A novel protein tyrosine kinase, JAK3, and a polynucleotide sequence encoding JAK3 polypeptide are disclosed herein. JAK3 is a new member of the JAK family of protein tyrosine kinases which are important in regulation of cellular proliferation and differentiation. Also disclosed are therapeutic methods utilizing JAK3 polypeptide and polynucleotide sequences.

BACKGROUND OF THE INVENTION 
1. Field of the Invention 
The present invention relates generally to a protein tyrosine kinases and 
specifically to JAK3, a novel protein tyrosine kinase of the JAK family. 
2. Description of Related Art 
Proliferation and differentiation of hematopoietic cells is dependent upon 
the binding of hematopoietic growth factors and cytokines to their 
respective cell surface receptors (Cross, et al., Cell, 64:271, 1991; 
Ogawa, M., Blood, 81:2844, 1993; Heimfeld, S., et al., Proc. Natl. Acad. 
Sci. USA, 88:9902, 1991). Some of these receptors transduce the signal at 
the cell surface to the cytoplasm through the activation of a tyrosine 
kinase domain in the cytoplasmic portion of the receptor (e.g., CSF1, 
c-kit, STK-1/FLT3/FLK2-) (Boyle, W. J., Current Opinion in Oncology, 
4:156, 1992, Chiba, T., et al, Nature, 362:646, 1993, Schlessinger, J., et 
al., Neuron, 9:383, 1992, Ullrich, A. and Schlessinger, J., Cell, 61:203, 
1990). Another group of hematopoietic receptors lack intrinsic kinase 
catalytic domains (e.g., IL-3, GM-CSF, G-CSF, and EPO receptors) 
(Miyajima, A., et al., Blood, 82:1960, 1993, Fukunaga, R., et al., EMBO, 
10:2855, 1991, Wojchowski, D. M., et al., Stem Cells, 11:381, 1993), 
however, upon binding of their ligands, these receptors activate protein 
tyrosine phosphorylation of second messengers and the subsequent signal 
pathways to the cell's nucleus (Kishimoto, T. et al, Science, 258:593, 
1992, Stahl, N., et al., Cell, 74:587, 1993). 
Tyrosine kinases often play pivotal roles in the proliferation and 
differentiation of many cell types. Many growth factor receptors contain a 
tyrosine kinase domain as part of their cytoplasmic tail such that binding 
by ligand directly activates their tyrosine kinase activity. However, many 
other receptors do not contain a tyrosine kinase domain in their 
cytoplasmic tail. Addition of ligand to many cell types expressing these 
receptors still results in increased levels of phosphotyrosine. The JAK 
family, a series of related intracellular tyrosine kinases, has recently 
been shown to link these receptors and other members of the signal 
transduction pathway. 
The JAK family members contain the highly conserved catalytic domain found 
in other tyrosine kinases (Firmbach-Kraft, I., et al., Oncogene, 5:1329, 
1990, Hanks, S. K., et al., Methods in Enzymology, 200: 38, 1991, Hunter, 
T., Methods in Enzymology, 200:3, 1991, Wilks, A. F., Proc. Natl. Acad. 
Sci. USA, 86:1603, 1989). One feature that distinguishes the JAK family 
from other tyrosine kinases is that each member also contains a second 
kinase-like domain of unknown function (Harpur, A. G., et al., Oncogene, 
7:1347, 1992). In addition, the JAK family members do not contain SH2 or 
SH3 domains, signal peptide sequences, or transmembrane domains, and are 
localized in the cytoplasm (Wilks, A. F., et al., Molecular and Cellular 
Biology, 11:2057, 1991). 
Three members of the JAK family, JAK1, JAK2, an TYK-2, have been 
functionally described. The first two members were isolated by a PCR 
approach utilizing degenerate oligonucleotide primers and TYK-2 was 
isolated by screening with a tyrosine kinase probe at reduced stringency 
(Silvennoinen, O. et al., Proc. Natl. Acad. Sci. USA, 90:8429,1993). To 
date, the JAK family members have been shown to be involved with the 
receptors for numerous cytokines and growth factors, including IFN 
.alpha..beta. and .gamma., IL-3, GM-CSF, EPO, GH, CNTF, LIF, OSM, IL-6, 
and PRL (Argetsinger, L. S., et al., Cell, 74:237, 1993, Luttichen, C., et 
al., Science, 263:89, 1994, Muller, M., et al., Nature, 366:129, 1993, 
Stahl, N., et al., Science, 263:92, 1994, Velazquez, L., et al., Cell, 
70:313, 1992, Watling, D., et al., Nature, 366:166, 1993, Witthuhn, B. A., 
et al., Cell, 74:227, 1993, Rui, H., et al., The Journal of Biological 
Chemistry, 269:5364, 1994). In most cases, the JAK family members seem to 
associate with the proximal membrane portion of the cytoplasmic domain of 
the receptor (e.g., gp130, LIFR.beta., EPO) as a constitutive complex 
(Narazaki, M., et al., Proc. Natl. Acad. Sci. USA, 91:2285, 1994). In 
other cases, the association is not evident until ligand binding takes 
place (e.g., GH receptor). In either case, ligand binding results in 
increased JAK kinase activity. 
