTTGCT ATTTACACCA CAAAGGAAAA AGCTGCACTG 3360

CTATACAAGA AAATTATGGA AAAATATTCT GTAACCTTTA TAAGTAGGCA TAACAGTTAT 3420

AATCATAACA TACTGTTTTT TCTTACTCCA CACAGGCATA GAGTGTCTGC TATTAATAAC 3480

TATGCTCAAA AATTGTGTAC CTTTAGCTTT TTAATTTGTA AAGGGGTTAA TAAGGAATAT 3540

TTGATGTATA GTGCCTTGAC TAGAGATC T AATCAGCCAT ACCACATTTG TAGAGGTTTT 3600

ACTTGCTTTA AAAAACCTCC CACACCTCCC CCTGAACCTG AAACATAAAA TGAATGCAAT 3660

TGTTGTTGTT AACTTGTTTA TTGCAGCTTA TAATGGTTAC AAATAAAGCA ATAGCATCAC 3720

AAATTTCACA AATAAAGCAT TTTTTTCACT GCATTCTAGT TGTGGTTTGT CCAAACTCAT 3780

CAATGTATCT TATCATGTCT GGATCGATCC CGCCATGGTA TCAACGCCAT ATTTCTATTT 3840

ACAGTAGGGA CCTCTTCGTT GTGTAGGTAC CGCTGTATTC CTAGGGAAAT AGTAGAGGCA 3900

CCTTGAACTG TCTGCATCAG CCATATAGCC CCCGCTGTTC GACTTACAAA CACAGGCACA 3960

GTACTGACAA ACCCATACAC CTCCTCTGAA ATACCCATAG TTGCTAGGGC TGTCTCCGAA 4020

CTCATTACAC CCTCCAAAGT CAGAGCTGTA ATTTCGCCAT CAAGGGCAGC GAGGGCTTCT 4080

CCAGATAAAA TAGCTTCTGC CGAGAGTCCC GTAAGGGTAG ACACTTCAGC TAATCCCTCG 4140

ATGAGGTCTA CTAGAATAGT CAGTGCGGCT CCCATTTTGA AAATTCACTT ACTTGATCAG 4200

CTTCAGAAGA TGGCGGAGGG CCTCCAACAC AGTAATTTTC CTCCCGACTC TTAAAATAGA 4260

AAATGTCAAG TCAGTTAAGC AGGAAGTGGA CTAACTGACG CAGCTGGCCG TGCGACATCC 4320

TCTTTTAATT AGTTGCTAGG CAACGCCCTC CAGAGGGCGT GTGGTTTTGC AAGAGGAAGC 4380

AAAAGCCTCT CCACCCAGGC CTAGAATGTT TCCACCCAAT CATTACTATG ACAACAGCTG 4440

TTTTTTTTAG TATTAAGCAG AGGCCGGGGA CCCCTGGCCC GCTTACTCTG GAGAAAAAGA 4500

AGAGAGGCAT TGTAGAGGCT TCCAGAGGCA ACTTGTCAAA ACAGGACTGC TTCTATTTCT 4560

GTCACACTGT CTGGCCCTGT CACAAGGTCC AGCACCTCCA TACCCCCTTT AATAAGCAGT 4620

TTGGGAACGG GTGCGGGTCT TACTCCGCCC ATCCCGCCCC TAACTCCGCC CAGTTCCGCC 4680

CATTCTCCGC CCCATGGCTG ACTAATTTTT TTTATTTATG CAGAGGCCGA GGCCGCCTCG 4740

GCCTCTGAGC TATTCCAGAA GTAGTGAGGA GGCTTTTTTG GAGGCCTAGG CTTTTGCAAA 4800

AAGCTAATTC 4810

(2) INFORMATION FOR SEQ ID' NO:8:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 228 amino acids

(B) TYPE: amino acid (D) TOPOLOGY: linear

(ii) MOLECULE TYPE: protein

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:8:

Met Ala Thr Gly Ser Arg Thr Ser Leu Leu Leu Ala Phe Gly Leu Leu 1 5 10 15

Cys Leu Pro Trp Leu Gin Glu Gly Ser Ala Ala Ala Ala Ala Asn Ser 20 25 30

Asp Pro Tyr Gin Val Arg Asn Ser Ser Gly Leu Tyr His Val Thr Asn 35 40 45

Asp Cys Pro Asn Ser Ser He Val Tyr Glu Ala Ala Asp Ala He Leu 50 55 60

His Thr Pro Gly Cys Val Pro Cys Val Arg Glu Gly Asn Ala Ser Arg 65 70 75 80

Cys Trp Val Ala Val Thr Pro Thr Val Ala Thr Arg Asp Gly Lys Leu 85 90 95

Pro Thr Thr Gin Leu Arg Arg His He Asp Leu Leu Val Gly Ser Ala 100 105 ^' 110

Thr Leu Cys Ser Ala Leu Tyr Val Gly Asp Leu Cys Gly Ser Val Phe 115 120 125

Leu Val Gly Gin Leu Phe Thr Phe Ser Pro Arg Arg His Trp Thr Thr 130 135 140

Gin Asp Cys Asn Cys Ser He Tyr Pro Gly His He Thr Gly His Arg 145 150 155 160

Met Ala Trp Asp Met Met Met Asn Trp Ser Pro Thr Ala Ala Leu Val 165 170 175

Val Ala Gin Leu Leu Arg He Pro Gin Ala He Leu Asp Met He Ala 180 185 190

Gly Ala His Trp Gly Val Leu Ala Gly He Ala Tyr Phe Ser Met Val 195 200 205

Gly Asn Trp Ala Lys Val Leu Val Val Leu Leu Leu Phe Ala Gly Val 210 215 220

Asp Ala Glu He 225

(2) INFORMATION FOR SEQ ID NO:9:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 5323 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: circular

