NOVEL ECTOPARASITE SALIVA PROTEINS AND APPARATUS TO COLLECT SUCH PROTEINS

FIELD OF THE INVENTION The present invention relates to a noveJ product and method for isolating ectoparasite saliva proteins, and a novel product and method for detecting and/or treating allergic dermatitis in an animal.

BACKGROUND OF THE INVENTION Bites from ectoparasites, in particular fleas, can cause a hypersensitive response m animals. In particular, hypersensitive responses to fleabites is manifested m a disease called flea allergy dermatitis (FAD) . Hypersensitivity refers to a state of altered reactivity in which an animal, having been previously exposed to a compound, exhibits an allergic response to the compound upon subsequent exposures. Hypersensitive responses include immediate and delayed-type hypersensitivity, and in particular Type I, Type II, Type III and Type IV hypersensitivities (described in detail in Janeway et al., Ixn unobiology, Garland Publishing, New York, 1994, which is incorporated in its entirety by this reference) .

Foreign compounds that induce symptoms of immediate and/or delayed hypersensitivity are herein referred to as allergens. The term "allergen" primarily refers to foreign compounds capable of causing an allergic response. The term can be used interchangeably with the term "antigen,"

especially with respect to a foreign compound capable of inducing symptoms of immediate and/or delayed hypersensitivity. Factors that influence an animal's susceptibility to an allergen can include a genetic component and/or environmental exposure to an allergen. Animals can be de-sensitized to an allergen by repeated injections of the allergen to which an animal is hypersensitive .

FAD can have manifestations of both immediate and delayed-type hypersensitivity (described m detail in Janeway et al . , ibid. ) . Effective treatment of FAD has been difficult if not impossible to achieve. FAD afflicts about 15% of cats and dogs in flea endemic areas and the frequency is increasing each year. In a geographical area, effective flea control requires treatment of all animals. One treatment investigators have proposed includes desensitization of animals using flea allergens. However, reliable, defined preparations of flea allergens are needed for such treatments. Until the discovery of the novel formulations of the present invention, flea allergens responsible for FAD had not been clearly defined. Whole flea antigen preparations have been used to diagnose and desensitize animals with FAD

(Benjammi et al . , 1960, pp. 214-222, Experimental Para si tol ogy, Vol. 10; Keep et al., 1967, pp. 425-426,

Australian Veterinary Journal , Vol. 43; Kristensen et al . , 1978, pp. 414-423, Nord. Vet -Med, Vol. 30; Van Winkle, 1981, pp. 343-354, J. Amer. Animal Hosp . Assoc , Vol. 17; Haliwell et al . , 1987, pp. 203-213, Veterinary Immunol ogy and Immunopathol ogy, Vol. 15; Greene et al., 1993, pp. 69- 74, Parasi te Immunology, Vol. 15); PCT Publication No. WO 93/18788 by Opdebeeck et al . ; and Van Winkle, pp. 343-354, 1981, J. Am . Ani . Hosp . Assoc , vol. 32. Available commercial whole flea extracts, however, are unpredictable and, therefore, have limited usefulness.

Prior investigators have suggested that products contained m flea saliva might be involved m FAD and have also suggested methods to isolate such products: Ben ^* jamιnι et al . , 1963, pp. 143-154, Experimental Parasi tol ogy, Vol. 13; Young et al., 1963, pp. 155-166, Experimental Parasi tology 13, Vol. 13; Michaeli et al . , 1965, pp. 162- 170, J. Immunol . , Vol. 95; and Michaeli et al . , 1996, pp. 402-406, J. Immunol . , Vol. 97. These investigators, however, have characterized the allergenic factors of flea saliva as being haptens having molecular weights of less than 6 kilodaltons (kD) . That they are not proteins is also supported by the finding that they are not susceptible to degradation when exposed to strong acids (e.g., 6 N hydrochloric acid) or heat. Some of the particular low molecular weight allergenic factors have also been

characterized as being a highly fluorescent aromatic fraction (Young et al., ibi d. ) . In addition, studies by such investigators have indicated that m order to be allergenic, such factors need to be associated with adjuvants and/or carriers, such as collagen or portions of the membrane used to collect the oral secretions. Moreover, the methods described to collect flea saliva factors were difficult and unpredictable. Furthermore the factors isolated by these methods were typically contaminated with material from the fleas, their culture medium or the skm- based membranes used to allow the fleas to feed.

Thus, there remains a need to more clearly define flea saliva allergens capable of inducing a hypersensitive response m animals. In addition, there remains a need to develop a method to collect substantially pure flea saliva allergens which provide predictable and less expensive preparations of allergens useful for desensitizing animals subject to, or having, FAD.

SUMMARY OF THE INVENTION One embodiment of the present invention is an isolated nucleic acid molecule that hybridizes under stringent conditions with a gene including a flea saliva gene comprising a nucleic acid sequence including SEQ ID NO: 52, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO:57, SEQ ID NO: 58, SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO: 63, SEQ ID NO: 64, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO: 69, SEQ ID N0*.71, SEQ ID

N0:73, SEQ ID NO:74, SEQ ID N0:76 and a nucleic acid sequence encoding an ammo acid sequence selected from the group consisting of SEQ ID NO: 78 and SEQ ID NO: 87.

The present invention also includes a nucleic acid molecule that hybridizes under stringent hybridization conditions with a nucleic acid molecule having a nucleic acid sequence encoding a protein comprising an amino acid sequence including SEQ ID NO: 53, SEQ ID NO: 62, SEQ ID NO: 65, SEQ ID NO:70, SEQ ID NO:72, SEQ ID NO:75, SEQ ID NO: 77, SEQ ID NO: 78 and SEQ ID NO: 87.

Another embodiment of the present invention includes an isolated protein encoded by a nucleic acid molecule that hybridizes under stringent hybridization conditions with a nucleic acid molecule having a nucleic acid sequence encoding a protein comprising an ammo acid sequence including SEQ ID NO:53, SEQ ID NO: 62, SEQ ID NO: 65, SEQ ID NO:70, SEQ ID NO:72, SEQ ID NO:75, SEQ ID NO:77, SEQ ID NO: 78 and SEQ ID NO: 87.

Also included in the present invention are recombinant molecules and cells having a nucleic acid molecule of the present invention.

Another aspect of the present invention includes an antibody capable of selectively binding to an ectoparasite protein, or mimetope. Yet another embodiment of the present invention is a therapeutic composition for treating allergic dermatitis

comprising a formulation comprising at least one isolated ectoparasite saliva protein, wherein said ectoparasite saliva protein comprises at least a portion of an ammo acid sequence, where said portion is encoded by a nucleic acid molecule that hybridizes under stringent hybridization conditions with a nucleic acid molecule having a nucleic acid sequence including SEQ ID NO: 52, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO: 63, SEQ ID NO: 64, SEQ ID NO: 66, SEQ ID NO:67, SEQ ID NO:69, SEQ ID NO:71, SEQ ID NO:73, SEQ ID NO:74, SEQ ID NO:76 and a nucleic acid sequence encoding an ammo acid sequence selected from the group consisting of SEQ ID NO: 78 and SEQ ID NO: 87. A preferred therapeutic composition of the present invention also includes an excipient, an adjuvant and/or a carrier. Also included in the present invention is a method to desensitize a host animal to allergic dermatitis. The method includes the step of administering to the animal a therapeutic composition of the present invention. Other embodiments of the present invention include methods to identify an animal susceptible to or having allergic dermatitis, using m vi vo or m vi tro methods. In one embodiment, an animal susceptible to or havmg allergic dermatitis is identified m vi vo by the method comprising: (a) administering to a site on the animal a formulation

comprising at least one isolated ectoparasite saliva protein, m which the ectoparasite saliva protein comprises an ammo acid sequence including SEQ ID NO: 53, SEQ ID NO: 62, SEQ ID NO: 65, SEQ ID NO:70, SEQ ID NO:72, SEQ ID NO: 75, SEQ ID NO: 77, SEQ ID NO: 78 and SEQ ID NO: 87; and (b) comparing a reaction resulting from administration of the formulation with a reaction resulting from administration of a control solution, in which the animal is determined to be susceptible to or to have allergic dermatitis if the reaction to the formulation is at least as large as said reaction to the positive control solution, and in which the animal is determined not to be susceptible to or not to have allergic dermatitis if the reaction to the formulation is about the same size as said reaction to the negative control solution.

In another embodiment, an animal susceptible to or having allergic dermatitis is identified m vi tro by measuring the presence of antibodies indicative of allergic dermatitis m the animal using the method comprising: (a) contacting a formulation with a body fluid from an animal under conditions sufficient for formation of an immunocomplex between the formulation and the antibodies, if present, in the body fluid, the formulation comprising at least one isolated ectoparasite saliva protein, in which the ectoparasite saliva protein comprises an ammo acid sequence including SEQ ID NO:53, SEQ ID NO: 62, SEQ ID

NO: 65, SEQ ID NO:70, SEQ ID NO:72, SEQ ID NO:75, SEQ ID NO: 77, SEQ ID NO: 78 and SEQ ID NO: 87; and (b) determining the amount of lmmunocomplex formed, which formation of the immunocomplex indicates that the animal is susceptible to or has allergic dermatitis.

The present invention further relates to an assay kit for testing if an animal is susceptible to or has allelic dermatitis, the kit comprising: (a) a formulation comprising at least one isolated ectoparasite saliva protein, in which the ectoparasite saliva protein comprises an amino acid sequence including SEQ ID NO: 53, SEQ ID NO: 62, SEQ ID NO: 65, SEQ ID NO:70, SEQ ID NO:72, SEQ ID NO:75, SEQ ID NO:77, SEQ ID NO:78 and SEQ ID NO: 87; and (b) a means for determining if the animal is susceptible to or has allergic dermatitis, in which the means comprises use of the formulation to identify animals susceptible to or having allergic dermatitis.

DETAILED DESCRIPTION OF THE INVENTION The present invention includes a novel product and method for diagnosing and treating allergic dermatitis of animals to ectoparasites.

According to the present invention, ectoparasites are external living parasites that attach and feed through the sk n of a host animal. Ectoparasites include parasites that live on a host animal and parasites that attach

temporarily to an animal m order to feed. Also, according to the present invention, ectoparasite saliva refers to the material released from the mouth of an ectoparasite when the ectoparasite attempts to feed in response to a temperature differential. Ectoparasite saliva includes ectoparasite saliva products.

One embodiment of the present invention is a formulation that contains ectoparasite saliva products that can be used to diagnose and/or treat animals susceptible to or having (i.e., suffering from) allergic dermatitis. Preferred types of allergic dermatitis to diagnose and/or treat using ectoparasite saliva products of the present invention include flea allergy dermatitis, Culi coides allergy dermatitis and mosquito allergy dermatitis. A preferred type of allergic dermatitis to diagnose and/or treat using ectoparasite saliva products of the present invention is flea allergy dermatitis. As used herem, an animal that is susceptible to allergic dermatitis refers to an animal that is genetically pre-disposed to developing allergic dermatitis and/or to an animal that has been primed with an antigen such a manner that re-exposure to the antigen results in symptoms of allergy that can be perceived by, for example, observing the animal or measuring antibody production by the animal to the antigen. As such, animals susceptible to allergic dermatitis can include animals having sub-clinical allergic dermatitis.

SuD-climcal allergic dermatitis refers to a condition m which allergy symptoms cannot be detected by simply observing an animal (i.e., manifestation of the disease can include the presence of anti-ectoparasite saliva protein antibodies withm an affected animal but no dermatitis) . For example, sub-clinical allergic dermatitis can be detected using in vi vo or m vi tro assays of the present invention, as described in detail below. Reference to animals having allergic dermatitis includes animals that do display allergy symptoms that can be detected by simply observing an animal and/or by using m vivo or m vi tro assays of the present invention, as described in detail below.

One embodiment of the present invention is a formulation that includes one or more isolated ectoparasite saliva proteins. According to the present invention, an isolated protein is a protein that has been removed from its natural milieu. An isolated ectoparasite saliva protein can, for example, be obtained from its natural source, be produced using recombinant DNA technology, or be synthesized chemically. As used herein, an isolated ectoparasite saliva protein can be a full-length ectoparasite saliva protein or any homologue of such a protein, such as an ectoparasite saliva protein in which ammo acids have been deleted (e.g., a truncated version of

the protein, such as a peptide), inserted, inverted, substituted and/or derivat zed (e.g., by glycosylation, phosphorylation, acetylation, myristylation, prenylation, palmitation, amidation and/or addition of glycosylphosphatidyl mositol) . A homologue of an ectoparasite saliva protein is a protein having an amino acid sequence that is sufficiently similar to a natural ectoparasite saliva protein ammo acid sequence that a nucleic acid sequence encoding the homologue is capable of hybridizing under stringent conditions to (i.e., with) a nucleic acid molecule encoding the natural ectoparasite saliva protein (i.e., the complement of a nucleic acid sequence encoding the natural ectoparasite saliva protein am o acid sequence) . A nucleic acid sequence complement of any nucleic acid sequence of the present invention refers to the nucleic acid sequence of the nucleic acid strand that is complementary to (i.e., can form a complete double helix with) the strand for which the sequence is cited. It is to be noted that a double-stranded nucleic acid molecule of the present invention for which a nucleic acid sequence has been determined for one strand that represented by a SEQ ID NO also comprises a complementary strand havmg a sequence that is a complement of that SEQ ID NO. As such, nucleic acid molecules of the present invention, which can be either double-stranded or smgle- stranded, include those nucleic acid molecules that form

stable hybrids under stringent hybridization conditions with either a given SEQ ID NO denoted herem and/or with the complement of that SEQ ID NO, which may or may not be denoted herein. Methods to deduce a complementary sequence are known to those skilled in the art.

As used herein, stringent hybridization conditions refer to standard hybridization conditions under which nucleic acid molecules, including oligonucleotides, are used to identify similar nucleic acid molecules. Such standard conditions are disclosed, for example, Sambrook et al., Molecular Cloning: A Laboratory Manual , Cold Spring Harbor Labs Press, 1989; Sambrook et al., ibi d. , is incorporated by reference herem in its entirety. Stringent hybridization conditions typically permit isolation of nucleic acid molecules havmg at least about 70% nucleic acid sequence identity with the nucleic ac d molecule being used to probe in the hybridization reaction. Formulae to calculate the appropriate hybridization and wash conditions to achieve hybridization permitting 30% or less mismatch of nucleotides are disclosed, for example, m Me koth et al., 1984, Anal . Bi ochem . 138, 267-284; Memkoth et al. , ibid. , is incorporated by reference herein m its entirety.

The minimal size of a protein homologue of the present invention is a size sufficient to be encoded by a nucleic

acid molecule capable of forming a stable hybrid with the complementary sequence of a nucleic acid molecule encoding the corresponding natural protein. As such, the size of the nucleic acid molecule encoding such a protein homologue is dependent on nucleic acid composition and percent homology between the nucleic acid molecule and complementary sequence as well as upon hybridization conditions per se (e.g., temperature, salt concentration, and formamide concentration) . The minimal size of such nucleic acid molecules is typically at least about 12 to about 15 nucleotides in length if the nucleic acid molecules are GC-rich and at least about 15 to about 17 bases m length if they are AT-πch. As such, the minimal size of a nucleic acid molecule used to encode an ectoparasite saliva protein homologue of the present invention is from about 12 to about 18 nucleotides length. There is no limit, other than a practical limit, on the maximal size of such a nucleic acid molecule that the nucleic acid molecule can include a portion of a gene, an entire gene, or multiple genes, or portions thereof. Similarly, the minimal size of an ectoparasite saliva protein homologue of the present invention is from about 4 to about 6 ammo acids in length, with preferred sizes depending on whether a full-length, multivalent (i.e., fusion protein having more than one domain each of which

has a function) , or functional portions of such proteins are desired.

Ectoparasite saliva protein homologues can be the result of allelic variation of a natural gene encoding an ectoparasite saliva protein. A natural gene refers to the form of the gene found most often m nature. Ectoparasite saliva protein homologues can be produced using techniques known in the art including, but not limited to, direct modifications to a gene encoding a protein using, for example, classic or recombinant DNA techniques to effect random or targeted mutagenesis.

Preferred ectoparasite saliva proteins of the present invention, including homologues thereof, are capable of detecting and/or treating allergic dermatitis resulting from the bites of ectoparasites. A preferred ectoparasite saliva protein homologue includes at least one epitope capable of eliciting a hypersensitive response to the natural ectoparasite saliva protein counterpart. An ectoparasite saliva protein homologue can also include an epitope capable of hyposensitiz g an animal to the natural form of the protein. The ability of an ectoparasite saliva protein homologue to detect and/or treat (i.e., immunomodulate or regulate by, for example, desensitizing) the hypersensitivity of an animal susceptible to or having allergic dermatitis, can be tested using techniques known to those skilled m the art. Such techniques include skm

tests and immunoabsorbent assays as described m detail below. Additional preferred ectoparasite saliva proteins of the present invention have other activities that include activities important for feeding and survival of the ectoparasite.

In one embodiment, a formulation of the present invention can comprise a protein having at least a portion of an isolated ectoparasite saliva protein. According to the present invention, "at least a portion of an ectoparasite saliva protein" refers to a portion of an ectoparasite saliva protein encoded by a nucleic acid molecule that is capable of hybridizing, under stringent conditions, with a nucleic acid encoding a full-length ectoparasite saliva protein of the present invention. Preferred portions of ectoparasite saliva proteins are useful for detecting and/or treating allergic dermatitis resulting from the bites of ectoparasites. Additional preferred portions have activities important for flea feeding and survival. Suitable sizes for portions of an ectoparasite saliva protein of the present invention are as disclosed for saliva protein homologues of the present invention.

As will be apparent to one of skill m the art, the present invention is intended to apply to all ectoparasites. A formulation of the present invention can include saliva products from any ectoparasites. A preferred

ectoparasite of the present invention from which to isolate saliva products (including proteins), and/or from which to identify proteins that can then be produced recombmantly or synthetically, include arachnids, insects and leeches. More preferred ectoparasites from which to obtain saliva products include fleas; ticks, including both hard ticks of the family Ixodidae (e.g., Ixodes and AmJblyomma) and soft ticks of the family Argasidae (e.g., Orm thodoros, such as 0. parkeπ and 0. tuπ ca ta ) ; flies, such as midges (e.g., Culi coides) , mosquitos, sand flies, black flies, horse flies, horn flies, deer flies, tsetse flies, stable flies, myiasis-causmg flies and biting gnats; ants; spiders, lice; mites; and true bugs, such as bed bugs and kissing bugs, including those carrying Chagas disease. Even more preferred ectoparasite saliva products include those from fleas, mosquitos, midqes, sandflies, blackflies, ticks and Rhodm us, with products from fleas, mosquitos and Culi coides being even more preferred.

A particularly preferred formulation of the present invention includes flea saliva proteins. Preferred flea saliva products include those from Ctenocephali des, Xenopsylla, Pulex, Tunga, Nosopsyll us, Diamanus, Ctopsyll us and Echidnophaga fleas, with saliva products from Ctenocephalides cams and Ctenocephalides feli s fleas being even more preferred. For the purposes of illustration, many

of the following embodiments discuss flea saliva proteins. Such discussion of flea saliva proteins is not intended, in any way, to limit the scope of the present invention.

In another embodiment, a formulation of the present invention includes at least a portion of an ectoparasite saliva protein homologue having at least a portion of one of the following ammo acid sequences: SEQ ID NO: 53, SEQ ID NO:62, SEQ ID NO: 65, SEQ ID NO:70, SEQ ID NO:72, SEQ ID NO: 75, SEQ ID NO:77, SEQ ID NO: 78 and SEQ ID NO: 87 and/or other sequences disclosed herem.

In one embodiment, a formulation of the present invention can include at least one isolated protein having (i.e., including) at least a portion of one of the ammo acid sequences identified m the Sequence ID Listing, and more specifically an ammo acid sequence selected from the group consisting of SEQ ID NO: 53, SEQ ID NO: 62, SEQ ID NO: 65, SEQ ID NO:70, SEQ ID NO:72, SEQ ID NO:75, SEQ ID NO: 77, SEQ ID NO: 78 and SEQ ID NO: 87.

It is to be appreciated that ectoparasite saliva proteins of the present invention include, but are not limited to, full-length proteins, hybrid proteins, fusion proteins, multivalent proteins, and proteins that are truncated homologues of, or are proteolytic products of, at least a portion of a protein havmg at least a portion of one of the following ammo acid sequences: SEQ ID NO:53, SEQ ID NO: 62, SEQ ID NO: 65, SEQ ID NO: 70, SEQ ID NO: 72, SEQ

ID NO:75, SEQ ID NO:77, SEQ ID NO:78, SEQ ID NO:87 and/or other sequences disclosed herein. As used herein, the term hybrid protein refers to a single protein produced from two different proteins. The foregoing SEQ ID NO's represent ammo acid sequences deduced according to methods disclosed m the Examples. It should be noted that since ammo acid sequencing technology is not entirely error-free, the foregoing SEQ ID NO's, at best, represent an apparent ammo acid sequence of the ectoparasite saliva proteins of the present invention. In addition, the variation seen in the foregoing SEQ ID NO's can also be due, at least part, to allelic variation since the proteins being sequenced were derived from populations of fleas. According to the present invention, a formulation of the present invention can include flea saliva proteins that have undergone post-translational modification. Such modification can include, for example, glycosylation. Glycosylation can include addition of N-lmked and/or 0- linked oligosaccharides. It is to be appreciated that post-translational modification of a protein of the present invention can contribute to an epitope's ability to induce an allergic response against the protein m an immediate or delayed hypersensitivity response. Another embodiment of the present invention is an isolated nucleic acid molecule capable of hybridizing,

under stringent conditions, with an ectoparasite saliva protein gene encoding an ectoparasite saliva protein of the present invention. In accordance with the present invention, an isolated nucleic acid molecule is a nucleic acid molecule that has been removed from its natural milieu (i.e., that has been subject to human manipulation) . As such, "isolated" does not reflect the extent to which the nucleic acid molecule has been purified. An isolated nucleic acid molecule can include DNA, RNA, or derivatives of either DNA or RNA.

An isolated nucleic acid molecule of the present invention can be obtained from its natural source either as an entire (i.e., complete) gene or a portion thereof capable of forming a stable hybrid with that gene. As used herein, the phrase "at least a portion of" an entity refers to an amount of the entity that is at least sufficient to have the functional aspects of that entity. For example, at least a portion of a nucleic acid sequence, as used herein, is an amount of a nucleic acid sequence capable of forming a stable hybrid with the corresponding gene under stringent hybridization conditions. An isolated nucleic acid molecule of the present invention can also be produced using recombinant DNA technology (e.g., polymerase chain reaction (PCR) amplification, cloning) or chemical synthesis. Isolated ectoparasite saliva protein nucleic acid molecules include natural nucleic acid molecules and homologues

thereof, including, but not limited to, natural allelic variants and modified nucleic acid molecules m which nucleotides have been inserted, deleted, substituted, and/or inverted m such a manner that such modifications do not substantially interfere with the nucleic acid molecule's ability to encode an ectoparasite saliva protein of the present invention or to form stable hybrids under stringent conditions with natural nucleic acid molecule isolates. An isolated nucleic acid molecule of the present invention can include a nucleic acid sequence that encodes at least one ectoparasite saliva protein of the present invention, examples of such proteins being disclosed herein. Although the phrase "nucleic acid molecule" primarily refers to the physical nucleic acid molecule and the phrase "nucleic acid sequence" primarily refers to the sequence of nucleotides on the nucleic acid molecule, the two phrases can be used interchangeably, especially with respect to a nucleic acid molecule, or a nucleic acid sequence, being capable of encoding an ectoparasite saliva protein. As heretofore disclosed, ectoparasite saliva proteins of the present invention include, but are not limited to, proteins having full-length ectoparasite saliva protein coding regions, portions thereof, and other ectoparasite saliva protein homologues.

It is to be appreciated that an ectoparasite saliva protein of the present invention can be encoded by a full- length nucleic acid sequence which encodes a polyprotem. The polyprotem can be post-translationally processed into multiple proteins which are found m saliva. As used herein, an ectoparasite saliva protein gene includes all nucleic acid sequences related to a natural ectoparasite saliva protein gene such as regulatory regions that control production of an ectoparasite saliva protein encoded by that gene (such as, but not limited to, transcription, translation or post-translation control regions) as well as the coding region itself. A nucleic acid molecule of the present invention can be an isolated natural ectoparasite saliva protein nucleic acid molecule or a homologue thereof. A nucleic acid molecule of the present invention can include one or more regulatory regions, full-length or partial coding regions, or combinations thereof. The minimal size of an ectoparasite saliva protein nucleic acid molecule of the present invention is the minimal size capable of forming a stable hybrid under stringent hybridization conditions with a corresponding natural gene.

An ectoparasite saliva protein nucleic acid molecule homologue can be produced using a number of methods known to those skilled m the art (see, for example, Sambrook et al . , ibid. ) . For example, nucleic acid molecules can be modified using a variety of techniques including, but not

limited to, classic mutagenesis techniques and recombinant DNA techniques, such as site-directed mutagenesis, chemical treatment of a nucleic acid molecule to induce mutations, restriction enzyme cleavage of a nucleic acid fragment, ligation of nucleic acid fragments, polymerase chain reaction (PCR) amplification and/or mutagenesis of selected regions of a nucleic acid sequence, synthesis of oligonucleotide mixtures and ligation of mixture groups to "build" a mixture of nucleic acid molecules and combinations thereof. Nucleic acid molecule homologues can be selected from a mixture of modified nucleic acids by screening for the function of the protein encoded by the nucleic acid (e.g., the ability of a homologue to elicit an allergic response in animals having allergic dermatitis or the ability of a homologue to act as an anti-coagulant) and/or by hybridization with isolated ectoparasite saliva protein nucleic acids under stringent conditions.

One embodiment of the present invention is an ectoparasite saliva protein nucleic acid molecule that encodes a protein havmg at least a portion of one or more of the following amino acid sequences: SEQ ID N0:1, as well as with the complements of any of these sequences or homologues thereof. Such preferred nucleic acid molecules can hybridize to the coding and/or complementary strand. A preferred nucleic acid molecule of the present invention is capable of hybridizing under stringent

conditions to the coding strand and/or to the strand complementary to the coding strand of a nucleic acid molecule that encodes at least a portion of such a flea saliva protein or homologue thereof. A particularly preferred nucleic acid sequence is a nucleic acid sequence having at least about 65 percent, preferably at least about 75 percent, more preferably at least about 85 percent, and even more preferably at least about 95 percent homology with a nucleic acid sequence encoding at least a portion of one or more of the following amino acid sequences .*SEQ ID NO: 53, SEQ ID NO: 62, SEQ ID NO: 65, SEQ ID NO: 70, SEQ ID NO:72, SEQ ID NO:75, SEQ ID NO:77, SEQ ID NO:78 and/or SEQ ID NO: 87.

Such nucleic acid molecules can be a full-length gene and/or a nucleic acid molecule encoding a full-length protein, a hybrid protein, a fusion protein, a multivalent protein or a truncation fragment. More preferred nucleic acid molecules of the present invention comprise isolated nucleic acid molecules having a nucleic acid sequence as represented by SEQ ID NO: 52, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO:57, SEQ ID NO: 58, SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO: 63, SEQ ID NO: 64, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO: 69, SEQ ID NO: 71, SEQ ID NO:73, SEQ ID NO:74, SEQ ID NO: 76, a nucleic acid sequence encoding ammo acid sequence SEQ ID NO:78 or SEQ ID NO: 87, or other sequences disclosed herein.

SEQ ID NO: 52, a nucleic acid sequence that includes about 595 nucleotides of the apparent gene encoding flea saliva protein fspG5 (denoted nfspG5 ) , encodes a protein of about 90 amino acids (denoted as PfspG5 ₉₀ ) , represented by SEQ ID NO: 53. The entire translation product of fspG5 is apparently about 71 amino acids and is denoted SEQ ID NO:56. SEQ ID NO: 61, a nucleic acid sequence that includes about 1007 nucleotides of the apparent gene encoding flea saliva protein fspl (denoted nfspl ₁₀₀₇ ) r encodes a protein of about 155 amino acids (denoted Pfspl ₁₅₅ ) , which is denoted SEQ ID NO: 62. SEQ ID NO: 64, a nucleic acid sequence that includes about 1205 nucleotides of the apparent gene encoding flea saliva protein fspNS (denoted nfspN5 _120b ) , encodes a protein of about 353 amino acids (denoted PfspN5 ₃₅₃ ) , which is denoted SEQ ID NO: 65. SEQ ID NO: 71, a nucleic acid sequence that includes about 406 nucleotides of the apparent gene encoding a fspN6 flea saliva protein

(denoted nfspN6 _40b ) , encodes a protein of about 135 amino acids (denoted PfspN6 ₁₃₅ ) , which is denoted SEQ ID NO: 72. SEQ ID NO: 74, a nucleic acid sequence that includes about 420 nucleotides of the apparent gene encoding a fspJ flea saliva protein, encodes a protein of about 72 amino acids, which is denoted SEQ ID NO:75.

Knowing a nucleic acid molecule of an ectoparasite saliva protein of the present invention allows one skilled in the art to make copies of that nucleic acid molecule as

well as to obtain a nucleic acid molecule including additional portions of ectoparasite saliva protein-encoding genes (e.g., nucleic acid molecules that include the translation start site and/or transcription and/or translation control regions), and/or ectoparasite saliva protein nucleic acid molecule homologues. Knowing a portion of an ammo acid sequence of an ectoparasite saliva protein of the present invention allows one skilled in the art to clone nucleic acid sequences encoding such an ectoparasite saliva protein. In addition, a desired ectoparasite saliva protein nucleic acid molecule can be obtained m a variety of ways including screening appropriate expression libraries with antibodies which bind to ectoparasite saliva proteins of the present invention; traditional cloning techniques using oligonucleotide probes of the present invention to screen appropriate libraries or DNA; and PCR amplification of appropriate libraries, or RNA or DNA using oligonucleotide primers of the present invention (genomic and/or cDNA libraries can be used) . To isolate flea saliva protein nucleic acid molecules, preferred cDNA libraries include cDNA libraries made from unfed whole flea, fed whole flea, fed flea idgut, unfed flea midgut, and flea salivary gland. Techniques to clone and amplify genes are disclosed, for example, m Sambrook et al., ibid. The Examples section includes examples of the isolation of cDNA

sequences encoding flea saliva proteins of the present invention.

The present invention also includes nucleic acid molecules that are oligonucleotides capable of hybridizing, under stringent conditions, with complementary regions of other, preferably longer, nucleic acid molecules of the present invention that encode at least a portion of one or more of the following am o acid sequences: SEQ ID NO: 53, SEQ ID NO: 62, SEQ ID NO: 65, SEQ ID NO:70, SEQ ID NO: 72, SEQ ID NO:75, SEQ ID NO:77, SEQ ID NO:78, SEQ ID NO:87, or homologues thereof, such oligonucleotides can hybridize to the coding or non-coding strand of a double-stranded nucleic acid molecule. Certain preferred oligonucleotides are capable of hybridizing to nucleic acid molecules including nucleic acid sequences represented by SEQ ID NO: 52, SEQ ID NO: 58, SEQ ID NO: 61, SEQ ID NO: 64, SEQ ID NO: 71, SEQ ID NO:74, a nucleic acid sequence that encodes SEQ ID NO: 78 or SEQ ID NO: 87, or complements thereof.

Oligonucleotides of the present invention can be RNA, DNA, or derivatives of either. The minimal size of such oligonucleotides is the size required to form a stable hybrid between a given oligonucleotide and the complementary sequence on another nucleic acid molecule of the present invention. Minimal size characteristics are disclosed herein. The size of the oligonucleotide must also be sufficient for the use of the oligonucleotide m

accordance with tne present invention. Oligonucleotides of the present invention can be used m a variety of applications including, but not limited to, as probes to identify additional nucleic acid molecules, as primers to amplify or extend nucleic acid molecules or in therapeutic applications to inhibit, for example, expression of saliva proteins by ectoparasites. Such therapeutic applications include the use of such oligonucleotides m, for example, antisense-, triplex formation-, ribozyme- and/or RNA drug- based technologies. The present invention, therefore, includes such oligonucleotides and methods to interfere with the production of ectoparasite saliva proteins by use of one or more of such technologies.

The present invention also includes a recombinant vector, which includes an ectoparasite saliva protein nucleic acid molecule of the present invention inserted into any vector capable of delivering the nucleic acid molecule into a host cell. Such a vector contains heterologous nucleic acid sequences, that is nucleic acid sequences that are not naturally found adjacent to ectoparasite saliva protein nucleic acid molecules of the present invention. The vector can be either RNA or DNA, either prokaryotic or eukaryotic, and typically is a virus or a plasmid. Recombinant vectors can be used the cloning, sequencing, and/or otherwise manipulating of ectoparasite saliva protein nucleic acid molecules of the

present invention. One type of recombinant vector, herein referred to as a recombinant molecule and described in more detail below, can be used in the expression of nucleic acid molecules of the present invention. Preferred recombinant vectors are capable of replicating in the transformed cell. A preferred nucleic acid molecule to include in a recombinant vector of the present invention is a nucleic acid molecule that encodes at least a portion of one or more of the following amino acid sequences: SEQ ID NO: 53, SEQ ID NO: 62, SEQ ID NO: 65, SEQ ID NO:70, SEQ ID NO:72, SEQ ID NO:75, SEQ ID NO:77, SEQ ID NO:78 and SEQ ID NO:87, or other sequences disclosed herein, or homologues thereof, and nucleic acid molecules including at least a portion of a nucleic acid sequence represented by SEQ ID NO: 52, SEQ ID NO: 58, SEQ ID NO: 61, SEQ ID NO:64, SEQ ID NO:71, SEQ ID NO: 74, a nucleic acid sequence that encodes SEQ ID NO: 78 or SEQ ID NO: 87, or other sequences disclosed herein, or complements thereof. A more preferred sequences to include in a recombinant vector include nfspG5 ₅₉₅ , nfspG5 ₂₇₀ nfspG5, ₁₃ , nfspl _]007 , nfspN5 ₁₂₀₅ , nfspN5 _10ϊg nfspN6 ₄₀₆ and nfspJ ₄₂₀ .

Preferred recombinant molecules of the present invention include pCro-nfspG5 ₃ and pCro-nfspl _{4 ϊ4} , the production of which are described in detail in the Examples section.

In one embodiment, an isolated ectoparasite saliva protein of the present invention is produced by culturing a cell capable of expressing the protein under conditions effective to produce the protein, and recovering the protein. A preferred cell to culture is a recombinant cell that is capable of expressing the ectoparasite saliva protein, the recombinant cell being produced by transforming a host cell with one or more nucleic acid molecules of the present invention. Transformation of a nucleic acid molecule into a cell can be accomplished by any method by which a nucleic acid molecule can be inserted into the cell. Transformation techniques include, but are not limited to, transfection, electroporation, microm ection, lipofection, adsorption, and protoplast fusion. A recombinant cell may remain unicellular or may grow into a tissue, organ or a multicellular organism. Transformed nucleic acid molecules of the present invention can remain extrachromosomal or can integrate into one or more sites withm a chromosome of the transformed (i.e., recombinant) cell in such a manner that their ability to be expressed is retained. Preferred nucleic acid molecules with which to transform a host cell include one or more nucleic acid molecules that are as disclosed herein for including m recombinant vectors of the present invention. Suitable host cells to transform include any cell that can be transformed and that can express the introduced

ectoparasite saliva protein. Such cells are, therefore, capable of producing ectoparasite saliva proteins of the present invention after being transformed with at least one nucleic acid molecule of the present invention. Host cells can be either untransformed cells or cells that are already transformed with at least one nucleic acid molecule. Suitable host cells of the present invention can include bacterial, fungal (including yeast), insect, animal and plant cells. Preferred host cells include bacterial, yeast, insect and mammalian cells, with bacterial (e.g., E. coli ) and insect (e.g., Spodoptera ) cells being particularly preferred.