The first evidence for the functional role of JAK family members was 
provided when it was shown that TYK-2 could rescue IFN.alpha./.beta. 
responsiveness in a cell line that had become unresponsive. In a similar 
fashion, JAK1 and JAK2 have been shown to function in the signalling of 
interferon pathways, as well. In each case, two different JAKS have been 
found to act with each type of IFN receptor; JAK2 and TYK-2 are involved 
exclusively with IFN .gamma. and IFN .alpha./.beta., respectively, whereas 
JAK1 is involved with both receptors. Stimulation of the IFN.alpha./.beta. 
receptors by the binding of their respective ligands results in the 
phosphorylation of p91 (STAT1) and p113 (STAT2), which are subunits of the 
ISGF3 transcription complex that binds the interferon-stimulated response 
element (ISRE). In the case of IFN.gamma., p91 alone is phosphorylated, 
which then binds gamma-activated sequences (GAS) of IFN.gamma. activated 
genes (Shual, K., et al., Nature, 366:580, 1993, Ihle, J. N., et al., 
Trends in Biological Science, 19:222, 1994). Because each of these 
receptors associate with JAK1 it has been suggested that JAK1 may directly 
phosphorylate p91 (Loh, J. E., et al., Molecular and Cellular Biology, 
14:2170, 1994). It has been recently reported that IL-6 (via gp130), which 
associates with JAK1 and TYK-2, also triggers the activation of p91 
(STAT1) (Yuan, J., et al., Molecular and Cellular Biology, 14:1657, 1994). 
The EPO, and IL-3 receptors are also believed to similarly activate STAT 
family members. As all of the hematopoietic receptors seem to utilize 
certain common proteins in their signal transduction pathways, some of the 
specificity of the pathways may reside in the cell specific expression of 
STAT family members and their activation by JAK family members (Metcalf, 
D., Blood, 82:3515, 1993, Darnell, J. E., et al., Science, 264:1415, 
1994). 
Additional pairs of JAK family members have been found to associate with 
other receptors (e.g., CNTF, LIF, IL-6) and both become tyrosine 
phosphorylated upon the stimulation of these receptors (Silvennoinen, O., 
et al., Nature, 366:583, 1993). It is possible that reciprocal tyrosine 
phosphorylation between two JAKs is required as phosphorylation of both 
associating JAKs is necessary for signal transduction to occur. Thus, JAK 
family members may act in pairs, possibly as heterodimers. 
Recently a Drosophila JAK family member, hop, was shown to be required 
maternally for normal embryonic development (Binari, et al., Genes & Dev., 
8:300, 1994). Mutants in hop showed defects in the expression of several 
paired-rule and segment polarity genes, implicating it in the control of 
transcription of these genes, a role that could be analogous to the defect 
in TYK-2, JAK1, or JAK2 in several cell lines that lost IFN 
responsiveness. 
The present invention provides a new member of the JAK protein tyrosine 
kinase family. The structural homology between the JAK3 of this invention 
and the other members of JAK family, indicates that JAK3 is a new member 
of this family of non-receptor tyrosine kinases. In analogy to the other 
JAK family members, JAK3 is likely involved in the signal transduction 
pathway of already characterized receptors which lack intrinsic activity. 
Because of its strong expression in the fraction enriched for CD34+ normal 
human bone marrow, JAK3 is likely to be important in stem/progenitor cell 
growth and/or differentiation, by transducing the signals of receptors 
which modulate these processes. In addition, JAK3 may also be involved in 
the signal transduction pathways of any of several non-tyrosine kinase 
receptors with which the other JAK members have not been shown to 
associate (e.g. IL-2, IL-4, IL-11). 