(ii) MOLECULE TYPE: DNA (genomic)

(ix) FEATURE:

(A) NAME/KEY: CDS

(B) LOCATION: 2227.-3423

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:9 :

GCGTAATCTG CTGCTTGCAA ACAAAAAAAC CACCGCTACC AGCGGTGGTT TGTTTGCCGG 60

ATCAAGAGCT ACCAACTCTT TTTCCGAAGG TAACTGGCTT CAGCAGAGCG CAGATACCAA 120

ATACTGTCCT TCTAGTGTAG CCGTAGTTAG GCCACCACTT CAAGAACTCT GTAGCACCGC 180

CTACATACCT CGCTCTGCTA ATCCTGTTAC CAGTGGCTGC TGCCAGTGGC GATAAGTCGT 240

GTCTTACCGG GTTGGACTCA AGACGATAGT TACCGGATAA GGCGCAGCGG TCGGGCTGAA 300

CGGGGGGTTC GTGCACACAG CCCAGCTTGG AGCGAACGAC CTACACCGAA CTGAGATACC 360

TACAGCGTGA GCATTGAGAA AGCGCCACGC TTCCCGAAGG GAGAAAGGCG GACAGGTATC 420

CGGTAAGCGG CAGGGTCGGA ACAGGAGAGC GCACGAGGGA GCTTCCAGGG GGAAACGCCT 480

GGTATCTTTA TAGTCCTGTC GGGTTTCGCC ACCTCTGACT TGAGCGTCGA TTTTTGTGAT 540

GCTCGTCAGG GGGGCGGAGC CTATGGAAAA ACGCCAGCAA CGCAAGCTAG CTTCTAGCTA 600

GAAATTGTAA ACGTTAATAT TTTGTTAAAA TTCGCGTTAA ATTTTTGTTA AATCAGCTCA 660

TTTTTTAACC AATAGGCCGA AATCGGCAAA ATCCCTTATA AATCAAAAGA ATAGCCCGAG 720

ATAGGGTTGA GTGTTGTTCC AGTTTGGAAC AAGAGTCCAC TATTAAAGAA CGTGGACTCC 780

AACGTCAAAG GGCGAAAAAC CGTCTATCAG GGCGATGGCC GCCCACTACG TGAACCATCA 840

CCCAAATCAA GTTTTTTGGG GTCGAGGTGC CGTAAAGCAC TAAATCGGAA CCCTAAAGGG 900

AGCCCCCGAT TTAGAGCTTG ACGGGGAAAG CCGGCGAACG TGGCGAGAAA GGAAGGGAAG 960

AAAGCGAAAG GAGCGGGCGC TAGGGCGCTG GCAAGTGTAG CGGTCACGCT GCGCGTAACC 1020

ACCACACCCG CCGCGCTTAA TGCGCCGCTA CAGGGCGCGT ACTATGGTTG CTTTGACGAG 1080

ACCGTATAAC GTGCTTTCCT CGTTGGAATC AGAGCGGGAG CTAAACAGGA GGCCGATTAA 1140

AGGGATTTTA GACAGGAACG GTACGCCAGC TGGATCACCG CGGTCTTTCT CAACGTAACA 1200

CTTTACAGCG GCGCGTCATT TGATATGATG CGCCCCGCTT CCCGATAAGG GAGCAGGCCA 1260

GTAAAAGCAT TACCCGTGGT GGGGTTCCCG AGCGGCCAAA GGGAGCAGAC TCTAAATCTG 1320

CCGTCATCGA CTTCGAAGGT TCGAATCCTT CCCCCACCAC CATCACTTTC AAAAGTCCGA 1380

AAGAATCTGC TCCCTGCTTG TGTGTTGGAG GTCGCTGAGT AGTGCGCGAG TAAAATTTAA 1440

GCTACAACAA GGCAAGGCTT GACCGACAAT TGCATGAAGA ATCTGCTTAG GGTTAGGCGT 1500

TTTGCGCTGC TTCGCGATGT ACGGGCCAGA TATACGCGTT GACATTGATT ATTGACTAGT 1560

TATTAATAGT AATCAATTAC GGGGTCATTA GTTCATAGCC CATATATGGA GTTCCGCGTT 1620

ACATAACTTA CGGTAAATGG CCCGCCTGGC TGACCGCCCA ACGACCCCCG CCCATTGACG 1680

TCAATAATGA CGTATGTTCC CATAGTAACG CCAATAGGGA CTTTCCATTG ACGTCAATGG 1740

GTGGACTATT TACGGTAAAC TGCCCACTTG GCAGTACATC AAGTGTATCA TATGCCAAGT 1800

ACGCCCCCTA TTGACGTCAA TGACGGTAAA TGGCCCGCCT GGCATTATGC CCAGTACATG 1860

ACCTTATGGG ACTTTCCTAC TTGGCAGTAC ATCTACGTAT TAGTCATCGC TATTACCATG 1920

GTGATGCGGT TTTGGCAGTA CATCAATGGG CGTGGATAGC GGTTTGACTC ACGGGGATTT 1980