A recombinant cell is preferably produced by transforming a host cell with one or more recombinant molecules, each comprising one or more nucleic acid molecules of the present invention operatively linked to an expression vector containing one or more transcription control sequences. The phrase operatively linked refers to insertion of a nucleic acid molecule into an expression vector in a manner such that the molecule is able to be expressed when transformed into a host cell. As used herein, an expression vector is a DNA or RNA vector that is capable of transforming a host cell and of effecting expression of a specified nucleic acid molecule. Preferably, the expression vector is also capable of

replicating withm the host cell. Expression vectors can be either prokaryotic or eukaryotic, and are typically viruses or plasmids. Expression vectors of the present invention include any vectors that function (i.e., direct gene expression) in recombinant cells of the present invention, including m bacterial, fungal, insect, animal, and/or plant cells. As such, nucleic acid molecules of the present invention can be operatively linked to expression vectors containing regulatory sequences such as promoters, operators, repressors, enhancers, termination sequences, origins of replication, and other regulatory sequences that are compatible with the recombinant cell and that control the expression of nucleic acid molecules of the present invention. As used herein, a transcription control sequence includes a sequence which is capable of controlling the initiation, elongation, and termination of transcription. Particularly important transcription control sequences are those which control transcription initiation, such as promoter, enhancer, operator and repressor sequences. Suitable transcription control sequences include any transcription control sequence that can function m at least one of the recombinant cells of the present invention. A variety of such transcription control sequences are known to those skilled in the art. Preferred transcription control sequences include those which function m bacterial, yeast, helminth, insect and

mammalian cells, such as, but not limited to, tac, l ac, trp, trc, oxy-pro, omp/lpp, rrnB, bacteriophage lambda (λ) (such as λp _L and λp _k and fusions that include such promoters), bacteriophage T7, T71ac, bacteriophage T3, bacteriophage SP6, bacteriophage SP01, metallothionem, alpha mating factor, Pi chia alcohol oxidase, alphavirus subgenomic promoters (such as S dbis virus subgenomic promoters) , baculovirus, Heliothis zea insect virus, vaccinia virus, herpesvirus, poxvirus, adenovirus, simian virus 40, retrovirus actm, retroviral long terminal repeat, Rous sarcoma virus, heat shock, phosphate and nitrate transcription control sequences as well as other sequences capable of controlling gene expression in prokaryotic or eukaryotic cells. Additional suitable transcription control sequences include tissue-specific promoters and enhancers as well as lymphokme-mducible promoters (e.g., promoters inducible by interferons or mterleukms) . Transcription control sequences of the present invention can also include naturally occurring transcription control sequences naturally associated with a DNA sequence encoding an ectoparasite saliva protein.

Expression vectors of the present invention may also contain secretory signals (i.e., signal segment nucleic acid sequences) to enable an expressed ectoparasite saliva protein to be secreted from the cell that produces the

protein. Suitable signal segments include an ectoparasite saliva protein signal segment or any heterologous signal segment capable of directing the secretion of an ectoparasite saliva protein, including fusion proteins, of the present invention. Preferred signal segments include, but are not limited to, tissue plasmmogen activator (t- PA) , mterferon, mterleukm, growth hormone, histocompatibility and viral envelope glycoprotein signal segments . Expression vectors of the present invention may also contain fusion sequences which lead to the expression of inserted nucleic acid molecules of the present invention as fusion proteins. Inclusion of a fusion sequence as part of an ectoparasite nucleic acid molecule of the present invention can enhance the stability during production, storage and/or use of the protein encoded by the nucleic acid molecule. Furthermore, a fusion segment can function as a tool to simplify purification of an ectoparasite saliva protein, such as to enable purification of the resultant fusion protein using affinity chromatography. A suitable fusion segment can be a domain of any size that has the desired function (e.g., increased stability and/or purification tool) . It is withm the scope of the present invention to use one or more fusion segments. Fusion segments can be joined to ammo and/or carboxyl termmi of an ectoparasite saliva protein. Linkages between fusion

segments and ectoparasite saliva proteins can be constructed to be susceptible to cleavage to enable straight-forward recovery of the ectoparasite saliva proteins. Fusion proteins are preferably produced by culturing a recombinant cell transformed with a fusion nucleic acid sequence that encodes a protein including the fusion segment attached to either the carboxyl and/or ammo terminal end of an ectoparasite saliva protein.

A recombinant molecule of the present invention is a molecule that can include at least one of any nucleic acid molecule heretofore described operatively linked to at least one of any transcription control sequence capable of effectalveoli regulating expression of the nucleic acid molecule (s) in the cell to be transformed. A preferred recombinant molecule includes one or more nucleic acid molecules that are as disclosed herein for including in a recombinant vector of the present invention.

A recombinant cell of the present invention includes any cells transformed with at least one of any nucleic acid molecules of the present invention. A preferred recombinant cell is a cell transformed with at least one nucleic acid molecule that encode a protein having at least a portion of one or more of the following ammo acid sequences: SEQ ID NO: 53, SEQ ID NO: 62, SEQ ID NO: 65, SEQ ID NO: 70, SEQ ID NO:72, SEQ ID NO:75, SEQ ID NO:77, SEQ ID NO: 78, SEQ ID NO: 87, or other sequences disclosed herem,

or homologues thereof, and nucleic acid molecules including at least a portion of a nucleic acid sequence represented by SEQ ID NO:52, SEQ ID NO:58, SEQ ID NO: 61, SEQ ID NO: 64, SEQ ID NO: 71, SEQ ID NO: 74, a nucleic acid sequence that encodes SEQ ID NO: 78 or SEQ ID NO: 87, or other sequences disclosed herein, or complements thereof. Particularly preferred recombinant cells include E. coli transformed with at least one of the aforementioned nucleic acid molecules. Preferred recombinant cells of the present invention include E. coli :pCro-nfspG5 ₂₁ , and E. coli :pCro- nfspl ₄₇₄ ,

It may be appreciated by one skilled in the art that use of recombinant DNA technologies can improve expression of transformed nucleic acid molecules by manipulating, for example, the number of copies of the nucleic acid molecules within a host cell, the efficiency with which those nucleic acid molecules are transcribed, the efficiency with which the resultant transcripts are translated, and the efficiency of post-translational modifications. Recombinant techniques useful for increasing the expression of nucleic acid molecules of the present invention include, but are not limited to, operatively linking nucleic acid molecules to high-copy number plasmids, integration of the nucleic acid molecules into one or more host cell chromosomes, addition of vector stability sequences to plasmids,

substitutions or modifications of transcription control signals (e.g., promoters, operators, enhancers), substitutions or modifications of translational control signals (e.g., ribosome binding sites, Sh e-Dalgarno sequences), modification of nucleic acid molecules of the present invention to correspond to the codon usage of the host cell, deletion of sequences that destabilize transcripts, and use of control signals that temporally separate recombinant cell growth from recombinant protein production during fermentation. The activity of an expressed recombinant protein of the present invention may be improved by fragmenting, modifying, or derivatizing the resultant protein.

In accordance with the present invention, recombinant cells can be used to produce an ectoparasite saliva protein of the present invention by culturing such cells under conditions effective to produce such a protein, and recovering the protein. Effective conditions to produce a protein include, but are not limited to, appropriate media, bioreactor, temperature, pH and oxygen conditions that permit protein production. An appropriate, or effective, medium refers to any medium in which a cell of the present invention, when cultured, is capable of producing an ectoparasite saliva protein. Such a medium is typically an aqueous medium comprising assimilable carbohydrate, nitrogen and phosphate sources, as well as appropriate

salts, minerals, metals and other nutrients, such as vitamins. The medium may comprise complex nutrients or may be a defined minimal medium.

Cells of the present invention can be cultured in conventional fermentation bioreactors, which include, but are not limited to, batch, fed-batch, cell recycle, and continuous fermentors. Culturing can also be conducted in shake flasks, test tubes, microtiter dishes, and petri plates. Culturing is carried out at a temperature, pH and oxygen content appropriate for the recombinant cell. Such culturing conditions are well withm the expertise of one of ordinary skill m the art.

Depending on the vector and host system used for production, resultant ectoparasite saliva proteins may either remain with the recombinant cell; be secreted into the fermentation medium; be secreted into a space between two cellular membranes, such as the periplasmic space in E. col i ; or be retained on the outer surface of a cell or viral membrane. The phrase "recovering the protein" refers simply to collecting the whole fermentation medium containing the protein and need not imply additional steps of separation or purification. Ectoparasite saliva proteins of the present invention can be purified using a variety of standard protein purification techniques, such as, but not limited to, affinity chromatography, ion exchange

chromatography, filtration, electrophoresis, hydrophobic interaction chromatography, gel filtration chromatography, reverse phase chromatography, chromatofocusmg and differential solubilization. Ectoparasite saliva proteins are preferably retrieved in "substantially pure" form. As used herein, "substantially pure" refers to a purity that allows for the effective use of the protein as a therapeutic composition or diagnostic. For example, an animal being administered dosages of ectoparasite saliva protein isolated from a recombinant cell of the present invention should exhibit no substantial toxicity from contaminants mixed with the protein.

Ectoparasite saliva that is substantially free of contaminating material can be collected using a saliva collection apparatus of the present invention (disclosed in related PCT Patent Publication No. WO 96/11,271, published April 18, 1996, by Frank et al.; this publication is incorporated by reference herein in its entirety) . The interior diameter of a preferred chamber of the present invention is preferably about 7.5 cm. The size of a collection means of the present invention is preferably larger than the open end of the 7.5 cm chamber, the size of tne collection means is more preferably about 8 cm. According to the present invention, ectoparasite saliva products can be extracted from a collection means

(described related PCT Patent Publication No. WO 96/11,271) by contacting a collection means with a Tris buffer containing sodium chloride, alcohol and Tris. A more preferred extraction buffer includes 2.5 M NaCl, 5? IPA and 20 mM Tris, about pH 8.0 to about pH 8.3. Suitable extraction times for elutmg proteins and other products from the collection means using the Tris buffer are described in detail in the Examples.

Further concentration of saliva proteins extracted from a collection means of the present invention can be performed by concentrating the extracted flea saliva product-containing solution using hydrophobic interaction chromatographic (HIC) resins. Suitable HIC resins include any resins that bind protein at high salt concentrations. Preferred HIC resins include, for example, butyl-, octyl- and phenyl-substrate conjugated resins. A more preferred resm includes a phenyl-sepharose resm. In a preferred embodiment, extracted flea saliva proteins contained m a Tr s buffer of the present invention can be contacted with a HIC resm to bind the flea saliva proteins to the resin.

In accordance with the present invention, a "mimetope" refers to any compound that is able to mimic the ability of an isolated ectoparasite saliva protein of the present invention to carry out its function (e.g., anti- coagulation, anti-complement, vasodialators, proteases, acid phosphatases or detecting and/or treating the

hypersensitivity of an animal susceptible to or having allergic dermatitis) . A mimetope can be a peptide that has been modified to decrease its susceptibility to degradation but that still retains the desired activity. Other examples of mimetopes include, but are not limited to, carbohydrate- based compounds, lipid-based compounds, nucleic acid-based compounds, natural organic compounds, synthetically derived organic compounds, anti-idiotypic antibodies and/or catalytic antibodies, or fragments thereof. Mimetopes of the present invention can also include non-proteinaceous portions of ectoparasite saliva products having allergenic and/or antigenic activity (e.g., carbohydrate moieties associated with ectoparasite saliva proteins) . A mimetope can be obtained by, for example, screening libraries of synthetic compounds for compounds capable of altering the ability of ectoparasites to feed, or of detecting and/or treating allergic dermatitis resulting from the bites of ectoparasites. A mimetope can also be obtained by, for example, rational drug design. In a rational drug design procedure, the three-dimensional structure of a compound of the present invention can be analyzed by, for example, nuclear magnetic resonance (NMR) or x-ray crystallography. The three-dimensional structure can then be used to predict structures of potential mimetopes by, for example, computer modeling. The predicted mimetope structures can then be produced by, for example, chemical synthesis, recombinant

DNA technology, or by isolating a mimetope from a natural source (e.g., plants, animals, bacteria and fungi) .

One embodiment of the present invention is an m vi vo test that is capable of detecting whether an animal is hypersensitive to ectoparasite saliva products. An m vi vo test of the present invention can initially be used to determine if an animal is hypersensitive to ectoparasite saliva products and then used to determine if an animal is hypersensitive to a particular ectoparasite saliva component, in particular to an ectoparasite saliva protein. An i n vi vo hypersensitivity test of the present invention is particularly useful for identifying animals susceptible to or having allergic dermatitis. An m vi vo hypersensitivity test of the present invention is even more useful for identifying animals susceptible to or having FAD. A suitable in vi vo hypersensitivity test of the present invention can be, but is not limited to, a skin test comprising administering (e.g., mtradermally injecting or superficial scratching) an effective amount of a formulation containing at least one ectoparasite saliva product, or a mimetope thereof. Methods to conduct sk tests of the present invention are known to those of skill in the art and are briefly disclosed herein.

Suitable formulations to use in an m vi vo skin test include one or more isolated ectoparasite saliva proteins of the present invention.

A suitable amount of ectoparasite saliva protein for use a skin test of the present invention can vary widely depending on the allergenicity of the product used m the test and on the site at which the product is delivered. Suitable amounts of ectoparasite saliva proteins for use in a skin test of the present invention include an amount capable of forming reaction, such as a detectable wheal or induration (hardness) resulting from an allergic reaction to the product. Preferred amounts of ectoparasite saliva proteins for use m a skin test of the present invention range from about 1 nanogra (ng) to about 500 micrograms (μg) , more preferably from about 5 ng to about 300 μg, and even more preferably from about 10 ng to about 50 μg of ectoparasite saliva proteins. It is to be appreciated by those of skill m the art that such amounts will vary depending upon the allergenicity of the protein (s) being administered.

According to the present invention, ectoparasite saliva proteins of the present invention can be combined with an immunopotentiator (e.g., carriers or adjuvants of the present invention as defined in detail below) . A novel aspect, however, of the present invention is that an ectoparasite saliva protein of the present invention can

induce a hypersensitive response m the absence of an lmmunopotentiator.

A skin test of the present invention further comprises administering a control solution to an animal. A control solution can include a negative control solution and/or a positive control solution. A positive control solution of the present invention contains an effective amount of at least one compound known to induce a hypersensitive response when administered to an animal. A preferred compound for use as positive control solution includes, but is not limited to, histamine. A negative control solution of the present invention can comprise a solution that is known not to induce a hypersensitive response when administered to an animal. As such, a negative control solution can comprise a solution having compounds essentially incapable of inducing a hypersensitive response or simply a buffer used to prepare the formulation, such as saline. An example of a preferred negative control solution is phenolated phosphate buffered saline (available from Greer Laboratories, Inc., Lenoir, NC) .

Hypersensitivity of an animal to one or more formulations of the present invention can be evaluated by measuring reactions (e.g., wheal size, induration or hardness; using techniques known to those skilled m the art) resulting from administration of one or more experimental sample (s) and control sample (s) into an animal

and comparing the reactions to the experimental sample (s) with reactions resulting from administration of one or more control solution. Preferred devices for mtradermal injections include individual syringes. Preferred devices for scratching include devices that permit the administration of a number of samples at one time. The hypersensitivity of an animal can be evaluated by determining if the reaction resulting from administration of a formulation of the present invention is larger than the reaction resulting from administration of a negative control, and/or by determining if the reaction resulting from administration of the formulation is at least about the same size as the reaction resulting from administration of a positive control solution. As such, if an experimental sample produces a reaction greater than or equal to the size of a wheal produced by administration of a positive control sample to an animal, then that animal is hypersensitive to the experimental sample. Conversely, if an experimental sample produces a reaction similar to the reaction produced by administration of a negative control sample to an animal, then that animal is not hypersensitive to the experimental sample.

Preferred wheal sizes for evaluation of the hypersensitivity of an animal range from about 16 mm to about 8 mm, more preferably from about 15 mm to about 9 mm,

and even more preferably from about 14 mm to about 10 mm in diameter.

Preferably, the ability or inability of an animal to exhibit an immediate hypersensitive response to a formulation of the present invention is determined by measuring wheal sizes from about 2 minutes to about 30 minutes after administration of a sample, more preferably from about 10 minutes to about 25 minutes after administration of a sample, and even more preferably about 15 minutes after administration of a sample.

Preferably, the ability or inability of an animal to exhibit a delayed hypersensitive response to a formulation of the present invention is determined by measuring induration and/or erythema from about 18 hours to about 30 hours after administration of a sample, more preferably from about 20 hours to about 28 hours after administration of a sample, and even more preferably at about 24 hours after administration of a sample. A delayed hypersensitivity response can also be measured using other techniques such as by determining, using techniques known to those of skill in the art, the extent of cell infiltrate at the site of administration during the time periods defined directly above.

In a preferred embodiment, a skin test of the present invention comprises mtradermally injecting into an animal at a given site an effective amount of a formulation that

includes at least one flea saliva protein of the present invention, and mtradermally injecting an effective amount of a control solution into the same animal at a different site. It is withm the scope of one of skill the art to use devices capable of delivering multiple samples simultaneously at a number of sites, preferably enabling concurrent evaluation of numerous formulations. One preferred formulation comprises flea saliva products collected in accordance with the present invention. Also preferred are formulations comprising one or more recombmantly produced flea saliva proteins.

Suitable flea saliva proteins for use with a skin test of the present invention include proteins having an ammo acid sequence such as is listed m the Sequence Listing herein, or homologues thereof. A preferred positive control sample can be a sample comprising histam e. A preferred negative control sample can be a sample comprising diluent.

Animals suitable and preferred to test for hypersensitivity to ectoparasite saliva proteins using a skin test of the present invention are disclosed herein. Particularly preferred animals to test with a skin test of the present invention include dogs, cats and horses, with dogs and cats being even more preferred.

Another embodiment of the present invention is an m vi tro immunoabsorbent test that is capable of detecting the presence of an antibody capable of binding to one or more ectoparasite saliva proteins of the present invention by contacting a putative antibody-contammg solution with a solution containing ectoparasite saliva proteins m such a manner that lmmunocomplexes can form and be detected. Thus, an in vi tro immunoabsorbent test of the present invention is particularly useful for identifying animals susceptible to or having allergic dermatitis by demonstrating that an animal has been previously exposed to an ectoparasite saliva antigen and, therefore may be hypersensitive to further exposure to an ectoparasite saliva antigen.

According to the present invention, an m vi tro hypersensitivity test of the present invention can be, but is not limited to, an immunoabsorbent test comprising: (a) contacting a formulation of the present invention with a body fluid from an animal under conditions sufficient for formation of an lmmunocomplex between the formulation and antibodies, if present, in the body fluid; and (b) determining the amount of lmmunocomplex formed, wherem formation of the lmmunocomplex indicates that the animal is susceptible to or has allergic dermatitis. The immunoabsorbent test is particularly useful for the detection of IgE antibodies m the body fluid, thereby

indicating immediate hypersensitivity in the animal. Determining the amount of lmmunocomplex formed can include the step of separating depending on the mode of detection. Immunoabsorbent assays can be a variety of protocols and can be set-up by those of skill in the art.

A preferred immunoabsorbent test of the present invention comprises a first step of coating one or more portions of a solid substrate with a suitable amount of one or more ectoparasite saliva proteins of the present invention or a mimetope thereof, and of coating one or more other portions of the (or another) solid substrate with a suitable amount of positive and/or negative control solutions of the present invention. A preferred solid substrate of the present invention can include, but is not limited to, an ELISA plate, a dipstick, a radioimmunoassay plate, agarose beads, plastic beads, immunoblot membranes and paper; a more preferred solid substrate includes an ELISA plate, a dipstick or a radioimmunoassay plate, with an ELISA plate and a dipstick being even more preferred. As used herein, a dipstick refers to any solid material having a surface to which antibodies can be bound, such solid material havmg a stick-like shape capable if being inserted into a test tube. Suitable and preferred flea saliva proteins for use with an m vi tro hypersensitivity test of the present invention are as disclosed for a skin test of the present invention.

A second step of a preferred m vi tro hypersensitivity test of the present invention comprises contacting the coated substrate with a body fluid, such as serum, plasma or whole blood, from an animal susceptible to allergic dermatitis in such a manner as to allow antibodies contained in the body fluid that are capable of binding to ectoparasite saliva products to bind to such products bound to the substrate to form immunocomplexes. Excess body fluid and antibodies are then washed from the substrate. In a preferred embodiment in which IgE antibodies the body fluid are to be measured, the body fluid can be pretreated to remove at least some of the other isotypes of immunoglobulin and/or other proteins, such as albumin, present in the fluid. Such removal can include, but is not limited to, contacting the body fluid with a material, such a Protein G, to remove IgG antibodies and/or affinity purifying the IgE antibodies from other components of the body fluid by exposing the fluid to, for example, Concanavalm A (Con-A) . A third step of a preferred m vi tro hypersensitivity test of the present invention comprises contacting the immunocomplexes bound to the substrate with a compound capable of binding to the immunocomplexes, such as a secondary antibody or other compound that is capable of binding to the heavy chain of allergy-related antibodies

produced by animals allergic to ectoparasites, such a manner that the compound (s) can bind to the immunocomplexes. Preferred binding compounds include, but are not limited to, secondary antibodies capable of binding to the heavy chain of IgE antibodies and Fc receptors (FcR) that bind to IgE antibodies (i.e., epsilon FcR), including single chains of an FcR (e.g., the alpha chain of an epsilon FcR) , as well as truncated forms with or without transmembrane domains. Preferred animals to test are disclosed herein. Compounds capable of binding to immunocomplexes are usually tagged with a label which enables the amount of compound bound to the antibody from the body fluid to be measured. Such labels include, but are not limited to, a radioactive label, an enzyme capable of producing a color reaction upon contact with a substrate, a fluorescent label, a chemiluminescent label, a chromophoric label or a compound capable of being bound by another compound. Preferred labels include, but are not limited to, fluorescein, radioisotopes, alkaline phosphatases, biotin, avidin, or peroxidases .

A fourth step of a preferred m vi tro hypersensitivity test of the present invention comprises measuring the amount of detectable label bound to the solid substrate using techniques known to those of skill in the art. It is withm the scope of the present invention that the amount of antibody from the body fluid bound to the substrate can

be determined using one or more layers of secondary antibodies or other binding compounds. For example, an untagged secondary antibody can be bound to a serum antibody and the untagged secondary antibody can then be bound by a tagged tertiary antibody.

A hypersensitive animal is identified by comparing the level of lmmunocomplex formation using samples of body fluid with the level of lmmunocomplex formation using control samples. An lmmunocomplex refers to a complex comprising an antibody and its ligand (i.e., antigen) . As such, immunocomplexes form using positive control samples and do not form using negative control samples. As such, if a body fluid sample results in lmmunocomplex formation greater than or equal to lmmunocomplex formation using a positive control sample, then the animal from which the fluid was taken is hypersensitive to the ectoparasite saliva product bound to the substrate. Conversely, if a body fluid sample results in lmmunocomplex formation similar to lmmunocomplex formation using a negative control sample, then the animal from which the fluid was taken is not hypersensitive to the ectoparasite saliva product bound to the substrate.

A preferred embodiment of an in vi tro hypersensitivity test of the present invention comprises the steps of: (a) contacting an ELISA plate, which is coated with a suitable amount of flea saliva extract (disclosed in related PCT

Patent Publication No. WO 96/11,271, published April 18, 1996, by Frank et al . ; this publication is incorporated by reference herein its entirety), including FS-1, FS-2, FS-3 and/or one or more flea saliva proteins (disclosed in related PCT Patent Publication No. WO 96/11,271 and disclosed herein) , with serum, plasma or whole blood from an animal being tested for susceptibility to allergic dermatitis; and (b) identifying whether immunocomplexes are formed by step (a) by assaying for the presence of such immunocomplexes by (i) contacting the plate with an antibody that specifically binds to IgE or other compounds capable of binding to such immunocomplexes, such as an epsilon Fc receptor, and (ii) determining whether such an antibody or other compound is bound thereto. It should be noted that citing of specific embodiments does not preclude the use of a variety of other immunoassay protocols, including those in which a compound that binds IgE is coated onto a substrate; the substrate is then contacted with serum, plasma or whole blood; and binding of IgE by the compound is detected by the ability to bind flea saliva extracts or proteins of the present invention.

One embodiment of the present invention is a kit useful for identification of an animal susceptible to or having allergic dermatitis. As used herein, a suspect animal is an animal to be tested. A kit of the present invention comprises a formulation of the present invention

and a means for determining if an animal is susceptible to or has allergic dermatitis, in which the formulation is used to identify animals susceptible to or having allergic dermatitis. A means for determining if an animal is susceptible to or has allergic dermatitis can include an in vi vo or in vi tro hypersensitivity test of the present invention as described in detail above. A kit of the present invention further comprises at least one control solution such as those disclosed herein. A preferred kit of the present invention comprises the elements useful for performing an immunoassay. A kit of the present invention can comprise one or more experimental samples {i.e., formulations of the present invention) and one or more control samples bound to at least one pre- packed dipstick or ELISA plate, and the necessary means for detecting immunocomplex formation (e.g., labeled secondary antibodies or other binding compounds and any necessary solutions needed to resolve such labels, as described in detail above) between antibodies contained in the bodily fluid of the animal being tested and the proteins bound to the dipstick or ELISA plate. It is within the scope of the invention that the kit can comprise simply a formulation of the present invention and that the detecting means can be provided in another way.

An alternative preferred kit of the present invention comprises elements useful for performing a skin test. A kit of the present invention can comprise at least one pre¬ packed syringe and needle apparatus containing one or more experimental samples and/or one or more control samples.

It is withm the scope of the present invention that two or more different m vi vo and/or in vi tro tests can be used m combination for diagnostic purposes. For example, the immediate hypersensitivity of an animal to an ectoparasite saliva allergen can be tested using an m vi tro immunoabsorbent test capable of detecting IgE antibodies specific for an ectoparasite saliva allergen m the animal's bodily fluid. While most animals that display delayed hypersensitivity to an ectoparasite saliva allergen also display immediate hypersensitivity to the allergen, a small number of animals that display delayed hypersensitivity to an allergen do not display immediate hypersensitivity to the allergen. In such cases, following negative results from the IgE-specific m vi tro test, the delayed hypersensitivity of the animal to an ectoparasite saliva allergen can be tested using an in vi vo test of the present invention.

Another aspect of the present invention includes treating animals susceptible to or havmg allergic dermatitis, with a formulation of the present invention.

According to the present invention, the term treatment can refer to the regulation of a hypersensitive response by an animal to bites from ectoparasites. Regulation can include, for example, lmmunomodulation of cells involved in the animal's hypersensitive response or alteration of the ability of an ectoparasite to introduce allergens into an animal, for example by inhibiting the anti-coagulation activity of a saliva enzyme, thereby impairing the ability of the arthropod to penetrate the dermis of an animal and feed. lmmunomodulation can include modulating the activity of molecules typically involved in an immune response

(e.g., antibodies, antigens, major histocompatibility molecules (MHC) and molecules co-reactive with MHC molecules) . In particular, lmmunomodulation refers to modulation of antigen: antibody interactions resulting m inflammatory responses, immunosuppression, and immunotolerization of cells involved in a hypersensitive response. Immunosuppression refers to inhibiting an immune response by, for example, killing particular cells involved in the immune response. Immunotolerization refers to inhibiting an immune response by anergiz g (i.e., diminishing reactivity of a T cell to an antigen) particular cells involved in the immune response. Suitable and preferred ectoparasites against which to treat an animal are disclosed herein. A particularly preferred formulation of the present invention is used to treat FAD.

One embodiment of the present invention is a therapeutic composition that, when administered to an animal in an effective manner, is useful for lmmunomodulating the immune response of the animal (i.e., lmmunomodulating the animal) so as to block (i.e., to inhibit, reduce or substantially prevent) a hypersensitive response by the animal upon subsequent exposure to allergenic components transmitted through bites from ectoparasites. Such a therapeutic composition is useful for lmmunomodulating animals known to be hypersensitive to ectoparasite saliva products and animals susceptible to hypersensitive responses against ectoparasite saliva products .

One embodiment of the present invention is a therapeutic composition that includes de-sensitizmg compounds capable of inhibiting an immune response to an ectoparasite saliva protein of the present invention. Such de-sensitizmg compounds include blocking compounds, toleragens and/or suppressor compounds. Blocking compounds comprise compounds capable of modulating antigen:antibody interactions that can result in inflammatory responses, toleragens are compounds capable of lmmunotoleπzmg an animal, and suppressor compounds are capable of lmmunosuppressmg an animal. A de-sensitizmg compound of the present invention can be soluble or membrane-bound. Membrane-bound de-sensitizmg compounds can be associated

with biomembranes, including cells, liposomes, planar membranes, cochleates or micelles. A soluble de¬ sensitizing compound of the present invention is useful for: (1) inhibiting a Type I hypersensitivity reaction by blocking IgE:antιgen mediated de-granulation of mast cells; (2) inhibiting a Type III hypersensitivity reaction by blocking IgG:antigen complex formation leading to complement destruction of cells; and (3) inhibiting a Type IV hypersensitivity reaction by blocking T helper cell stimulation of cytokme secretion by macrophages. A membrane-bound de-sensitizmg compound of the present invention is useful for: (1) inhibiting a Type II hypersensitivity reaction by blocking IgG:antigen complex formation on the surface of cells leading to complement destruction of cells; (2) inhibiting a Type II hypersensitivity reaction by blocking IgG regulated signal transduction in immune cells; and (3) inhibiting a Type IV hypersensitivity reaction • by blocking T cytotoxic cell killing of antigen-bearing cells. A de-sensitiz g compound of the present invention can also be covalently linked to a ligand molecule capable of targeting the de-sensitizmg compound to a specific cell involved in a hypersensitive response to ectoparasite saliva products. Appropriate ligands with which to link a de-sensitizmg compound include, for example, at least a portion of an immunoglobulin molecule, cytokmes, lectins,

heterologous allergens, CD8 molecules, CD4 molecules or major histocompatibility molecules (e.g., MHC class I or MHC class II molecules) . Preferred portions of immunoglobulin molecules to link to a de-sensitizmg compound include variable regions capable of binding to immune cell specific surface molecules and constant regions capable of binding to Fc receptors on immune cells, m particular IgE constant regions. Preferred CD8 molecules include at least the extracellular functional domains of the β chain of CD8. Preferred CD4 molecules include at least the extracellular functional domains of CD4. An immune cell refers to a cell involved in an immune response, m particular, cells having MHC class I or MHC class II molecules. Preferred immune cells include antigen presenting cells, T cells and B cells.

In one embodiment, a therapeutic composition of the present invention includes ectoparasite saliva products of the present invention, or mimetopes thereof. Preferred therapeutic compositions include formulations comprising ectoparasite saliva extracts or at least one ectoparasite saliva product (preferably protein) of the present invention or mimetopes thereof.

Suitable therapeutic compositions of the present invention for treating flea allergy dermatitis include flea saliva extracts (such as those disclosed m related PCT Patent Publication No. WO 96/11,271) and other formulations

including at least one flea saliva protein, or a mimetope thereof. Preferred therapeutic compositions include FS-1, FS-2 and/or FS-3 (such as those disclosed related PCT Patent Publication No. WO 96/11,271) as well as at least a portion of at least one flea saliva protein that can be isolated from FS-1, FS-2 and/or FS-3. As such, preferred formulations for use as therapeutic compositions include FS-1, FS-2, FS-3, and/or at least a portion of one or more of the proteins havmg an ammo acid sequence including SEQ ID NO: 53, SEQ ID NO: 62, SEQ ID NO: 65, SEQ ID NO: 70, SEQ ID NO:72, SEQ ID NO: 75, SEQ ID NO:77, SEQ ID NO: 78 and SEQ ID NO: 87.

In another embodiment, a therapeutic composition can include ectoparasite products of the present invention associated with a suitable excipient. A therapeutic composition of the present invention can be formulated m an excipient that the animal to be treated can tolerate. Preferred excipients are capable of maintaining a product of the present invention a form that is capable of being bound by cells involved in an allergic response in an animal such that the cells are stimulated to initiate or enhance an immune response. Examples of such excipients include water, saline, Ringer's solution, dextrose solution, Hank's solution, and other aqueous physiologically balanced salt solutions. Nonaqueous vehicles, such as fixed oils, sesame oil, ethyl oleate, or

tnglycerides may also be used. Other useful formulations include suspensions containing viscosity enhancing agents, such as sodium carboxymethylcellulose, sorbitol, or dextran. Excipients can also contain minor amounts of additives, such as substances that enhance isotonicity and chemical stability. Examples of buffers include phosphate buffer, bicarbonate buffer and Tris buffer, while examples of preservatives include thimerosal, m- or o-cresol, formalin and benzyl alcohol. Standard formulations can either be liquid mjectables or solids which can be taken up m a suitable liquid as a suspension or solution for injection. Thus, in a non-liquid formulation, the excipient can comprise dextrose, human serum albumin, preservatives, etc., to which sterile water or saline can be added prior to administration.

In another embodiment, a therapeutic composition of the present invention can also comprise a carrier or adjuvant, although it is to be appreciated that an advantage of saliva products of the present invention is that adjuvants and/or carriers are not required for administration. Adjuvants are typically substances that generally enhance the immune response of an animal to a specific antigen. Suitable adjuvants include, but are not limited to, cytokmes, chemokmes, and compounds that induce the production of cytokmes and chemokmes (e.g., granulocyte macrophage colony stimulating factor [GM-CSF] ,

macrophage colony stimulating factor [M-CSF] , granulocyte colony stimulating factor [G-CSF] , colony stimulating factor [CSF], erythropoietm [EPO], mterleukm-2 [IL-2], mterleukιn-3 [IL-3], mterleukm-5 [IL-5], mterleukm-6 [IL-6], mterleukm-7 [IL-7], mterleukm-8 [IL-8], mterleukm-10 [IL-10], mterleukm-12 [IL-12], gamma mterferon [IFN-γ], mterferon gamma inducing factor [IGIF], transforming growth factor beta, RANTES [regulated upon activation, normal T cell expressed and presumably secreted], macrophage inflammatory proteins [e.g., MlPlα and MlPlβ], and Leishmania elongation initiating factor

[LeIF] ; bacterial components (e.g., endotoxms, in particular superantigens, exotoxms and cell wall components); aluminum-based salts; calcium-based salts; silica; polynucleotides; toxoids; serum proteins, viral coat proteins; block copolymer adjuvants (e.g., Hunter's Tιtermax ^,M adjuvant [Vaxcel™, Inc. Norcross, GA] , Ribi adjuvants [Ribi ImmunoChem Research, Inc., Hamilton, MT] ; and sapomns and their derivatives (e.g., Quil A [Superfos Biosector A/S, Denmark] . Protein adjuvants of the present invention can be delivered m the form of the protein themselves or of nucleic acid molecules encoding such proteins using the methods described herem.

Carriers are typically compounds that increase the half-life of a therapeutic composition m the treated animal. Suitable carriers include, but are not limited to,

polymeric controlled release formulations, biodegradable implants, liposomes, bacteria, viruses, oils, esters, ana glycols .

One embodiment of the present invention is a controlled release formulation that is capable of slowly releasing a therapeutic composition of the present invention into the bloodstream of an animal. Suitable controlled release formulations include, but are not limited to, biocompatible (including biodegradable) polymers, other polymeric matrices, capsules, microcapsules, microparticles, bolus preparations, osmotic pumps, diffusion devices, liposomes, lipospheres, and transdermal delivery systems. Other controlled release formulations of the present invention include liquids that, upon administration to an animal, form a solid or a gel m si t u .

The present invention also includes a recombinant virus particle therapeutic composition. Such a composition includes a recombinant molecule of the present invention that is packaged in a viral coat and that can be expressed an animal after administration. Preferably, the recombinant molecule is packaging-deficient. A number of recombinant virus particles can be used, including, but not limited to, those based on alphaviruses, poxviruses, adenoviruses, herpesviruses, and retroviruses . Preferred

recombinant particle viruses are those based on alphaviruses (such as Sindbis virus) , herpesviruses and poxviruses. Methods to produce and use recombinant virus particle vaccines are disclosed in U.S. Patent Application Serial No. 08/015/414, filed February 8, 1993, entitled "Recombinant Virus Particle Vaccines", U.S. Patent No. 5,266,313, by Esposito et al. , issued November 30, 1993 and U.S. Patent Application Serial No. 08/602,010, by Haanes et al . , filed January 15, 1996, entitled "Recombinant Canine Herpesvirus", each of the patents and patent application referred to m this section is incorporated by reference herein in its entirety.