SUMMARY OF THE INVENTION 
The present invention provides a novel protein tyrosine kinase JAK3, a 
polynucleotide sequence which encodes JAK3 and antibodies which are 
immunoreactive with the protein. The amino acid sequence of JAK3 indicates 
that it is a new member of the JAK family of non-receptor tyrosine 
kinases. JAK3 is highly expressed in the CD34+/lin- fraction in normal 
human bone marrow which is highly enriched in hematopoietic 
stem/progenitor cells. Therefore, by analogy to other JAK family member, 
it is likely that JAK3 plays a role in the growth factor modulated 
differentiation/proliferation of the stem/progenitor cells. 
JAK3 is expressed in mammalian tissues, and particularly human tissue. For 
example, JAK3 is expressed in human hematopoietic tissues, (e.g., bone 
marrow), and non-hematopoietic human tissues, such as liver, lung, kidney, 
spleen and intestine. In particular, JAK-3 is most highly expressed in the 
stem/progenitor cell enriched fraction of normal human bone marrow. JAK-3 
is further expressed in a wide range of leukemic derived cell lines 
including AMLs (KG1, TF-1, HEL), B lineage ALLs (PB697, Nalm-16, and 
Nalm-6), and T-ALLs (Molt-16, and Molt-3). 
JAK3 is localized to chromosome 19, band p12-13.1, where the another member 
of the JAK family, TYK-2 is co-localized. Several other genes containing 
tyrosine kinase domains are tandemly linked and may have evolved by cis 
duplications. Examples include the genes for the receptors of c-fms (CSF-1 
receptor) and PDGFR .beta. on chromosome 5 bands q31-q33, c-kit and PDGFR 
.alpha. on chromosome 4 bands q11-q13, as well as FLT1 and STK-1/FLT3/FLK2 
on chromosome 13 band q12. 
In another embodiment, the invention provides a method for ameliorating a 
cell proliferative disorder associated with JAK-3. In another embodiment, 
the invention provides a method for stimulating stem/progenitor cell 
proliferation/survival and differentiation in vitro.

DETAILED DESCRIPTION OF THE INVENTION 
The present invention provides a novel protein tyrosine kinase, JAK3, and a 
polynucleotide sequence encoding JAK3 polypeptide. The amino acid sequence 
of JAK3 indicates that it is a new member of the JAK family of 
non-receptor tyrosine kinases. In normal human bone marrow, JAK3 is highly 
expressed in the CD34+/lin- fraction which is enriched in hematopoietic 
stem/progenitor cells. As JAK kinases have been shown to be involved in 
the signal transduction pathways of various hematopoietic growth factors, 
it is likely that JAK3 plays a role in the growth factor modulated 
differentiation/proliferation/survival of hematopoietic stem/progenitor 
cells. 
In a first embodiment, the invention provides a substantially pure JAK3 
polypeptide consisting essentially of the amino acid sequence of SEQ ID 
NO:2. The full-length JAK3 polypeptide sequence has 1082 amino acids with 
a molecular weight of approximately 121 kD. JAK3 has 48% identity and 67% 
similarity with JAK2 (murine), 41% identity and 61% similarity with JAK1 
(human), and 40% identity and 60% similarity with TYK-2 (human). 
Comparison of human JAK3 with the rat JAK3 shows 77% identity and 84% 
similarity (Takahashi, T. and Shirasawa, T., FEBS Letters, 342:124, 1994). 
The term "substantially pure" or "isolated" as used herein, refers to JAK3 
polypeptide which is substantially free of other proteins, lipids, 
carbohydrates, nucleic acids, or other materials with which it is 
naturally associated. One skilled in the art can purify JAK3 using 
standard techniques for protein purification. The substantially pure 
polypeptide will yield a single major band on a non-reducing 
polyacrylamide gel. The purity of the JAK3 polypeptide can also be 
determined by amino-terminal amino acid sequence analysis. 