CCAAGTCTCC ACCCCATTGA CGTCAATGGG AGTTTGTTTT GGCACCAAAA TCAACGGGAC 2040

TTTCCAAAAT GTCGTAACAA. CTCCGCCCCA TTGACGCAAA TGGGCGGTAG GCGTGTACGG 2100

TGGGAGGTCT ATATAAGCAG AGCTCTCTGG CTAACTAGAG AACCCACTGC TTAACTGGCT 2160

TATCGAAATT AATACGACTC ACTATAGGGA GACCGGAAGC TTGGTACCGA GCTCGGATCT 2220

GCCACC ATG GCA ACA GGA TCA AGA ACA TCA CTG CTG CTG GCA TTT GGA 2268 Met Ala Thr Gly Ser Arg Thr Ser Leu Leu Leu Ala Phe Gly 1 5 10

CTG CTG TGT CTG CCA TGG CTG CAA GAA GGA TCA GCA GCA GCA GCA GCG 2316 Leu Leu Cys Leu Pro Trp Leu Gin Glu Gly Ser Ala Ala Ala Ala Ala 15 20 25 30

AAT TCA GAA ACC CAC GTC ACC GGG GGA AGT GCC GGC CAC ACC ACG GCT 2364 Asn Ser Glu Thr His Val Thr Gly Gly Ser Ala Gly His Thr Thr Ala 35 40 45

GGG CTT GTT CGT CTC CTT TCA CCA GGC GCC AAG CAG AAC ATC CAA CTG 2412 Gly Leu Val Arg Leu Leu Ser Pro Gly Ala Lys Gin Asn He Gin Leu 50 55 60

ATC AAC ACC AAC GGC AGT TGG CAC ATC AAT AGC ACG GCC TTG AAC TGC 2 60 He Asn Thr Asn Gly Ser Trp His He Asn Ser Thr Ala Leu Asn Cys 65 70 75

AAT GAA AGC CTT AAC ACC GGC TGG TTA GCA GGG CTC TTC TAT CAC CAC 2508 Asn Glu Ser Leu Asn Thr Gly Trp Leu Ala Gly Leu Phe Tyr His His 80 85 90

AAA TTC AAC TCT TCA GGT TGT CCT GAG AGG TTG GCC AGC TGC CGA CGC 2556 Lys Phe Asn Ser Ser Gly Cys Pro Glu Arg Leu Ala Ser Cys Arg Arg 95 100 105 110

CTT ACC GAT TTT GCC CAG GGC GGG GGT CCT ATC AGT TAC GCC AAC GGA 2604 Leu Thr Asp Phe Ala Gin Gly Gly Gly Pro He Ser Tyr Ala Asn Gly 115 120 125

AGC GGC CTC GAT GAA CGC CCC TAC TGC TGG CAC TAC CCT CCA AGA CCT 2652 Ser Gly Leu Asp Glu Arg Pro Tyr Cys Trp His Tyr Pro Pro Arg Pro 130 135 140 ^"

TGT GGC ATT GTG CCC GCA AAG AGC GTG TGT GGC CCG GTA TAT TGC TTC 2700 Cys Gly He Val Pro Ala Lys Ser Val Cys Gly Pro Val Tyr Cys Phe 145 150 155

ACT CCC AGC CCC GTG GTG GTG GGA ACG ACC GAC AGG TCG GGC GCG CCT 2748 Thr Pro Ser Pro Val Val Val Gly Thr Thr Asp Arg Ser Gly Ala Pro 160 165 170

ACC TAC AGC TGG GGT GCA AAT GAT ACG GAT GTC TTT GTC CTT AAC AAC 2796 Thr Tyr Ser Trp Gly Ala Asn Asp Thr Asp Val Phe Val Leu Asn Asn 175 180 185 190

ACC AGG CCA CCG CTG GGC AAT TGG TTC GGT TGC ACC TGG ATG AAC TCA 2844 Thr Arg Pro Pro Leu Gly Asn Trp Phe Gly Cys Thr Trp Met Asn Ser 195 200 205

ACT GGA TTC ACC AAA GTG TGC GGA GCG CCC CCT TGT GTC ATC GGA GGG 2892 Thr Gly Phe Thr Lys Val Cys Gly Ala Pro Pro Cys Val He Gly Gly 210 215 220

GTG GGC AAC AAC ACC TTG CTC TGC CCC ACT GAT TGC TTC CGC AAG CAT 2940 Val Gly Asn Asn Thr Leu Leu Cys Pro Thr Asp Cys Phe Arg Lys His 225 230 235