When administered to an animal, a recombinant virus particle therapeutic composition of the present invention infects cells withm the immunized animal and directs the production of a protective protein or RNA nucleic acid molecule that is capable of protecting the animal from allergic dermatitis caused by the bites of ectoparasites. For example, a recombinant virus particle comprising a nucleic acid molecule encoding one or more ectoparasite saliva protein of the present invention is administered according to a protocol that results m the tolerization of an animal against ectoparasite saliva allergens.

According to one embodiment, a nucleic acid molecule of the present invention can be delivered to an animal as a naked (i.e., not packaged m a viral coat or cellular

membrane) nucleic acid vaccine (e.g., as naked DNA or RNA molecules, such as is taught, for example m Wolff et al . , 1990, Sci ence 247, 1465-1468) . A naked nucleic acid vaccine of the present invention includes a nucleic acid molecule of the present invention and preferably includes a recombinant molecule of the present invention that preferably is replication, or otherwise amplification, competent. A naked nucleic acid vaccine of the present invention can comprise one or more nucleic acid molecules of the present invention m the form of, for example, a dicistromc recombinant molecule. Preferred naked nucleic acid vaccines include at least a portion of a viral genome (i.e., a viral vector) . Preferred viral vectors include those based on alphaviruses, poxviruses, adenoviruses, herpesviruses, and retroviruses, with those based on alphaviruses (such as Smdbis or Semliki virus), species- specific herpesviruses and species-specific poxviruses Demg particularly preferred. Any suitable transcription control sequence can be used, including those disclosed as suitable for protein production. Particularly preferred transcription control sequence include cytomegalovirus intermediate early (preferably in conjunction with Intron- A) , Rous Sarcoma Virus long terminal repeat, and tissue- specific transcription control sequences, as well as transcription control sequences endogenous to viral vectors

if viral vectors are used. The incorporation of "strong" poly (A) sequences are also preferred.

Naked nucleic acid vaccines of the present invention can be administered in a variety of ways, with intramuscular, subcutaneous, mtradermal, transdermal, tranasal and oral routes of administration being preferred. An example of one embodiment is disclosed in PCT Patent Publication No. WO 95/05853, published March 2, 1995. A preferred single dose of a naked nucleic acid vaccine ranges from about 1 nanogram (ng) to about 100 μg, depending on the route of administration and/or method of delivery, as can be determined by those skilled the art. Suitable delivery methods include, for example, by injection, as drops, aerosolized, oral and/or topical. Naked DNA of the present invention can be contained m an aqueous excipient (e.g., phosphate buffered saline) alone or a carrier (e.g., lipid-based vehicles) .

Therapeutic compositions of the present invention can be sterilized by conventional methods which do not result in protein degradation (e.g., filtration) and/or lyophilized.

A therapeutic composition of the present invention can be administered to any animal susceptible to ectoparasite infestation as herein described. Acceptable protocols by which to administer therapeutic compositions of the present invention m an effective manner can vary according to

individual dose size, number of doses, frequency of dose administration, and mode of administration. Determination of such protocols can be accomplished by those skilled in the art. An effective dose refers to a dose capable of treating an animal against hypersensitivity to ectoparasite saliva allergens. Effective doses can vary depending upon, for example, the therapeutic composition used, the arthropod from which the composition was derived, and the size and type of the recipient animal. Effective doses to immunomodulate an animal against ectoparasite saliva allergens include doses administered over time that are capable of alleviating a hypersensitive response by an animal to ectoparasite saliva allergens. For example, a first tolerizing dose can comprise an amount of a therapeutic composition of the present invention that causes a minimal hypersensitive response when administered to a hypersensitive animal. A second tolerizing dose can comprise a greater amount of the same therapeutic composition than the first dose. Effective tolerizing doses can comprise increasing concentrations of the therapeutic composition necessary to tolerize an animal such that the animal does not have a hypersensitive response to the bite of an ectoparasite. An effective dose to desensitize an animal can comprise a concentration of a therapeutic composition of the present invention sufficient to block an animal from havmg a hypersensitive response to the bite of

an ectoparasite. Effective desensitizing doses can include repeated doses having concentrations of a therapeutic composition that cause a minimal hypersensitive response when administered to a hypersensitive animal. A suitable single dose is a dose that is capable of treating an animal against hypersensitivity to ectoparasite saliva allergens when administered one or more times over a suitable time period. For example, a preferred single dose of an ectoparasite saliva product, or mimetope therapeutic composition is from about 0.5 ng to about 1 g of the therapeutic composition per kilogram body weight of the animal. Further treatments with the therapeutic composition can be administered from about 1 hour to 1 year after the original administration. Further treatments with the therapeutic composition preferably are administered when the animal is no longer protected from hypersensitive responses to ectoparasite. Particular administration doses and schedules can be developed by one of skill in the art based upon the parameters discussed above. Modes of administration can include, but are not limited to, subcutaneous, mtradermal, intravenous, nasal, oral, transdermal and intramuscular routes.

A therapeutic composition of the present invention can be used in conjunction with other compounds capable of modifying an animal's hypersensitivity to ectoparasite bites. For example, an animal can be treated with compounds

capable of modifying the function of a cell involved in a hypersensitive response, compounds that reduce allergic reactions, such as by systemic agents or anti-mflammatory agents (e.g., anti-histammes, anti-steroid reagents, anti- inflammatory reagents and reagents that drive immunoglobulin heavy chain class switching from IgE to IgG) . Suitable compounds useful for modifying the function of a cell involved in a hypersensitive response include, but are not limited to, antlhistamines, cromolyn sodium, theophyllme, cyclosporm A, adrenalin, cortisone, compounds capable of regulating cellular signal transduction, compounds capable of regulating adenosme 3',5'-cyclic phosphate (cAMP) activity, and compounds that block IgE activity, such as peptides from IgE or IgE specific Fc receptors, antibodies specific for peptides from IgE or IgE-specific Fc receptors, or antibodies capable of blocking binding of IgE to Fc receptors.

Another aspect of the present invention includes a method for prescribing treatment for animals susceptible to or having allergic dermatitis, using a formulation of the present invention. A preferred method for prescribing treatment for flea allergy dermatitis, for example, comprises: (1) mtradermally injecting nto an animal at one site an effective amount of a formulation containing at least one flea saliva antigen of the present invention, or a mimetope thereof (suitable and preferred formulations are

disclosed herein) ; (2) mtradermally injecting into the animal at a second site an effective amount of a control solution; (3) evaluating if the animal has flea allergy dermatitis by measuring and comparing the wheal size resulting from injection of the formulation with the wheal size resulting from injection of the control solution; and (4) prescribing a treatment for the flea allergy dermatitis.

An alternative preferred method for prescribing treatment for flea allergy dermatitis comprises: (1) contacting a first portion of a sample of bodily fluid obtained from an animal to be tested with an effective amount of a formulation containing at least one flea saliva antigen, or a mimetope thereof (suitable and preferred formulations are disclosed herein) to form a first lmmunocomplex solution; (2) contacting a positive control antibody to form a second lmmunocomplex solution; (3) evaluating if the animal has flea allergy dermatitis by measuring and comparing the amount of lmmunocomplex formation in the first and second lmmunocomplex solutions; and (4) prescribing a treatment for the flea allergy dermatitis. It is to be noted that similar methods can be used to prescribe treatment for allergies caused by other ectoparasites using ectoparasite saliva product formulations as disclosed herein.

Another aspect of the present invention includes a method for monitoring animals susceptible to or havmg allergic dermatitis, using a formulation of the present invention. In vi vo and in vi tro tests of the present invention can be used to test animals for allergic dermatitis prior to and following any treatment for allergic dermatitis. A preferred method to monitor treatment of flea allergy dermatitis (which can also be adapted to monitor treatment of other ectoparasite allergies) comprises: (1) tradermally injecting an animal at one site with an effective amount of a formulation containing at least one flea saliva protein, or a mimetope thereof (suitable and preferred formulations are disclosed herein); (2) mtradermally injecting an effective amount of a control solution into the animal at a second site; and (3) determining if the animal is desensitized to flea saliva antigens by measuring and comparing the wheal size resulting from injection of the formulation with the wheal size resulting from injection of the control solution. An alternative preferred method to monitor treatment of flea allergy dermatitis (which can be adapted to monitor treatments of other ectoparasite allergies) comprises: (1) contacting a first portion of a sample of bodily fluid obtained from an animal to be tested with an effective amount of a formulation containing at least one flea saliva protein or mimetope thereof (suitable and preferred

formulations are disclosed herein) to form a first lmmunocomplex solution; (2) contacting a positive control antibody to form a second lmmunocomplex solution; and (3) determining if the animal is desensitized to flea saliva antigens by measuring and comparing the amount of lmmunocomplex formation m the first and second lmmunocomplex solutions.

The present invention also includes antibodies capable of selectively binding to an ectoparasite saliva protein, or mimetope thereof. Such an antibody is herein referred to as an anti-ectoparasite saliva protein antibody. As used herein, the term "selectively binds to" refers to the ability of such an antibody to preferentially bind to ectoparasite saliva proteins and mimetopes thereof. In particular, the present invention includes antibodies capable of selectively binding to flea saliva proteins. Binding can be measured using a variety of methods known to those skilled m the art including immunoblot assays, immunoprecipitation assays, enzyme immunoassays (e.g., ELISA) , radioimmunoassays, immunofluorescent antibody assays and lmmunoelectron microscopy; see, for example, Sambrook et al . , ibid.

Antibodies of the present invention can be either polyclonal or monoclonal antibodies. Antibodies of the present invention include functional equivalents such as antibody fragments and genetically-engineered antibodies,

including single chain antibodies, that are capable of selectively binding to at least one of the epitopes of the protein or mimetope used to obtain the antibodies. Preferably, an antibody of the present invention has a single site binding affinity of from about IO ¹ M ^~ to about IO ¹ - M ^~* for a flea saliva product of the present invention. A preferred method to produce antibodies of the present invention includes administering to an animal an effective amount of an ectoparasite saliva protein or mimetope thereof to produce the antibody and recovering the antibodies. Antibodies raised against defined proteins or mimetopes can be advantageous because such antibodies are not substantially contaminated with antibodies against other substances that might otherwise cause interference a diagnostic assay or side effects if used in a therapeutic composition.

Antibodies of the present invention have a variety of potential uses that are withm the scope of the present invention. For example, such antibodies can be used (a) as vaccines to passively immunize an animal m order to protect the animal from allergic dermatitis, (b) as positive controls in test kits, and/or (c) as tools to recover desired ectoparasite saliva proteins from a mixture of proteins and other contaminants.

The following examples are provided for the purposes of illustration and are not intended to limit the scope of the present invention.

EXAMPLES It s to be noted that the Examples include a number of molecular biology, microbiology, immunology and biochemistry techniques considered to be known to those skilled m the art. Disclosure of such techniques can be found, for example, in Sambrook et al., ibid. , Borovsky, Arch . Insect Bi ochem . and Phys. , 7:187-210, 1988, and related references. Examples 1 through 16, and the SEQ ID NO's cited therein, of related PCT Publication WO 96/11,271, published April 18, 1996, are incorporated herein by this reference in their entirety. Example 1

This example describes the ammo acid sequence analysis of additional isolated flea saliva proteins from FS-1 extract and eluted from DE-81 filters.

FS-1 flea saliva extract and flea saliva product eluted from DE-81 filters were collected using techniques described m Example 2 of related PCT Publication No. WO 96/11,271. Using standard purification techniques (e.g., C4 reverse phase chromatography; SDS-PAGE gel electrophoresis and blotting; and/or flow through electrophoresis), several proteins were isolated from peak

M and partial ammo acid sequences were determined as described Example 4 of related PCT Publication No. WO 96/11,271. Partial N-termmal ammo acid sequencing indicated that peak M contained fspJ, fspL and fspN proteins (as described m Example 4 of related PCT

Publication No. WO 96/11,271) as well as newly identified proteins referred to herein as fspM(G), fspM(H), fspM(I), fspM(J), fspM(K), fspM(L) and fspM(M) . Flea saliva protein fspM(G), having a molecular weight of about 37 kD, had an N-termmal partial ammo acid sequence of M R G N H V F L

E D G M A D M T G G Q Q M G R D L Y, denoted SEQ ID N0:1. Flea saliva protein fspM(H), having a molecular weight of about 34 kD, had an N-termmal partial ammo acid sequence of K Y R N (Y/D) X T N D P Q Y, denoted SEQ ID NO: 2. Flea saliva protein fspM(I), having a molecular weight of about

10 kD had an N-termmal partial ammo acid sequence of E I K R N D R E P G N L S K I R T V M D K V I K Q T Q, denoted SEQ ID NO: 3. Flea saliva protein fspM(J), having a molecular weight of about 25 kD, had an N-termmal partial ammo acid sequence of L K D N D I Y (A/H) (A/H) R D I N E

I L R V L D P S K, denoted SEQ ID NO: 4. Flea saliva protein fspM(K), havmg a molecular weight of about 30 kD, had an N-termmal partial ammo acid sequence of N Y G R V Q I E D Y T X S N H K D X E E K D Q I N G L, denoted SEQ ID NO : 5. Flea saliva protein fspM(L), having a molecular weight of about 37 kD, had an N-termmal partial am o acid

sequence of K Y R N X Y T N D P Q L K L L D E G, denoted SEQ ID NO: 6. Flea saliva protein fspM(M) was recovered from peak M and subjected to ammo acid sequence analysis as described in Example 4 of related PCT Publication No. WO 96/11,271. Flea saliva protein fsp(M), having a molecular weight of about 31 kD, had an N-terminal partial ammo acid sequence of Y F N D Q I K S V M E P X V F K Y P X A X L, denoted SEQ ID NO:7. A Genbank homology search revealed no significant homology between known ammo acid sequences and those determined for fspM(G), fspM(H), fspM(I), fspM(J), fspM(K), fspM(L) and fspM(M) . Example 2

This example describes the isolation of nucleic acid molecules encoding at least a portion of a fspG flea saliva protein. This example also describes expression of a fspG protein by bacteria.

A. Isolation of fspG4 nucleic acid molecules The partial N-termmal ammo acid sequence of fspG2 (i.e., SEQ ID NO: 29 of related PCT Publication No. WO 96/11,271) was used to synthesize degenerate antisense Primer G2-2, having the nucleic acid sequence 5' TGR TTT CCW ATR AAR TCT TC 3 ' , denoted SEQ ID NO: 8. Primer G2-2 was used combination with the M13 reverse primer (SEQ ID NO: 40; described m Example 7 of related PCT Publication No. WO 96/11,271) , to PCR amplify, using standard techniques, the 5'-terminal portion of the fspG4 gene from

a salivary gland cDNA expression library as described above in Example 6A of related PCT Publication No. WO 96/11,271. The resulting PCR product was approximately 225-bp when visualized on a 1% agarose gel. The nucleotide sequence of the 225-bp PCR fragment was obtained, named nfspG4_ is presented as SEQ ID NO: 9.

The nucleic acid sequence of nfspG4 ₂₂₅ was used to synthesize sense Primer G5, havmg nucleic acid sequence 5' AAT TCG GCA CGA GTG 3', denoted SEQ ID NO: 10. Primer G5 was used in combination with the M13 universal primer (SEQ ID NO: 19; described m Example 6 of related PCT Publication No. WO 96/11,271), to PCR amplify, as described above, the 3'-terminal portion of the fspG4 gene from the salivary gland cDNA expression library described above in Example 6A of related PCT Publication No. WO 96/11,271) . The resulting PCR product, denoted nfspG4 _61(J was approximately 610-bp when visualized on a 1% agarose gel. The nucleotide sequence of the 610-bp PCR fragment was obtained, 565 nucleotides of which are presented as SEQ ID NO: 11. The nucleic acid molecule containing nucleic acid sequence SEQ ID NO: 11 is referred to herein as nfspG4 _bj . Translation of SEQ ID NO: 11 suggests that nucleic acid molecule nfspG4 , _t encodes a full-length fspG protein of about 90 ammo acids, referred to herein as PfspG4 ₉₀ , assuming an open reading frame having a start codon spanning from about nucleotide 45 through about nucleotide

47 of SEQ ID NO: 11 and a stop codon spanning from about nucleotide 315 through about nucleotide 317 of SEQ ID NO: 11. This open reading frame, excluding the stop codon, comprises nucleic acid molecule nfspG4 _/G of the present invention, the nucleic acid sequence of which is represented herem by SEQ ID NO: 13. PfspG4 ₉₀ is denoted herem as SEQ ID N0:12. Residues 20-42 of SEQ ID NO:12 appear to be identical to SEQ ID NO:29 of related PCT Publication No. WO 96/11,271 (N-termmal partial ammo acid sequence of fspG2), except that residue 37 of SEQ ID NO: 12 is a glutamic acid rather than a lysine. In addition, residues 38-57 of SEQ ID NO: 12 appear to be identical to SEQ ID NO:30 of related PCT Publication No. WO 96/11,271 (N-terminal partial ammo acid sequence of fspG3) . These similarities support the likelihood of a family of fspG proteins m flea saliva.

Analysis of SEQ ID NO: 11 suggests that the sequence includes a leader segment of about 19 ammo acids followed by a mature protein. The leader sequence is apparently cleaved to form a mature protein termed PfspG4 _71ι denoted SEQ ID NO: 12. PfspG4 ₇₁ has a calculated molecular weight of 7536 daltons and calculated pi of about 9.0. PfspG4 ₉₀ has a calculated molecular weight of 9657 daltons and calculated pi of about 9.26. A Genbank homology search revealed no significant homology between SEQ ID NO: 11 or SEQ ID NO: 12

and known nucleic acid sequences or known ammo acid sequences, respectively. B. Expression

An about 216-bp DNA fragment of nfspG4 was PCR amplified from nucleic acid molecule nfspG4, using: Primer

G7, a sense primer having the nucleic acid sequence 5' AGT

GGA TCC GTC AAA AAT GGT CAC TG 3', denoted as (SEQ ID NO: 15

(BamHI site bold) ; and Primer G8, an antisense primer having the nucleic acid sequence 5' CCG GAA TTC GGT TAT TCG CAA TAA CAG T 3' (£coRI site m bold), denoted SEQ ID

NO: 16. The PCR product, a fragment of about 216 nucleotides, denoted nfspG4 ₂₁₆ , was digested with BamHI and

EcoRI restriction endonucleases, gel purified, and subcloned into expression vector P _R /T ² oπ/S10HIS-RSET-A9 (described in Example 16 of related PCT Publication No. WO

96/11,271) that had been digested with BamHI and EcoRI to produce recombinant molecule pHis-nfspG4 _2U .

The recombinant molecule was transformed into E . coli to form recombinant cell E . coli :pHιs-nfspG4 ₂₁₆ . The recombinant cell was cultured and induced as described m Example 11A of related PCT Publication No. WO 96/11,271 to produce fusion protein PHIS-fspG4 _7> . The recombinant fusion protein was detected by immunoblot analysis using the T7 Tag monoclonal antibody as described in Example 11A of related PCT Publication No. WO 96/11,271.

Exampl e 3

This example describes the isolation of nucleic acid sequences encoding at least a portion of flea saliva proteins fsp (A) , fspM(B), fspM(C), fspM(D), fspM(E), and fspM(F) .

A. nfspM (A) ₈₉₇ and nfspM(B) ₂₇₀₆

A flea salivary gland cDNA library (prepared as described in Example 6 of related PCT Publication No. WO

96/11,271) was immunoscreened with antiserum collected from a rabbit that was immunized with the proteins in peak M2 of the HPLC separation of flea saliva extract described

Example 3 of related PCT Publication No. WO 96/11,271

(i.e., fspM2 proteins) . Immunoscreenmg was performed as described m Example 12 of related PCT Publication No. WO 96/11,271.

A nucleotide sequence for a nfspM nucleic acid molecule named nfspM (A) ₈₉₇ is denoted as SEQ ID NO: 17. Translation of SEQ ID NO: 17 suggests that nucleic acid molecule nfspM (A) ₈₉₇ encodes a full-length fspM protein of about 157 ammo acids, referred to herem as PfspM(A) ₁₅₇ , assuming an open reading frame having a start codon spanning from about nucleotide 97 through about nucleotide 99 of SEQ ID NO: 17 and a stop codon spanning from about nucleotide 568 through about nucleotide 570 of SEQ ID NO: 17. This open reading frame, excluding the stop codon, comprises nucleic acid molecule nfspM (A) _47] of the present

invention, the nucleic acid sequence of which is represented herein by SEQ ID NO: 19. The ammo acid sequence of PfspM (A) _]5/ is denoted SEQ ID NO: 18. PfspM(A) , has a calculated molecular weight of about 18,291.68 daltons and calculated pi of about 10.3. A Genbank homology search revealed no significant homology between SEQ ID NO: 17 or SEQ ID NO: 18 and known nucleic acid or ammo acid sequences, respectively.

A nucleotide sequence for another nfspM nucleic acid molecule named nfspM(B) _27Qf is denoted as SEQ ID NO:20. Translation of SEQ ID NO:20 suggests that nucleic acid molecule nfspM(B) ₂₇₀₆ encodes a non-full-length fspM protein of about 900 ammo acids, referred to herein as PfspM(B) ₉₀₀ , assuming an open reading frame having a start codon spanning from about nucleotide 5 through about nucleotide 7 of SEQ ID NO:20. The ammo acid sequence of PfspM(B) ₉₀₀ is denoted SEQ ID NO:21. PfspM(B) ₉₀₀ has a calculated molecular weight of about 104,647 daltons and calculated pi of about 5.8. The nucleic acid and ammo acid sequences of the nfspM(B) _270o nucleic acid molecule and PfspM(B) ₉₀₀ protein, respectively, were compared to known nucleic acid and ammo acid sequences using a Genbank homology search. SEQ ID NO:21 was found to be similar to the ammo acid sequence of RhoA-b dmg alpha kmase (ROK) . The most highly conserved region of continuous similarity between SEQ ID NO:21 and

ROK ammo acid sequences spans from about ammo acid 32 through about amino acid 351 of SEQ ID NO:21 and from about ammo acid 1 through about amino acid 900 of the ROK, there being about 75% identity between the two regions. Comparison of the nucleic acid sequence encoding ammo acids from about 326 through about 1285 of the ROK kmase with the corresponding regions, spanning nucleotides from about 98 through about 1075 of nfspM(B) _270f indicate that those regions are about 71% identical. B. nfspM (C) ₄₁₄ and nfspM(D) ₂₇ ι

A flea salivary gland cDNA library (prepared as described m Example 6 of related PCT Publication No. WO 96/11,271) was lmmunoscreened with antiserum collected from a rabbit that was immunized with the proteins in peak Ml of the HPLC separation of flea saliva extract described in

Example 3 of related PCT Publication No. WO 96/11,271

(i.e., fspMl proteins) . Immunoscreening was performed as described m Example 12 of related PCT Publication No. WO

96/11,271. Nucleotide sequence for a nfspM nucleic acid molecule named nfspM(C) ₄₁₄ is denoted as SEQ ID NO:22. Translation of SEQ ID NO:22 suggests that nucleic acid molecule nfspM(C) ₄₁₄ encodes a non-full-length fspM protein of about 137 ammo acids, referred to herein as PfspM(C) ₁₃₇ , assuming the first residue spans from about nucleotide 2 through about nucleotide 4 of SEQ ID NO:22. The ammo acid

sequence of PfspMfC),^ is denoted SEQ ID NO: 23. PfspM(C) , has a calculated molecular weight of about 14,452 daltons and calculated pi of about 2.81. A Genbank homology search revealed no significant homology between SEQ ID NO:22 or SEQ ID NO: 23 and known nucleic acid sequences or known ammo acid sequences, respectively.

A nucleotide sequence for another nfspM nucleic acid molecule named nfspM(D) ₂₇₃ is denoted as SEQ ID NO: 24. Translation of SEQ ID NO: 24 suggests that nucleic acid molecule nfspM(D) _{2 3} encodes a non-full-length fspM protein of about 90 ammo acids, referred to herein as PfspM(D) ₉₀ , assuming the first residue spans from about nucleotide 3 through about nucleotide 5 of SEQ ID NO: 24. The am o acid sequence of PfspM(D) ₉₀ ιs denoted SEQ ID NO:25. PfspM(D) ₉₀ has a calculated molecular weight of about 9, 503 daltons and calculated pi of about 3.01. SEQ ID NO: 24 and SEQ ID NO:25 appear to be substantially similar to SEQ ID NO:22 and SEQ ID NO:23, respectively, suggesting a family of fspM proteins m flea saliva. C. nfspM(E) ₁₇₀₄ and nfspM(F) -, _7S8

A flea salivary gland cDNA library (prepared as described in Example 6 as described of related PCT Publication No. WO 96/11,271) was lmmunoscreened with antiserum collected from a rabbit that was immunized with the proteins in peak M2 of the HPLC separation of flea saliva extract described m Example 3 of related PCT

Publication No. WO 96/11,271 (i.e., fspM2 proteins) . Immunoscreenmg was performed as described m Example 12 of related PCT Publication No. WO 96/11,271.

A nucleotide sequence for another nfspM nucleic acid molecule named nfspM (E) ₁₇₀₄ is denoted as SEQ ID NO:26. Translation of SEQ ID NO:26 suggests that nucleic acid molecule nfspM (E) ₁₇₀₄ encodes a full-length fspM protein of about 461 ammo acids, referred to herem as PfspM(E) ₄₆₁ , assuming the first residue spans from about nucleotide 24 through about nucleotide 26 of SEQ ID NO: 26 and a stop codon spanning from about nucleotide 1407 through about nucleotide 1409 of SEQ ID NO:26. This open reading frame, excluding the stop codon, comprises nucleic acid molecule nfspM(E) _ι383 of the present invention, the nucleic acid sequence of which is represented herein by SEQ ID NO: 28. The ammo acid sequence of PfspM(E) _4bl is denoted SEQ ID NO: 27. PfspM(E) ₄₆₁ has a calculated molecular weight of about 54,139 daltons and calculated pi of about 7.00. A Genbank homology search revealed no significant homology between SEQ ID NO:26 or SEQ ID NO:27 and known nucleic acid sequences or known ammo acid sequences, respectively.

A nucleotide sequence for another nfspM nucleic acid molecule named nfspM (F) _{17 H} is denoted as SEQ ID NO:29. Translation of SEQ ID NO:29 suggests that nucleic acid molecule nfspM(F) _175tj encodes a non-full-length fspM protein of about 586 ammo acids, referred to herein as PfspM(F) ₅₈₆ ,

assuming an open reading frame having a start codon spanning from about nucleotide 1 through about nucleotide 3 of SEQ ID NO:29. The ammo acid sequence of PfspM(F) _8β is denoted SEQ ID NO: 30. PfspM(F) ₅₈₆ has a calculated molecular weight of about 66,547 daltons and calculated pi of about 4.80. A Genbank homology search revealed no significant homology between SEQ ID NO:29 or SEQ ID NO: 30 and known nucleic acid sequences or known ammo acid sequences, respectively. Example 4

This Example demonstrates the expression of a fspM protein in E. Coli cells.

Flea saliva protein PHIS-PfspM(D) ₉₀ fusion protein was produced m the following manner. An about 305-bp DNA fragment, referred to herein as nfspM(D) ₃₀ , was isolated from nfspM(D) ₂₉₃ (denoted SEQ ID NO: 31) subcloned into pBluescript plasmid by digesting the nfspM(D) -containing plasmid with BamHI and X ol restriction endonucleases. The digestion product was gel purified and subcloned into expression vector pTrcHisB that had been digested with

BamHI and Xhol, and dephosphorylated. The resultant recombinant molecule, referred to herein as pHis-nfspM(D) _J05 , was transformed into E. coli HB101 competent cells

(available from Gibco BRL, Gaithersburg, MD) to form recombinant cell E . coli :pHis-nfspM(D) ₃₀₅ . The recombinant

cell was cultured and expression of nfspM _J0 induced using conditions described m Example 11A of related PCT Publication No. WO 96/11,271. Immunoblot analysis of recombinant cell E. coli :pHιs-nfspM (D) ₃₀ lysates using a T7 tag monoclonal antibody (Novagen, Ine) directed against the fusion portion of the recombinant PHis-nfspM(D) ₃₀₅ fusion protein identified a protein of the appropriate size, namely an about 15,851 kD protein. Example 5 This example describes the isolation of nucleic acid sequences encoding at least a portion of flea saliva proteins fspN(C), fspN(D), fspN(E), fspN(F), fspN(G), fspN(H), fspN(I), fspN(J), fspN(K), fspN(L), fspN(M), fspN(N) and fspN(O) . A. Preparation of IgE enriched antiserum

Serum was obtained from the artificially sensitized dog CQQ2 (described in Example 8 of related PCT Publication No. WO 96/11,271) . About 10 ml of antiserum was incubated with protein G-Sepharose (5 ml) over night at 4°C. B. Immunoscreenmg with IgE enriched antiserum

About 2.4 ml of Escherichia coli (XL1 Blue, O.D. _60C =0.5) was incubated with 6.48 x 10 ^r pfu of phage from a flea salivary gland ZAP-cDNA library (1.8 x 10 ⁷ pfu/ml), at 37°C for 15 mm and plated m 12 Luπa-Bertani (LB) medium agar plates (150 mm) . The plates were incubated at 37°C over

night. Each plate was then overlaid with an IPTG (lOmM) treated nitrocellulose filters for about 4 hours at 37 C. The filters were then removed and washed with TBST (20 mM Tπs-HCl pH 7.5, 150 mM NaCl, 0.05 Tween-20) . The filters were blocked with 5% dry milk in TBST for 2 hours at room temperature. Different filters were then incubated first with either IgE enriched CQQ2 antiserum or antiserum obtained from dogs infected with Dirofilari a immi ti s ) at 4°C, overnight, then with a monoclonal anti-canme IgE antibody (D-9; gift from the laboratory of Dr. D.J. DeBoer, School of Veterinary Medicine, University of Wisconsin, Madison, WI) , and then with a donkey anti-mouse IgG antibody conjugated to horseradish peroxidase (available from Jackson ImmunoResearch, West Grove, PN) for 2 hours at room temperature at each step. All of the filters were washed with TBST (3 x 15 mm/wash) between each incubation. All of the filters were then treated to a final wash in TBS. Immunocomplexed plaques were identified by immersing the filters into the developing solution (TMB Peroxidase Substrate/TMB Peroxidase Solution/TMB Membrane Enhancer from Kirkegaard & Perry Laboratories) at 1/1/0.1 volume ratio to produce a color reaction. Eighteen plaques were identified and further plaque purified under the same immunoscreenmg condition as described above. C. nfspN(C) _i , nfspN(D) _39l , nfspN(E) , _β- nfspN(F) _{7 f} nfspN(G) _33q , nfspN(G) ₄₉₃ ,

Single plaque of purified clones were isolated and stored in SM phage buffer (50mM Tris, pH 7.4, 0.58 NaCl, 0.2c MgCl 7HO and 0.01% Gelatin) . The m vi vo excision of the pBluescript phagemid from each positive clone was prepared by using ExAssιst™/SOLR ^','M system (Stratagene; . The pBluescript plasmid was purified by plasmid midi kit (Qiagen), and denatured with NaOH (0.4 N) at 37 ^C C for 15 mm. The denatured plasmid was precipitated by ethanol and nucleic acid sequence obtained. A nucleotide sequence for a nfspN nucleic acid molecule named nfspN(C) ₃₃₅ is denoted as SEQ ID NO: 32. A Genbank homology search revealed some similarity between SEQ ID NO: 32 and πbosomal protein S6.

A nucleotide sequence for another nfspN nucleic acid molecule named nfspN(D) ₃₉₆ is denoted as SEQ ID NO: 33. A Genbank homology search revealed some similarity between SEQ ID NO: 33 and erythropoiet .

A nucleotide sequence for another nfspN nucleic acid molecule named nfspN(E) ₂₈ , is denoted as SEQ ID NO: 34. A Genbank homology search revealed some similarity between

SEQ ID NO: 34 and glutamic acid-rich protein or heat-shock protein, HSP81.

A nucleotide sequence for another nfspN nucleic acid molecule named nfspN(F) _22B is denoted as SEQ ID NO:35. Nucleic acid sequence for portions of another nfspN nucleic acid molecule, denoted herem as nfspN(G), were

obtained. The nucleic acid molecule representing a 5' portion of nfspN(G) named nfspN(G) , _(ϋ is denoted as SEQ ID NO: 36. Translation of SEQ ID NO: 36 suggests that nucleic acid molecule nfspN (G) _λ39 encodes a non-full-length fspN(G) protein of about 113 ammo acids, referred to herein as PfspN(G) _n3 , assuming the first residue spans from about nucleotide 1 through about nucleotide 3 of SEQ ID NO: 36. The ammo acid sequence of PfspN(G) ₁₁₃ is denoted SEQ ID NO:37. The nucleic acid molecule representing a 3' portion of nfspN(G) named nfspN(G) ₄₉₃ is denoted as SEQ ID NO: 38. Translation of SEQ ID NO: 38 suggests that nucleic acid molecule nfspN(G) ₄₉₃ encodes a non-full-length fspN(G) protein of about 130 ammo acids, referred to herem as PfspN(G) ₁₃₀ , assuming the first residue spans from about nucleotide 1 through about nucleotide 3 of SEQ ID NO: 38 and a stop codon spanning from about nucleotide 391 through about nucleotide 393 of SEQ ID NO: 38. The ammo acid sequence of Pfsp (G) _o is denoted SEQ ID NO:39. A Genbank homology search revealed some similarity between SEQ ID NO: 36 and SEQ ID NO: 38 and vitellogenm.

A nucleotide sequence for another nfspN nucleic acid molecule named nfspN(H) _30t is denoted as SEQ ID NO: 40.

A nucleotide sequence for another nfspN nucleic acid molecule named nfspN(I) _4QC is denoted as SEQ ID NO: 41.

A nucleotide sequence for another nfspN nucleic acid molecule named nfspN(J) _bU is denoted as SEQ ID NO: 2.

A nucleotide sequence for another nfspN nucleic acid molecule named nfspN(K) _4/ is denoted as SEQ ID NO: 43. A nucleotide sequence for another nfspN nucleic acid molecule named nfspN (L) ₂₉₅ is denoted as SEQ ID NO:44.

A nucleotide sequence for another nfspN nucleic acid molecule named nfspN(M) _37^ is denoted as SEQ ID NO: 45.

Nucleic acid sequence for portions of another nfspN nucleic acid molecule, denoted herein as nfspN(N), were obtained. The nucleic acid molecule representing a 5' portion of nfspN(N) named nfspN(N) ₂₅₂ is denoted as SEQ ID NO:46. The nucleic acid molecule representing a 3' portion of nfspN(N) named nfspN (N) _6]3 is denoted as SEQ ID NO:47. Nucleic acid sequence for portions of another nfspN nucleic acid molecule, denoted herein as nfspN(O), were obtained. The nucleic acid molecule representing a 5' portion of nfspN(O) named nfspN(0) _S38 is denoted as SEQ ID NO: 48. Translation of SEQ ID NO: 48 suggests that nucleic acid molecule nfspN (0) ₅₃₈ encodes a non-full-length fspN(O) protein of about 178 ammo acids, referred to herein as PfspN(0) ₁₇₈ , assuming the first residue spans from about nucleotide 1 through about nucleotide 3 of SEQ ID NO: 48. The ammo acid sequence of PfspN(N) _ι;a is denoted SEQ ID NO: 9.

The nucleic acid molecule representing a 3' portion of nfspN(O) named nfspN(0) ₄ , is denoted as SEQ ID NO:50. Translation of SEQ ID NO: 50 suggests that nucleic acid molecule nfspN(0) _4i , encodes a non-full-length fspN(O) protein of about 129 ammo acids, referred to herein as PfspN(O) _:29 , assuming the first residue spans from about nucleotide 1 through about nucleotide 3 of SEQ ID NO: 50 and a stop codon spanning from about nucleotide 388 through about nucleotide 390 of SEQ ID NO: 50. The ammo acid sequence of PfspN (0) _{1 9} is denoted SEQ ID NO: 51 Example 6

This example describes studies confirming the specificity of IgE enriched antiserum from CQQ2 to fspN protein. Three different petπ dishes (100 mm) were overlaid with 300 microliter per plate of E. coli (XL1 Blue, O.D. _b00 =500) . A drop (about 100 pfu/drop) of each of the eighteen isolated phage clones was dropped onto each plate (18 phage clones/plate) . Using the methods described in Example 5 above, the plates were incubated, filter lifted and the filters immunoscreened with IgE enriched antiserum from CQQ2, antiserum from a D. Immi ti s infected dog and antiserum from rabbits injected with flea saliva product from peak N (as described Example 3 of related PCT Publication No. WO 96/11,271) .