The invention includes a functional polypeptide, JAK3, and functional 
fragments thereof. As used herein, the term "functional polypeptide" 
refers to a polypeptide which possesses a biological function or activity 
which is identified through a defined functional assay and 
The invention includes a functional polypeptide, JAK3, and functional 
fragments thereof. As used herein, the term "functional polypeptide" 
refers to a polypeptide whic possesses a biological function or activity 
which is identified through a defined functional assay and which is 
associated with a particular biologic, morphologic, or phenotypic 
alteration in the cell. Functional fragments of the JAK3 polypeptide, 
includes fragments of JAK3 as long as the activity, e.g., protein tyrosine 
kinase activity, of JAK3 remains. Smaller peptides containing the 
biological activity of JAK3 are included in the invention. The biological 
function, for example, can vary from a polypeptide fragment as small as an 
epitope to which an antibody molecule can bind to a large polypeptide 
which is capable of participating in the characteristic induction or 
programming of phenotypic changes within a cell. An enzymatically 
functional JAK3 polypeptide or fragment thereof possesses JAK3 tyrosine 
kinase activity. A "functional polynucleotide" denotes a polynucleotide 
which encodes a functional polypeptide as described herein. 
Minor modifications of the JAK3 primary amino acid sequence may result in 
proteins which have substantially equivalent activity as compared to the 
JAK3 polypeptide described herein. Such modifications may be deliberate, 
as by site-directed mutagenesis, or may be spontaneous. All of the 
polypeptides produced by these modifications are included herein as long 
as the tyrosine kinase activity of JAK3 is present. Further, deletion of 
one or more amino acids can also result in a modification of the structure 
of the resultant molecule without significantly altering its kinase 
activity. This can lead to the development of a smaller active molecule 
which may have broader utility. For example, it is possible to remove 
amino or carboxyl terminal amino acids which may not be required for JAK3 
kinase activity. 
The JAK3 polypeptide of the invention also includes conservative variations 
of the polypeptide sequence. The term "conservative variation" as used 
herein denotes the replacement of an amino acid residue by another, 
biologically similar residue. Examples of conservative variations include 
the substitution of one hydrophobic residue such as isoleucine, valine, 
leucine or methionine for another, or the substitution of one polar 
residue for another, such as the substitution of arginine for lysine, 
glutamic for aspartic acids, or glutamine for asparagine, and the like. 
The term "conservative variation" also includes the use of a substituted 
amino acid in place of an unsubstituted parent amino acid provided that 
antibodies raised to the substituted polypeptide also immunoreact with the 
unsubstituted polypeptide. 
The invention also provides an isolated polynucleotide sequence consisting 
essentially of a polynucleotide sequence encoding a polypeptide having the 
amino acid sequence of SEQ ID NO:2. As used herein, "polynucleotide" 
refers to a polymer of deoxyribonucleotides or ribonucleotides, in the 
form of a separate fragment or a larger construct. The term "isolated" as 
used herein includes polynucleotides substantially free of other nucleic 
acids, proteins, lipids, carbohydrates or other materials with which it is 
naturally associated. Polynucleotide sequences of the invention include 
DNA, cDNA and RNA sequences which encode JAK3. It is understood that all 
polynucleotides encoding all or a portion of JAK3 are also included 
herein, as long as they encode a polypeptide with JAK3 kinase activity. 
Such polynucleotides include naturally occurring, synthetic, and 
intentionally manipulated polynucleotides. For example, JAK3 
polynucleotide may be subjected to site-directed mutagenesis. The 
polynucleotide sequence for JAK3 also includes antisense sequences. The 
polynucleotides of the invention include sequences that are degenerate as 
a result of the genetic code. There are 20 natural amino acids, most of 
which are specified by more than one codon. Therefore, all degenerate 
nucleotide sequences are included in the invention as long as the amino 
acid sequence of JAK3 polypeptide encoded by the nucleotide sequence is 
functionally unchanged. In addition, the invention also includes a 
polynucleotide consisting essentially of a polynucleotide sequence 
encoding a polypeptide having an amino acid sequence of SEQ ID NO:2 and 
having at least one epitope for an antibody immunoreactive with JAK3 
polypeptide. 
Specifically disclosed herein is a cDNA sequence which encodes JAK3 which 
comprising a 3,807 base pair (bp) predicted coding region for JAK3, 167 
base pairs of 5' untranslated and 394 base pairs of 3' untranslated 
sequence (SEQ. ID NO: 1). The cDNA includes an open reading frame of 3,246 
base pairs encoding a protein of about 1082 amino acids, having a 
molecular weight of about 121 kD. The putative initiating methionine shows 
the strongest homology with the Kozak consensus sequence (Kozak, M., 
Nucleic Acids Research, 15:8125, 1987). At the 3' end, an in frame stop 
codon defines the C-terminus of the JAK3 protein at position 3242. 