CCG GAA GCC ACA TAC TCT CGG TGC GGC TCC GGT CCC TGG ATT ACA CCC 2988 Pro Glu Ala Thr Tyr Ser Arg Cys Gly Ser Gly Pro Trp He Thr Pro 240 245 250

AGG TGC ATG GTC GAC TAC CCG TAT AGG CTT TGG CAC TAT CCT TGT ACC 3036 Arg Cys Met Val Asp Tyr Pro Tyr Arg Leu Trp His Tyr Pro Cys Thr 255 260 265 270

ATC AAT TAC ACC ATA TTC AAA GTC AGG ATG TAC GTG GGA GGG GTC GAG 3084 He Asn Tyr Thr He Phe Lys Val Arg Met Tyr Val Gly Gly Val Glu 275 280 285

CAC AGG CTG GAA GCG GCC TGC AAC TGG ACG CGG GGC GAA CGC TGT GAT 3132 His Arg Leu Glu Ala Ala Cys Asn Trp Thr Arg Gly Glu Arg Cys Asp 290 295 300

CTG GAA GAC AGG GAC AGG TCC GAG CTC AGC CCG TTA CTG CTG TCC ACC 3180 Leu Glu Asp Arg Asp Arg Ser Glu Leu Ser Pro Leu Leu Leu Ser Thr 305 310 315

ACG CAG TGG CAG GTC CTT CCG TGT TCT TTC ACG ACC CTG CCA GCC TTG 3228 Thr Gin Trp Gin Val Leu Pro Cys Ser Phe Thr Thr Leu Pro Ala Leu 320 325 330

TCC ACC GGC CTC ATC CAC CTC CAC CAG AAC ATT GTG GAC GTG CAG TAC 3276 Ser Thr Gly Leu He His Leu His Gin Asn He Val Asp Val Gin Tyr 335 340 345 350

TTG TAC GGG GTA GGG TCA AGC ATC GCG TCC TGG GCT ATT AAG TGG GAG 3324 Leu Tyr Gly Val Gly Ser Ser He Ala Ser Trp Ala He Lys Trp Glu 355 360 365

TAC GAC GTT CTC CTG TTC CTT CTG CTT GCA GAC GCG CGC GTT TGC TCC 3372 Tyr Asp Val Leu Leu Phe Leu Leu Leu Ala Asp Ala Arg Val Cys Ser 370 375 380

TGC TTG TGG ATG ATG TTA CTC ATA TCC CAA GCG GAG GCG GCT TTG GAG 3420 Cys Leu Trp Met Met Leu Leu He Ser Gin Ala Glu Ala Ala Leu Glu 385 390 395