The results of the experiment indicate that ooth the IgE enriched CQQ2 antiserum and the antiserum specific for peak N flea saliva product bind to the products of the purified phage clones significantly better than the antiserum from a D. Immi ti s infected dog. Example 7

This example describes the isolation of nucleic acid molecules encoding a fspG flea saliva protein. This example also describes expression of a fspG protein by bacteria.

A DNA probe labeled with ^? P comprising nucleotides from nfspG4 ₆₁₀ (described m Example 2) was used to screen a flea salivary gland cDNA library (described m Example 6 of related PCT Publication No. WO 96/11,706) using standard hybridization techniques. A clone was isolated havmg about a 595 nucleotide insert, referred to herein as nfspG5 _5g having a nucleic acid sequence of the coding strand which is denoted herein as SEQ ID NO: 52. Translation of SEQ ID NO: 52 suggests that nucleic acid molecule nfspG5 _5g5 encodes a full-length flea salivary protein of about 90 ammo acids, referred to herein as PfspG5 ₉₀ , having ammo acid sequence SEQ ID NO: 53, assuming an open reading frame which the initiation codon spans from about nucleotide 46 through about nucleotide 48 of SEQ ID NO: 52 and the termination codon spans from about nucleotide 316 through about nucleotide 318 of SEQ ID NO: 52. The complement of

SEQ ID NO: 52 is represented herein by SEQ ID NO: 54. The coding region encoding PfspG5 _qo , is represented by nucleic acid molecule nfspG5_ ₇ , having a coding strand with the nucleic acid sequence represented by SEQ ID NO: 55 and a complementary strand with nucleic acid sequence SEQ ID NO: 57. The am o acid sequence of PfspG5 ₉₀ (i.e., SEQ ID NO: 53) predicts that PfspG5 ₉₀ has an estimated molecular weight of about 9.6 kD and an estimated pi of about 9.28.

Analysis of SEQ ID NO: 53 suggests the presence of a signal peptide encoded by a stretch of ammo acids spanning from about ammo acid 1 through about ammo acid 19. The proposed mature protein, denoted herein as PfsG5 ₇₁ , contains about 71 ammo acids which is represented herein as SEQ ID NO: 59. The complement of SEQ ID NO: 58 is represented by

SEQ ID NO: 60. The ammo acid sequence of PfspG5 ₇₁ (i.e., SEQ

ID NO: 59) predicts that PfspG5 _π has an estimated molecular weight of about 7.48 kD, and an estimated pi of about 8.28.

Comparison of ammo acid sequence SEQ ID NO: 53 with ammo acid sequences reported m GenBank indicates that SEQ ID NO: 53 showed the most homology, i.e., about 38% identity between SEQ ID NO: 53 and a Ctenocephalides feli s fl ea sal i vary pro tein FS-H precursor (GenBank accession

U63544) . Comparison of nucleic acid sequence SEQ ID NO: 52 with nucleic acid sequences reported m GenBank indicates

that SEQ ID NO: 52 showed the most homology, i.e., about 63% identity between SEQ ID NO: 52 and a Ctenocephal i des felis flea sali vary protein FS-H precursor gene (GenBank accession U63544) . Flea salivary protein PfspG5 ₇₁ was produced in the following manner. An about 213 bp nucleic acid molecule, referred to herein as nfspG5 _2]3 (designed to encode an apparently mature flea salivary protein) was PCR amplified from nfspG5 ₅₉₅ using sense primer G7 having the nucleotide sequence 5' A GTG GAT CCG TCA AAA ATG GTC ACT G-3'

(containing an BamHI-site shown m bold; denoted SEQ ID

NO: 79) and anti-sense primer G8 having the nucleotide sequence 5' CC GGA ATT CGG TTA TTC GCA ATA ACA GT-3'

(containing a £coRI shown m bold; denoted SEQ ID NO: 80) . The resulting PCR product nfspG5 _?)3 was digested with BamHI and £coRI restriction endonucleases, gel purified, and subcloned into expression vector lambdaP _R /T ⁷ orι/SlOHIS-RSET- A9, that had been digested with BamHI and £co.RI and dephosphorylated. The resultant recombinant molecule, referred to herein as pCro-nfspG5 _2]3 , was transformed into E . coli BL-21 competent cells (available from Novagen, Madison, WI) to form recombinant cell E. coli:pCro-nfspG5 ₂₁₃ . The recombinant cell was cultured and induced as described in Example 11A of related PCT Publication No. WO 96/11,271. Immunoblot analysis of the proteins using a T7 antibody

showed expression of an about 12 kD protein m the induced sample but not in the unmduced sample. Example 8

This example describes the further sequencing of a nucleic acid sequence encoding a fspl flea saliva protein. This example also describes expression of a fspl protein by bacteria.

The nucleic acid molecule denoted nfspl<, ₇₃ described in Example 6 of related PCT Publication No. WO 96/11,706 was further sequenced using standard nucleotide sequencing methods. A nucleic acid molecule was identified of about 1007 nucleotides, referred to herein as nfspl _1007/ the coding strand is denoted herein as SEQ ID NO: 61. Translation of SEQ ID NO: 61 suggests that SEQ ID NO: 61 encodes a non-full- length flea salivary protein of about 155 ammo acids, referred to herein as Pfspl ₁₅₅ , having ammo acid sequence SEQ ID NO: 62, assuming the first codon spans from about nucleotide 1 through about nucleotide 3 of SEQ ID NO: 61 and the termination codon spans from about nucleotide 466 through about nucleotide 468 of SEQ ID NO: 61. The complement of SEQ ID NO: 61 is represented herein by SEQ ID NO: 63.

Flea salivary protein Pfspl ₁₅₆ was produced m the following manner. An about 474-bp nucleic acid molecule, referred to herein as nfspl ₄₇₄ (designed to encode an apparently mature flea salivary protein) was PCR amplified

from nfspl _i0υ7 using sense primer II havmg the nucleotide sequence 5' GCG CGG ATC CGC ATA TGG AAG ACA TCT GGA AAG TTA ATA

AAA AAT GTA CAT CAG-3 ¹ (containing an BamHI-site shown m bold as well as nucleic acid sequence encoding three am o acids, Glu-Asp-Isoleucme, shown in italics; denoted SEQ ID NO: 81) and anti-sense primer 12 having the nucleotide sequence 5' CCG GAA TTC TTA TTT ATT TTT TGG TCG ACA ATA ACA AAA GTT TCC-3' (containing a coRI shown m bold; denoted SEQ ID NO: 82) . The resulting PCR product nfspl ₄₇₄ which contained the nucleic acid sequences incorporated into primer II that encode three ammo acids, was digested with BamHI and £coRI restriction endonucleases, gel purified, and subcloned into expression vector lambdaP _R /T or.-/S10HIS- RSET-A9, that had been digested with BamHI and Xbal and dephosphorylated. The resultant recombinant molecule, referred to herein as pCro-nfspI ₄₇₄ , was transformed into E . coli BL-21 competent cells (available from Novagen, Madison, WI) to form recombinant cell E. coli :pCro-nfspl _4?4 . The recombinant cell was cultured and protein production resolved using the methods described in Example 11A of related PCT Publication No. WO 96/11,271. Immunoblot analysis of the proteins using a T7 antibody showed expression of an about 30 kD protein m the induced sample but not in the unmduced sample.

Example 9

This example describes the isolation of nucleic acid molecules encoding a fspN flea saliva protein.

A DNA probe comprising nucleotides from nfspN(B) ₆ (SEQ ID NO:52 of related PCT Publication No. WO 96/11,706) was labeled with ⁱ P and used to screen the flea salivary gland cDNA library using standard hybridization techniques. A clone was isolated having about a 1205 nucleotide insert, referred to herein as nfspN5 _12o;> havmg a nucleic acid sequence of the coding strand which is denoted herein as SEQ ID NO: 64. Translation of SEQ ID NO: 64 suggests that nucleic ac d molecule nfspN5 ₁₂₀ encodes a non-full-length flea salivary protein of about 353 ammo acids, referred to herein as PfspN5 _3b3 , having amino acid sequence SEQ ID NO: 65, assuming an open reading frame m which the initiation codon spans from about nucleotide 4 through about nucleotide 6 of SEQ ID NO: 64 and the termination codon spans from about nucleotide 1060 through about nucleotide 1062 of SEQ ID NO: 64. The complement of SEQ ID NO: 64 is represented herem by SEQ ID NO: 66. The coding region encoding PfspN5 ₃₅₃ , is represented by nucleic acid molecule nfspN5 ₁₀₅₉ , having a coding strand with the nucleic acid sequence represented by SEQ ID NO: 67 and a complementary strand with nucleic acid sequence SEQ ID NO: 69. The ammo acid sequence of PfspN5 _3s , (i.e., SEQ ID NO: 65) predicts that

PfspN5 _< has an estimated molecular weight of about 39.7 kD and an estimated pl of about 9.45.

Comparison of ammo acid sequence SEQ ID NO: 65 with ammo acid sequences reported m GenBank indicates that SEQ ID NO: 65 showed the most homology, i.e., about 32° identity between SEQ ID NO: 65 and a Human prostatic acid phosphatase precursor protein (GenBank accession P15309) . A GenBank homology search revealed no significant homology between SEQ ID NO: 64 and known nucleic acid sequences. Example 10

This example describes the isolation of nucleic acid molecules encoding a fspN flea saliva protein identified using IgE antibodies isolated from a dog having clinical flea allergy dermatitis. A pool of sera (referred to herein as Pool #4) was collected from numerous known to have clinic flea allergy dermatitis (FAD) . Pool #4 sera was used to identify flea saliva antigens that bind specifically to IgE antibodies m the FAD dog sera as follows. Flea saliva extract was collected using the general methods described in Examples 1 and 2 of related PCT Publication No. WO 96/11,706, except a carboxymethyl cation exchange (CM) membrane (available from Schleicher and Scheull, Keene, NH) was used rather than a Durapore® membrane. In addition, flea saliva extract was eluted from the membrane by contacting the membrane m an extraction buffer of 2.5 M NaCl, 5° ₀

isopropyl alcohol (IPA) and 20 mM Tris, pH 8.0. The membrane was eluted overnight at room temperature. The flea saliva extract was resolved by high pressure liquid chromatography (HPLC) using the method generally described in Example 2 of related PCT Publication No. WO 96/11,706. Proteins contained the HPLC fractions were resolved on a 16°o Tris-glycme SDS PAGE gel. Proteins on the gel were then blotted to an Immobilon P™ filter (available from Millipore Co., Bedford, MA) using standard Western Blot techniques. IgE antibodies bound to protein on the blot was then detected as follows. The blot was first incubated with about a 1:200 dilution of Pool #4 sera using standard antibody hybridization techniques, washed, and then incubated with about a 1:500 dilution of a 145 μg/milliliter solution of biotmylated human Fc R alpha chain protein using standard Western Blot techniques. Following washing, the blot was incubated with about a 1:5,000 dilution of streptavid conjugated to alkaline phosphatase (available from Sigma, St. Louis, MO) . About 10 milliliter of BCIP/NBT substrate (available from Gibco BRL, Gaithersburg, MD) was then added to the blot, incubated until visible bands appeared, at room temperature, and then the blot was rinsed in water to stop the reaction. Protein bands were detected m samples containing Fractions 34, 37, 38, 47, 49, 51, 52 and 53.

Ammo (N-) terminal ammo acid sequencing analysis was performed on protein contained in the about 40 kD protein band identified m the sample containing Fraction 52, using standard procedures known to those in the art (see, for example, Geisow et al . , 1989, m Protein Sequencing: A Practi cal Approach, JBC Fmdlay and MJ Geisow (eds.), IRL Press, Oxford, England, pp. 85-98; Hewick et al. , 1981, J. Bi ol . Chem. , Vol. 256, pp. 7990-7997) . The N-termmal partial ammo acid sequence of the protein was determined to be X Glu Leu Lys Phe Val Phe Val Met Val Lys Gly Pro Asp His Glu Ala Cys Asn Tyr Ala Gly Gly X Gin (denoted herein as SEQ ID NO: 70; wherem "X" represents any ammo acid residue) .

Synthetic oligonucleotide primers were designed using SEQ ID NO: 70 and used to isolate a nucleic acid molecule encoding SEQ ID NO: 70 as follows. Sense primer 1 having the nucleotide sequence 5' AAA TTT GTA(T) TTT GTA(T) ATG GTA(T) AAA GGA(T) CCA(T) GAT CAT GAA GC -3' (denoted SEQ ID NO: 83) was used m combination with the M13 forward universal standard primer 5' GTAAAACGACGGCCAGT 3' (denoted SEQ ID NO: 84) to produce a PCR product from the a flea salivary gland cDNA library described above in Example 9. PCR amplification was conducted using standard techniques. The resulting PCR amplification product was a fragment of about 406 nucleotides, denoted herein as nfspN6 _40b . The PCR product

was cloned into the InVitrogen, Corp., TA™ cloning vector (procedures provided by InVitrogen, Corp.) and subjected to DNA sequence analysis using standard techniques.

The nucleic acid sequence of the coding strand of nfspN6 _40o is denoted herein as SEQ ID NO: 71. Translation of SEQ ID NO: 71 suggests that nucleic acid molecule nfspN6 _40t encodes a non-full-length flea salivary protein of about 135 ammo acids, referred to herem as PfspN6 ₁₃ , having amino acid sequence SEQ ID NO: 72, assuming the first codon spans from about nucleotide 1 through about nucleotide 3 of SEQ ID NO: 71 and the last codon spans from about nucleotide 403 through about nucleotide 405 of SEQ ID NO: 71. The complement of SEQ ID NO: 71 is represented herein by SEQ ID NO: 73. A GenBank homology search revealed no significant homology between amino acid sequence SEQ ID NO:72 and nucleic acid sequence SEQ ID NO: 71 and known amino acid sequences or nucleic acid sequences, respectively. Example 11 This example describes the isolation of nucleic acid molecules encoding a fspj flea saliva protein.

Degenerate oligonucleotide primers were designed from the ammo acid sequence deduced for fspJ (described m Example 4 of related PCT Publication No.WO 96/11,706) and were used to isolate a fspJ nucleic acid molecule as follows. Two synthetic oligonucleotides were synthesized

that corresponded to the region of fspJ spanning from about residues 7 through about 26 of SEQ ID NO: 8 of related PCT Publication No.WO 96/11,706. Primer 1, a "sense" primer corresponding to ammo acid residues fro about residue 7 to about 16 of SEQ ID NO: 8 of related PCT Publication No. O 96/11,706, has the nucleotide sequence 5' CAT GAA CCA(T) GGA(T) AAT ACA(T) CGA(T) AAA(G) ATA(C/T) A(C)G 3' (denoted herein as SEQ ID NO: 84) . Primer 2, a "sense" primer corresponding to ammo acid residues form about residue 17 through about 26 of SEQ ID NO: 8 of related PCT Publication No. WO 96/11,706, has the nucleic acid sequence 5' GAA GTA(T) ATG GAC(T) AAA TTA(G) AGA(G) CAA(G) GC -3' (denoted herein as SEQ ID NO: 86) .

PCR amplification of fragments from the flea salivary gland cDNA library described above m Example 9 was conducted using standard techniques. PCR amplification products were generated using a combination of Primer 1 and M13 primer (denoted SEQ ID NO: 85) . The resultant PCR products were used for a nested PCR amplification using Primer 2 and the T7 standard primer 5' GTA ATA CGA CTC ACT ATA TAG GGC 3' (denoted SEQ ID NO: 88) . The resultant PCR product, a fragment of about 420 nucleotides, denoted herein as nfspJ ₄₂₀ . The PCR product was cloned into the InVitrogen, Corp., TA ^11*1 cloning vector (procedures provided by InVitrogen, Corp.) and subjected to DNA sequence analysis using standard techniques.

The nucleic acid sequence of the coding strand of nfspJ ₄ - ₀ is denoted herein as SEQ ID NO: 74. Translation of SEQ ID NO: 74 suggests that nucleic acid molecule nfspJ _42ϋ encodes a non-full-length flea salivary protein of about 72 ammo acids, referred to herein as PfspJ^, having amino acid sequence SEQ ID NO: 75, assuming the first codon spans from about nucleotide 1 through about nucleotide 3 of SEQ ID NO:74 and the last codon spans from about nucleotide 214 through about nucleotide 216 of SEQ ID NO: 74. The complement of SEQ ID NO: 74 is represented herein by SEQ ID NO: 76.

A GenBank homology search revealed no significant homology between amino acid sequence SEQ ID NO:75 and nucleic acid sequence SEQ ID NO: 74 and known ammo acid sequences or nucleic acid sequences, respectively. Example 12

This example describes the ammo acid sequence analysis of an isolated and HPLC purified fspN7 flea saliva protein. Fractions of flea saliva proteins described above m

Example 10 were tested for the ability to stimulate T cell clones that respond specifically to the flea saliva extract described m Example 10 (FS-specific T cells) . T cell activation were performed using standard methods such as those described in Current Protocols m Immunol ogy, Vol. 1, Chapter 3 [3.13.2], ed. J.E. Coligan et al . , pub. Wiley

Interscience, 1993. Briefly, about 10' FS-1-specxfic I cells (clone CP02-7; isolated from dog CP02 described m Example 8 of related PCT Patent Publication No. WO 96/11,271) were added to individual wells of a 96 well tissue culture plate, in the presence of about 2 x 104 autologous antigen presenting cells (isolated by ficoll gradient from dog CP02) and about 100 units/milliliter of recombinant human mterleukm-2 (Proleukm®; available from Chiron Inc., Emeryville, CA) . About 1 microliter of each fraction of protein resolved by HPLC was to added to each well m triplicate. The cells were incubated for about 4 to about 6 days. About 16 hours prior to harvesting, about 1 μCi of tritiated thymid e (available from Amersham Inc., Arlington Heights, IL) was added to each well. The cells were then harvested and the amount of tritium incorporated into the cellular protein was determined. The results indicated that protein contained m a HPLC fraction containing fspN protein (Fraction 51) stimulated the FS- specific T cells. Ammo (N-) terminal amino acid sequencing analysis was performed on protein contained in Fraction 51 using standard procedures known to those in the art (see, for example, Geisow et al . , ibid. ; Hewick et al . , 1981, ibid. ) . The N-termmal partial ammo acid sequence of the band was determined to be Asn Asp Lys Leu Gin Phe Val Phe Val Met

Ala Arg Gly Pro Asp His Glu Ala Cys Asn Tyr Pro Gly Gly Pro (denoted herein as SEQ ID NO:78) . Example 13

This example describes the ammo acid sequence analysis of an isolated and HPLC purified fspM2 flea saliva protein.

Proteins contained withm Fraction 47 described above m Example 10 were resolved on a 16% Tris-glycme SDS PAGE gel. A major band at about 34 kD was identified. Ammo (N-) terminal ammo acid sequencing analysis was performed on protein contained in the about 34 kD using standard procedures known to those m the art (see, for example, Geisow et al., ibid. ; Hewick et al., 1981, ibid. ) . The N- termmal partial ammo acid sequence of the band was determined to be Tyr Phe Asn Lys leu Val Gin Ser Trp Thr Glu Pro Met Val Phe Lys Tyr Pro Tyr (denoted herein as SEQ ID NO:87) .

SEQUENCE LISTING

The following Sequence Listing is submitted pursuant to 37 CFR §1.821. A copy in computer readable form is also submitted herewith.

Applicants assert pursuant to 37 CFR §1.821 (f) that the content of the paper and computer readable copies of SEQ ID NO:l through SEQ ID NO: 88 submitted herewith are the same.

(1) GENERAL INFORMATION:

(i) APPLICANT: Frank, Glenn R.

Wu Hunter, Shirley allenfels, Lynda

(ll) TITLE OF INVENTION: NOVEL ECTOPARASITE SALIVA PROTEINS AND APPARATUS TO COLLECT SUCH PROTEINS (in) NUMBER OF SEQUENCES: 88

(iv) CORRESPONDENCE ADDRESS:

(A) ADDRESSEE: SHERIDAN ROSS P.C.

(B) STREET: 1700 LINCOLN ST., SUITE 3500 (C) CITY: DENVER

(D) STATE: CO

(E) COUNTRY: U.S.A.

(F) ZIP: 80203 (v) COMPUTER READABLE FORM:

(A) MEDIUM TYPE: Floppy disk

(B) COMPUTER: IBM PC compatible

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

(D) SOFTWARE: Patentln Release #1.0, Version #1.30

(vi) CURRENT APPLICATION DATA:

(A) APPLICATION NUMBER:

(B) FILING DATE:

(C) CLASSIFICATION:

(viii) ATTORNEY/AGENT INFORMATION:

(A) NAME: Connell, Gary J.

(B) REGISTRATION NUMBER: 32,020

(C) REFERENCE/DOCKET NUMBER: 2618-17-C4

(IX) TELECOMMUNICATION INFORMATION:

(A) TELEPHONE: 303/863-9700

(B) TELEFAX: 303/863-0223

(2) INFORMATION FOR SEQ ID NO:l:

(l) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 26 amino acids

(B) TYPE: amino acid

(C) STRANDEDNESS:

(D) TOPOLOGY: linear

(n) MOLECULE TYPE: protein

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

Met Arg Gly Asn His Val Phe Leu Glu Asp Gly Met Ala Asp Met Thr 1 5 10 15

Gly Gly Gin Gin Met Gly Arg Asp Leu Tyr 20 25

(2) INFORMATION FOR SEQ ID NO: 2:

(l) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 12 amino acids

(B) TYPE: amino acid

(C) STRANDEDNESS: (D) TOPOLOGY: linear

(li) MOLECULE TYPE: protein

(IX) FEATURE: (A) NAME/KEY: Xaa = Tyr or Asp

(B) LOCATION: 5

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:2: Lys Tyr Arg Asn Xaa Xaa Thr Asn Asp Pro Gin Tyr

1 5 10

(2) INFORMATION FOR SEQ ID NO:3:

(l) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 27 amino acids

(B) TYPE: amino acid

(C) STRANDEDNESS: (D) TOPOLOGY: linear

(ii) MOLECULE TYPE: protein

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

Glu lie Lys Arg Asn Asp Arg Glu Pro Gly Asn Leu Ser Lys lie Arg 1 5 10 15

Thr Val Met Asp Lys Val lie Lys Gin Thr Gin 20 25

(2) INFORMATION FOR SEQ ID NO: 4 : (l) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 23 amino acids

(B) TYPE: amino acid

(C) STRANDEDNESS:

(D) TOPOLOGY: linear

(11) MOLECULE TYPE: protein

(ix) FEATURE:

(A) NAME/KEY- Xaa = Ala or His

(B) LOCATION: 8

(ix) FEATURE:

(A) NAME/KEY: Xaa = Ala or His

(B) LOCATION: 9

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

Leu Lys Asp Asn Asp lie Tyr Xaa Xaa Arg Asp lie Asn Glu lie Leu 1 5 10 15

Arg Val Leu Asp Pro Ser Lys 20

(2) INFORMATION FOR SEQ ID NO:5:

(l) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 27 amino acids (B) TYPE: amino acid

(C) STRANDEDNESS:

(D) TOPOLOGY: linear

(n) MOLECULE TYPE: protein

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

Asn Tyr Gly Arg Val Gin lie Glu Asp Tyr Thr Xaa Ser Asn His Lys

1 5 10 15

Asp Xaa Glu Glu Lys Asp Gin lie Asn Gly Leu 20 25

(2) INFORMATION FOR SEQ ID NO: 6:

(l) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 18 amino acids

(B) TYPE: amino acid (C) STRANDEDNESS:

(D) TOPOLOGY: linear

(ll) MOLECULE TYPE: protein (XI) SEQUENCE DESCRIPTION' SEQ ID NO: 6:

Lys Tyr Arg Asn Xaa Tyr Thr Asn Asp Pro Gin Leu Lys Leu Leu Asp

1 5 10 15 Glu Gly

(2) INFORMATION FOR SEQ ID NO:7: (l) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 22 amino acids

(B) TYPE: amino acid

(C) STRANDEDNESS:

(D) TOPOLOGY: linear

(n) MOLECULE TYPE: protein

(XI) SEQUENCE DESCRIPTION: SEQ ID NO: :

Tyr Phe Asn Asp Gin lie Lys Ser Val Met Glu Pro Xaa Val Phe Lys 1 5 10 15

Tyr Pro Xaa Ala Xaa Leu 20

(2) INFORMATION FOR SEQ ID NO: 8 : (ι) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 20 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(n) MOLECULE TYPE: DNA (genomic)

( x) FEATURE:

(A) NAME/KEY: mιsc_feature (B) LOCATION: 1..20

(D) OTHER INFORMATION: /label= primer

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:8: TGRTTTCCWA TRAARTCTTC 20

(2) INFORMATION FOR SEQ ID NO: 9: (i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 225 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(li) MOLECULE TYPE: DNA (genomic)

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:9: GAATTCGGCA CGAGTGAAAT TCAATATTTT GTTTTACATT AAATTTTTCA AATTCGATAT 60

GAAATTTTTA CTGGCAATTT GCGTGTTGTG TGTTTTATTA AATCAAGTAT CTATGTCAAA 120

AATGGTCACT GAAAAGTGTA AGTCAGGTGG AAATAATCCA AGTACAGAAG AGGTGTCAAT 180

ACCATCTGGG AAGCTTACTA TTGAAGATTT TTGTATTGGA AATCA 225

(2) INFORMATION FOR SEQ ID NO:10: (l) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 15 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(li) MOLECULE TYPE: DNA (genomic)

(ix) FEATURE:

(A) NAME/KEY: mιsc_feature (B) LOCATION: 1..15

(D) OTHER INFORMATION: /label= primer

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

AATTCGGCAC GAGTG 15

(2) INFORMATION FOR SEQ ID NO:11:

(l) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 565 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: single (D) TOPOLOGY: linear

(n) MOLECULE TYPE: cDNA

(ix) FEATURE: (A) NAME/KEY: CDS

(B) LOCATION: 45..314

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 11:

TGAAATTCAA TATTTTGTTT TACATTAAAT TTTTCAAATT CGAT ATG AAA TTT TTA 56

Met Lys Phe Leu

1 CTG GCA ATT TGC GTG TTG TGT GTT TTA TTA AAT CAA GTA TCT ATG TCA 104 Leu Ala lie Cys Val Leu Cys Val Leu Leu Asn Gin Val Ser Met Ser 5 10 15 20

AAA ATG GTC ACT GAA AAG TGT AAG TCA GGT GGA AAT AAT CCA AGT ACA 152 Lys Met Val Thr Glu Lys Cys Lys Ser Gly Gly Asn Asn Pro Ser Thr

25 30 35

GAA GAG GTG TCA ATA CCA TCT GGG AAG CTT ACT ATT GAA GAT TTT TGT 200 Glu Glu Val Ser lie Pro Ser Gly Lys Leu Thr lie Glu Asp Phe Cys 40 45 50

ATT GGA AAT CAT CAA AGT TGC AAA ATA TTT TAC AAA AGT CAA TGT GGA 248 lie Gly Asn His Gin Ser Cys Lys He Phe Tyr Lys Ser Gin Cys Gly 55 60 65

TTT GGA GGT GGT GCT TGT GGA AAC GGT GGT TCA ACA CGA CCA AAT CAA 296 Phe Gly Gly Gly Ala Cys Gly Asn Gly Gly Ser Thr Arg Pro Asn Gin 70 75 80 AAA CAC TGT TAT TGC GAA TAACCATATT CCGGATGAAA GACCAAATTG 344

Lys His Cys Tyr Cys Glu 85 90

ATATAAATTA CTAAAATTAT GCTAGATAGC AATCATAAAA TTTTGAAGTT TTCAATGATC 404

CTAACATGTT TTGCCTCCAA TTTATTTTAA CAGCAAATTG CTGGAACTTA CCGTACCGTA 464

ACTAAATGTT CAAGAAATAC TGAATGTTTA CAAATAGATT ATTATAAATA TTGTAACATT 524 GTCTAA ATT TATAAGAATT ATATAAACTG AATTGCAAAA A 565

(2) INFORMATION FOR SEQ ID NO: 12: (l) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 90 amino acids

(B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein

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

Met Lys Phe Leu Leu Ala He Cys Val Leu Cys Val Leu Leu Asn Gin

1 5 10 15

Val Ser Met Ser Lys Met Val Thr Glu Lys Cys Lys Ser Gly Gly Asn

20 25 30

Asn Pro Ser Thr Glu Glu Val S e r He Pro Ser Gly Lys Leu Thr He

35 40 45

Glu Asp Phe Cys He Gly Asn His Gin Ser Cys Lys He Phe Tyr Lys

50 55 60

Ser Gin Cys Gly Phe Gly Gly Gly Ala Cys Gly Asn Gly Gly Ser Thr 65 70 75 SO

Arg Pro Asn Gin Lys His Cys Tyr Cys Glu 85 90

(2) INFORMATION FOR SEQ ID NO: 13:

(l) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 270 base pairs

(B) TYPE: nucleic acid (C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: cDNA (ix) FEATURE:

(A) NAME/KEY: CDS

(B) LOCATION: 1..270

( i) SEQUENCE DESCRIPTION: SEQ ID NO: 13:

ATG AAA TTT TTA CTG GCA ATT TGC GTG TTG TGT GTT TTA TTA AAT CAA 48

Met Lys Phe Leu Leu Ala He Cys Val Leu Cys Val Leu Leu Asn Gin 1 5 10 15

GTA TCT ATG TCA AAA ATG GTC ACT GAA AAG TGT AAG TCA GGT GGA AAT 96 Val Ser Met Ser Lys Met Val Thr Glu Lys Cys Lys Ser Gly Gly Asn 20 25 30 AAT CCA AGT ACA GAA GAG GTG TCA ATA CCA TCT GGG AAG CTT ACT ATT 144 Asn Pro Ser Thr Glu Glu Val Ser He Pro Ser Gly Lys Leu Thr He 35 40 45

GAA GAT TTT TGT ATT GGA AAT CAT CAA AGT TGC AAA ATA TTT TAC AAA 192 Glu Asp Phe Cys He Gly Asn His Gin Ser Cys Lys He Phe Tyr Lys 50 55 60

AGT CAA TGT GGA TTT GGA GGT GGT GCT TGT GGA AAC GGT GGT TCA ACA 240 Ser Gin Cys Gly Phe Gly Gly Gly Ala Cys Gly Asn Gly Gly Ser Thr 65 70 75 80

CGA CCA AAT CAA AAA CAC TGT TAT TGC GAA 270

Arg Pro Asn Gin Lys His Cys Tyr Cys Glu 85 90

(2) INFORMATION FOR SEQ ID NO: 14:

(l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 90 amino acids

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

(11) MOLECULE TYPE: protein

( XI ) SEQUENCE DESCRI PTION : SEQ ID NO : 14 :

Met Lys Phe Leu Leu Al a He Cys Val Leu Cys Val Leu Leu Asn Gin

1 5 10 15

Val Ser Met Ser Lys Met Val Thr Glu Lys Cys Lys Ser Gly Gly Asn 20 25 30

Asn Pro Ser Thr Glu Glu Val Ser He Pro Ser Gly Lys Leu Thr He 35 40 45

Glu Asp Phe Cys He Gly Asn His Gin Ser Cys Lys He Phe Tyr Lys 50 55 60 Ser Gin Cys Gly Phe Gly Gly Gly Ala Cys Gly Asn Gly Gly Ser Thr 65 70 75 80

Arg Pro Asn Gin Lys His Cys Tyr Cys Glu 85 90

(2) INFORMATION FOR SEQ ID NO: 15:

(l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 26 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear (11) MOLECULE TYPE: DNA (genomic)

(ix) FEATURE:

(A) NAME/KEY: mιsc_feature

(B) LOCATION: 1..26 (D) OTHER INFORMATION: /label= primer

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 15: AGTGGATCCG TCAAAAATGG TCACTG 26

(2) INFORMATION FOR SEQ ID NO: 16:

(1) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 28 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: single (D) TOPOLOGY: linear

(n) MOLECULE TYPE: DNA (genomic)

(ix) FEATURE: (A) NAME/KEY: πusc_feature

(B) LOCATION: 1..28

(D) OTHER INFORMATION: /label= primer

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 16: CCGGAATTCG GTTATTCGCA ATAACAGT 28

(2) INFORMATION FOR SEQ ID NO: 17:

(1) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 897 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY : linear ( li ) MOLECULE TYPE : cDNA

( ix ) FEATURE :

(A) NAME/KEY: CDS

(B) LOCATION: 97..568

( i) SEQUENCE DESCRIPTION: SEQ ID NO: 17: CCGAAATCTC CTATCACAGT GTACGGAGTG TAAAATATTG TTGAAGTATT TTGAAATTTA 60

TTAATTTATT CGAAAAGGAG ATTTCATTAA ATAAAA ATG GTT TAC GAA AGT GAC 114

Met Val Tyr Glu Ser Asp 1 5

TTT TAC ACG ACC CGT CGG CCC TAC AGT CGT CCG GCT TTG TCT TCA TAC 162 Phe Tyr Thr Thr Arg Arg Pro Tyr Ser Arg Pro Ala Leu Ser Ser Tyr 10 15 20

TCC GTA ACG GCA CGT CCA GAG CCG GTT CCT TGG GAC AAA TTG CCG TTC 210 Ser Val Thr Ala Arg Pro Glu Pro Val Pro Trp Asp Lys Leu Pro Phe 25 30 35 GTC CCC CGT CCA AGT TTG GTA GCA GAT CCC ATA ACA GCA TTT TGC AAG 258 Val Pro Arg Pro Ser Leu Val Ala Asp Pro He Thr Ala Phe Cys Lys 40 ^' 45 50

CGA AAA CCT CGC CGA GAA GAA GTT GTT CAA AAA GAG TCC ATT GTT CGA 306 Arg Lys Pro Arg Arg Glu Glu Val Val Gin Lys Glu Ser He Val Arg 55 60 65 70

AGG ATC AAT TCT GCA GGA ATT AAA CCC AGC CAG AGA GTT TTA TCG GCT 354 Arg He Asn Ser Ala Gly He Lys Pro Ser Gin Arg Val Leu Ser Ala

75 80 85

CCA ATA AGA GAA TAC GAA TCC CCA AGG GAC CAG ACC AGG CGT AAA GTT 402 Pro He Arg Glu Tyr Glu Ser Pro Arg Asp Gin Thr Arg Arg Lys Val 90 95 100

TTG GAA AGC GTC AGA AGA CAA GAA GCT TTT CTG AAC CAA GGA GGA ATT 450

Leu Glu Ser Val Arg Arg Gin Glu Ala Phe Leu Asn Gin Gly Gly He 105 110 115

TGT CCA TTG ACC ACC AGA AAT GAT GAC ATG GAT AGA CTT CTA CCC CGT 498 Cys Pro Leu Thr Thr Arg Asn Asp Asp Met Asp Arg Leu Leu Pro Arg 120 125 130 CTC CAC AGT TCA CAC ACA ACA CCT TCT GCG GAT AGG AAA GTT TTG TTG 546 Leu His Ser Ser His Thr Thr Pro Ser Ala Asp Arg Lys Val Leu Leu 135 140 145 150

ACC ACT TTT CAC AGA AGA TAC T GATTAAAAAT GAAAGTTAAG AAATTTGTTG 598 Thr Thr Phe His Arg Arg Tyr

155

AAGTCATGTG GTGTTTTTTA TACATTCTTT ATTAATCGAT ATTCCTAACG AACGATACGA 658 TAACTTTCGA TAACTTTTTC TGGTTAATTT TGACAAAATA TGCATTTGCA AGCATAACAT 718

TCATTTTCAA GGCAAACGCT TTCTGATGAT TATCTTGTTA AAAGTGTGGA AACAAGCGTA 778

GTGTTAACAA ATGCATTGCT TGTTTTGATT ATTTATTTAT CTATTATATA TTCCATATTG 838

TATTGTAGGT GGTGTACTTG GTATTACTAA TACACGTACT TTGTGAAAAA AAAAAAAAA 897

INFORMATION FOR SEQ ID NO: 18:

(l) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 157 anu.no acids

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

(n) MOLECULE TYPE: protein

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 18: Met Val Tyr Glu Ser Asp Phe Tyr Thr Thr Arg Arg Pro Tyr Ser Arg 1 5 10 15

Pro Ala Leu Ser Ser Tyr Ser Val Thr Ala Arg Pro Glu Pro Val Pro 20 25 30

Trp Asp Lys Leu Pro Phe Val Pro Arg Pro Ser Leu Val Ala Asp Pro 35 40 45

He Thr Ala Phe Cys Lys Arg Lys Pro Arg Arg Glu Glu Val Val Gin 50 55 60

Lys Glu Ser He Val Arg Arg He Asn Ser Ala Gly He Lys Pro Ser

65 70 75 80

Gin Arg Val Leu Ser Ala Pro He Arg Glu Tyr Glu Ser Pro Arg Asp 85 90 95

Gin Thr Arg Arg Lys Val Leu Glu Ser Val Arg Arg Gin Glu Ala Phe 100 105 110

Leu Asn Gin Gly Gly He Cys Pro Leu Thr Thr Arg Asn Asp Asp Met 115 120 125

Asp Arg Leu Leu Pro Arg Leu His Ser Ser His Thr Thr Pro Ser Ala 130 135 140

Asp Arg Lys Val Leu Leu Thr Thr Phe His Arg Arg Tyr 145 150 155

(2) INFORMATION FOR SEQ ID NO: 19:

(l) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 471 base pairs (B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: cDNA

(XI) SEQUENCE DESCRIPTION: SEQ ID NO: 19:

ATGGTTTACG AAAGTGACTT TTACACGACC CGTCGGCCCT ACAGTCGTCC GGCTTTGTCT 60 TCATACTCCG TAACGGCACG TCCAGAGCCG GTTCCTTGGG ACAAATTGCC GTTCGTCCCC 120

CGTCCAAGTT TGGTAGCAGA TCCCATAACA GCATTTTGCA AGCGAAAACC TCGCCGAGAA 180

GAAGTTGTTC AAAAAGAGTC CATTGTTCGA AGGATCAATT CTGCAGGAAT TAAACCCAGC 240

CAGAGAGTTT TATCGGCTCC AATAAGAGAA TACGAATCCC CAAGGGACCA GACCAGGCGT 300

AAAGTTTTGG AAAGCGTCAG AAGACAAGAA GCTTTTCTGA ACCAAGGAGG AATTTGTCCA 360 TTGACCACCA GAAATGATGA CATGGATAGA CTTCTACCCC GTCTCCACAG TTCACACACA 420

ACACCTTCTG CGGATAGGAA AGTTTTGTTG ACCACTTTTC ACAGAAGATA C 471

(2) INFORMATION FOR SEQ ID NO:20:

(l) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 2706 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(11) MOLECULE TYPE: cDNA

(IX) FEATURE:

(A) NAME/KEY: CDS

(B) LOCATION: 5..2706 (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 20:

GCGG ATG AAG AGC ATC GAG GCT TAT ACA AAC AGA TAT GAA ATC ATA GCT 49 Met Lys Ser He Glu Ala Tyr Thr Asn Arg Tyr Glu He He Ala 1 5 10 15

TCT GAA ATA GTT AAT CTT CGA ATG AAA CCA GAT GAT TTT AAT TTA ATA 97 Ser Glu He Val Asn Leu Arg Met Lys Pro Asp Asp Phe Asn Leu He 20 25 30

AAA GTT ATT GGT CGA GGA GCA TTT GGT GAA GTA CAG TTA GTG CGA CAC 145 Lys Val He Gly Arg Gly Ala Phe Gly Glu Val Gin Leu Val Arg His 35 40 45

AAA TCA ACT GCA CAA GTT TTT GCT ATG AAA CGC CTA TCA AAA TTT GAA 193 Lys Ser Thr Ala Gin Val Phe Ala Met Lys Arg Leu Ser Lys Phe Glu 50 55 60 ATG ATT AAG AGA CCA GAC TCT GCA TTT TTT TGG GAA GAA CGT CAT ATA 241

Met He Lys Arg Pro Asp Ser Ala Phe Phe Trp Glu Glu Arg His He 65 70 75

ATG GCT CAT GCA AAA TCA GAA TGG ATT GTA CAA TTA CAT TTT GCT TTT 289 Met Ala His Ala Lys Ser Glu Trp He Val Gin Leu His Phe Ala Phe

80 85 90 95

CAA GAT CAA AAA TAT CTT TAT ATG GTC ATG GAT TAT ATG CCG GGG GGT 337 Gin Asp Gin Lys Tyr Leu Tyr Met Val Met Asp Tyr Met Pro Gly Gly 100 105 110

GAC TTG GTG AGT CTT ATG TCC GAT TAT GAA ATT CCA GAA AAA TGG GCA 385 Asp Leu Val Ser Leu Met Ser Asp Tyr Glu He Pro Glu Lys Trp Ala 115 120 125

ATG TTC TAT ACA ATG GAA GTG GTG CTA GCA CTT GAT ACA ATT CAC TCC 433 Met Phe Tyr Thr Met Glu Val Val Leu Ala Leu Asp Thr He His Ser 130 135 140 ATG GGA TTT GTA CAT CGT GAT GTT AAA CCT GAT AAT ATG CTT CTA GAC 481 Met Gly Phe Val His Arg Asp Val Lys Pro Asp Asn Met Leu Leu Asp 145 150 155

AAA TAT GGT CAT TTA AAG TTA GCT GAC TTT GGA ACC TGT ATG AAA ATG 529 Lys Tyr Gly His Leu Lys Leu Ala Asp Phe Gly Thr Cys Met Lys Met 160 165 170 175

GAT ACA GAT GGT TTG GTA CGT TCT AAT AAT GCT GTT GGA ACG CCT GAT 577 Asp Thr Asp Gly Leu Val Arg Ser Asn Asn Ala Val Gly Thr Pro Asp 180 185 190

TAC ATT TCT CCC GAA GTT TTG CAG TCC CAA GGT GGT GAA GGA GTT TAC 625 Tyr He Ser Pro Glu Val Leu Gin Ser Gin Gly Gly Glu Gly Val Tyr 195 200 205

GGT CGT GAA TGC GAT TGG TGG TCT GTG GGA ATT TTT TTG TAT GAA ATG 673

Gly Arg Glu Cys Asp Trp Trp Ser Val Gly He Phe Leu Tyr Glu Met 210 215 220 TTA TTT GGA GAA ACA CCT TTT TAT GCA GAC AGT TTG GTT GGA ACT TAC 721

Leu Phe Gly Glu Thi Pro Phe Tyr Ala Asp Ser Leu Val Gly Thr Tyr

225 230 235

AGT AAA ATT ATG GAT CAC AGA AAC TCA TTA ACT TTT CCT CCA GAA GTG 769 Ser Lys He Met Asp His Arg Asn Ser Leu Thr Phe Pro Pro Glu Val 240 245 250 255

GAA ATA AGC CAA TAT GCC CGA TCT TTG ATA CAA GGA TTT TTA ACA GAC 817

Glu He Ser Gin Tyr Ala Arg Ser Leu He Gin Gly Phe Leu Thr Asp 260 265 270

AGA ACA CAG CGT TTA GGC AGA AAT GAA GTG GAA GAA ATT AAA CGA CAT 865

Arg Thr Gin Arg Leu Gly Arg Asn Glu Val Glu Glu He Lys Arg His 275 280 285

CCA TTT TTC ATA AAT GAT CAA TGG ACT TTT GAC AAT TTA AGA GAC TCT 913

Pro Phe Phe He Asn Asp Gin Trp Thr Phe Asp Asn Leu Arg Asp Ser 290 295 300 GCC CCA CCT GTA GTG CCA GAG CTG AGT GGT GAT GAT GAT ACA AGG AAC 961

Ala Pro Pro Val Val Pro Glu Leu Ser Gly Asp Asp Asp Thr Arg Asn

305 310 315

TTT GAT GAT ATT GAA CGT GAT GAA ACA CCT GAA GAG AAT TTT CCT ATA 1009 Phe Asp Asp He Glu Arg Asp Glu Thr Pro Glu Glu Asn Phe Pro He

320 325 330 335

CCA AAA ACT TTT GCT GGT AAT CAT CTG CCA TTT GTT GGA TTC ACA TAT 105

Pro Lys Thr Phe Ala Gly Asn His Leu Pro Phe Val Gly Phe Thr Tyr 340 345 350

AAT GGT GAT TAC CAA TTA TTA ACA AAT GGA GGT GTT AGA AAT AGT GAT 1105 Asn Gly Asp Tyr Gin Leu Leu Thr Asn Gly Gly Val Arg Asn Ser Asp 355 360 365

ATG GTT GAT ACA AAA TTA AAC AAC ATT TGT GTT TCA AGT AAG GAT GAT 1153

Met Val Asp Thr Lys Leu Asn Asn He Cys Val Ser Ser Lys Asp Asp 370 375 380 GTG TTA AAT TTA CAA AAT TTA TTA GAA CAA GAG AAA GGT AAC AGT GAA 1201

Val Leu Asn Leu Gin Asn Leu Leu Glu Gin Glu Lys Gly Asn Ser Glu

385 390 395

AAT TTG AAA ACA AAC ACC CAA TTA TTA AGT AAT AAA TTA GAT GAA CTA 1249 Asn Leu Lys Thr Asn Thr Gin Leu Leu Ser Asn Lys Leu Asp Glu Leu 400 405 410 415

GGT CAG AGA GAA TGT GAA TTA AGG AAT CAG GCT GGA GAT TAT GAG AAA 1297

Gly Gin Arg Glu Cys Glu Leu Arg Asn Gin Ala Gly Asp Tyr Glu Lys 420 425 430

GAA TTG ACT AAA TTC AAA TTA TCG TGC AAA GAA TTA CAA CGT AAG GCA 1345

Glu Leu Thr Lys Phe Lys Leu Ser Cys Lys Glu Leu Gin Arg Lys Ala 435 440 ^" 445

GAA TTT GAG AAT GAA TTA CGG CGT AAA ACT GAG TCC TTA CTA GTT GAA 1393 Glu Phe Glu Asn Glu Leu Arg Arg Lys Thr Glu Ser Leu Leu Val Glu 450 455 460 ACA AAG AAA AGA CTA GAC GAA GAG CAG AAT AAA AGA ACT AGA GAA ATG 1441 Thr Lys Lys Arg Leu Asp Glu Glu Gin Asn Lys Arg Thr Arg Glu Met 465 470 475

AAT AAT AAT CAA CAG CAC AAT GAC AAA ATA AAT ATG TTA GAA AAA CAA 1489

Asn Asn Asn Gin Gin His Asn Asp Lys He Asn Met Leu Glu Lys Gin 480 485 490 495

ATT AAT GAT TTA CAA GAA AAA TTG AAA GGT GAA TTA GAG CAC AAT CAG 1537 He Asn Asp Leu Gin Glu Lys Leu Lys Gly Glu Leu Glu His Asn Gin

500 505 510

AAA TTA AAG AAG CAA GCT GTT GAG CTT AGA GTT GCT CAG TCT GCT ACT 1585 Lys Leu Lys Lys Gin Ala Val Glu Leu Arg Val Ala Gin Ser Ala Thr 515 520 525

GAA CAA CTG AAT AAT GAA TTA CAG GAA ACT ATG CAG GGT TTA CAA ACA 1633

Glu Gin Leu Asn Asn Glu Leu Gin Glu Thr Met Gin Gly Leu Gin Thr 530 535 540

CAA AGA GAT GCT TTA CAA CAA GAA GTA GCA TCT CTC CAA GGC AAA CTT 1681

Gin Arg Asp Ala Leu Gin Gin Glu Val Ala Ser Leu Gin Gly Lys Leu

545 550 555 TCT CAA GAG AGG AGC TCT AGA TCA CAG GCT TCT GAT ATG CAG ATA GAA 1729 Ser Gin Glu Arg Ser Ser Arg Ser Gin Ala Ser Asp Met Gin He Glu 560 565 570 575

CTA GAA GCA AAA TTG CAG GCT CTC CAT ATT GAA CTG GAG CAT GTC AGA 1777 Leu Glu Ala Lys Leu Gin Ala Leu His He Glu Leu Glu His Val Arg

580 585 590

AAT TGT GAA GAC AAA GTT ACC CAA GAC AAC AGA CAA CTA TTG GAA AGG 1825 Asn Cys Glu Asp Lys Val Thr Gin Asp Asn Arg Gin Leu Leu Glu Arg 595 600 605

ATA TCA ACA TTG GAG AAA GAA TGT GCT TCT CTA GAA TTA GAA TTG AAA 1873 He Ser Thr Leu Glu Lys Glu Cys Ala Ser Leu Glu Leu Glu Leu Lys 610 615 620

GCA ACA CAA AAC AAA TAT GAG CAA GAG GTC AAA GCA CAT CGC GAA ACT 1921 Ala Thr Gin Asn Lys Tyr Glu Gin Glu Val Lys Ala His Arg Glu Thr 625 630 635 GAA AAA TCA AGA CTG GTC AGT AAA GAA GAA GCA AAT ATG GAG GAA GTT 1969 Glu Lys Ser Arg Leu Val Ser Lys Glu Glu Ala Asn Met Glu Glu Val 640 645 650 655

AAA GCA CTC CAA ATA AAA TTA AAT GAA GAG AAA TCT GCT CGA CAG AAA 2017 Lys Ala Leu Gin He Lys Leu Asn Glu Glu Lys Ser Ala Arg Gin Lys

660 665 670

TCT GAT CAG AAT TCT CAA GAA AAG GAA CGA CAA ATT TCT ATG TTA TCT 2065 Ser Asp Gin Asn Ser Gin Glu Lys Glu Arg Gin He Ser Met Leu Ser 675 680 685

GTG GAT TAT CGT CAA ATC CAA CAG CGT TTG CAA AAG CTA GAA GGA GAA 2113 Val Asp Tyr Arg Gin He Gin Gin Arg Leu Gin Lys Leu Glu Gly Glu 690 695 700

TAT AGG CAA GAG AGT GAA AAA GTT AAA GCT CTC CAC AGT CAG ATT GAG 2161

Tyr Arg Gin Glu Ser Glu Lys Val Lys Ala Leu His Ser Gin He Glu

705 710 715 CAA GAG CAA CTA AAA AAA TCA CAA TTA CAA AGC GAA TTG GGT GTT CAA 2209 Gin Glu Gin Leu Lys Lys Ser Gin Leu Gin Ser Glu Leu Gly Val Gin 720 725 730 735

AGG TCT CAG ACT GCA CAT TTA ACA GCC AGG GAA GCT CAG CTA GTT GGA 2257 Arg Ser Gin Thr Ala His Leu Thr Ala Arg Glu Ala Gin Leu Val Gly

740 745 750

GAA GTT GCT CAT CTT AGA GAT GCT AAA AGA AAT GTT GAA GAA GAG TTA 2305

Glu Val Ala His Leu Arg Asp Ala Lys Arg Asn Val Glu Glu Glu Leu 755 760 765

CAC AAG TTA AAA ACT GCT CGA TCA GTG GAT AAT GCT CAG ATG AAA GAG 2353 His Lys Leu Lys Thr Ala Arg Ser Val Asp Asn Ala Gin Met Lys Glu

770 775 780

CTT CAA GAA CAA GTT GAA GCC GAG CAA GTT TTC TCG ACT CTT TAT AAA 2401

Leu Gin Glu Gin Val Glu Ala Glu Gin Val Phe Ser Thr Leu Tyr Lys 785 790 795

ACA CAT TCT AAT GAA CTT AAG GAA GAA CTT GAG GAA AAA TCT CGT CAT 2449

Thr His Ser Asn Glu Leu Lys Glu Glu Leu Glu Glu Lys Ser Arg His 800 805 810 815

ATT CAA GAA ATG GAA GAA GAA AGA GAA AGT TTG GTT CAT CAG CTA CAA 2497

He Gin Glu Met Glu Glu Glu Arg Glu Ser Leu Val His Gin Leu Gin 820 825 830 ATT GCA TTA GCT AGA GCT GAT TCA GAG GCA TTG GCG AGA TCA ATA GCT 2545

He Ala Leu Ala Arg Ala Asp Ser Glu Ala Leu Ala Arg Ser He Ala 835 840 845

GAT GAA AGT ATA GCT GAT TTA GAA AAG GAA AAG ACT ATG AAG GAA TTA 2593 Asp Glu Ser He Ala Asp Leu Glu Lys Glu Lys Thr Met Lys Glu Leu

850 855 860

GAA CTA AAA GAA TTA TTA AAC AAA AAT CGT ACT GAA CTT TCC CAG AAA 2641

Glu Leu Lys Glu Leu Leu Asn Lys Asn Arg Thr Glu Leu Ser Gin Lys 865 870 875

GAC ATT TCA ATA AGT GCA TTG CGT GAA CGA GAA AAT GAA CAG AAG AAA 2689

Asp He Ser He Ser Ala Leu Arg Glu Arg Glu Asn Glu Gin Lys Lys 880 885 890 895

CTT TTA GAA CAA ATC TC 2706

Leu Leu Glu Gin He 900

(2) INFORMATION FOR SEQ ID NO:21:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 900 airu.no acids (B) TYPE: amino acid

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO:21:

Met Lys Ser He Glu Ala Tyr Thr Asn Arg Tyr Glu He He Ala Ser 1 5 10 15 Glu He Val Asn Leu Arg Met Lys Pro Asp Asp Phe Asn Leu He Lys

20 25 30

Val He Gly Arg Gly Ala Phe Gly Glu Val Gin Leu Val Arg His Lys 35 40 45

Ser Thr Ala Gin Val Phe Ala Met Lys Arg Leu Ser Lys Phe Glu Met 50 55 60

He Lys Arg Pro Asp Ser Ala Phe Phe Trp Glu Glu Arg His He Met 65 70 75 80

Ala His Ala Lys Ser Glu Trp He Val Gin Leu His Phe Ala Phe Gin 85 90 95

Asp Gin Lys Tyr Leu Tyr Met Val Met Asp Tyr Met Pro Gly Gly Asp 100 105 110

Leu Val Ser Leu Met Ser Asp Tyr Glu He Pro Glu Lys Trp Ala Met

115 120 125

Phe Tyr Thr Met Glu Val Val Leu Ala Leu Asp Thr He His Ser Met 130 135 140

Gly Phe Val His Arg Asp Val Lys Pro Asp Asn Met Leu Leu Asp Lys 145 150 155 160

Tyr Gly His Leu Lys Leu Ala Asp Phe Gly Thr Cys Met Lys Met Asp 165 170 175

Thr Asp Gly Leu Val Arg Ser Asn Asn Ala Val Gly Thr Pro Asp Tyr 180 185 190

He Ser Pro Glu Val Leu Gin Ser Gin Gly Gly Glu Gly Val Tyr Gly 195 200 205

Arg Glu Cys Asp Trp Trp Ser Val Gly He Phe Leu Tyr Glu Met Leu 210 215 220 Phe Gly Glu Thr Pro Phe Tyr Ala Asp Ser Leu Val Gly Thr Tyr Ser 225 230 235 240

Lys He Met Asp His Arg Asn Ser Leu Thr Phe Pro Pro Glu Val Glu 245 250 255

He Ser Gin Tyr Ala Arg Ser Leu He Gin Gly Phe Leu Thr Asp Arg 260 265 270

Thr Gin Arg Leu Gly Arg Asn Glu Val Glu Glu He Lys Arg His Pro 275 280 285

Phe Phe He Asn Asp Gin Trp Thr Phe Asp Asn Leu Arg Asp Ser Ala 290 295 300

Pro Pro Val Val Pro Glu Leu Ser Gly Asp Asp Asp Thr Arg Asn Phe 305 310 315 320

Asp Asp He Glu Arg Asp Glu Thr Pro Glu Glu Asn Phe Pro He Pro 325 330 335

Lys Thr Phe Ala Gly Asn His Leu Pro Phe Val Gly Phe Thr Tyr Asn 340 345 350

Gly Asp Tyr Gin Leu Leu Thr Asn Gly Gly Val Arg Asn Ser Asp Met 355 360 365

Val Asp Thr Lys Leu Asn Asn He Cys Val Ser Ser Lys Asp Asp Val

370 375 380 Leu Asn Leu Gin Asn Leu Leu Glu Gin Glu Lys Gly Asn Ser Glu Asn

385 390 395 400

Leu Lys Thr Asn Thr Gin Leu Leu Ser Asn Lys Leu Asp Glu Leu Gly 405 410 415

Gin Arg Glu Cys Glu Leu Arg Asn Gin Ala Gly Asp Tyr Glu Lys Glu 420 425 430

Leu Thr Lys Phe Lys Leu Ser Cys Lys Glu Leu Gin Arg Lys Ala Glu 435 440 445

Phe Glu Asn Glu Leu Arg Arg Lys Thr Glu Ser Leu Leu Val Glu Thr 450 455 460

Lys Lys Arg Leu Asp Glu Glu Gin Asn Lys Arg Thr Arg Glu Met Asn 465 470 475 480

Asn Asn Gin Gin His Asn Asp Lys He Asn Met Leu Glu Lys Gin He

485 490 495

Asn Asp Leu Gin Glu Lys Leu Lys Gly Glu Leu Glu His Asn Gin Lys 500 505 510

Leu Lys Lys Gin Ala Val Glu Leu Arg Val Ala Gin Ser Ala Thr Glu 515 520 525

Gin Leu Asn Asn Glu Leu Gin Glu Thr Met Gin Gly Leu Gin Thr Gin 530 535 540

Arg Asp Ala Leu Gin Gin Glu Val Ala Ser Leu Gin Gly Lys Leu Ser 545 550 555 560

Gin Glu Arg Ser Ser Arg Ser Gin Ala Ser Asp Met Gin He Glu Leu 565 570 575

Glu Ala Lys Leu Gin Ala Leu His He Glu Leu Glu His Val Arg Asn 580 585 590 Cys Glu Asp Lys Val Thr Gin Asp Asn Arg Gin Leu Leu Glu Arg He 595 600 605

Ser Thr Leu Glu Lys Glu Cys Ala Ser Leu Glu Leu Glu Leu Lys Ala 610 615 620

Thr Gin Asn Lys Tyr Glu Gin Glu Val Lys Ala His Arg Glu Thr Glu 625 630 635 640

Lys Ser Arg Leu Val Ser Lys Glu Glu Ala Asn Met Glu Glu Val Lys 645 650 655

Ala Leu Gin He Lys Leu Asn Glu Glu Lys Ser Ala Arg Gin Lys Ser 660 665 670

Asp Gin Asn Ser Gin Glu Lys Glu Arg Gin He Ser Met Leu Ser Val 675 680 685

Asp Tyr Arg Gin He Gin Gin Arg Leu Gin Lys Leu Glu Gly Glu Tyr

690 695 700

Arg Gin Glu Ser Glu Lys Val Lys Ala Leu His Ser Gin He Glu Gin

705 710 715 720

Glu Gin Leu Lys Lys Ser Gin Leu Gin Ser Glu Leu Gly Val Gin Arg 725 730 735

Ser Gin Thr Ala His Leu Thr Ala Arg Glu Ala Gin Leu Val Gly Glu 740 745 750 Val Ala His Leu Arg Asp Ala Lys Arg Asn Val Glu Glu Glu Leu His 755 760 765

Lys Leu Lys Thr Ala Arg Ser Val Asp Asn Ala Gin Met Lys Glu Leu 770 775 780

Gin Glu Gin Val Glu Ala Glu Gin Val Phe Ser Thr Leu Tyr Lys Thr 785 790 795 800 His Ser Asn Glu Leu Lys Glu Glu Leu Glu Glu Lys Ser Arg His He

805 810 815

Gin Glu Met Glu Glu Glu Arg Glu Ser Leu Val His Gin Leu Gin He 820 825 830

Ala Leu Ala Arg Ala Asp Ser Glu Ala Leu Ala Arg Ser He Ala Asp 835 840 845

Glu Ser He Ala Asp Leu Glu Lys Glu Lys Thr Met Lys Glu Leu Glu 850 855 860

Leu Lys Glu Leu Leu Asn Lys Asn Arg Thr Glu Leu Ser Gin Lys Asp 865 870 875 880 He Ser He Ser Ala Leu Arg Glu Arg Glu Asn Glu Gin Lys Lys Leu

885 890 895

Leu Glu Gin He 900

INFORMATION FOR SEQ ID NO:22:

(l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 414 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear (n) MOLECULE TYPE: cDNA

(IX) FEATURE:

(A) NAME/KEY: CDS

(B) LOCATION: 3..414

(xi ) SEQUENCE DESCRIPTION: SEQ ID NO : 22 :

GA GCT GAT GAG AAT GGA AAT GTG ATT AGC ATT ACT GAT GAA AAT GGA 47 Ala Asp Glu Asn Gly Asn Val He Ser He Thr Asp Glu Asn Gly

1 5 10 15

AAC ATT ATT AGT ACT ACT GAT GAG AAT GGA AAT GTG ATT AGC ATT ACT 95 Asn He He Ser Thr Thr Asp Glu Asn Gly Asn Val He Ser He Thr 20 25 30

GAT GAG AAT GGA AAC ATT ATT AGT ACT ACT GAT GAG AAT GGA AAT GTG 143

Asp Glu Asn Gly Asn He He Ser Thr Thr Asp Glu Asn Gly Asn Val 35 40 45

ATT AGC ATT ACT GAT GAA AAT GGA AAC ATT ATT AGT ACT ACT GAT GAG 191 He Ser He Thr Asp Glu Asn Gly Asn He He Ser Thr Thr Asp Glu 50 55 60 AAT GGA AAT GTG ATT AGC ATT ACT GAT GAG AAT GGA AAT GTG ATT AGC 239 Asn Gly Asn Val He Ser He Thr Asp Glu Asn Gly Asn Val He Ser 65 70 75

ATT ACT GAT GAA AAT GGA AAC TCG AAT AGC ACT ACT AGT GTT TTC AAT 287

He Thr Asp Glu Asn Gly Asn Ser Asn Ser Thr Thr Ser Val Phe Asn 80 85 90 95

GAA ACT GAA AAT ATG ACT GGT GCT GCT GAT ACA AAT GAA TAT TCA ATT 335 Glu Thr Glu Asn Met Thr Gly Ala Ala Asp Thr Asn Glu Tyr Ser He

100 105 110

GGT TCT ACT GAC GGA AAT GGA AAT TTT ATA AGT ACT TTT AGT GAT CAT 383 Gly Ser Thr Asp Gly Asn Gly Asn Phe He Ser Thr Phe Ser Asp His 115 120 125

GAT TAC GTA AGT AAT ACT GAA GAA AAT GAA A 414

Asp Tyr Val Ser Asn Thr Glu Glu Asn Glu 130 135

(2) INFORMATION FOR SEQ ID NO: 23:

(l) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 137 amino acids

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

(n) MOLECULE TYPE: protein

(XI) SEQUENCE DESCRIPTION: SEQ ID NO: 23:

Ala Asp Glu Asn Gly Asn Val He Ser He Thr Asp Glu Asn Gly Asn 1 5 10 15

He He S ex Thr Thr Asp Glu Asn Gly Asn Val He Ser He Thr Asp 20 25 30

Glu Asn Gly Asn He He Ser Thr Thr Asp Glu Asn Gly Asn Val He 35 40 45

Ser He Thr Asp Glu Asn Gly Asn He He Ser Thr Thr Asp Glu Asn

50 55 60 Gly Asn Val He Ser He Thr Asp Glu Asn Gly Asn Val He Ser He

65 70 75 80

Thr Asp Glu Asn Gly Asn Ser Asn Ser Thr Thr Ser Val Phe Asn Glu 85 90 95

Thr Glu Asn Met Thr Gly Ala Ala Asp Thr Asn Glu Tyr Ser He Gly 100 105 110

Ser Thr Asp Gly Asn Gly Asn Phe He Ser Thr Phe Ser Asp His Asp 115 120 125

Tyr Val Ser Asn Thr Glu Glu Asn Glu

130 135

(2) INFORMATION FOR SEQ ID NO:24:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 273 base pairs (B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(n) MOLECULE TYPE: cDNA

(ix) FEATURE:

(A) NAME/KEY: CDS

(B) LOCATION: 3..273

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:24:

AT GAG AAT GGA AAT GTG ATT AGC TAT ACT GAT GAA AAT GGA AAC ATT 47

Glu Asn Gly Asn Val He Ser Tyr Thr Asp Glu Asn Gly Asn He 1 5 10 15

ATC AGT ACT ACT GAT GAG AAT GGA AAT GTG ATT AGC ATT ACT GAT GAA 95

He Ser Thr Thr Asp Glu Asn Gly Asn Val He Ser He Thr Asp Glu 20 25 30

AAT GGA AAT GTG ATT AGC ATT ACT GAT GAA AAT GGA AAC ATT ATC AGT 143 Asn Gly Asn Val He Ser He Thr Asp Glu Asn Gly Asn He He Ser 35 40 45

ACT ACT GAT GAG AAT GGA AAT GTG ATT AGC ATT ACT GAT GAA AAT GGA 191 Thr Thr Asp Glu Asn Gly Asn Val He Ser He Thr Asp Glu Asn Gly 50 55 60 AAT GTG ATT AGC ATT ACT GAT GAA AAT GGA AAC ATT ATT AGT ACT ACT 239 Asn Val He Ser He Thr Asp Glu Asn Gly Asn He He Ser Thr Thr 65 70 75

GAT GAG AAT GGA AAT GTG ATT AGC AAT ACT CGA G 273 Asp Glu Asn Gly Asn Val He Ser Asn Thr Arg 80 85 90

(2) INFORMATION FOR SEQ ID NO:25:

(l) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 90 amino acids

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

(n) MOLECULE TYPE: protein

(Xl) SEQUENCE DESCRIPTION: SEQ ID NO: 25:

Glu Asn Gly Asn Val He Ser Tyr Thr Asp Glu Asn Gly Asn He He

1 5 10 15

Ser Thr Thr Asp Glu Asn Gly Asn Val He Ser He Thr Asp Glu Asn 20 25 30

Gly Asn Val He Ser He Thr Asp Glu Asn Gly Asn He He Ser Thr

35 40 45

Thr Asp Glu Asn Gly Asn Val He Ser He Thr Asp Glu Asn Gly Asn 50 55 60

Val He Ser He Thr Asp Glu Asn Gly Asn He He Ser Thr Thr Asp 65 70 75 80 Glu Asn Gly Asn Val He Ser Asn Thr Arg

85 90

(2) INFORMATION FOR SEQ ID NO:26:

(l) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 1704 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: single (D) TOPOLOGY: linear

(ll) MOLECULE TYPE: cDNA

(ix) FEATURE: (A) NAME/KEY: CDS

(B) LOCATION: 24..1406

(Xi) SEQUENCE DESCRIPTION: SEQ ID NO:26:

CAGAAACCCG ACATTCTCAA AAT ATG GAA CCT CAA TCG CTG TCT TGG CAA 50

Met Glu Pro Gin Ser Leu Ser Trp Gin 1 5 CTT CCG ACT CAA GTA GTT CAG CCA GTT TTT GAA CAA CAA ATG CAG ATT 98

Leu Pro Thr Gin Val Val Gin Pro Val Phe Glu Gin Gin Met Gin He 10 15 20 25

CCT GGA TAT AAT ATG CAA ATT CAA TCT AAT TAT TAT CAA ATT CAC CCA 146

Pro Gly Tyr Asn Met Gin He Gin Ser Asn Tyr Tyr Gin He His Pro 30 35 40

GAA ATG TTG GAT CCA AAT TTG AAC AAT CCT CAG CAG TTA ATG TTT AAT 1 4 Glu Met Leu Asp Pro Asn Leu Asn Asn Pro Gin Gin Leu Met Phe Asn

45 50 55

TAT ATG CAA TTA CAA CAA TTG CAG GAA CTA CAA CAT TTA AGT CAA CAA 242 Tyr Met Gin Leu Gin Gin Leu Gin Glu Leu Gin His Leu Ser Gin Gin 60 65 70

CAG CCA ATG CAT CAT GAA TTT GAA CAT CAT ATC CCC ATT CCA CAA GAA 290

Gin Pro Met His His Glu Phe Glu His His He Pro He Pro Gin Glu 75 80 85

GCA ACT TCA ACT AAT TAC GGT CCA TCC GGA CAG TAT ATT ACT AGT GAC 338

Ala Thr Ser Thr Asn Tyr Gly Pro Ser Gly Gin Tyr He Thr Ser Asp 90 95 100 105 GCA ACA TCT TAT CAA TCA ATT GCC CAA CAA TTT GTA CCA CAA CCA CCA 386 Ala Thr Ser Tyr Gin Ser He Ala Gin Gin Phe Val Pro Gin Pro Pro 110 115 120

ATT GAA ACT ACC ACC ACG AAA ATA CCT GAA ACT GAA ATT CAA ATT GGC 434 He Glu Thr Thr Thr Thr Lys He Pro Glu Thr Glu He Gin He Gly

125 130 135

GTT TCG AAT CAA TAT GCC CAA AAT ATA ACT TAT AAT TCA AAT ATC AGT 482 Val Ser Asn Gin Tyr Ala Gin Asn He Thr Tyr Asn Ser Asn He Ser 140 145 150

CCT GAA GTG ATT GGA TTC CGA GAA CAT TAT GTT GCG GAA CAG CCT TCT 530

Pro Glu Val He Gly Phe Arg Glu His Tyr Val Ala Glu Gin Pro Ser

155 160 165

GGT GAC GTG CTT CAC AAA AGT CAT TTA ACA GAA CAA CCA GCA GAT AAA 578

Gly Asp Val Leu His Lys Ser His Leu Thr Glu Gin Pro Ala Asp Lys

170 175 180 185

AGC ACA CGT GGT GAT CAG GAA CCT GTT AGT GAG ACA GGC TCT GGT TTT 626 Ser Thr Arg Gly Asp Gin Glu Pro Val Ser Glu Thr Gly Ser Gly Phe 190 195 200 TCG TAT GCA CAA ATT TTA TCA CAG GGA CTT AAG CCT ACC CAG CCA TCC 674 Ser Tyr Ala Gin He Leu Ser Gin Gly Leu Lys Pro Thr Gin Pro Ser 205 210 215

AAC TCA GTT AAT TTG CTT GCA GAT CGA TCG AGA TCA CCT CTA GAT ACG 722 Asn Ser Val Asn Leu Leu Ala Asp Arg Ser Arg Ser Pro Leu Asp Thr 220 225 230

AAA ACG AAA GAA AAT TAT AAA TCT CCT GGT CGT GTG CAG GAT ATC ACG 770 Lys Thr Lys Glu Asn Tyr Lys Ser Pro Gly Arg Val Gin Asp He Thr 235 240 245

AAA ATA ATA GAT GAG AAA CAA AAG TCG TCA AAA GAC ACA GAG TGG CAT 818 Lys He He Asp Glu Lys Gin Lys Ser Ser Lys Asp Thr Glu Trp His 250 255 260 265

AAT AAG AAA GTG AAA GAA CAT AAA AAA GTG AAA GAT ATC AAA CCT GAT 866 Asn Lys Lys Val Lys Glu His Lys Lys Val Lys Asp He Lys Pro Asp 270 275 280 TTC GAA TCT TCT CAA AGG AAT AAG AAA AGC AAG AAT ATT CCT AAG CAA 914 Phe Glu Ser Ser Gin Arg Asn Lys Lys Ser Lys Asn He Pro Lys Gin 285 290 295

ATT GAA AAT ATC ACA CCT CAA CTT GAC AGC TTA CGA TCA CGA GAT ATA 962

He Glu Asn He Thr Pro Gin Leu Asp Ser Leu Arg Ser Arg Asp He 300 305 310

GTA ATT AAG GGA GAA TTA CTA ACA AAA GAT ACT ACA AAA AGT TTA ACT 1010 Val He Lys Gly Glu Leu Leu Thr Lys Asp Thr Thr Lys Ser Leu Thr 315 320 325