The polynucleotide encoding JAK3 includes the nucleotide sequence in FIG. 1 
(SEQ ID NO: 1), as well as nucleic acid sequences complementary to that 
sequence. A complementary sequence may include an antisense nucleotide. 
When the sequence is RNA, the deoxynucleotides A, G, C, and T of FIG. 1 
are replaced by ribonucleotides A, G, C, and U, respectively. Also 
included in the invention are fragments of the above-described nucleic 
acid sequences that are at least 15 bases in length, which is sufficient 
to permit the fragment to selectively hybridize to DNA that encodes the 
protein of FIG. 1 (SEQ ID NO: 2) under physiological conditions. 
DNA sequences of the invention can be obtained by several methods. For 
example, the DNA can be isolated using hybridization techniques which are 
well known in the art. These include, but are not limited to: 1) 
hybridization of genomic or cDNA libraries with probes to detect 
homologous nucleotide sequences; 2) antibody screening of expression 
libraries to detect cloned DNA fragments with shared structural features; 
and 3) PCR amplification of a desired nucleotide sequence using 
oligonucleotide primers. 
Preferably the JAK3 polynucleotide of the invention is derived from a 
mammalian organism, and most preferably from human. Screening procedures 
which rely on nucleic acid hybridization make it possible to isolate any 
gene sequence from any organism, provided the appropriate probe is 
available. Oligonucleotide probes, which correspond to a part of the 
sequence encoding the protein in question, can be synthesized chemically. 
This requires that short, oligopeptide stretches of amino acid sequence 
must be known. The DNA sequence encoding the protein can be deduced from 
the genetic code, however, the degeneracy of the code must be taken into 
account. It is possible to perform a mixed addition reaction when the 
sequence is degenerate. This includes a heterogeneous mixture of denatured 
double-stranded DNA. For such screening, hybridization is preferably 
performed on either single-stranded DNA or denatured double-stranded DNA. 
Hybridization is particularly useful in the detection of cDNA clones 
derived from sources where an extremely low amount of mRNA sequences 
relating to the polypeptide of interest are present. In other words, by 
using stringent hybridization conditions directed to avoid non-specific 
binding, it is possible, for example, to allow the autoradiographic 
visualization of a specific cDNA clone by the hybridization of the target 
DNA to that single probe in the mixture which is its complete complement 
(Wallace, et al., Nucl. Acid Res., 9:879, 1981). 
The development of specific DNA sequences encoding JAK3 can also be 
obtained by: 1) isolation of double-stranded DNA sequences from the 
genomic DNA; 2) chemical manufacture of a DNA sequence to provide the 
necessary codons for the polypeptide of interest; and 3) in vitro 
synthesis of a double-stranded DNA sequence by reverse transcription of 
mRNA isolated from a eukaryotic donor cell. In the latter case, a 
double-stranded DNA complement of mRNA is eventually formed which is 
generally referred to as cDNA. 
Of the three above-noted methods for developing specific DNA sequences for 
use in recombinant procedures, the isolation of genomic DNA isolates is 
the least common. This is especially true when it is desirable to obtain 
the microbial expression of mammalian polypeptides due to the presence of 
introns. 
The synthesis of DNA sequences is frequently the method of choice when the 
entire sequence of amino acid residues of the desired polypeptide product 
is known. When the entire sequence of amino acid residues of the desired 
polypeptide is not known, the direct synthesis of DNA sequences is not 
possible and the method of choice is the synthesis of cDNA sequences. 
Among the standard procedures for isolating cDNA sequences of interest is 
the formation of plasmid- or phage-carrying cDNA libraries which are 
derived from reverse transcription of mRNA which is abundant in donor 
cells that have a high level of genetic expression. When used in 
combination with polymerase chain reaction technology, even rare 
expression products can be cloned. In those cases where significant 
portions of the amino acid sequence of the polypeptide are known, the 
production of labeled single or double-stranded DNA or RNA probe sequences 
duplicating a sequence putatively present in the target cDNA may be 
employed in DNA/DNA hybridization procedures which are carried out on 
cloned copies of the cDNA which have been denatured into a single-stranded 
form (Jay, et al., Nucl. Acid Res., 11:2325, 1983). 
A preferred method for obtaining genomic DNA for example is Polymerase 
Chain Reaction (PCR), which relies on an in vitro method of nucleic acid 
synthesis by which a particular segment of DNA is specifically replicated. 