AAC TAATCTAGAG GGCCCTATTC TATAGTGTCA CCTAAATGCT AGAGGATCTT 3473

Asn

TGTGAAGGAA CCTTACTTCT GTGGTGTGAC ATAATTGGAC AAACTACCTA CAGAGATTTA 3533

AAGCTCTAAG GTAAATATAA AATTTTTAAG TGTATAATGT GTTAAACTAC TGATTCTAAT 3593

TGTTTGTGTA TTTTAGATTC CAACCTATGG AACTGATGAA TGGGAGCAGT GGTGGAATGC 3653

CTTTAATGAG GAAAACCTGT TTTGCTCAGA AGAAATGCCA TCTAGTGATG ATGAGGCTAC 3713

TGCTGACTCT CAACATTCTA CTCCTCCAAA AAAGAAGAGA AAGGTAGAAG ACCCCAAGGA 3773

CTTTCCTTCA GAATTGCTAA GTTTTTTGAG TCATGCTGTG TTTAGTAATA GAACTCTTGC 3833

TTGCTTTGCT ATTTACACCA CAAAGGAAAA AGCTGCACTG CTATACAAGA AAATTATGGA 3893

AAAATATTCT GTAACCTTTA TAAGTAGGCA TAACAGTTAT AATCATAACA TACTGTTTTT 3953

TCTTACTCCA CACAGGCATA GAGTGTCTGC TATTAATAAC TATGCTCAAA AATTGTGTAC 4013

CTTTAGCTTT TTAATTTGTA AAGGGGTTAA TAAGGAATAT TTGATGTATA GTGCCTTGAC 4073

TAGAGATCAT AATCAGCCAT ACCACATTTG TAGAGGTTTT ACTTGCTTTA AAAAACCTCC 4133

CACACCTCCC CCTGAACCTG AAACATAAAA TGAATGCAAT TGTTGTTGTT AACTTGTTTA 4193

TTGCAGCTTA TAATGGTTAC AAATAAAGCA ATAGCATCAC AAATTTCACA AATAAAGCAT 4253

TTTTTTCACT GCATTCTAGT TGTGGTTTGT CCAAACTCAT CAATGTATCT TATCATGTCT 4313

GGATCGATCC CGCCATGGTA TCAACGCCAT ATTTCTATTT ACAGTAGGGA CCTCTTCGTT 4373

GTGTAGGTAC CGCTGTATTC CTAGGGAAAT AGTAGAGGCA CCTTGAACTG TCTGCATCAG 4433

CCATATAGCC CCCGCTGTTC GACTTACAAA CACAGGCACA GTACTGACAA ACCCATACAC 4493

CTCCTCTGAA ATACCCATAG TTGCTAGGGC TGTCTCCGAA CTCATTACAC CCTCCAAAGT 4553

CAGAGCTGTA A.TTTCGCCAT CAAGGGCAGC GAGGGCTTCT CCAGATAAAA TAGCTTCTGC 4613

CGAGAGTCCC GTAAGGGTAG ACACTTCAGC TAATCCCTCG ATGAGGTCTA CTAGAATAGT 4673

CAGTGCGGCT CCCATTTTGA AAATTCACTT ACTTGATCAG CTTCAGAAGA TGGCGGAGGG 4733

CCTCCAACAC AGTAATTTTC CTCCCGACTC TTAAAATAGA AAATGTCAAG TCAGTTAAGC 4793

AGGAAGTGGA CTAACTGACG CAGCTGGCCG TGCGACATCC TCTTTTAATT AGTTGCTAGG 4853

CAACGCCCTC CAGAGGGCGT GTGGTTTTGC AAGAGGAAGC AAAAGCCTCT CCACCCAGGC 4913

CTAGAATGTT TCCACCCAAT CATTACTATG ACAACAGCTG TTTTTTTTAG TATTAAGCAG 4973

AGGCCGGGGA CCCCTGGCCC GCTTACTCTG GAGAAAAAGA AGAGAGGCAT TGTAGAGGCT 5033

TCCAGAGGCA ACTTGTCAAA ACAGGACTGC TTCTATTTCT GTCACACTGT CTGGCCCTGT 5093

CACAAGGTCC AGCACCTCCA TACCCCCTTT AATAAGCAGT TTGGGAACGG GTGCGGGTCT 5153

TACTCCGCCC ATCCCGCCCC TAACTCCGCC CAGTTCCGCC CATTCTCCGC CCCATGGCTG 5213

ACTAATTTTT TTTATTTATG CAGAGGCCGA GGCCGCCTCG GCCTCTGAGC TATTCCAGAA 5273

GTAGTGAGGA GGCTTTTTTG GAGGCCTAGG CTTTTGCAAA AAGCTAATTC 5323

(2) INFORMATION FOR SEQ ID NO:10:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 399 amino acids

(B) TYPE: amino acid (D) TOPOLOGY: linear

(ii) MOLECULE TYPE: protein

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 10:

Met Ala Thr Gly Ser Arg Thr Ser Leu Leu Leu Ala Phe Gly Leu Leu 1 5 10 15

Cys Leu Pro Trp Leu Gin Glu Gly Ser Ala Ala Ala Ala Ala Asn Ser 20 25 30

Glu Thr His Val Thr Gly Gly Ser Ala Gly His Thr Thr Ala Gly Leu 35 40 45

Val Arg Leu Leu Ser Pro Gly Ala Lys Gin Asn He Gin Leu He Asn 50 55 60

Thr Asn Gly Ser Trp His He Asn Ser Thr Ala Leu Asn Cys Asn Glu 65 70 75 80

Ser Leu Asn Thr Gly Trp Leu Ala Gly Leu Phe Tyr His His Lys Phe 85 90 95

Asn Ser Ser Gly Cys Pro Glu Arg Leu Ala Ser Cys Arg Arg Leu Thr 100 105 110

Asp Phe Ala Gin Gly Gly Gly Pro He Ser Tyr Ala Asn Gly Ser Gly 115 120 125

Leu Asp Glu Arg Pro Tyr Cys Trp His Tyr Pro Pro Arg Pro Cys Gly 130 135 140

He Val Pro Ala Lys Ser Val Cys Gly Pro Val Tyr Cys Phe Thr Pro 145 150 155 160

Ser Pro Val Val Val Gly Thr Thr Asp Arg Ser Gly Ala Pro Thr Tyr 165 170 175

Ser Trp Gly Ala Asn Asp Thr Asp Val Phe Val Leu Asn Asn Thr Arg 180 185 190

Pro Pro Leu Gly Asn Trp Phe Gly Cys Thr Trp Met Asn Ser Thr Gly 195 20G 205

Phe Thr Lys Val Cys Gly Ala Pro Pro Cys Val He Gly Gly Val Gly 210 215 220

Asn Asn Thr Leu Leu Cys Pro Thr Asp Cys Phe Arg Lys His Pro Glu 225 230 235 240

Ala Thr Tyr Ser Arg Cys Gly Ser Gly Pro Trp He Thr Pro Arg Cys 245 250 255

Met Val Asp Tyr Pro Tyr Arg Leu Trp His Tyr Pro Cys Thr He Asn 260 265 270

Tyr Thr He Phe Lys Val Arg Met Tyr Val Gly Gly Val Glu His Arg 275 280 285

Leu Glu Ala Ala Cys Asn Trp Thr Arg Gly Glu Arg Cys Asp Leu Glu 290 295 300

Asp Arg Asp Arg Ser Glu Leu Ser Pro Leu Leu Leu Ser Thr Thr Gin 305 310 315 320

Trp Gin Val Leu Pro Cys Ser Phe Thr Thr Leu Pro Ala Leu Ser Thr 325 330 335

Gly Leu He His Leu His Gin Asn He Val Asp Val Gin Tyr Leu Tyr 340 345 350

Gly Val Gly Ser Ser He Ala Ser Trp Ala He Lys Trp Glu Tyr Asp 355 360 365

Val Leu Leu Phe Leu Leu Leu Ala Asp Ala Arg Val Cys Ser Cys Leu 370 375 380

Trp Met Met Leu Leu He Ser Gin Ala Glu Ala Ala Leu Glu Asn 385 390 395

(2) INFORMATION FOR SEQ ID NO:11:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 5125 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: circular