ACT GTT AAT GTT GAT AGT GAA TTA GAT AGT GTA AAA CCT AAA GAT GAA 1058

Thr Val Asn Val Asp Ser Glu Leu Asp Ser Val Lys Pro Lys Asp Glu 330 335 340 345

AAA CCT GAA CCT TCT GAA CCT AGT AAA ACG TTT ATT GAT ACT TCA GTT 1106

Lys Pro Glu Pro Ser Glu Pro Ser Lys Thr Phe He Asp Thr Ser Val 350 355 360

GCA AAG GAT GTT GAT AAT TCT ACA CAG GCG AAC CAT AAA AAG AAG AAA 1154 Ala Lys Asp Val Asp Asn Ser Thr Gin Ala Asn His Lys Lys Lys Lys 365 370 375 AGT AAA TCT AAG CCG AGG AAA ACG GAA CCG GAA GAT GAA ATT GAA AAA 1202 Ser Lys Ser Lys Pro Arg Lys Thr Glu Pro Glu Asp Glu He Glu Lys 380 385 390

GCT TTG AAA GAA ATT CAA GCT AGT GAG AAA AAA CTT ACG AAG TCT ATC 1250 Ala Leu Lys Glu He Gin Ala Ser Glu Lys Lys Leu Thr Lys Ser He 395 400 405

GAT AAC ATT GTG AAT AAA TTT AAT ACA CCA CTT GCT AGT GTT AAA GCC 1298 Asp Asn He Val Asn Lys Phe Asn Thr Pro Leu Ala Ser Val Lys Ala 410 415 420 425

GAT GAT TCC AAT TCT ACC AAG GAT AAT GTA CCA GCA AAG AAG AAA AAA 1346 Asp Asp Ser Asn Ser Thr Lys Asp Asn Val Pro Ala Lys Lys Lys Lys 430 435 440

CCT TCG AAG TCA TCT GTT TCT TTA CCT GAG AAT GTA GTA CAA AAT CTA 1394 Pro Ser Lys Ser Ser Val Ser Leu Pro Glu Asn Val Val Gin Asn Leu 445 450 455 TTG ATA CTA ACA TAA CTACTAGTAG CGACAAGATT GAAAACATGC CGCAACCGCA 1449 Leu He Leu Thr 460

ACCAAAAAGA GAAGATTTAC AAGATGCAGC TAAGGAAGTA TTGACTTCAA TAGAGTCAGT 1509

AATGATGCAG TCTGTTGAGA CTATTCCTAT TACGAAGAAA AGAGTAAATA AGAAAAAGAA 1569

TACCACTCAA CAGACGAAGG AATTTGTGGA ACACGAAATA TGCGATACAT CAAAAAATGA 1629 AACTTTAAAA AATATTGAAA AAGAATCGCA TGAGAATATG GCTATATTGC AAACAAGTCC 1689

GAAACCGCCA CTAAG 1704

(2) INFORMATION FOR SEQ ID NO:27:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 461 amino acids

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

(ii) MOLECULE TYPE: protein

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 27:

Met Glu Pro Gin Ser Leu Ser Trp Gin Leu Pro Thr Gin Val Val Gin

1 5 10 15

Pro Val Phe Glu Gin Gin Met Gin He Pro Gly Tyr Asn Met Gin He 20 25 30

Gin Ser Asn Tyr Tyr Gin He His Pro Glu Met Leu Asp Pro Asn Leu 35 40 45

Asn Asn Pro Gin Gin Leu Met Phe Asn Tyr Met Gin Leu Gin Gin Leu 50 55 60

Gin Glu Leu Gin His Leu Ser Gin Gin Gin Pro Met His His Glu Phe 65 70 75 80

Glu His His He Pro He Pro Gin Glu Ala Thr Ser Thr Asn Tyr Gly 85 90 95

Pro Ser Gly Gin Tyr He Thr Ser Asp Ala Thr Ser Tyr Gin Ser He 100 105 110

Ala Gin Gin Phe Val Pro Gin Pro Pro He Glu Thr Thr Thr Thr Lys 115 120 125

He Pro Glu Thr Glu He Gin He Gly Val Ser Asn Gin Tyr Ala Gin 130 135 140

Asn He Thr Tyr Asn Ser Asn He Ser Pro Glu Val He Gly Phe Arg 145 150 155 160

Glu His Tyr Val Ala Glu Gin Pro Ser Gly Asp Val Leu His Lys Ser 165 170 175

His Leu Thr Glu Gin Pro Ala Asp Lys Ser Thr Arg Gly Asp Gin Glu 180 185 190 Pro Val Ser Glu Thr Gly Ser Gly Phe Ser Tyr Ala Gin He Leu Ser 195 200 205

Gin Gly Leu Lys Pro Thr Gin Pro Ser Asn Ser Val Asn Leu Leu Ala

210 215 220

Asp Arg Ser Arg Ser Pro Leu Asp Thr Lys Thr Lys Glu Asn Tyr Lys

225 230 235 240

Ser Pro Gly Arg Val Gin Asp He Thr Lys He He Asp Glu Lys Gin 245 250 255

Lys Ser Ser Lys Asp Thr Glu Trp His Asn Lys Lys Val Lys Glu His 260 265 270 Lys Lys Val Lys Asp He Lys Pro Asp Phe Glu Ser Ser Gin Arg Asn 275 280 285

Lys Lys Ser Lys Asn He Pro Lys Gin He Glu Asn He Thr Pro Gin

290 295 300

Leu Asp Ser Leu Arg Ser Arg Asp He Val He Lys Gly Glu Leu Leu

305 310 315 320

Thr Lys Asp Thr Thr Lys Ser Leu Thr Thr Val Asn Val Asp Ser Glu 325 330 335

Leu Asp Ser Val Lys Pro Lys Asp Glu Lys Pro Glu Pro Ser Glu Pro 340 345 350 Ser Lys Thr Phe He Asp Thr Ser Val Ala Lys Asp Val Asp Asn Ser 355 360 365

Thr Gin Ala Asn His Lys Lys Lys Lys Ser Lys Ser Lys Pro Arg Lys 370 375 380

Thr Glu Pro Glu Asp Glu He Glu Lys Ala Leu Lys Glu He Gin Ala

385 390 395 400

Ser Glu Lys Lys Leu Thr Lys Ser He Asp Asn He Val Asn Lys Phe

405 410 415

Asn Thr Pro Leu Ala Ser Val Lys Ala Asp Asp Ser Asn Ser Thr Lys

420 425 430

Asp Asn Val Pro Ala Lys Lys Lys Lys Pro Ser Lys Ser Ser Val Ser 435 440 445

Leu Pro Glu Asn Val Val Gin Asn Leu Leu He Leu Thr 450 455 460

(2) INFORMATION FOR SEQ ID NO:28:

(1) SEQUENCE CHARACTERISTICS: (A) LENGTH: 1383 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear (n) MOLECULE TYPE: cDNA

(XI) SEQUENCE DESCRIPTION: SEQ ID NO:28:

ATGGAACCTC AATCGCTGTC TTGGCAACTT CCGACTCAAG TAGTTCAGCC AGTTTTTGAA 60

CAACAAATGC AGATTCCTGG ATATAATATG CAAATTCAAT CTAATTATTA TCAAATTCAC 120

CCAGAAATGT TGGATCCAAA TTTGAACAAT CCTCAGCAGT TAATGTTTAA TTATATGCAA 180 TTACAACAAT TGCAGGAACT ACAACATTTA AGTCAACAAC AGCCAATGCA TCATGAATTT 240

GAACATCATA TCCCCATTCC ACAAGAAGCA ACTTCAACTA ATTACGGTCC ATCCGGACAG 300

TATATTACTA GTGACGCAAC ATCTTATCAA TCAATTGCCC AACAATTTGT ACCACAACCA 360

CCAATTGAAA CTACCACCAC GAAAATACCT GAAACTGAAA TTCAAATTGG CGTTTCGAAT 420

CAATATGCCC AAAATATAAC TTATAATTCA AATATCAGTC CTGAAGTGAT TGGATTCCGA 480 GAACATTATG TTGCGGAACA GCCTTCTGGT GACGTGCTTC ACAAAAGTCA TTTAACAGAA 540

CAACCAGCAG ATAAAAGCAC ACGTGGTGAT CAGGAACCTG TTAGTGAGAC AGGCTCTGGT 600

TTTTCGTATG CACAAATTTT ATCACAGGGA CTTAAGCCTA CCCAGCCATC CAACTCAGTT 660

AATTTGCTTG CAGATCGATC GAGATCACCT CTAGATACGA AAACGAAAGA AAATTATAAA 720

TCTCCTGGTC GTGTGCAGGA TATCACGAAA ATAATAGATG AGAAACAAAA GTCGTCAAAA 780 GACACAGAGT GGCATAATAA GAAAGTGAAA GAACATAAAA AAGTGAAAGA TATCAAACCT 840

GATTTCGAAT CTTCTCAAAG GAATAAGAAA AGCAAGAATA TTCCTAAGCA AATTGAAAAT 900

ATCACACCTC AACTTGACAG CTTACGATCA CGAGATATAG TAATTAAGGG AGAATTACTA 960

ACAAAAGATA CTACAAAAAG TTTAACTACT GTTAATGTTG ATAGTGAATT AGATAGTGTA 1020

AAACCTAAAG ATGAAAAACC TGAACCTTCT GAACCTAGTA AAACGTTTAT TGATACTTCA 1080 GTTGCAAAGG ATGTTGATAA TTCTACACAG GCGAACCATA AAAAGAAGAA AAGTAAATCT 1140

AAGCCGAGGA AAACGGAACC GGAAGATGAA ATTGAAAAAG CTTTGAAAGA AATTCAAGCT 1200

AGTGAGAAAA AACTTACGAA GTCTATCGAT AACATTGTGA ATAAATTTAA TACACCACTT 1260

GCTAGTGTTA AAGCCGATGA TTCCAATTCT ACCAAGGATA ATGTACCAGC AAAGAAGAAA 1320

AAACCTTCGA AGTCATCTGT TTCTTTACCT GAGAATGTAG TACAAAATCT ATTGATACTA 1380 ACA 1383

(2) INFORMATION FOR SEQ ID NO: 29: (l) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 1758 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(ll) MOLECULE TYPE: cDNA

(ix) FEATURE:

(A) NAME/KEY: CDS (B) LOCATION: 1...1758

(ix) FEATURE:

(A) NAME/KEY: W = A or T (B) LOCATION: 1136

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:29: CTA GAG ATG GCT AAA TTT CTG ACG GAA ACA TTA GAC GAC ATG ACT CTA 48 Leu Glu Met Ala Lys Phe Leu Thr Glu Thr Leu Asp Asp Met Thr Leu 1 5 10 15

CAA CAC AAA GAT CAC AGA TCA GAA TTG GCT AAA GAG TTT TCA ATT TGG 96 Gin His Lys Asp His Arg Ser Glu Leu Ala Lys Glu Phe Ser He Trp

20 25 30

TTT ACG AAA ATG AGA CAG TCT GGC GCT CAA GCC AGT AAC GAA GAA ATC 144 Phe Thr Lys Met Arg Gin Ser Gly Ala Gin Ala Ser Asn Glu Glu He 35 40 45

ATG AAA TTT TCA AAA TTG TTT GAA GAT GAA ATC ACT CTT GAC TCG CTG 192 Met Lys Phe Ser Lys Leu Phe Glu Asp Glu He Thr Leu Asp Ser Leu 50 55 60

GCG AGG CCG CAA CTT GTT GCT TTG TGC AGG GTA CTA GAA ATC AGT ACT 240

Ala Arg Pro Gin Leu Val Ala Leu Cys Arg Val Leu Glu He Ser Thr

65 70 75 80 TTA GGA ACA ACA AAT TTC TTA AGG TTT CAA CTG CGA ATG AAA CTG CGT 288

Leu Gly Thr Thr Asn Phe Leu Arg Phe Gin Leu Arg Met Lys Leu Arg 85 90 95

TCA TTA GCT GCT GAT GAT AAA ATG ATT CAA AAA GAA GGC ATA GTT TCT 336 Ser Leu Ala Ala Asp Asp Lys Met He Gin Lys Glu Gly He Val Ser

100 105 110

ATG ACT TAT TCG GAG GTG CAA CAG GCC TGC AGA GCT CGT GGA ATG CGA 384

Met Thr Tyr Ser Glu Val Gin Gin Ala Cys Arg Ala Arg Gly Met Arg 115 120 125

GCT TAT GGT ATG CCT GAA CAT AGG TTG AGG AGG CAA TTG GAA GAC TGG 432

Ala Tyr Gly Met Pro Glu His Arg Leu Arg Arg Gin Leu Glu Asp Trp 130 135 140

ATT AAT TTA AGC TTG AAT GAA AAG GTT CCA CCA TCA TTA TTG CTT TTG 480 He Asn Leu Ser Leu Asn Glu Lys Val Pro Pro Ser Leu Leu Leu Leu 145 150 155 160

TCA AGG GCG CTG ATG TTG CCC GAG AAT GTT CCA GTG TCT GAT AAA CTT 528 Ser Arg Ala Leu Met Leu Pro Glu Asn Val Pro Val Ser Asp Lys Leu 165 170 175 AAA GCA ACA ATA AAT GCT CTT CCT GAA ACT ATT GTA ACT CAG ACA AAG 576 Lys Ala Thr He Asn Ala Leu Pro Glu Thr He Val Thr Gin Thr Lys 180 185 190

GCT GCT ATT GGA GAA AGA GAA GGA AAG ATT GAC AAT AAG ACC AAA ATT 624 Ala Ala He Gly Glu Arg Glu Gly Lys He Asp Asn Lys Thr Lys He 195 200 205

GAG GTC ATC AAA GAG GAA GAA CGC AAA ATT CGC GAA GAG CGC CAA GAA 672 Glu Val He Lys Glu Glu Glu Arg Lys He Arg Glu Glu Arg Gin Glu 210 215 220

GCA CGT GAG GAA GAG GAA CAA CGC AAG CAA GCC GAA CTT GCT CTT AAT 720

Ala Arg Glu Glu Glu Glu Gin Arg Lys Gin Ala Glu Leu Ala Leu Asn 225 230 235 240

GCC AGT TCT GCA GCA GCT GAG GCC TCT TCA GCT CAG GAA CTT TTG ATA 768 Ala Ser Ser Ala Ala Ala Glu Ala Ser Ser Ala Gin Glu Leu Leu He 245 250 255 GAT ACA GCT CCT GTA ATA GAT GCA GAA AAG ACA CCA AAG GTG GCA ACA 816 Asp Thr Ala Pro Val He Asp Ala Glu Lys Thr Pro Lys Val Ala Thr 260 265 270

TCA CCT GTT GAA TCA CCA TTG GCA CCA CCA GAA GTT CTG ATT ATG GGT 864 Ser Pro Val Glu Ser Pro Leu Ala Pro Pro Glu Val Leu He Met Gly 275 280 285

GCT CCT AAA ACA CCT GTT GCA ACC GAA GTG GAT AAG AAT GCT GAT GAG 912 Ala Pro Lys Thr Pro Val Ala Thr Glu Val Asp Lys Asn Ala Asp Glu 290 295 300

GTG GAA TTC ACC AAG AAA GAT CTT GAG GTT GTT GAA GAT GCA TTG GAT 960

Val Glu Phe Thr Lys Lys Asp Leu Glu Val Val Glu Asp Ala Leu Asp 305 310 315 320

ACA CTA TCG AAA GAC AAA AAT AAT TTG GTG ATT GAA AAG GAA GTT ATT 1008 Thr Leu Ser Lys Asp Lys Asn Asn Leu Val He Glu Lys Glu Val He 325 330 335 AAA GAC ATT AAG GAA GAA ATT GCT GAT TAC CAA GAA GAT GTA GAA GAA 1056 Lys Asp He Lys Glu Glu He Ala Asp Tyr Gin Glu Asp Val Glu Glu 340 345 350

TTG AAA GAA GCC ATA GTT GCT GCT GAG AAA CCA AAG GAT GAG ATA AAA 1104 Leu Lys Glu Ala He Val Ala Ala Glu Lys Pro Lys Asp Glu He Lys 355 360 365

GAA ACT AAA GGA GCT CAA CGA TTG TTG AAG AWG GTT AAC AAG ATG ATA 1152 Glu Thr Lys Gly Ala Gin Arg Leu Leu Lys Xaa Val Asn Lys Met He 370 375 380

ACG AAA ATG GAT ACT GTT GTA CAA GAA ATT GAA AGC AAA GAA TCT GAG 1200 Thr Lys Met Asp Thr Val Val Gin Glu He Glu Ser Lys Glu Ser Glu 385 390 395 400

AAG AAA GCC AAA ACA TTG CCA CTT GAA GCT CCT AGG AGC GCT ACT GAA 1248 Lys Lys Ala Lys Thr Leu Pro Leu Glu Ala Pro Arg Ser Ala Thr Glu 405 410 415 ACT CAA GAA TTA GAT GTA AGG AAA GAA AGA GGA GAG ATT TTA ATT GAC 1296 Thr Gin Glu Leu Asp Val Arg Lys Glu Arg Gly Glu He Leu He Asp 420 425 430

GAA TTA ATG GAC GCT ATT AAG AAA GTT AAA AAT GTG CCA GAC GAA AAT 1344

Glu Leu Met Asp Ala He Lys Lys Val Lys Asn Val Pro Asp Glu Asn 435 440 445

CGC TTG AAA TTA ATT GAG AAC ATT TTG GGC AGG ATC GAT ACT GAC AAA 1392 Arg Leu Lys Leu He Glu Asn He Leu Gly Arg He Asp Thr Asp Lys 450 455 460

GAT AGG CAT ATC AAA GTT GAA GAT GTA TTG AAG GTT ATT GAC ATT GTG 1440 Asp Arg His He Lys Val Glu Asp Val Leu Lys Val He Asp He Val 465 470 475 480

GAA AAA GAA GAT GGT ATC ATG AGT ACA AAA CAA TTA GAT GAG TTG GTT 1488

Glu Lys Glu Asp Gly He Met Ser Thr Lys Gin Leu Asp Glu Leu Val

485 490 495

CAG CTT TTG AAA AAG GAG GAA GTT ATT GAA TTG GAA GAA AAG AAA GAA 1536

Gin Leu Leu Lys Lys Glu Glu Val He Glu Leu Glu Glu Lys Lys Glu

500 505 510 AAG CAA GAG TCT CAA CAG AAA AGT TTT GTA CCA CCA AGT GAA ACT TTG 1584 Lys Gin Glu Ser Gin Gin Lys Ser Phe Val Pro Pro Ser Glu Thr Leu 515 520 525

CAT CTT GAA TCA TCA CAG CAG AAG AGT ACA GTT CCT AGC TCG GGA CAT 1632 His Leu Glu Ser Ser Gin Gin Lys Ser Thr Val Pro Ser Ser Gly His 530 535 540

GAA GCT AAG GTG TCC GAA GAT GAC TTA AAT GTT AAA AAT AAA AAT TTG 1680 Glu Ala Lys Val Ser Glu Asp Asp Leu Asn Val Lys Asn Lys Asn Leu 545 550 555 560

GAA GAA TCG ACC AAA ACT GAA TGT GGA GCA ATT GAC GAA GAG CAC AGA 1728 Glu Glu Ser Thr Lys Thr Glu Cys Gly Ala He Asp Glu Glu His Arg 565 570 575

AGA GAG CAT TGC CAG TAC CCA GAC ATT ACA 1758

Arg Glu His Cys Gin Tyr Pro Asp He Thr 580 585

(2) INFORMATION FOR SEQ ID NO:30:

(l) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 586 amino acids (B) TYPE: amino acid

(D) TOPOLOGY: linear

(n) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO:30:

Leu Glu Met Ala Lys Phe Leu Thr Glu Thr Leu Asp Asp Met Thr Leu 1 5 10 15 Gin His Lys Asp His Arg Ser Glu Leu Ala Lys Glu Phe Ser He Trp

20 25 30

Phe Thr Lys Met Arg Gin Ser Gly Ala Gin Ala Ser Asn Glu Glu He 35 40 45

Met Lys Phe Ser Lys Leu Phe Glu Asp Glu He Thr Leu Asp Ser Leu 50 55 60

Ala Arg Pro Gin Leu Val Ala Leu Cys Arg Val Leu Glu He Ser Thr 65 70 75 80

Leu Gly Thr Thr Asn Phe Leu Arg Phe Gin Leu Arg Met Lys Leu Arg 85 90 95

Ser Leu Ala Ala Asp Asp Lys Met He Gin Lys Glu Gly He Val Ser

100 105 110

Met Thi Tyr Ser Glu Val Gin Gin Ala Cys Arg Ala Arg Gly Met Arg 115 120 125

Ala Tyr Gly Met Pro Glu His Arg Leu Arg Arg Gin Leu Glu Asp Trp

130 135 140

He Asn Leu Ser Leu Asn Glu Lys Val Pro Pro Ser Leu Leu Leu Leu 145 150 155 160

Ser Arg Ala Leu Met Leu Pro Glu Asn Val Pro Val Ser Asp Lys Leu 165 170 175

Lys Ala Thr He Asn Ala Leu Pro Glu Thr He Val Thr Gin Thr Lys 180 185 190

Ala Ala He Gly Glu Arg Glu Gly Lys He Asp Asn Lys Thr Lys He 195 200 205

Glu Val He Lys Glu Glu Glu Arg Lys He Arg Glu Glu Arg Gin Glu 210 215 220

Ala Arg Glu Glu Glu Glu Gin Arg Lys Gin Ala Glu Leu Ala Leu Asn 225 230 235 240

Ala Ser Ser Ala Ala Ala Glu Ala Ser Ser Ala Gin Glu Leu Leu He 245 250 255

Asp Thr Ala Pro Val He Asp Ala Glu Lys Thr Pro Lys Val Ala Thr 260 265 270

Ser Pro Val Glu Ser Pro Leu Ala Pro Pro Glu Val Leu He Met Gly 275 280 285

Ala Pro Lys Thr Pro Val Ala Thr Glu Val Asp Lys Asn Ala Asp Glu 290 295 300 Val Glu Phe Thr Lys Lys Asp Leu Glu Val Val Glu Asp Ala Leu Asp 305 310 315 320

Thr Leu Ser Lys Asp Lys Asn Asn Leu Val He Glu Lys Glu Val He 325 330 335

Lys Asp He Lys Glu Glu He Ala Asp Tyr Gin Glu Asp Val Glu Glu 340 345 350

Leu Lys Glu Ala He Val Ala Ala Glu Lys Pro Lys Asp Glu He Lys 355 360 365

Glu Thr Lys Gly Ala Gin Arg Leu Leu Lys Xaa Val Asn Lys Met He 370 375 380 Thr Lys Met Asp Thr Val Val Gin Glu He Glu Ser Lys Glu Ser Glu 385 390 395 400

Lys Lys Ala Lys Thr Leu Pro Leu Glu Ala Pro Arg Ser Ala Thr Glu

405 410 415

Thr Gin Glu Leu Asp Val Arg Lys Glu Arg Gly Glu He Leu He Asp

420 425 430

Glu Leu Met Asp Ala He Lys Lys Val Lys Asn Val Pro Asp Glu Asn 435 440 445

Arg Leu Lys Leu He Glu Asn He Leu Gly Arg He Asp Thr Asp Lys

450 455 460

Asp Arg His He Lys Val Glu Asp Val Leu Lys Val He Asp He Val

465 470 475 480

Glu Lys Glu Asp Gly He Met Ser Thr Lys Gin Leu Asp Glu Leu Val

485 490 495

Gin Leu Leu Lys Lys Glu Glu Val He Glu Leu Glu Glu Lys Lys Glu

500 505 510 Lys Gin Glu Ser Gin Gin Lys Ser Phe Val Pro Pro Ser Glu Thr Leu 515 520 525

His Leu Glu Ser Ser Gin Gin Lys Ser Thr Val Pro Ser Ser Gly His 530 535 540

Glu Ala Lys Val Ser Glu Asp Asp Leu Asn Val Lys Asn Lys Asn Leu 545 550 555 560

Glu Glu Ser Thr Lys Thr Glu Cys Gly Ala He Asp Glu Glu His Arg 565 570 575

Arg Glu His Cys Gin Tyr Pro Asp He Thr 580 585

(2) INFORMATION FOR SEQ ID NO:31:

(l) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 293 base pairs (B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(ll) MOLECULE TYPE: cDNA (XI) SEQUENCE DESCRIPTION: SEQ ID NO:31:

CCCGGGCTGC AGGAATTCGG CACGAGATGA GAATGGAAAT GTGATTAGCT ATACTGATGA 60 AAATGGAAAC ATTATCAGTA CTACTGATGA GAATGGAAAT GTGATTAGCA TTACTGATGA 120

AAATGGAAAT GTGATTAGCA TTACTGATGA AAATGGAAAC ATTATCAGTA CTACTGATGA 180

GAATGGAAAT GTGATTAGCA TTACTGATGA AAATGGAAAT GTGATTAGCA TTACTGATGA 240

AAATGGAAAC ATTATTAGTA CTACTGATGA GAATGGAAAT GTGATTAGCA ATA 293

(2) INFORMATION FOR SEQ ID NO: 32:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 335 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: single (D) TOPOLOGY: linear ill) MOLECULE TYPE: cDNA

( i) SEQUENCE DESCRIPTION: SEQ ID NO: 32:

TTGGAAACAG CTATGACCAT GATTACCCCA AGCTCGAAAG TTAAVCCCTC ACTHARAGGG 60

GAACAAAAGT CTGGAGCTCC ACCCGCGGAT GGCGGCCGCB TCTAGAACCT AGTGGACTCC 120 CCCGGSGCTG CAGGAATTCG GGCACGAGCT CCAGCTAGCC ATATACATTC ATCCAAAATG 180

AAGTTGSAAT GTGTCCTACC CGGCAACGGG ATGCCAGAAA TTGTKTCGAA ATKTGTGGAC 240

GAGCACAAGC TTCGTGTCTK TCTATGAAAA ACGTATGGGA GCAGAAGTCG AGGGCCGACA 300

TCCTCGGCGA TGAATGGARA GGTTATGTGC TCCGA 335

(2) INFORMATION FOR SEQ ID NO: 33:

(l) SEQUENCE CHARACTERISTICS'

(A) LENGTH: 396 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: single (D) TOPOLOGY: linear

(n) MOLECULE TYPE: cDNA

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:33:

ATAGCTTTTA ATATTTTTAA TTGATGTATT GCTCAATGGT GATTTCTGTT TATTAAACTG 60

AGTTACCAAT ATGCTCGCTT CAATAGACAT AGCAAATGAA AGCATTCCGT ATCCTCAAGC 120 GTTACCAAAC TAACATTAAG GAGTTAAATA AATGTTGTTT CCAATAAATA TAATGGGAAA 180

AACATTTAAT ATTTGTTCCA ATTTGTATTT ATTTTTACTA CAATTATATA CAATAAAATA 240

TTTTTATAT TATTTTATAA AGTTTATGAT GCAGGAGAGA AAATAATGTT AAGAATATAG 300

GTAATGTGTA TATATAAATG TTTGACAAGC ATGTTCTAGT TAAATAATAA ATACAATGTT 360

AAATCTACTT AAAAAAAAAA AAAAAAAAAA AAAAAA 396

(2) INFORMATION FOR SEQ ID NO:34:

(l) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 285 base pairs (B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(n) MOLECULE TYPE: cDNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:34:

GGAAAGCGAA GAATGAAAAG GGGAAACAAA AAAAGAAAAG ACGAAGGAGT GGAGAGATAA 60 AACGGAGGCA AAGAAGAAAA TGAGGATGCA AAAGAAAGGT AATAAAAGAG ATGAAAAGAA 120

GGAAAAAGGA AATAAGAAAG AAAGAGTGAG GGAAAAATAA AGACAGAGGC GAAGCAAAAA 180

AGGAGGAGAA ATAGAGATTA AAAAAGAAAT ACAGCGAAGA AACCAGGAAA GCGATAAAGA 240

AAAAAAAAGA AAAAAAGAGA GCAGTGAAAA AAAAAAAAAA AAAAA 285

(2) INFORMATION FOR SEQ ID NO: 35:

(l) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 228 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: single (D) TOPOLOGY: linear

(n) MOLECULE TYPE: cDNA

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:35:

CAGATATTTA CTAAAYATTG TGAAAYAAAT CATTTTCAAA ATGGTSTCCA AAGTGTTTGT 60

TGCTCTTGCC ATCAATGGCT TTATAGGGGG CTSCACAAGY CTTTTTTCGA ACAAGATGMC 120 GTCTTAGATA ASATSGTAGA TRACATCTCT GRCTSMATAT GAGAACARCA TTGSMAGAAT 180

TAGCCAAGGR TNGCRAAATT GATATGMTTS CYGCTGTAAT TCGAAAAA 228

(2) INFORMATION FOR SEQ ID NO: 36 :

(l) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 339 base pairs

(B) TYPE: nucleic acid (C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(11) MOLECULE TYPE: cDNA (ix) FEATURE:

(A) NAME/KEY: CDS

(B) LOCATION: 1..339

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:36:

CTT CGT GTC AAC CGC TGG GTC AGA CCT GTT ATT GCT ATG CAC CCA ACC 48

Leu Arg Val Asn Arg Trp Val Arg Pro Val He Ala Met His Pro Thr

1 5 10 15 ATG ACT CTT GCT GAA CGT CTC GGC AAA AAA GCT TTG CGC GAC CAA TAT 96

Met Thr Leu Ala Glu Arg Leu Gly Lys Lys Ala Leu Arg Asp Gin Tyr 20 25 30

GCT CCC GTT TGC TCC ATT GGA CAA CGT AAC ATC AAC ACC TTT GAC AAC 144 Ala Pro Val Cys Ser He Gly Gin Arg Asn He Asn Thr Phe Asp Asn 35 40 45

ATG ACC TTC CCC GCT CAA TTC GGA AAA TGC TGG CAC GCT TTG TTG CAA 192

Met Thr Phe Pro Ala Gin Phe Gly Lys Cys Trp His Ala Leu Leu Gin 50 55 60

ACT GTT CCC CAA AAG TAT TCC GAA GAA CGT GAA TAC AGC GAA GAA CAA 240

Thr Val Pro Gin Lys Tyr Ser Glu Glu Arg Glu Tyr Ser Glu Glu Gin

65 70 75 80

CAA TAC GAC CGT CAA ATG TCC GTC CTC GTT CGT GAA AAC GGC GAA GAA 288 Gin Tyr Asp Arg Gin Met Ser Val Leu Val Arg Glu Asn Gly Glu Glu 85 90 95

AAA AGA CGT TAT GAT TGT CTT GGG CAA CCG TTA CAA CAA TTG AAT TGC 336 Lys Arg Arg Tyr Asp Cys Leu Gly Gin Pro Leu Gin Gin Leu Asn Cys 100 105 110

AAT 339 Asn

(2) INFORMATION FOR SEQ ID NO:37:

(l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 113 amino acids

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

(n) MOLECULE TYPE: protein

( xi ) SEQUENCE DESCRI PTION : SEQ ID NO : 37 :

Leu Arg Val Asn Arg Trp Val Arg Pro Val He Ala Met His Pro Thr 1 5 10 15

Met Thr Leu Ala Glu Arg Leu Gly Lys Lys Ala Leu Arg Asp Gin Tyr 20 25 30

Ala Pro Val Cys Ser He Gly Gin Arg Asn He Asn Thr Phe Asp Asn 35 40 45

Met Thr Phe Pro Ala Gin Phe Gly Lys Cys Trp His Ala Leu Leu Gin 50 55 60 Thr Val Pro Gin Lys Tyr Ser Glu Glu Arg Glu Tyr Ser Glu Glu Gin 65 70 75 80

Gin Tyr Asp Arg Gin Met Ser Val Leu Val Arg Glu Asn Gly Glu Glu 85 90 95

Lys Arg Arg Tyr Asp Cys Leu Gly Gin Pro Leu Gin Gin Leu Asn Cys 100 105 110

Asn

(2) INFORMATION FOR SEQ ID NO: 38:

(i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 493 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear (n) MOLECULE TYPE: cDNA

(ix) FEATURE:

(A) NAME/KEY: CDS

(B) LOCATION: 1..390

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:38:

TCC AGC TCC TCC AGC TCC AGC AGT GAC TCT TCC AGC TCC AGC AGC TCT 48 Ser Ser Ser Ser Ser Ser Ser Ser Asp Ser Ser Ser Ser Ser Ser Ser 1 5 10 15

TCC TCT TCC AGC TCC AGC AGC TCC TCT TCT GAA TCT TCC GAA GAA AAA 96 Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Glu Ser Ser Glu Glu Lys 20 25 30

ACC TCC CAC AAA AAA TCC GAA AAG AAG GAA CAC AAA TCC TGC TCC ATC 144 Thr Ser His Lys Lys Ser Glu Lys Lys Glu His Lys Ser Cys Ser He 35 40 45 AAG AAG CAA GTA CAA TTC GTA GAA AAA GAC GGT AAA CTC TGC TTC AGC 192 Lys Lys Gin Val Gin Phe Val Glu Lys Asp Gly Lys Leu Cys Phe Ser 50 55 60

ATC CGT CCC TTG GCC GCT TGC CAA AAA CAC TGC AAA GCC ACT GAA ACC 240 He Arg Pro Leu Ala Ala Cys Gin Lys His Cys Lys Ala Thr Glu Thr 65 70 75 80

ACT CAA ATG GAA GTC GAA GTA TAC TGC CCC TCT GGC AGC CTT GCT GAA 288 Thr Gin Met Glu Val Glu Val Tyr Cys Pro Ser Gly Ser Leu Ala Glu 85 90 95

CTT TAC AAA CAA AAG ATC CTT AAG GGA GCC AAC CCC GAC TTG AGC GAC 336 Leu Tyr Lys Gin Lys He Leu Lys Gly Ala Asn Pro Asp Leu Ser Asp 100 105 110

AAG ACT CCT TCC AGA ATC TTG AAA TTC AAG GTT CCC AAA GCT TGC ACC 384 Lys Thr Pro Ser Arg He Leu Lys Phe Lys Val Pro Lys Ala Cys Thr 115 120 125

GCT TAC TAAATCTGAA ATAAATTACA TGGATTAGTT CATTTCTGAT GTAGTGCAAT 440 Ala Tyr 130

TAGTTCGATA ATAAATTATT CAATGAGCAT TTAAAAAAAA AAAAAAAAAA AAC 493

(2) INFORMATION FOR SEQ ID NO:39:

(i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 130 amino acids

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

(ii) MOLECULE TYPE: protein

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:39:

Ser Ser Ser Ser Ser Ser Ser Ser Asp Ser Ser Ser Ser Ser Ser Ser

1 5 10 15

Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Glu Ser Ser Glu Glu Lys 20 25 30

Thr Ser His Lys Lys Ser Glu Lys Lys Glu His Lys Ser Cys Ser He 35 40 45

Lys Lys Gin Val Gin Phe Val Glu Lys Asp Gly Lys Leu Cys Phe Ser

50 55 60

He Arg Pro Leu Ala Ala Cys Gin Lys His Cys Lys Ala Thr Glu Thr 65 70 75 80

Thr Gin Met Glu Val Glu Val Tyr Cys Pro Ser Gly Ser Leu Ala Glu 85 90 95

Leu Tyr Lys Gin Lys He Leu Lys Gly Ala Asn Pro Asp Leu Ser Asp 100 105 110

Lys Thr Pro Ser Arg He Leu Lys Phe Lys Val Pro Lys Ala Cys Thr 115 120 125

Ala Tyr 130

(2) INFORMATION FOR SEQ ID NO: 40:

(l) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 306 base pairs (B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(n) MOLECULE TYPE: cDNA

(XI) SEQUENCE DESCRIPTION: SEQ ID NO:40:

GTAGTGCCAT CATTCGTAAA CSTTYTGACG GTKGGGCGCT GTATWGGTGC TGCCTGGAAA 60 TTGCATCGAT GCACTWCCGT GTCGGGCGCA WATAGTGCKK TGGSCCCTGT CTGMTTATAG 120