Two oligonucleotide primers that flank the DNA fragment to be amplified 
are utilized in repeated cycles of heat denaturation of the DNA, annealing 
of the primers to their complementary sequences, and extension of the 
annealed primers with DNA polymerase. These primers hybridize to opposite 
strands of the target sequence and are oriented so that DNA synthesis by 
the polymerase proceeds across the region between the primers. Since the 
extension products themselves are also complementary to and capable of 
binding primers, successive cycles of amplification essentially double the 
amount of the target DNA synthesized in the previous cycle. The result is 
an exponential accumulation of the specific target fragment, approximately 
2.sup.n, where n is the number of cycles of amplification performed (see 
PCR Protocols, Eds. Innis, et al., Academic Press, Inc., 1990, 
incorporated herein by reference). 
A cDNA expression library, such as lambda gt11, can be screened indirectly 
for JAK3 peptides having at least one epitope, using antibodies specific 
for JAK3. Such antibodies can be either polyclonally or monoclonally 
derived and used to detect expression product indicative of the presence 
of JAK3 cDNA. 
The polynucleotide sequence for JAK3 also includes sequences complementary 
to the polynucleotide encoding JAK3 (antisense sequences). Antisense 
nucleic acids are DNA or RNA molecules that are complementary to at least 
a portion of a specific mRNA molecule (Weintraub, Scientific American, 
262:40, 1990). The invention embraces all antisense polynucleotides 
capable of inhibiting production of JAK3 polypeptide. In the cell, the 
antisense nucleic acids hybridize to the corresponding mRNA, forming a 
double-stranded molecule. The antisense nucleic acids may interfere with 
the translation of the mRNA since the cell will not translate a mRNA that 
is double-stranded. Antisense oligomers of about 15 nucleotides are 
preferred, since they are easily synthesized and are less likely to cause 
problems than larger molecules when introduced into the target 
JAK3-producing cell. The use of antisense methods to inhibit the 
translation of genes is well known in the art (Marcus-Sakura, Anal. 
Biochem., 172:289, 1988). 
In addition, ribozyme nucleotide sequences for JAK3 are included in the 
invention. Ribozymes are RNA molecules possessing the ability to 
specifically cleave other single-stranded RNA in a manner analogous to DNA 
restriction endonucleases. Through the modification of nucleotide 
sequences which encode these RNAs, it is possible to engineer molecules 
that recognize specific nucleotide sequences in an RNA molecule and cleave 
it (Cech, J. Amer. Med. Assn., 260:3030, 1988). A major advantage of this 
approach is that, because they are sequence-specific, only mRNAs with 
particular sequences are inactivated. 
There are two basic types of ribozymes namely, tetrahymena-type 
(Hasselhoff, Nature, 334:585, 1988) and "hammerhead"-type. 
Tetrahymena-type ribozymes recognize sequences which are four bases in 
length, while "hammerhead"-type ribozymes recognize base sequences 11-18 
bases in length. The longer the recognition sequence, the greater the 
likelihood that sequence will occur exclusively in the target mRNA 
species. Consequently, hammerhead-type ribozymes are preferable to 
tetrahymena-type ribozymes for inactivating a specific mRNA species and 
18-based recognition sequences are preferable to shorter recognition 
sequences. 
DNA sequences encoding JAK3 can be expressed in vitro by DNA transfer into 
a suitable host cell. "Host cells" are cells in which a vector can be 
propagated and its DNA expressed. The term also includes any progeny of 
the subject host cell. It is understood that all progeny may not be 
identical to the parental cell since there may be mutations that occur 
during replication. However, such progeny are included when the term "host 
cell" is used. Methods of stable transfer, meaning that the foreign DNA is 
continuously maintained in the host, are known in the art. 
In the present invention, the JAK3 polynucleotide sequences may be inserted 
into a recombinant expression vector. The term "recombinant expression 
vector" refers to a plasmid, virus or other vehicle known in the art that 
has been manipulated by insertion or incorporation of the JAK3 genetic 
sequences. Such expression vectors contain a promoter sequence which 
facilitates the efficient transcription of the inserted genetic sequence 
of the host. The expression vector typically contains an origin of 
replication, a promoter, as well as specific genes which allow phenotypic 
selection of the transformed cells. Vectors suitable for use in the 
present invention include, but are not limited to the T7-based expression 
vector for expression in bacteria (Rosenberg, et al., Gene, 56:125, 1987), 
the pMSXND expression vector for expression in mammalian cells (Lee and 
Nathans, J. Biol. Chem., 263:3521, 1988) and baculovirus-derived vectors 
for expression in insect cells. The DNA segment can be present in the 
vector operably linked to regulatory elements, for example, a promoter 
(e.g., T7, metallothionein I, or polyhedrin promoters). 