(ii) MOLECULE TYPE: DNA (genomic)

(ix) FEATURE:

(A) NAME/KEY: CDS

(B) LOCATION: 2227. -3225

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:11: GCGTAATCTG CTGCTTGCAA ACAAAAAAAC CACCGCTACC AGCGGTGGTT TGTTTGCCGG 60 ATCAAGAGCT ACCAACTCTT TTTCCGAAGG TAACTGGCTT CAGCAGAGCG CAGATACCAA 120 ATACTGTCCT TCTAGTGTAG CCGTAGTTAG GCCACCACTT CAAGAACTCT GTAGCACCGC 180 CTACATACCT CGCTCTGCTA ATCCTGTTAC CAGTGGCTGC TGCCAGTGGC GATAAGTCGT 240 GTCTTACCGG GTTGGACTCA AGACGATAGT TACCGGATAA GGCGCAGCGG TCGGGCTGAA 300 CGGGGGGTTC GTGCACACAG CCCAGCTTGG AGCGAACGAC CTACACCGAA CTGAGATACC 360

TACAGCGTGA GCATTGAGAA AGCGCCACGC TTCCCGAAGG GAGAAAGGCG GACAGGTATC 420

CGGTAAGCGG CAGGGTCGGA ACAGGAGAGC GCACGAGGGA GCTTCCAGGG GGAAACGCCT 480 GGTATCTTTA TAGTCCTGTC GGGTTTCGCC ACCTCTGACT TGAGCGTCGA TTTTTGTGAT 540 GCTCGTCAGG GGGGCGGAGC CTATGGAAAA ACGCCAGCAA CGCAAGCTAG CTTCTAGCTA 600

GAAATTGTAA ACGTTAATAT TTTGTTAAAA TTCGCGTTAA ATTTTTGTTA AATCAGCTCA 660 TTTTTTAACC AATAGGCCGA AATCGGCAAA ATCCCTTATA AATCAAAAGA ATAGCCCGAG 720 ATAGGGTTGA GTGTTGTTCC AGTTTGGAAC AAGAGTCCAC TATTAAAGAA CGTGGACTCC 780 AACGTCAAAG GGCGAAAAAC CGTCTATCAG GGCGATGGCC GCCCACTACG TGAACCATCA 840 CCCAAATCAA GTTTTTTGGG GTCGAGGTGC CGTAAAGCAC TAAATCGGAA CCCTAAAGGG 900 AGCCCCCGAT TTAGAGCTTG ACGGGGAAAG CCGGCGAACG TGGCGAGAAA GGAAGGGAAG 960 AAAGCGAAAG GAGCGGGCGC TAGGGCGCTG GCAAGTGTAG CGGTCACGCT GCGCGTAACC 1020 ACCACACCCG CCGCGCTTAA TGCGCCGCTA CAGGGCGCGT ACTATGGTTG CTTTGACGAG 1080 ACCGTATAAC GTGCTTTCCT CGTTGGAATC AGAGCGGGAG CTAAACAGGA GGCCGATTAA 1140

AGGGATTTTA GACAGGAACG GTACGCCAGC TGGATCACCG CGGTCTTTCT CAACGTAACA 1200 CTTTACAGCG GCGCGTCATT TGATATGATG CGCCCCGCTT CCCGATAAGG GAGCAGGCCA 1260 GTAAAAGCAT TACCCGTGGT GGGGTTCCCG AGCGGCCAAA GGGAGCAGAC TCTAAATCTG 1320 CCGTCATCGA CTTCGAAGGT TCGAATCCTT CCCCCACCAC CATCACTTTC AAAAGTCCGA 1380 AAGAATCTGC TCCCTGCTTG TGTGTTGGAG GTCGCTGAGT AGTGCGCGAG TAAAATTTAA 1440 GCTACAACAA GGCAAGGCTT GACCGACAAT TGCATGAAGA ATCTGCTTAG GGTTAGGCGT 1500 TTTGCGCTGC TTCGCGATGT ACGGGCCAGA TATACGCGTT GACATTGATT ATTGACTAGT 1560