ACATTCAGGG CGCSGGSAKT AGCCATGTTC ATGGCTCMCA AWMTGCATTC ACAGTGGGGT 180

CACATTTCAG TCGCATGATT KMTCAARTTA GTATMWGADA TATATTTTTA TCATACTAAG 240

TAGTGAGCDA ATAACACGCG ARWWACRAAC ACCGAATATC TTKAGTTTTT GCACAGATAT 300

KTGTAA 306

(2) INFORMATION FOR SEQ ID NO: 41:

(l) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 490 base pairs (B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

( ii ) MOLECULE TYPE : cDNA

( i ) SEQUENCE DESCRIPTION : SEQ ID NO : 41 :

ACCGGATACG TTGCCAATGA CTACGTCACC ACCAATGTTG TTTCCACTCC AGTTACTGGA 60 TACACCACCG GACATCTTGC TAATGACTAC GTCACCACCA ATGTTGTATC CACTCCAGTT 120

ACTGGATACA CCACCGGACA TCTTGCCAAT GACTACGTCA CCACCAACGT AGTTTCCGCA 180

CCAGTCACCA CTGGATACAC CACTGGCTAT ACCACCGGTA ATGTCGGATA CACCACCGGA 240

GTTACTGGTT ACACCAACGG AGTTAGTGGA TATACCAATG GACTTAATGG TTATACCACT 300

GGTAGCTATG TCAGCTCCCC AGGATACACT TCTTCTGGAC TTGTCAACGT TTTCTAGATT 360 TATGATTTCG TCTGCCCTCA ATGATGATGA CCACACTTTT TACTTTTTAT GATATTTGGA 420

AAAAATAAAT AACTGGAAGA ATATATAATA ATTTCAAAAT AAAAAAAAAA AAAAAAAAAA 480

CTCGAGGGGG 490 (2) INFORMATION FOR SEQ ID NO: 2:

( l ) SEQUENCE CHARACTERISTICS :

(A) LENGTH : 616 base pairs ( B) TYPE : nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(ll) MOLECULE TYPE: cDNA

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 42:

AAAAAATCGA AAGAAGGCGT AAAACCAAAA TGGGCACAGA AGGATATTCG GGATTTTAGT 60 GATGCCGACA TGGAGAGGTT ACTGGATCAA TGGGAAGAAG ATGAAGACCC CCTTCCAGAA 120

GACGAATTGC CCGAACATCT CAGACCTGAT CCAAAGATCG ACATAAGCAA CATCGATATG 180

AGCAATCCCB AAAACATACT AAAGGCTTCC AAAAAAGGCA AGACTTTGAT GGCATTCGTA 240

CAAGTCAGTG GAAATCCAAC ACAAGAAGAA GCCGAAACCA TCACTAAATT GTGGCAAGGC 300

AGTCTATGGA ATAGTCATAT ACAAGCCGAA AGATATATGG TTAGCGATGA CAGGGCTATA 360 TTTATGTTTA AAGATGGTTC TCAAGCTTGG CCTGCTAAAG ACTTTTTAGT GGAACAAGAA 420

AGGTGTAAAG ATGTTACAAT TGAAAATAAA ATATATCCTG GTAAATATTC TTCGACTAAA 480

GAAGAATTAT AATATAATAT ATTATAATTA TAATCTATAA AATAGATTTG AAATTCTACA 540

TTCATGATCT ACTATGTATG ATATTAATTT ATTAAAAATA ATGTTTTTTC AAGTAAAAAA 600

AAAAAAAAAA AAAAAA 616

(2) INFORMATION FOR SEQ ID NO: 43:

(l) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 475 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(il) MOLECULE TYPE: cDNA

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 43:

CTCGTGCGGG ACAGATATAG GACCGGATTC GTTAATTGAT TTGAGTGAAG TGGCTTCTGG 60 TGGTTCTGAT ATTGACACAA AATTTTCCAA TTTAAAAATA GATAAAAAGC CTGTTGCAAC 120

TTCACAACAA GGAATTGATG AATTTGATAT GTTTGCACAA TCGAGAAACA TTTCTAGTGA 180

GGGATCAACC AGTGCTATGA AGGAAGGACA CGGTTTGGAC TTATTATCAA ATACACATAA 240

AAATGTACCA CCAACAATTC CACAAGCCGG ACAACTTCCA AGGGATTCTG AGTTTGATGA 300

AATTGCTGCT TGGCTTGATG AAAAGGTTGA AGACAAAGCC CAAGTTCCCG AAGACAGTAT 360 TACAAGCAGT GAATTTGATA AATTCCTGGC AGAACGGGCA GCTGTTGCTG AAACTTTGCC 420

AAATATTCCA CCGACTACAC AAAGTAATCA TTCAAATATT GAAGCAAACG ATAAA 475

(2) INFORMATION FOR SEQ ID NO: 44:

(l) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 295 base pairs

(B) TYPE: nucleic acid (C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(li) MOLECULE TYPE: cDNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 4:

CCGGCACGGG AGGTAGTGAC GAAAAATAAC GATACGGGAC TCATCCGAGG CCCCGTAATC 60 GGAATGAGTA CACTTTAAAT CCTTTAACGA GGATCTATTA GAGGGCCAGT CTGTGTGCCA 120

GCAGCCGCGG TAATTCCAGC TCTAATAGCG TATATTAAAG TTGTTGCGGT TAAAAAGCTC 180

GTAGTTGAAT CTGTGTCCCA CACTGTYGGT TCACCGCTCG CGGTGTTCAA CTGGCATGTC 240 TGTGGGACGT CCTACCGGTG GGCTTAGCCC GTCAAAAGGC GGCCCAACTC AAAAT 295

(2) INFORMATION FOR SEQ ID NO: 5: (l) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 372 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(ll) MOLECULE TYPE: cDNA

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 45: CTGACTAATC CCAGGACTCC TTTATCCTGT TTGCGCAATG TCGATACCCA TCTCACAATG 60

GTTAATGATT TATCGGCTAA ACAGAAGAGT CCTAAGAAGG TTGTTAAAGG TGTTTCTAGA 120

ATACCGACTT TTAGACCCAA GGCTATGAAT GCTGATGTTG AGAATTTTGA TTCGATGAGG 180

TGCGATGTTT GGRACAAAGA CACCAGTGTT GTTATATAAT TACTAAAGCA ATCCACATGT 240

AGCTAATTTT TTTTTTACAA TTTTATTTGT AACTATGTGT ATTTATATGA ATTCTTGTGG 300 AATATAATTT TAAGTTTTTA AATGAAATAT AGATATTATT CTAAAAAAAA AAAACAAAAA 360

AAAAAAAAAA AA 372

(2) INFORMATION FOR SEQ ID NO: 46 :

(l) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 252 base pairs

(B) TYPE: nucleic acid (C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(ll) MOLECULE TYPE: cDNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 6:

GGATTCGGCA CGAGAATTTA TTAAGCGCAT TATTTGCAAG TGTAATTTGC TCCTTTAACG 60 CGGAAGTACA AAATCGAATC GTTGGTGGCA ATGATGTAAG TATTTCAAAA ATTGGGTGGC 120

AAGTATCTAT TCAAAGTAAT AACCAACATT TCTGTGGTGG TTCAATCATT GCTAAAGATT 180

GGGTACTGAC TTCTTCTCAA TGCGTCGTGG ACAAACAAAG TCCACCGAAG GATTTAACTG 240 TTCGTGTTGG AA 252

(2) INFORMATION FOR SEQ ID NO: 47: (l) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 613 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(ll) MOLECULE TYPE: cDNA

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 7: ATTCCTGCTG TTAATAGTAC TAATGCAGTA ATTGCTGCHA GCTGCTGCAC AGAGGTTTTT 60

AAAATGGCAA CAAGTTGTTA CACCCACATG AACAACTACA TGGTATTCAA TGATACCGAT 120

GGGATTTATA CATATACTTA CGAAGCTGAA AGAAAACCTG ACTGTTTAGC TTGTTCACAA 180

ATTCCAAAAA CTATAGAAGT TTCTAATCCT GAAAATATGA CTCTCCAAGA CTTGATTACT 240

TTGTTGTGTG AAGGGGCTGA ATATCAAATG AAGAGCCCAG GTATTGTAGC CTCAATCGAA 300 GGCAAAAACA AAACCTTATA CATGTCAACA GTAGCAAGTA TAGAAGAAAA GACTAAACAG 360

AATCTAACAA AGTCTCTAAA AGAATTAAAT CTAGAAAATG GAATGGAACT GATGGTTGCA 420

GATGTGACGA CACCAAACAC AATATTACTT AAATTAAAAT ATAAGAATGT AATTGAAAAC 480

GATGTTGAGA TGACTTGATA TTTACTTAAA AATGTTATCT TACAATAATT GATAATTTAT 540

ATTTAATACT TTTGGAACTT TGTATTTAAT GATAATAAAT TATTATAAGA ATTAAAAAAA 600 AAAAAAAAAA AAA 613

(2) INFORMATION FOR SEQ ID NO: 48:

(1) SEQUENCE CHARACTERISTICS.

(A) LENGTH: 538 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: single (D) TOPOLOGY: linear

(11) MOLECULE TYPE: cDNA

(ix) FEATURE: (A) NAME/KEY: CDS

(B) LOCATION: 3..538

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 48: TT GAT ATT TGC TCT GTT GAG GGT GCC TTA GGA TTT TTA GTG GAA ATG 47

Asp He Cys Ser Val Glu Gly Ala Leu Gly Phe Leu Val Glu Met 1 5 10 15

TTA AAA TAT AAG GCC CCA AGT AAA ACT CTA GCT ATT GTA GAG AAT GCT 95 Leu Lys Tyr Lys Ala Pro Ser Lys Thr Leu Ala He Val Glu Asn Ala

20 25 30

GGT GGA ATA TTA CGA AAT GTA TCT AGT CAT ATA GCC CTT AGA GAG GAC 143 Gly Gly He Leu Arg Asn Val Ser Ser His He Ala Leu Arg Glu Asp 35 40 45

TAC AGA GAA ATA CTT CGA CAT CAT AAT TGC TTA ACA ATA TTA CTA CAA 191 Tyr Arg Glu He Leu Arg His His Asn Cys Leu Thr He Leu Leu Gin 50 55 60

CAA TTA AAA TCA CCA AGC CTC ATA ATT GTC AGT AAT GCT TGT GGG ACA 239 Gin Leu Lys Ser Pro Ser Leu He He Val Ser Asn Ala Cys Gly Thr 65 70 75 TTA TGG AAT TTA TCT GCT AGG AAT TCA ACA GAT CAA CAA TTT TTA TGG 287

Leu Trp Asn Leu Ser Ala Arg Asn Ser Thr Asp Gin Gin Phe Leu Trp

80 85 90 95

GAG AAT GGT GCT GTC CCT TTA TTA AGA AGT TTG ATA TAT TCT AAG CAT 335 Glu Asn Gly Ala Val Pro Leu Leu Arg Ser Leu He Tyr Ser Lys His

100 105 110

AAA ATG ATA TCT ATG GGA TCA AGT GCA GCT CTC AAA AAT TTG TTA AAT 383 Lys Met He Ser Met Gly Ser Ser Ala Ala Leu Lys Asn Leu Leu Asn 115 120 125

GCA AAA CCT GAG TGC ATC AAT TTC TTA AGT GAT TCT TCT TCT AAA GGA 431 Ala Lys Pro Glu Cys He Asn Phe Leu Ser Asp Ser Ser Ser Lys Gly 130 135 140

GTT CCA AAT CTA ACT ACA TTG GGT GTA AGA AAA CAA AAA TCT CTA CAT 479 Val Pro Asn Leu Thr Thr Leu Gly Val Arg Lys Gin Lys Ser Leu His 145 150 155 GAG TTA ATA GAT CAA AAT CTT TCA GAA ACT TGT GAT AAT ATA GAT AGT 527 Glu Leu He Asp Gin Asn Leu Ser Glu Thr Cys Asp Asn He Asp Ser 160 165 170 175

GTG GCC GCT AA 538 Val Ala Ala

(2) INFORMATION FOR SEQ ID NO:49:

(l) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 178 amino acids

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

(11) MOLECULE TYPE: protein

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 49:

Asp He Cys Ser Val Glu Gly Ala Leu Gly Phe Leu Val Glu Met Leu 1 5 10 15

Lys Tyr Lys Ala Pro Ser Lys Thr Leu Ala He Val Glu Asn Ala Gly 20 25 30

Gly He Leu Arg Asn Val Ser Ser His He Ala Leu Arg Glu Asp Tyr 35 40 45

Arg Glu He Leu Arg His His Asn Cys Leu Thr He Leu Leu Gin Gin 50 55 60

Leu Lys Ser Pro Ser Leu He He Val Ser Asn Ala Cys Gly Thr Leu 65 70 75 80 Trp Asn Leu Ser Ala Arg Asn Ser Thr Asp Gin Gin Phe Leu Trp Glu

85 90 95

Asn Gly Ala Val Pro Leu Leu Arg Ser Leu He Tyr Ser Lys His Lys 100 105 110

Met He Ser Met Gly Ser Ser Ala Ala Leu Lys Asn Leu Leu Asn Ala 115 120 125

Lys Pro Glu Cys He Asn Phe Leu Ser Asp Ser Ser Ser Lys Gly Val 130 135 140

Pro Asn Leu Thr Thr Leu Gly Val Arg Lys Gin Lys Ser Leu His Glu 145 150 155 160

Leu He Asp Gin Asn Leu Ser Glu Thr Cys Asp Asn He Asp Ser Val

165 170 175

Ala Ala

(2) INFORMATION FOR SEQ ID NO: 50:

(l) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 432 base pairs (B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(ll) MOLECULE TYPE: cDNA

(ix) FEATURE:

(A) NAME/KEY: CDS

(B) LOCATION: 1..388 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:50:

GTT CTT CTT AAA CAG TTG GAC TCT GGA TTG TTA CTT GTT ACA GGT CCC 48

Val Leu Leu Lys Gin Leu Asp Ser Gly Leu Leu Leu Val Thr Gly Pro

1 5 10 15

TTC TTA ATC AAT GCA TGC CCA TTG CGT CGC ATT TCC CAA AAC TAT GTC 96

Phe Leu He Asn Ala Cys Pro Leu Arg Arg He Ser Gin Asn Tyr Val

20 25 30 ATT GCC ACC TCT ACC CGA TTA GAC GTT AGT GGA GTT AAA TTA CCA GAA 144

He Ala Thr Ser Thr Arg Leu Asp Val Ser Gly Val Lys Leu Pro Glu

35 40 45

CAC ATC AAT GAT GAT TAT TTC AAA AGG CAA AAG AAC AAG CGT GCA AAG 192

H s He Asn Asp Asp Tyr Phe Lys Arg Gin 50 55

AAA GAG GAA GGT GAT ATT TTT GCT GCC AAG 240

Lys Glu Glu Gly Asp He Phe Ala Ala Lys

65 70

ACT GAG CAA AGG AAG AAT GAC CAA AAG CTT

Thr Glu Gin Arg Lys Asn Asp Gin Lys Leu

85 90

GGA GTA ATC AAG AAG CAC CCA GAC CAC AAA 336

Gly Val He Lys Lys His Pro Asp His Lys 100 105

TCA GCT ATG TTT GGT TTG AAA TCT TCC CAA 384

Ser Ala Met Phe Gly Leu Lys Ser Ser Gin

115 120

TTC T AAATACTATA TTCATAAAAT AAATTGAACT TCTCAAAAAA AAAA 432 Phe

(2) INFORMATION FOR SEQ ID NO: 51:

(l) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 129 amino acids

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

(ii) MOLECULE TYPE: protein

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 51:

Val Leu Leu Lys Gin Leu Asp Ser Gly Leu Leu Leu Val Thr Gly Pro

1 5 10 15

Phe Leu He Asn Ala Cys Pro Leu Arg Arg He Ser Gin Asn Tyr Val 20 25 30

He Ala Thr Ser Thr Arg Leu Asp Val Ser Gly Val Lys Leu Pro Glu 35 40 45 His He Asn Asp Asp Tyr Phe Lys Arg Gin Lys Asn Lys Arg Ala Lys 50 55 60

Lys Glu Glu Gly Asp He Phe Ala Ala Lys Lys Glu Ala Tyr Lys Pro 65 70 75 80

Thr Glu Gin Arg Lys Asn Asp Gin Lys Leu Val Asp Lys Met Val Leu 85 90 95

Gly Val He Lys Lys His Pro Asp His Lys Leu Leu Tyr Thr Tyr Leu 100 105 110

Ser Ala Met Phe Gly Leu Lys Ser Ser Gin Tyr Pro His Arg Met Lys

115 120 125

Phe

(2) INFORMATION FOR SEQ ID NO: 52: (l) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 595 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(11) MOLECULE TYPE: cDNA (ix) FEATURE:

(A) NAME/KEY: CDS

(B) LOCATION: 47..315

( xi ) SEQUENCE DESCRIPTION : SEQ ID NO : 52 :

TGGAAATTCA ATATTTTGTT TTAACATTAA ATTTTTCAAA TTCGAT ATG AAA TTT 55

Met Lys Phe

1

TTA CTG GCA ATT TGC GTG TTG TGT GTT TTA TTA AAT CAA GTA TCT ATG 103 Leu Leu Ala He Cys Val Leu Cys Val Leu Leu Asn Gin Val Ser Met 5 10 15 TCA AAA ATG GTC ACT GAA AAG TGT AAA TCG GGA GGA AAT AAT CCA AGT 151

Ser Lys Met Val Thr Glu Lys Cys Lys Ser Gly Gly Asn Asn Pro Ser

20 25 30 35

ACA AAA GAG GTG TCA ATA CCA TCT GGG AAG CTT ACT ATT GAA GAT TTT 199 Thr Lys Glu Val Ser He Pro Ser Gly Lys Leu Thr He Glu Asp Phe

40 45 50

TGT ATT GGA AAT CAT CAA AGT TGC AAA ATA TTT TGC AAA AGT CAA TGT 247 Cys He Gly Asn His Gin Ser Cys Lys He Phe Cys Lys Ser Gin Cys 55 60 65

GGA TTT GGA GGT GGT GCT TGT GGA AAC GGT GGT TCA ACA CGA CCA AAT 295 Gly Phe Gly Gly Gly Ala Cys Gly Asn Gly Gly Ser Thr Arg Pro Asn 70 75 80

CAA AAA CAC TGT TAT TGC GA ATAACCATAT TCCGGATGAA AGACCAAATT 345

Gin Lys His Cys Tyr Cys 85 GATATAAATT ACTAAAATTA TGCTAGATAG CAATCATAAA ATTTTGAAGT TTTCAATGAT 405

CCTAACATGT TTTGCCTCCA ATTTATTTTA ACAGCAAATT GCTGGGAACT TACCGTACCG 465

TAACAAAATG TTCAAGAAAT ACTGAATGTT TACAAATAGA TTATTATAAA TATTGTAACA 525

TTGTCTAATA TTTATAAGAA TTATATAAAC TGAATTGCAA AAGTTGAAAA AAAAAAAAAA 585

AAAAAAAAAA 595

(2) INFORMATION FOR SEQ ID NO:53:

(i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 89 amino acids

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

(il) MOLECULE TYPE: protein

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:53:

Met Lys Phe Leu Leu Ala He Cys Val Leu Cys Val Leu Leu Asn Gin 1 5 10 15

Val Ser Met Ser Lys Met Val Thr Glu Lys Cys Lys Ser Gly Gly Asn 20 25 30

Asn Pro Ser Thr Lys Glu Val Ser He Pro Ser Gly Lys Leu Thr He

35 40 45

Glu Asp Phe Cys He Gly Asn His Gin Ser Cys Lys He Phe Cys Lys

50 55 60

Ser Gin Cys Gly Phe Gly Gly Gly Ala Cys Gly Asn Gly Gly Ser Thr

65 70 75 80

Arg Pro Asn Gin Lys His Cys Tyr Cys 85

(2) INFORMATION FOR SEQ ID NO:54:

(1) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 595 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: single (D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic)

(XI) SEQUENCE DESCRIPTION: SEQ ID NO:54: _{ττττττττττ ττττττττττ} TTTT _C AA _C TT TT _G CAATTCA _G TTTATATAA TTCTTATAAA 60

TATTAGACAA TGTTACAATA TTTATAATAA TCTATTTGTA AACATTCAGT ATTTCTTGAA 120 CATTTTGTTA CGGTACGGTA AGTTCCCAGC AATTTGCTGT TAAAATAAAT TGGAGGCAAA 180

ACATGTTAGG ATCATTGAAA ACTTCAAAAT TTTATGATTG CTATCTAGCA TAATTTTAGT 240

AATTTATATC AATTTGGTCT TTCATCCGGA ATATGGTTAT TCGCAATAAC AGTGTTTTTG 300

ATTTGGTCGT GTTGAACCAC CGTTTCCACA AGCACCACCT CCAAATCCAC ATTGACTTTT 360

GCAAAATATT TTGCAACTTT GATGATTTCC AATACAAAAA TCTTCAATAG TAAGCTTCCC 420 AGATGGTATT GACACCTCTT TTGTACTTGG ATTATTTCCT CCCGATTTAC ACTTTTCAGT 480

GACCATTTTT GACATAGATA CTTGATTTAA TAAAACACAC AACACGCAAA TTGCCAGTAA 540

AAATTTCATA TCGAATTTGA AAAATTTAAT GTTAAAACAA AATATTGAAT TTCCA 595

(2) INFORMATION FOR SEQ ID NO:55:

(1) SEQUENCE CHARACTERISTICS: (A) LENGTH: 270 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear (ll) MOLECULE TYPE: cDNA

(ix) FEATURE:

(A) NAME/KEY: CDS

(B) LOCATION: 1..270

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:55:

ATG AAA TTT TTA CTG GCA ATT TGC GTG TTG TGT GTT TTA TTA AAT CAA 48 Met Lys Phe Leu Leu Ala He Cys Val Leu Cys Val Leu Leu Asn Gin 1 5 10 15

GTA TCT ATG TCA AAA ATG GTC ACT GAA AAG TGT AAA TCG GGA GGA AAT 96 Val Ser Met Ser Lys Met Val Thr Glu Lys Cys Lys Ser Gly Gly Asn 20 25 30

AAT CCA AGT ACA AAA GAG GTG TCA ATA CCA TCT GGG AAG CTT ACT ATT 144 Asn Pro Ser Thr Lys Glu Val Ser He Pro Ser Gly Lys Leu Thr He 35 40 45 GAA GAT TTT TGT ATT GGA AAT CAT CAA AGT TGC AAA ATA TTT TGC AAA 192 Glu Asp Phe Cys He Gly Asn His Gin Ser Cys Lys He Phe Cys Lys 50 55 60

AGT CAA TGT GGA TTT GGA GGT GGT GCT TGT GGA AAC GGT GGT TCA ACA 240 Ser Gin Cys Gly Phe Gly Gly Gly Ala Cys Gly Asn Gly Gly Ser Thr 65 70 75 80

CGA CCA AAT CAA AAA CAC TGT TAT TGC GAA 270

Arg Pro Asn Gin Lys His Cys Tyr Cys Glu 85 90

(2) INFORMATION FOR SEQ ID NO:56: (l) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 90 amino acids

(B) TYPE: amino acid (D) TOPOLOGY: linear (n) MOLECULE TYPE: protein

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:56:

Met Lys Phe Leu Leu Ala He Cys Val Leu Cys Val Leu Leu Asn Gin 1 5 10 15

Val Ser Met Ser Lys Met Val Thr Glu Lys Cys Lys Ser Gly Gly Asn 20 25 30 Asn Pro Ser Thr Lys Glu Val Ser He Pro Ser Gly Lys Leu Thr He

35 40 45

Glu Asp Phe Cys He Gly Asn His Gin Ser Cys Lys He Phe Cys Lys 50 55 60

Ser Gin Cys Gly Phe Gly Gly Gly Ala Cys Gly Asn Gly Gly Ser Thr 65 70 75 80

Arg Pro Asn Gin Lys His Cys Tyr Cys Glu 85 90

(2) INFORMATION FOR SEQ ID NO:57: (l) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 270 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(n) MOLECULE TYPE: DNA (genomic)

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:57: TTCGCAATAA CAGTGTTTTT GATTTGGTCG TGTTGAACCA CCGTTTCCAC AAGCACCACC 60

TCCAAATCCA CATTGACTTT TGCAAAATAT TTTGCAACTT TGATGATTTC CAATACAAAA 120

ATCTTCAATA GTAAGCTTCC CAGATGGTAT TGACACCTCT TTTGTACTTG GATTATTTCC 180

TCCCGATTTA CACTTTTCAG TGACCATTTT TGACATAGAT ACTTGATTTA ATAAAACACA 240

CAACACGCAA ATTGCCAGTA AAAATTTCAT 270

(2) INFORMATION FOR SEQ ID NO: 58:

(l) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 213 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(n) MOLECULE TYPE: cDNA

(ix) FEATURE:

(A) NAME/KEY: CDS

(B) LOCATION: 1..213 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:58:

TCA AAA ATG GTC ACT GAA AAG TGT AAA TCG GGA GGA AAT AAT CCA AGT 48 Ser Lys Met Val Thr Glu Lys Cys Lys Ser Gly Gly Asn Asn Pro Ser 1 5 10 15

ACA AAA GAG GTG TCA ATA CCA TCT GGG AAG CTT ACT ATT GAA GAT TTT 96 Thr Lys Glu Val Ser He Pro Ser Gly Lys Leu Thr He Glu Asp Phe 20 25 30 TGT ATT GGA AAT CAT CAA AGT TGC AAA ATA TTT TGC AAA AGT CAA TGT 144 Cys He Gly Asn His Gin Ser Cys Lys He Phe Cys Lys Ser Gin Cys 35 40 45

GGA TTT GGA GGT GGT GCT TGT GGA AAC GGT GGT TCA ACA CGA CCA AAT 192 Gly Phe Gly Gly Gly Ala Cys Gly Asn Gly Gly Ser Thr Arg Pro Asn 50 55 60

CAA AAA CAC TGT TAT TGC GAA 213

Gin Lys His Cys Tyr Cys Glu 65 70

(2) INFORMATION FOR SEQ ID NO:59: (1) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 71 amino acids

(B) TYPE: amino acid (D) TOPOLOGY: linear (ll) MOLECULE TYPE: protein

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:59:

Ser Lys Met Val Thr Glu Lys Cys Lys Ser Gly Gly Asn Asn Pro Ser 1 5 10 15

Thr Lys Glu Val Ser He Pro Ser Gly Lys Leu Thr He Glu Asp Phe 20 25 30 Cys He Gly Asn His Gin Ser Cys Lys He Phe Cys Lys Ser Gin Cys 35 40 45

Gly Phe Gly Gly Gly Ala Cys Gly Asn Gly Gly Ser Thr Arg Pro Asn 50 55 60

Gin Lys His Cys Tyr Cys Glu 65 70

(2) INFORMATION FOR SEQ ID NO:60:

(l) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 213 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(11) MOLECULE TYPE: DNA (genomic)

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 60:

TTCGCAATAA CAGTGTTTTT GATTTGGTCG TGTTGAACCA CCGTTTCCAC AAGCACCACC 60 TCCAAATCCA CATTGACTTT TGCAAAATAT TTTGCAACTT TGATGATTTC CAATACAAAA 120

ATCTTCAATA GTAAGCTTCC CAGATGGTAT TGACACCTCT TTTGTACTTG GATTATTTCC 180

TCCCGATTTA CACTTTTCAG TGACCATTTT TGA 213

(2) INFORMATION FOR SEQ ID NO: 61:

(i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 1007 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA

(ix) FEATURE:

(A) NAME/KEY: CDS

(B) LOCATION: 1..465

(XI) SEQUENCE DESCRIPTION: SEQ ID NO: 61:

TGG AAA GTT AAT AAA AAA TGT ACA TCA GGT GGA AAA AAT CAA GAT AGA 48 Trp Lys Val Asn Lys Lys Cys Thr Ser Gly Gly Lys Asn Gin Asp Arg 1 5 10 15

AAA CTC GAT CAA ATA ATT CAA AAA GGC CAA CAA GTT AAA ATC CAA AAT 96 Lys Leu Asp Gin He He Gin Lys Gly Gin Gin Val Lys He Gin Asn 20 25 30

ATT TGC AAA TTA ATA CGA GAT AAA CCA CAT ACA AAT CAA GAG AAA GAA 144 He Cys Lys Leu He Arg Asp Lys Pro His Thr Asn Gin Glu Lys Glu 35 40 45

AAA TGT ATG AAA TTT TGC AAA AAA GTT TGC AAA GGT TAT AGA GGA GCT 192 Lys Cys Met Lys Phe Cys Lys Lys Val Cys Lys Gly Tyr Arg Gly Ala 50 55 60

TGT GAT GGC AAT ATT TGC TAC TGC AGC AGG CCA AGT AAT TTA GGT CCT 240 Cys Asp Gly Asn He Cys Tyr Cys Ser Arg Pro Ser Asn Leu Gly Pro 65 70 75 80 GAT TGG AAA GTA AGC AAA GAA TGC AAA GAT CCC AAT AAC AAA GAT TCT 288 Asp Trp Lys Val Ser Lys Glu Cys Lys Asp Pro Asn Asn Lys Asp Ser 85 90 95

CGT CCT ACG GAA ATA GTT CCA TAT CGA CAA CAA TTA GCA AAT CCA AAT 336 Arg Pro Thr Glu He Val Pro Tyr Arg Gin Gin Leu Ala Asn Pro Asn

100 105 110

ATT TGC AAA CTA AAA AAT TCA GAG ACC AAT GAA GAT TCC AAA TGC AAA 384 He Cys Lys Leu Lys Asn Ser Glu Thr Asn Glu Asp Ser Lys Cys Lys 115 120 125

AAA CAT TGC AAA GAA AAA TGT CGT GGT GGA AAT GAT GCT GGA TGT GAT 432 Lys His Cys Lys Glu Lys Cys Arg Gly Gly Asn Asp Ala Gly Cys Asp 130 135 140

GGA AAC TTT TGT TAT TGT CGA CCA AAA AAT AAA TAATAATTAT AATAAATAAA 485 Gly Asn Phe Cys Tyr Cys Arg Pro Lys Asn Lys 145 150 155

TTGTTATAGT TATTAGTTAT CCCATCACAT ATTAGAAAAG TGGCTTATAA TTTATGAACA 545

ATATAACACA TAAATTAGTT GTGTAATTTC GAATGTTTTT TTCAAATATA AGGCGTTTTT 605

CTAGAATATC TTGATATTAG AAACTAACTT AGATTATTTT GTTGTGTATA AAATATTCAA 665

ATACGTAAGT TATATTGAAC AAAGCATTTA GAAGCTACAT TAGATATACT AAATAAGTGC 725

AAAATTGCAT GGAAACCCTT ACTGGATTTA CTACATATTT TCTTCCTAAA TATTGTCTTG 785 GTATTACTCT TATTATATAA AAATTAATAT AAAATTGTAG ACAGAGACGA ATTGGGGTAT 845

TGTTATATAT AAAAAAGTAG TGGATTATTT AATTCTAAAA AAGTTTGCAA AATGTTTCAT 905

ACATAATAAC CGAATATTTT CAAATATATA AATATTGTAA TGAATAAATG CGCATCTGTA 965

TGCTTAATAT AAAAAAAAAA AAAAAAAAAA AAAAAAAAAA AA 1007

(2) INFORMATION FOR SEQ ID NO: 62:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 155 amino acids

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

(ii) MOLECULE TYPE: protein

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 62:

Trp Lys Val Asn Lys Lys Cys Thr Ser Gly Gly Lys Asn Gin Asp Arg 1 5 10 15

Lys Leu Asp Gin He He Gin Lys Gly Gin Gin Val Lys He Gin Asn 20 25 30

He Cys Lys Leu He Arg Asp Lys Pro His Thr Asn Gin Glu Lys Glu 35 40 45

Lys Cys Met Lys Phe Cys Lys Lys Val Cys Lys Gly Tyr Arg Gly Ala 50 55 60

Cys Asp Gly Asn He Cys Tyr Cys Ser Arg Pro Ser Asn Leu Gly Pro

65 70 75 80 Asp Trp Lys Val Ser Lys Glu Cys Lys Asp Pro Asn Asn Lys Asp Ser

85 90 95

Arg Pro Thr Glu He Val Pro Tyr Arg Gin Gin Leu Ala Asn Pro Asn

100 105 110

He Cys Lys Leu Lys Asn Ser Glu Thr Asn Glu Asp Ser Lys Cys Lys

115 120 125

Lys His Cys Lys Glu Lys Cys Arg Gly Gly Asn Asp Ala Gly Cys Asp 130 135 140

Gly Asn Phe Cys Tyr Cys Arg Pro Lys Asn Lys 145 150 155

INFORMATION FOR SEQ ID NO: 63:

(l) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 1007 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(n) MOLECULE TYPE: DNA (genomic)

(Xl) SEQUENCE DESCRIPTION: SEQ ID NO: 63: _{ττττττττττ τχττττττττ χτττττττττ} TTATATTAAG CATACAGATG CGCATTTATT 60

CATTACAATA TTTA ATATT TGAAAATATT CGGTTAT AT GTATGAAACA TTTTGCAAAC 120

TTTTTTAGAA TTAAATAATC CACTACTTTT TTATATATAA CAATACCCCA ATTCGTCTCT 180 GTCTACAATT TTATATTAAT TTTTATATAA TAAGAGTAAT ACCAAGACAA TATTTAGGAA 240

GAAAATATGT AGTAAATCCA GTAAGGGTTT CCATGCAATT TTGCACTTAT TTAGTATATC 300

TAATGTAGCT TCTAAATGCT TTGTTCAATA TAACTTACGT ATTTGAATAT TTTATACACA 360

ACAAAATAAT CTAAGTTAGT TTCTAATATC AAGATATTCT AGAAAAACGC CTTATATTTG 420

AAAAAAACAT TCGAAATTAC ACAACTAATT TATGTGTTAT ATTGTTCATA AATTATAAGC 480 CACTTTTCTA ATATGTGATG GGATAACTAA TAACTATAAC AATTTATTTA TTATAATTAT 540

TATTTATTTT TTGGTCGACA ATAACAAAAG TTTCCATCAC ATCCAGCATC ATTTCCACCA 600

CGACATTTTT CTTTGCAATG TTTTTTGCAT TTGGAATCTT CATTGGTCTC TGAATTTTTT 660

AGTTTGCAAA TATTTGGAAT TGCTAATTGT TGTCGATATG GAACTATTTC CGTAGGACGA 720

GAATCTTTGT TATTGGGATC TTTGCATTCT TTGCTTACTT TCCAATCAGG ACCTAAATTA 780 CTTGGCCTGC TGCAGTAGCA AATATTGCCA TCACAAGCTC CTCTATAACC TTTGCAAACT 840

TTTTTGCAAA ATTTCATACA TTTTTCTTTC TCTTGATTTG TATGTGGTTT ATCTCGTATT 900

AATTTGCAAA TATTTTGGAT TTTAACTTGT TGGCCTTTTT GAATTATTTG ATCGAGTTTT 960

CTATCTTGAT TTTTTCCACC TGATGTACAT TTTTTATTAA CTTTCCA 1007

(2) INFORMATION FOR SEQ ID NO: 64:

(l) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 1205 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: single (D) TOPOLOGY: linear

(n) MOLECULE TYPE: cDNA

(ix) FEATURE: (A) NAME/KEY: CDS

(B) LOCATION: 4..1062

( i) SEQUENCE DESCRIPTION: SEQ ID NO: 64: GCA GAA TTG AAA TTT GTG TTT GCG ACT GCA CGA GGT ATG TCA CAT ACA 48 Glu Leu Lys Phe Val Phe Ala Thr Ala Arg Gly Met Ser His Thr 1 5 10 15

CCT TGT GAT TAT CCA GGC GGT CCA AAA ATT ACA CAC AAG TCT GAA GAT 96

Pro Cys Asp Tyr Pro Gly Gly Pro Lys He Thr His Lys Ser Glu Asp 20 25 30 TCA AGC CAA TTG ACA CCG GCA GGT CAA GAA GAG GCA TTA AAA ATT GGC 144 Ser Ser Gin Leu Thr Pro Ala Gly Gin Glu Glu Ala Leu Lys He Gly 35 40 45

AAA TTA TTA TCC GAA CAT TAC AGA ACT AAT TTA AAA GTT GAC AAA TGG 192 Lys Leu Leu Ser Glu His Tyr Arg Thr Asn Leu Lys Val Asp Lys Trp 50 55 60

GAT TCA AAT AAA AAT TAT TGG ACA TTA GCT AGT GCT ACG AGA AGA TCT 240 Asp Ser Asn Lys Asn Tyr Trp Thr Leu Ala Ser Ala Thr Arg Arg Ser 65 70 75

CAA GAA GGA GCG CTT ATC ATT GGT TCT GGT CTA GAA GAA AAG GAA AAG 288 Gin Glu Gly Ala Leu He He Gly Ser Gly Leu Glu Glu Lys Glu Lys 80 85 90 95

GCA GTT TGG ACA AAA GAG AAA GGA GAT AAA ACC ATA TTT TCT TCG TTT 336

Ala Val Trp Thr Lys Glu Lys Gly Asp Lys Thr He Phe Ser Ser Phe

100 105 110 GGT GAA TAT GCT AAA TTT TAT AGT CCA AAA ACT TGT CCA AAC TTC ATA 384 Gly Glu Tyr Ala Lys Phe Tyr Ser Pro Lys Thr Cys Pro Asn Phe He 115 120 125

GCA CAA CAG AAA ATA GCA GTA AGA GAC TTG TTA ACA AAA AGT GCA AAA 432 Ala Gin Gin Lys He Ala Val Arg Asp Leu Leu Thr Lys Ser Ala Lys 130 135 140

GAT TAT AAA AAT TCA CTT GCA AAA TTA AAA GAA GCG TAT AAA ATA GAT 480 Asp Tyr Lys Asn Ser Leu Ala Lys Leu Lys Glu Ala Tyr Lys He Asp 145 150 155

GCG ACG ACA AGC CCT CAG AAT GTT TGG CTG GCA TAT GAA ACT TTG AAT 528

Ala Thr Thr Ser Pro Gin Asn Val Trp Leu Ala Tyr Glu Thr Leu Asn 160 165 170 175

TTA CAA AGC AAG CAA AAT AAC GCT CCA ACA TGG TGG AAT ACT GTA AAC 576

Leu Gin Ser Lys Gin Asn Asn Ala Pro Thr Trp Trp Asn Thr Val Asn 180 185 190 AAA GAT CTA AAA CAA TTC TCT GAG AAA TAT TTA TGG ACC GCC TTG ACT 624

Lys Asp Leu Lys Gin Phe Ser Glu Lys Tyr Leu Trp Thr Ala Leu Thr 195 200 205

TCT AAT GAT AAT CTT AGA AAG ATG TCA GGA GGT CGT ATG ATT AAC GAT 672 Ser Asn Asp Asn Leu Arg Lys Met Ser Gly Gly Arg Met He Asn Asp 210 215 220

ATA TTG AAC GAT ATC GAA AAC ATA AAG AAA GGA GAG GGA CAA CCG GGT 720 He Leu Asn Asp He Glu Asn He Lys Lys Gly Glu Gly Gin Pro Gly 225 230 235

GCT CCA GGA GGA AAG GAA AAC AAA TTA TCA GTG CTG ACC GTT CCT CAA 768 Ala Pro Gly Gly Lys Glu Asn Lys Leu Ser Val Leu Thr Val Pro Gin 240 245 250 255

GCT ATC TTA GCA GCA TTT GTT TCA GCA TTT GCT CCC GAA GGT ACA AAA 816 Ala He Leu Ala Ala Phe Val Ser Ala Phe Ala Pro Glu Gly Thr Lys 260 265 270 ATT GAA AAT AAG GAC CTT GAT CCG TCT ACT TTA TAT CCT GGC CAA GGA 864 He Glu Asn Lys Asp Leu Asp Pro Ser Thr Leu Tyr Pro Gly Gin Gly 275 280 285

GCA CTT CAC GTT ATT GAA CTA CAC CAA GAT AAG AGC GAT TGG AGC ATA 912

Ala Leu His Val He Glu Leu His Gin Asp Lys Ser Asp Trp Ser He 290 295 300 AAA GTT CTC TAT AGA AAC AAT GAC CAA ATG AAG CTG AAA CCA ATG AAA 960

Lys Val Leu Tyr Arg Asn Asn Asp Gin Met Lys Leu Lys Pro Met Lys 305 310 315

CTT GCA CAA TGC GGT GAC AAG TGT TCT TAT GGT ACT TTC AAA TCA ATG 1008 Leu Ala Gin Cys Gly Asp Lys Cys Ser Tyr Gly Thr Phe Lys Ser Met 320 325 330 335

CTA CAA AAA TAT AAC ATG GAG AAG GAA GCT CAT GAT AAA TTA TGT AAA 1056

Leu Gin Lys Tyr Asn Met Glu Lys Glu Ala His Asp Lys Leu Cys Lys 340 345 350

ACG TCG TAAAAATTAA AAATAAAAAC TTTTCAATAT ATTTTCCGCT AAAATAAATA 1112 Thr Ser

AATATGTTTG TATATTTAAA CTTATCAAAA TAATAGTAGT GTTTTAATAA AGATTTTAAA 1172 TAAATAATTG TAAAAAAAAA AAAAAAAAAA AAA 1205

(2) INFORMATION FOR SEQ ID NO: 65:

(i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 353 amino acids

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

(ii) MOLECULE TYPE: protein

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 65:

Glu Leu Lys Phe Val Phe Ala Thr Ala Arg Gly Met Ser His Thr Pro 1 5 10 15

Cys Asp Tyr Pro Gly Gly Pro Lys He Thr His Lys Ser Glu Asp Ser 20 25 30

Ser Gin Leu Thr Pro Ala Gly Gin Glu Glu Ala Leu Lys He Gly Lys 35 40 45

Leu Leu Ser Glu His Tyr Arg Thr Asn Leu Lys Val Asp Lys Trp Asp 50 55 60 Ser Asn Lys Asn Tyr Trp Thr Leu Ala Ser Ala Thr Arg Arg Ser Gin 65 70 75 80

Glu Gly Ala Leu He He Gly Ser Gly Leu Glu Glu Lys Glu Lys Ala

85 90 95

Val Trp Thr Lys Glu Lys Gly Asp Lys Thr He Phe Ser Ser Phe Gly

100 105 110

Glu Tyr Ala Lys Phe Tyr Ser Pro Lys Thr Cys Pro Asn Phe He Ala 115 120 125

Gin Gin Lys He Ala Val Arg Asp Leu Leu Thr Lys Ser Ala Lys Asp

130 135 140 Tyr Lys Asn Ser Leu Ala Lys Leu Lys Glu Ala Tyr Lys He Asp Ala

145 150 155 160

Thr Thr Ser Pro Gin Asn Val Trp Leu Ala Tyr Glu Thr Leu Asn Leu

165 170 175

in Ser Lys Gin Asn Asn Ala Pro Thr Trp Trp Asn Thr Val Asn Lys 180 185 190 sp Leu Lys Gin Phe Ser Glu Lys Tyr Leu Trp Thr Ala Leu Thr Ser 195 200 205

Asn Asp Asn Leu Arg Lys Met Ser Gly Gly Arg Met He Asn Asp He 210 215 220 Leu Asn Asp He Glu Asn He Lys Lys Gly Glu Gly Gin Pro Gly Ala 225 230 235 240

Pro Gly Gly Lys Glu Asn Lys Leu Ser Val Leu Thr Val Pro Gin Ala 245 250 255

He Leu Ala Ala Phe Val Ser Ala Phe Ala Pro Glu Gly Thr Lys He 260 265 270

Glu Asn Lys Asp Leu Asp Pro Ser Thr Leu Tyr Pro Gly Gin Gly Ala 275 280 285

Leu His Val He Glu Leu His Gin Asp Lys Ser Asp Trp Ser He Lys

290 295 300

Val Leu Tyr Arg Asn Asn Asp Gin Met Lys Leu Lys Pro Met Lys Leu 305 310 315 320

Ala Gin Cys Gly Asp Lys Cys Ser Tyr Gly Thr Phe Lys Ser Met Leu 325 330 335

Gin Lys Tyr Asn Met Glu Lys Glu Ala His Asp Lys Leu Cys Lys Thr 340 345 350

Ser

(2) INFORMATION FOR SEQ ID NO:66:

(l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 1205 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic)

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 66: _{ττττττττττ χτττττττττ} TTACAATTAT TTATTTAAAA TCTTTATTAA AACACTA _C TA 60

TTATTTTGAT AAGTTTAAAT ATACAAACAT ATTTATTTAT TTTAGCGGAA AATATATTGA 120

AAAGTTTTTA TTTTTAATTT TTACGACGTT TTACATAATT TATCATGAGC TTCCTTCTCC 180 ATGTTATATT TTTGTAGCAT TGATTTGAAA GTACCATAAG AACACTTGTC ACCGCATTGT 240

GCAAGTTTCA TTGGTTTCAG CTTCATTTGG TCATTGTTTC TATAGAGAAC TTTTATGCTC 300

CAATCGCTCT TATCTTGGTG TAGTTCAATA ACGTGAAGTG CTCCTTGGCC AGGATATAAA 360

GTAGACGGAT CAAGGTCCTT ATTTTCAATT TTTGTACCTT CGGGAGCAAA TGCTGAAACA 420

AATGCTGCTA AGATAGCTTG AGGAACGGTC AGCACTGATA ATTTGTTTTC CTTTCCTCCT 480 GGAGCACCCG GTTGTCCCTC TCCTTTCTTT ATGTTTTCGA TATCGTTCAA TATATCGTTA 540

ATCATACGAC CTCCTGACAT CTTTCTAAGA TTATCATTAG AAGTCAAGGC GGTCCATAAA 600

TATTTCTCAG AGAATTGTTT TAGATCTTTG TTTACAGTAT TCCACCATGT TGGAGCGTTA 660

TTTTGCTTGC TTTGTAAATT CAAAGTTTCA TATGCCAGCC AAACATTCTG AGGGCTTGTC 720

GTCGCATCTA TTTTATACGC TTCTTTTAAT TTTGCAAGTG AATTTTTATA ATCTTTTGCA 780

CTTTTTGTTA ACAAGTCTCT TACTGCTATT TTCTGTTGTG CTATGAAGTT TGGACAAGTT 840

TTTGGACTAT AAAATTTAGC ATATTCACCA AACGAAGAAA ATATGGTTTT ATCTCCTTTC 900

TCTTTTGTCC AAACTGCCTT TTCCTTTTCT TCTAGACCAG AACCAATGAT AAGCGCTCCT 960

TCTTGAGATC TTCTCGTAGC ACTAGCTAAT GTCCAATAAT TTTTATTTGA ATCCCATTTG 1020

TCAACTTTTA AATTAGTTCT GTAATGTTCG GATAATAATT TGCCAATTTT TAATGCCTCT 1080 TCTTGACCTG CCGGTGTCAA TTGGCTTGAA TCTTCAGACT TGTGTGTAAT TTTTGGACCG 1140

CCTGGATAAT CACAAGGTGT ATGTGACATA CCTCGTGCAG TCGCAAACAC AAATTTCAAT 1200

TCTGC 1205

(2) INFORMATION FOR SEQ ID NO: 67:

(l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 1059 base pairs

( B) TYPE : nucleic acid

(C ) STRANDEDNESS : single

(D) TOPOLOGY : linear (ll) MOLECULE TYPE: cDNA

(ix) FEATURE:

(A) NAME/KEY: CDS

(B) LOCATION: 1..1059

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 67:

GAA TTG AAA TTT GTG TTT GCG ACT GCA CGA GGT ATG TCA CAT ACA CCT 48 Glu Leu Lys Phe Val Phe Ala Thr Ala Arg Gly Met Ser His Thr Pro 1 5 10 15

TGT GAT TAT CCA GGC GGT CCA AAA ATT ACA CAC AAG TCT GAA GAT TCA 96 Cys Asp Tyr Pro Gly Gly Pro Lys He Thr His Lys Ser Glu Asp Ser 20 25 30

AGC CAA TTG ACA CCG GCA GGT CAA GAA GAG GCA TTA AAA ATT GGC AAA 144 Ser Gin Leu Thr Pro Ala Gly Gin Glu Glu Ala Leu Lys He Gly Lys 35 40 45 TTA TTA TCC GAA CAT TAC AGA ACT AAT TTA AAA GTT GAC AAA TGG GAT 192 Leu Leu Ser Glu His Tyr Arg Thr Asn Leu Lys Val Asp Lys Trp Asp 50 55 60

TCA AAT AAA AAT TAT TGG ACA TTA GCT AGT GCT ACG AGA AGA TCT CAA 240 Ser Asn Lys Asn Tyr Trp Thr Leu Ala Ser Ala Thr Arg Arg Ser Gin 65 70 75 80

GAA GGA GCG CTT ATC ATT GGT TCT GGT CTA GAA GAA AAG GAA AAG GCA 288 Glu Gly Ala Leu He He Gly Ser Gly Leu Glu Glu Lys Glu Lys Ala 85 90 95

GTT TGG ACA AAA GAG AAA GGA GAT AAA ACC ATA TTT TCT TCG TTT GGT 336 Val Trp Thr Lys Glu Lys Gly Asp Lys Thr He Phe Ser Ser Phe Gly 100 105 110

GAA TAT GCT AAA TTT TAT AGT CCA AAA ACT TGT CCA AAC TTC ATA GCA 384 Glu Tyr Ala Lys Phe Tyr Ser Pro Lys Thr Cys Pro Asn Phe He Ala 115 120 125

CAA CAG AAA ATA GCA GTA AGA GAC TTG TTA ACA AAA AGT GCA AAA GAT 43?

Gin Gin Lys He Ala Val Arg Asp Leu Leu Thr Lys Ser Ala Lys Asp

130 135 140 TAT AAA AAT TCA CTT GCA AAA TTA AAA GAA GCG TAT AAA ATA GAT GCG 480

Tyr Lys Asn Ser Leu Ala Lys Leu Lys Glu Ala Tyr Lys He Asp Ala 145 150 155 160

ACG ACA AGC CCT CAG AAT GTT TGG CTG GCA TAT GAA ACT TTG AAT TTA 528 Thr Thr Ser Pro Gin Asn Val Trp Leu Ala Tyr Glu Thr Leu Asn Leu

165 170 175

CAA AGC AAG CAA AAT AAC GCT CCA ACA TGG TGG AAT ACT GTA AAC AAA 576

Gin Ser Lys Gin Asn Asn Ala Pro Thr Trp Trp Asn Thr Val Asn Lys 180 185 190

GAT CTA AAA CAA TTC TCT GAG AAA TAT TTA TGG ACC GCC TTG ACT TCT 624

Asp Leu Lys Gin Phe Ser Glu Lys Tyr Leu Trp Thr Ala Leu Thr Ser 195 200 205

AAT GAT AAT CTT AGA AAG ATG TCA GGA GGT CGT ATG ATT AAC GAT ATA 672

Asn Asp Asn Leu Arg Lys Met Ser Gly Gly Arg Met He Asn Asp He

210 215 220 TTG AAC GAT ATC GAA AAC ATA AAG AAA GGA GAG GGA CAA CCG GGT GCT 720

Leu Asn Asp He Glu Asn He Lys Lys Gly Glu Gly Gin Pro Gly Ala 225 230 235 240

CCA GGA GGA AAG GAA AAC AAA TTA TCA GTG CTG ACC GTT CCT CAA GCT 768 Pro Gly Gly Lys Glu Asn Lys Leu Ser Val Leu Thr Val Pro Gin Ala

245 250 255

ATC TTA GCA GCA TTT GTT TCA GCA TTT GCT CCC GAA GGT ACA AAA ATT 816

He Leu Ala Ala Phe Val Ser Ala Phe Ala Pro Glu Gly Thr Lys He 260 265 270

GAA AAT AAG GAC CTT GAT CCG TCT ACT TTA TAT CCT GGC CAA GGA GCA 864

Glu Asn Lys Asp Leu Asp Pro Ser Thr Leu Tyr Pro Gly Gin Gly Ala 275 280 285

CTT CAC GTT ATT GAA CTA CAC CAA GAT AAG AGC GAT TGG AGC ATA AAA 912

Leu His Val He Glu Leu His Gin Asp Lys Ser Asp Trp Ser He Lys 290 295 300 GTT CTC TAT AGA AAC AAT GAC CAA ATG AAG CTG AAA CCA ATG AAA CTT 960 Val Leu Tyr Arg Asn Asn Asp Gin Met Lys Leu Lys Pro Met Lys Leu 305 310 315 320

GCA CAA TGC GGT GAC AAG TGT TCT TAT GGT ACT TTC AAA TCA ATG CTA 1008 Ala Gin Cys Gly Asp Lys Cys Ser Tyr Gly Thr Phe Lys Ser Met Leu

325 330 335

CAA AAA TAT AAC ATG GAG AAG GAA GCT CAT GAT AAA TTA TGT AAA ACG 1056 Gin Lys Tyr Asn Met Glu Lys Glu Ala His Asp Lys Leu Cys Lys Thr 340 345 350

TCG 1059

Ser

(2) INFORMATION FOR SEQ ID NO: 68:

(l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 353 amino acids

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

(n) MOLECULE TYPE- protein

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 68:

Glu Leu Lys Phe Val Phe Ala Thr Ala Arg Gly Met Ser His Thr Pro

1 5 10 15

Cys Asp Tyr Pro Gly Gly Pro Lys He Thr His Lys Ser Glu Asp Ser 20 25 30

Ser Gin Leu Thr Pro Ala Gly Gin Glu Glu Ala Leu Lys He Gly Lys 35 40 45

Leu Leu Ser Glu His Tyr Arg Thr Asn Leu Lys Val Asp Lys Trp Asp 50 55 60 Ser Asn Lys Asn Tyr Trp Thr Leu Ala Ser Ala Thr Arg Arg Ser Gin 65 70 75 80

Glu Gly Ala Leu He He Gly Ser Gly Leu Glu Glu Lys Glu Lys Ala 85 90 95

Val Trp Thr Lys Glu Lys Gly Asp Lys Thr He Phe Ser Ser Phe Gly 100 105 110

Glu Tyr Ala Lys Phe Tyr Ser Pro Lys Thr Cys Pro Asn Phe He Ala 115 120 125

Gin Gin Lys He Ala Val Arg Asp Leu Leu Thr Lys Ser Ala Lys Asp 130 135 140 Tyr Lys Asn Ser Leu Ala Lys Leu Lys Glu Ala Tyr Lys He Asp Ala 145 150 155 160

Thr Thr Ser Pro Gin Asn Val Trp Leu Ala Tyr Glu Thr Leu Asn Leu 165 170 175

Gin Ser Lys Gin Asn Asn Ala Pro Thr Trp Trp Asn Thr Val Asn Lys 180 185 190

Asp Leu Lys Gin Phe Ser Glu Lys Tyr Leu Trp Thr Ala Leu Thr Ser 195 200 205

Asn Asp Asn Leu Arg Lys Met Ser Gly Gly Arg Met He Asn Asp He 210 215 220 Leu Asn Asp He Glu Asn He Lys Lys Gly Glu Gly Gin Pro Gly Ala 225 230 235 240

Pro Gly Gly Lys Glu Asn Lys Leu Ser Val Leu Thr Val Pro Gin Ala

245 250 255

He Leu Ala Ala Phe Val Ser Ala Phe Ala Pro Glu Gly Thr Lys He

260 265 270

Glu Asn Lys Asp Leu Asp Pro Ser Thr Leu Tyr Pro Gly Gin Gly Ala 275 280 285

Leu His Val He Glu Leu His Gin Asp Lys Ser Asp Trp Ser He Lys 290 295 300 Val Leu Tyr Arg Asn Asn Asp Gin Met Lys Leu Lys Pro Met Lys Leu

305 310 315 320

Ala Gin Cys Gly Asp Lys Cys Ser Tyr Gly Thr Phe Lys Ser Met Leu 325 330 335

Gin Lys Tyr Asn Met Glu Lys Glu Ala His Asp Lys Leu Cys Lys Thr 340 345 350

Ser

(2) INFORMATION FOR SEQ ID NO:69:

(l) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 1059 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(n) MOLECULE TYPE: DNA (genomic)

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 69:

CGACGTTTTA CATAATTTAT CATGAGCTTC CTTCTCCATG TTATATTTTT GTAGCATTGA 60 TTTGAAAGTA CCATAAGAAC ACTTGTCACC GCATTGTGCA AGTTTCATTG GTTTCAGCTT 120

CATTTGGTCA TTGTTTCTAT AGAGAACTTT TATGCTCCAA TCGCTCTTAT CTTGGTGTAG 180

TTCAATAACG TGAAGTGCTC CTTGGCCAGG ATATAAAGTA GACGGATCAA GGTCCTTATT 240

TTCAATTTTT GTACCTTCGG GAGCAAATGC TGAAACAAAT GCTGCTAAGA TAGCTTGAGG 300

AACGGTCAGC ACTGATAATT TGTTTTCCTT TCCTCCTGGA GCACCCGGTT GTCCCTCTCC 360 TTTCTTTATG TTTTCGATAT CGTTCAATAT ATCGTTAATC ATACGACCTC CTGACATCTT 420

TCTAAGATTA TCATTAGAAG TCAAGGCGGT CCATAAA AT TTCTCAGAGA ATTGTTTTAG 480

ATCTTTGTTT ACAGTATTCC ACCATGTTGG AGCGTTATTT TGCTTGCTTT GTAAATTCAA 540

AGTTTCATAT GCCAGCCAAA CATTCTGAGG GCTTGTCGTC GCATCTATTT TATACGCTTC 600

TTTTAATTTT GCAAGTGAAT TTTTATAATC TTTTGCACTT TTTGTTAACA AGTCTCTTAC 660 TGCTATTTTC TGTTGTGCTA TGAAGTTTGG ACAAGTTTTT GGACTATAAA ATTTAGCATA 720

TTCACCAAAC GAAGAAAATA TGGTTTTATC TCCTTTCTCT TTTGTCCAAA CTGCCTTTTC 780

CTTTTCTTCT AGACCAGAAC CAATGATAAG CGCTCCTTCT TGAGATCTTC TCGTAGCACT 840

AGCTAATGTC CAATAATTTT TATTTGAATC CCATTTGTCA ACTTTTAAAT TAGTTCTGTA 900

ATGTTCGGAT AATAATTTGC CAATTTTTAA TGCCTCTTCT TGACCTGCCG GTGTCAATTG 960 GCTTGAATCT TCAGACTTGT GTGTAATTTT TGGACCGCCT GGATAATCAC AAGGTGTATG 1020

TGACATACCT CGTGCAGTCG CAAACACAAA TTTCAATTC 1059

(2) INFORMATION FOR SEQ ID NO: 0:

(l) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 25 amino acids

(B) TYPE: amino acid (C) STRANDEDNESS:

(D) TOPOLOGY: linear

( n ) MOLECULE TYPE : peptide

(xi ) SEQUENCE DESCRI PTION : SEQ ID NO : 70 :

Xaa Glu Leu Lys Phe Val Phe Val Met Val Lys Gly Pro Asp His Glu

1 5 10 15

Ala Cys Asn Tyr Ala Gly Gly Xaa Gin 20 25

(2) INFORMATION FOR SEQ ID NO: 1:

(l) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 406 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(ll) MOLECULE TYPE: cDNA

(ix) FEATURE:

(A) NAME/KEY: CDS

(B) LOCATION: 1..405

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 71:

AAA GGT CCA GAT CAC GAA GCT TGT AAC 48 Lys Gly Pro Asp His Glu Ala Cys Asn 5 10

ACT ACT CTT CAA GAA AAA GAT AGT GTT 96 Thr Thr Leu Gin Glu Lys Asp Ser Val 20 25

GAA GCA TAC GAA TTG GGA AAA CTT TTG 144 Glu Ala Tyr Glu Leu Gly Lys Leu Leu 35 40

TTA AAA GTT GAC AAA TGG GAT GCC ACG 192

Leu Lys Val Asp Lys Trp Asp Ala Thr

55

ACA AAA GCT ATG CGT ACT AAA GAA GCA 240

Thr Lys Ala Met Arg Thr Lys Glu Ala

70 75

TTG GAA AAT AAT CCT GCA AAA GCT AAA 288 Leu Glu Asn Asn Pro Ala Lys Ala Lys 85 90

CTC GAT TCA ACA CAT TTT GAT GCG ATG 336 Leu Asp Ser Thr His Phe Asp Ala Met 100 105

AAT CCT CAA CAA TGT CCG GCA TAT TTC 384 Asn Pro Gin Gin Cys Pro Ala Tyr Phe 115 120

CAG AAA ATA AAG AAA T 406

Gin Lys He Lys Lys 135

(2) INFORMATION FOR SEQ ID NO:72:

(l) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 135 amino acids

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

(n) MOLECULE TYPE: protein

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 72:

Met Val Lys Gly Pro Asp His Glu Ala Cys Asn Tyi Ala Gly Gly Pro

1 '5 10 15

Gin Leu Thr Thr Leu Gin Glu Lys Asp Ser Val Leu Thr Glu Asp Gly

20 25 30

Lys Thr Glu Ala Tyr Glu Leu Gly Lys Leu Leu Asp Lys Val Tyr Lys

35 40 45

Lys Gin Leu Lys Val Asp Lys Trp Asp Ala Thr Lys Thr Tyr Trp Ala 50 55 60

Val Ser Thr Lys Ala Met Arg Thr Lys Glu Ala Ala Leu He Val Gly 65 70 75 80

Ala Gly Leu Glu Asn Asn Pro Ala Lys Ala Lys Gly Asn Trp Thr Gin 85 90 95

Gin Gin Leu Asp Ser Thr His Phe Asp Ala Met Pro Gly Phe Ser Arg 100 105 110

Phe Trp Asn Pro Gin Gin Cys Pro Ala Tyr Phe Arg Ala Leu Ser Leu

115 120 125

Gin Asn Gin Lys He Lys Lys 130 135

(2) INFORMATION FOR SEQ ID NO:73:

(l) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 407 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: single (D) TOPOLOGY: linear

(n) MOLECULE TYPE: DNA (genomic) (XI) SEQUENCE DESCRIPTION: SEQ ID NO: 73:

AATTTCTTTA TTTTCTGATT TTGTAGCGAG AGCGCTCTGA AATATGCCGG ACATTGTTGA 60

GGATTCCAAA ATCTAGAAAA GCCAGGCATC GCATCAAAAT GTGTTGAATC GAGCTGTTGT 120 TGTGTCCAAT TACCTTTAGC TTTTGCAGGA TTATTTTCCA ATCCTGCTCC TACAATTAAG 180

GCTGCTTCTT TAGTACGCAT AGCTTTTGTG GACACAGCCC AGTAGGTTTT CGTGGCATCC 240

CATTTGTCAA CTTTTAATTG TTTTTTATAT ACCTTGTCCA AAAGTTTTCC CAATTCGTAT 300

GCTTCTGTCT TGCCATCTTC AGTTAGAACA CTATCTTTTT CTTGAAGAGT AGTTAACTGA 360

GGACCTCCTG CATAGTTACA AGCTTCGTGA TCTGGACCTT TAACCAT 407

(2) INFORMATION FOR SEQ ID NO: 74:

(l) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 420 base pairs (B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(n) MOLECULE TYPE: cDNA

(ix) FEATURE:

(A) NAME/KEY: CDS

(B) LOCATION- 1..216

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 74'

GAA GTT ATG GAT AAA TTG CGA AAA CAG GCA CCT CCT AAA ACT GAT GGC 48 Glu Val Met Asp Lys Leu Arg Lys Gin Ala Pro Pro Lys Thr Asp Gly 1 5 10 15

AAT CCT CCA AAA ACA ACC ATA ATG AGT ACA CTT CAA AAG CAA CAA ATA 96 Asn Pro Pro Lys Thr Thr He Met Ser Thr Leu Gin Lys Gin Gin He 20 25 30

AGT TGC ACA GAA GTG AAA GCG GTT AAC TTA GAA AGT CAT GTT TGT GCT 144 Ser Cys Thr Glu Val Lys Ala Val Asn Leu Glu Ser His Val Cys Ala 35 40 45 TAT GAT TGT AGT CAA CCT GAA ACT GCA GGA ATT ACA TGC AAA GGA AAT 192 Tyr Asp Cys Ser Gin Pro Glu Thr Ala Gly He Thr Cys Lys Gly Asn 50 55 60

AAG TGT GAT TGT CCT AAA AAA CGC TAAAAATTTA TTCAAAACAT TTACATTTTT 246 Lys Cys Asp Cys Pro Lys Lys Arg 65 70

TATTAATATT CAACTATCAA AAATTCTGTG TTGATTGTTA TTATATTTAT CATAGTTACT 306 AGAAATAAAA TTTTATAACA TTGTTAATTC GAAATTGAAT ACACATAATA TTATAATTAG 366

TGAGGTTAAA AGAAATAAAC CGAATATCCA AATCAAAAAA AAAAAAAAAA AAAA 420

(2) INFORMATION FOR SEQ ID NO: 75:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 72 a ino acids (B) TYPE: a ino acid

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: protein (XI) SEQUENCE DESCRIPTION: SEQ ID NO: 75:

Glu Val Met Asp Lys Leu Arg Lys Gin Ala Pro Pro Lys Thr Asp Gly

1 5 10 15 Asn Pro Pro Lys Thr Thr He Met Ser Thr Leu Gin Lys Gin Gin He

20 25 30

Ser Cys Thr Glu Val Lys Ala Val Asn Leu Glu Ser His Val Cys Ala 35 40 45

Tyr Asp Cys Ser Gin Pro Glu Thr Ala Gly He Thr Cys Lys Gly Asn 50 55 60

Lys Cys Asp Cys Pro Lys Lys Arg 65 70

(2) INFORMATION FOR SEQ ID NO:76: (l) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 420 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(n) MOLECULE TYPE: DNA (genomic)

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

_TTTT TTTTTT TTTTTTTTTT GATTTGGATA TTCGGTTTAT TTCTTTTAAC CTCACTAATT 60

ATAATATTAT GTGTATTCAA TTTCGAATTA ACAATGTTAT AAAATTTTAT TTCTAGTAAC 120

TATGATAAAT ATAATAACAA TCAACACAGA ATTTTTGATA GTTGAATATT AATAAAAAAT 180

GTAAATGTTT TGAATAAATT TTTAGCGTTT TTTAGGACAA TCACACTTAT TTCCTTTGCA 240

TGTAATTCCT GCAGTTTCAG GTTGACTACA ATCATAAGCA CAAACATGAC TTTCTAAGTT 300

AACCGCTTTC ACTTCTGTGC AACTTATTTG TTGCTTTTGA AGTGTACTCA TTATGGTTGT 360

TTTTGGAGGA TTGCCATCAG TTTTAGGAGG TGCCTGTTTT CGCAATTTAT CCATAACTTC 420

(2) INFORMATION FOR SEQ ID NO: 77:

(l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 71 amino acids

(B) TYPE: amino acid

(C) STRANDEDNESS:

(D) TOPOLOGY: linear (li) MOLECULE TYPE: peptide

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:77:

Ser Lys Met Val Thr Glu Lys Cys Lys Ser Gly Gly Asn Asn Pro Ser 1 5 10 15

Thr Lys Glu Val Ser He Pro Ser Gly Lys Leu Thr He Glu Asp Phe 20 25 30 Cys He Gly Asn His Gin Ser Cys Lys He Phe Cys Lys Ser Gin Cys

35 40 45

Gly Phe Gly Gly Gly Ala Cys Gly Asn Gly Gly Ser Thr Arg Pro Asn 50 55 60

Gin Lys His Cys Tyr Cys Glu 65 70

(2) INFORMATION FOR SEQ ID NO:78:

(l) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 25 amino acids (B) TYPE: amino acid

(C) STRANDEDNESS:

(D) TOPOLOGY: linear

(n) MOLECULE TYPE: peptide

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

Asn Asp Lys Leu Gin Phe Val Phe Val Met Ala Arg Gly Pro Asp His

1 5 10 15

Glu Ala Cys Asn Tyr Pro Gly Gly Pro 20 25

(2) INFORMATION FOR SEQ ID NO:79:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 26 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY, linear

(ll) MOLECULE TYPE: DNA (genomic)

(ix) FEATURE:

(A) NAME/KEY: mιsc_feature

(B) LOCATION: 1..26

(D) OTHER INFORMATION: /label= primer

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:79: AGTGGATCCG TCAAAAATGG TCACTG 26

(2) INFORMATION FOR SEQ ID NO:80:

(l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 28 base pairs (B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic)

(ix) FEATURE:

(A) NAME/KEY: mιsc_feature

(B) LOCATION: 1..28

(D) OTHER INFORMATION: /label- primer

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:80: CCGGAATTCG GTTATTCGCA ATAACAGT 28

(2) INFORMATION FOR SEQ ID NO:81:

(l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 54 base pairs (B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(n) MOLECULE TYPE: DNA (genomic)

(ix) FEATURE:

(A) NAME/KEY: mιsc_feature

(B) LOCATION: 1..54

(D) OTHER INFORMATION: /label= primer

(XI) SEQUENCE DESCRIPTION: SEQ ID NO:81: GCGCGGATCC GCATATGGAA GACATCTGGA AAGTTAATAA AAAATGTACA TCAG 54

(2) INFORMATION FOR SEQ ID NO:82:

(i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 45 base pairs (B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(li) MOLECULE TYPE: DNA (genomic)

(IX) FEATURE'

(A) NAME/KEY: nusr eature

(B) LOCATION: 1..45

(D) OTHER INFORMATION: /label= primer

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:82: CCGGAATTCT TATTTATTTT TTGGTCGACA ATAACAAAAG TTTCC 45

(2) INFORMATION FOR SEQ ID NO:83:

(l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 46 base pairs (B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(n) MOLECULE TYPE: DNA (genomic)

(ix) FEATURE:

(A) NAME/KEY: mιsc_feature

(B) LOCATION: 1..46

(D) OTHER INFORMATION: /label= primer

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 83: AAATTTGTAT TTTGTATATG GTATAAAGGA TCCATGATCA TGAAGC 46

(2) INFORMATION FOR SEQ ID NO: 84:

(l) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 37 base pairs

(B) TYPE: nucleic acid (C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(n) MOLECULE TYPE: DNA (genomic) (ix) FEATURE:

(A) NAME/KEY: mιsc_feature

(B) LOCATION: 1..37

(D) OTHER INFORMATION: /label= primer

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 84: CATGAACCAT GGATAATACA TCGATAAAGA TACTACG 37

(2) INFORMATION FOR SEQ ID NO:85:

(l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 17 base pairs (B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(n) MOLECULE TYPE: DNA (genomic)

(ix) FEATURE:

(A) NAME/KEY: mιsc_feature

(B) LOCATION: 1..17

(D) OTHER INFORMATION: /label= primer

(xi) SEQUENCE DESCRIPTION SEQ ID NO: 85

GTAAAACGAC GGCCAGT 17

(2) INFORMATION FOR SEQ ID NO:86:

(l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 31 base pairs (B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic)

(IX) FEATURE:

(A) NAME/KEY: mιsc_feature

(B) LOCATION: 1..31

(D) OTHER INFORMATION: /label= primer

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:86: GAAGTATATG GACTAAATTA GAGAGCAAGG C 31

(2) INFORMATION FOR SEQ ID NO:87:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 19 amino acids

(B) TYPE: amino acid

(C) STRANDEDNESS: (D) TOPOLOGY: linear

(li) MOLECULE TYPE: peptide

(ix) FEATURE: (A) NAME/KEY: Peptide

(B) LOCATION: 1..19

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 87: Tyr Phe Asn Lys Leu Val Gin Ser Trp Thr Glu Pro Met Val Phe Lys

1 5 10 15

Tyr Pro Tyr

(2) INFORMATION FOR SEQ ID NO:88:

(l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 24 base pairs (B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(n) MOLECULE TYPE: DNA (genomic)

(ix) FEATURE:

(A) NAME/KEY: mιsc_feature

(B) LOCATION: 1..24

(D) OTHER INFORMATION: /label= primer

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:88: GTAATACGAC TCACTATATA GGGC 24

While various embodiments of the present invention have been described in detail, it is apparent that modifications and adaptations of those embodiments will occur to those skilled in the art. It is to be expressly understood, however, that such modifications and adaptations are within the scope of the present invention, as set forth in the following claims.