TATTAATAGT AATCAATTAC GGGGTCATTA GTTCATAGCC CATATATGGA GTTCCGCGTT 1620

ACATAACTTA CGGTAAATGG CCCGCCTGGC TGACCGCCCA ACGACCCCCG CCCATTGACG 1680

TCAATAATGA CGTATGTTCC CATAGTAACG CCAATAGGGA CTTTCCATTG ACGTCAATGG 1740

GTGGACTATT TACGGTAAAC TGCCCACTTG GCAGTACATC AAGTGTATCA TATGCCAAGT 1800

ACGCCCCCTA TTGACGTCAA TGACGGTAAA TGGCCCGCCT GGCATTATGC CCAGTACATG 1860

ACCTTATGGG ACTTTCCTAC TTGGCAGTAC ATCTACGTAT TAGTCATCGC TATTACCATG 1920

GTGATGCGGT TTTGGCAGTA CATCAATGGG CGTGGATAGC GGTTTGACTC ACGGGGATTT 1980

CCAAGTCTCC ACCCCATTGA CGTCAATGGG AGTTTGTTTT GGCACCAAAA TCAACGGGAC 2040

TTTCCAAAAT GTCGTAACAA CTCCGCCCCA TTGACGCAAA TGGGCGGTAG GCGTGTACGG 2100

TGGGAGGTCT ATATAAGCAG AGCTCTCTGG CTAACTAGAG AACCCACTGC TTAACTGGCT 2160

TATCGAAATT AATACGACTC ACTATAGGGA GACCGGAAGC TTGGTACCGA GCTCGGATCT 2220

GCCACC ATG GCA ACA GGA TCA AGA ACA TCA CTG CTG CTG GCA TTT GGA 2268 Met Ala Thr Gly Ser Arg Thr Ser Leu Leu Leu Ala Phe Gly 1 5 10

CTG CTG TGT CTG CCA TGG CTG CAA GAA GGA TCA GCA GCA GCA GCA GCG 2316 Leu Leu Cys Leu Pro Trp Leu Gin Glu Gly Ser A.la Ala Ala Ala Ala 15 20 25 30

AAT TCA GAA ACC CAC GTC ACC GGG GGA AGT GCC GGC CAC ACC ACG GCT 2364 Asn Ser Glu Thr His Val Thr Gly Gly Ser Ala Gly His Thr Thr Ala 35 40 45

GGG CTT GTT CGT CTC CTT TCA CCA GGC GCC AAG CAG AAC ATC CAA CTG 2412 Gly Leu Val Arg Leu Leu Ser Pro Gly Ala Lys Gin Asn He Gin Leu 50 55 60

ATC AAC ACC AAC GGC AGT TGG CAC ATC AAT AGC ACG GCC TTG AAC TGC 2460 He Asn Thr Asn Gly Ser Trp His He Asn Ser Thr Ala Leu Asn Cys 65 70 75

AAT GAA AGC CTT AAC ACC GGC TGG TTA GCA GGG CTC TTC TAT CAC CAC 2508 Asn Glu Ser Leu Asn Thr Gly Trp Leu Ala Gly Leu Phe Tyr His His 80 85 90

AAA TTC AAC TCT TCA GGT TGT CCT GAG AGG TTG GCC AGC TGC CGA CGC 2556 Lys Phe Asn Ser Ser Gly Cys Pro Glu Arg Leu Ala Ser Cys Arg Arg 95 100 105 110

CTT ACC GAT TTT GCC CAG GGC GGG GGT CCT ATC AGT TAC GCC AAC GGA 2604 Leu Thr Asp Phe Ala Gin Gly Gly Gly Pro He Ser Tyr Ala Asn Gly 115 120 125

AGC GGC CTC GAT GAA CGC CCC TAC TGC TGG CAC TAC CCT CCA AGA CCT 2652

Ser Gly Leu Asp Glu Arg Pro Tyr Cys Trp His Tyr Pro Pro Arg Pro 130 135 140

TGT GGC ATT GTG CCC GCA AAG AGC GTG TGT GGC CCG GTA TAT TGC TTC 2700 Cys Gly He Val Pro Ala Lys Ser Val Cys Gly Pro Val Tyr Cys Phe 145 150 155

ACT CCC AGC CCC GTG GTG GTG GGA ACG ACC GAC AGG TCG GGC GCG CCT 2748 Thr Pro Ser Pro Val Val Val Gly Thr Thr Asp Arg Ser Gly Ala Pro 160 165 170

ACC TAC AGC TGG GGT GCA AAT GAT ACG GAT GTC TTT GTC CTT AAC AAC 2796 Thr Tyr Ser Trp Gly Ala Asn Asp Thr Asp Val Phe Val Leu Asn Asn 175 180 185 190

ACC AGG CCA CCG CTG GGC AAT TGG TTC GGT TGC ACC TGG ATG AAC TCA 2844 Thr Arg Pro Pro Leu Gly Asn Trp Phe Gly Cys Thr Trp Met Asn Ser 195 200 205

ACT GGA TTC ACC AAA GTG TGC GGA GCG CCC CCT TGT GTC ATC GGA GGG 2892 Thr Gly Phe Thr Lys Val Cys Gly Ala Pro Pro Cys Val He Gly Gly 210 215 220

GTG GGC AAC AAC ACC TTG CTC TGC CCC ACT GAT TGC TTC CGC AAG CAT 2940 Val Gly Asn Asn Thr Leu Leu Cys Pro Thr Asp Cys Phe Arg Lys His 225 230 235

CCG GAA GCC ACA TAC TCT CGG TGC GGC TCC GGT CCC TGG ATT ACA CCC 2988 Pro Glu Ala Thr Tyr Ser Arg Cys Gly Ser Gly Pro Trp He Thr Pro 240 245 250

AGG TGC ATG GTC GAC TAC CCG TAT AGG CTT TGG CAC TAT CCT TGT ACC 3036 Arg Cys Met Val Asp Tyr Pro Tyr Arg Leu Trp His Tyr Pro Cys Thr 255 260 265 270

ATC AAT TAC ACC ATA TTC AAA GTC AGG ATG TAC GTG GGA GGG GTC GAG 3084 He Asn Tyr Thr He Phe Lys Val Arg Met Tyr Val Gly Gly Val Glu 275 280 285

CAC AGG CTG GAA GCG GCC TGC AAC TGG ACG CGG GGC GAA CGC TGT GAT 3132 His Arg Leu Glu Ala Ala Cys Asn Trp Thr Arg Gly Glu Arg Cys Asp 290 295 300

CTG GAA GAC AGG GAC AGG TCC GAG CTC AGC CCG TTA CTG CTG TCC ACC 3180 Leu Glu Asp Arg Asp Arg Ser Glu Leu Ser Pro Leu Leu Leu Ser Thr 305 310 315

ACG CAG TGG CAG GTC CTT CCG TGT TCT TTC ACG ACC CTG CCA GCC 3225

Thr Gin Trp Gin Val Leu Pro Cys Ser Phe Thr Thr Leu Pro Ala 320 325 330

TAATCTAGAG GGCCCTATTC TATAGTGTCA CCTAAATGCT AGAGGATCTT TGTGAAGGAA 3285

CCTTACTTCT GTGGTGTGAC ATAATTGGAC AAACTACCTA CAGAGATTTA AAGCTCTAAG 3345

TTTATTTATG CAGAGGCCGA GGCCGCCTCG GCCTCTGAGC TATTCCAGAA GTAGTGAGGA 5085 GGCTTTTTTG GAGGCCTAGG CTTTTGCAAA AAGCTAATTC 5125

(2) INFORMATION FOR SEQ ID NO:12:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 333 amino acids

(B) TYPE: amino acid (D) TOPOLOGY: linear

(ii) MOLECULE TYPE: protein

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 12:

Met Ala Thr Gly Ser Arg Thr Ser Leu Leu Leu Ala Phe Gly Leu Leu 1 5 10 15

Cys Leu Pro Trp Leu Gin Glu Gly Ser Ala Ala Ala Ala Ala Asn Ser 20 25 30

Glu Thr His Val Thr Gly Gly Ser Ala Gly His Thr Thr Ala Gly Leu 35 40 45

Val Arg Leu Leu Ser Pro Gly Ala Lys Gin Asn He Gin Leu He Asn 50 55 60

Thr Asn Gly Ser Trp His He Asn Ser Thr Ala Leu Asn Cys Asn Glu 65 70 75 80

Ser Leu Asn Thr Gly Trp Leu Ala Gly Leu Phe Tyr His His Lys Phe 85 90 95

Asn Ser Ser Gly Cys Pro Glu A.rg Leu Ala Ser Cys Arg Arg Leu Thr 100 105 110

Asp Phe Ala Gin Gly Gly Gly Pro He Ser Tyr Ala Asn Gly Ser Gly 115 120 125

Leu Asp Glu Arg Pro Tyr Cys Trp His Tyr Pro Pro Arg Pro Cys Gly 130 135 140

He Val Pro Ala Lys Ser Val Cys Gly Pro Val Tyr Cys Phe Thr Pro 145 150 155 160

Ser Pro Val Val Val Gly Thr Thr Asp Arg Ser Gly Ala Pro Thr Tyr 165 170 175

Ser Trp Gly Ala Asn Asp Thr Asp Val Phe Val Leu Asn Asn Thr Arg 180 185 190

Pro Pro Leu Gly Asn Trp Phe Gly Cys Thr Trp Met Asn Ser Thr Gly 195 200 205

Phe Thr Lys Val Cys Gly Ala Pre Pro Cys Val He Gly Gly Val Gly 210 215 220

Asn Asn Thr Leu Leu Cys Pro Thr Asp Cys Phe Arg Lys His Pro Glu 225 230 235 240

Ala Thr Tyr Ser Arg Cys Gly Ser Gly Pro Trp He Thr Pro Arg Cys 245 250 255

Met Val Asp Tyr Pro Tyr Arg Leu Trp His Tyr Pro Cys Thr He Asn 260 265 270

Tyr Thr He Phe Lys Val Arg Met Tyr Val Gly Gly Val Glu His Arg 275 280 285

Leu Glu Ala Ala Cys Asn Trp Thr Arg Gly Glu Arg Cys Asp Leu Glu 290 295 300

Asp Arg Asp Arg Ser Glu Leu Ser Pro Leu Leu Leu Ser Thr Thr Gin 305 310 315 320

Trp Gin Val Leu Pro Cys Ser Phe Thr Thr Leu Pro Ala 325 330