Polynucleotide sequences encoding JAK3 can be expressed in either 
prokaryotes or eukaryotes. Hosts can include microbial, yeast, insect and 
mammalian organisms. Methods of expressing DNA sequences having eukaryotic 
or viral sequences in prokaryotes are well known in the art. Biologically 
functional viral and plasmid DNA vectors capable of expression and 
replication in a host are known in the art. Such vectors are used to 
incorporate DNA sequences of the invention. 
Methods which are well known to those skilled in the art can be used to 
construct expression vectors containing the JAK3 coding sequence and 
appropriate transcriptional/translational control signals. These methods 
include in vitro recombinant DNA techniques, synthetic techniques, and in 
vivo recombination/genetic techniques. See, for example, the techniques 
described in Maniatis, et al., 1989 Molecular Cloning A Laboratory Manual, 
Cold Spring Harbor Laboratory, N.Y. 
A variety of host-expression vector systems may be utilized to express the 
JAK3 coding sequence. These include but are not limited to microorganisms 
such as bacteria transformed with recombinant bacteriophage DNA, plasmid 
DNA or cosmid DNA expression vectors containing the JAK3 coding sequence; 
yeast transformed with recombinant yeast expression vectors containing the 
JAK3 coding sequence; plant cell systems infected with recombinant virus 
expression vectors (e.g., cauliflower mosaic virus, CaMV; tobacco mosaic 
virus, TMV) or transformed with recombinant plasmid expression vectors 
(e.g., Ti plasmid) containing the JAK3 coding sequence; insect cell 
systems infected with recombinant virus expression vectors (e.g., 
baculovirus) containing the JAK3 coding sequence; or animal cell systems 
infected with recombinant virus expression vectors (e.g., retroviruses, 
adenovirus, vaccinia virus) containing the JAK3 coding sequence, or 
transformed animal cell systems engineered for stable expression. Since 
JAK3 has not been confirmed to contain carbohydrates, both bacterial 
expression systems as well as those that provide for translational and 
post-translational modifications may be used; e.g., mammalian, insect, 
yeast or plant expression systems. 
Depending on the host/vector system utilized, any of a number of suitable 
transcription and translation elements, including constitutive and 
inducible promoters, transcription enhancer elements, transcription 
terminators, etc. may be used in the expression vector (see e.g., Bitter, 
et al., 1987, Methods in Enzymology, 153:516-544). For example, when 
cloning in bacterial systems, inducible promoters such as pL of 
bacteriophage .gamma., plac, ptrp, ptac (ptrp-lac hybrid promoter) and the 
like may be used. When cloning in mammalian cell systems, promoters 
derived from the genome of mammalian cells (e.g., metallothionein 
promoter) or from mammalian viruses (e.g., the retrovirus long terminal 
repeat; the adenovirus late promoter; the vaccinia virus 7.5K promoter) 
may be used. Promoters produced by recombinant DNA or synthetic techniques 
may also be used to provide for transcription of the inserted JAK3 coding 
sequence. 
In bacterial systems a number of expression vectors may be advantageously 
selected depending upon the use intended for the expressed. For example, 
when large quantities of JAK3 are to be produced, vectors which direct the 
expression of high levels of fusion protein products that are readily 
purified may be desirable. Those which are engineered to contain a 
cleavage site to aid in recovering are preferred. Such vectors include but 
are not limited to the E. coli expression vector pUR278 (Ruther, et al., 
EMBO J., 2:1791, 1983), in which the JAK3 coding sequence may be ligated 
into the vector in frame with the lac Z coding region so that a hybrid 
-lac Z protein is produced; pIN vectors (Inouye and Inouye, Nucleic Acids 
Res., 13:3101, 1985; Van Heeke and Schuster, J. Biol. Chem. 264:5503, 
1989) and the like. 
In yeast, a number of vectors containing constitutive or inducible 
promoters may be used. For a review see, Current Protocols in Molecular 
Biology, Vol. 2, 1988, Ed. Ausubel, et al., Greene Publish. Assoc. & Wiley 
Interscience, Ch. 13; Grant, et al., 1987, Expression and Secretion 
Vectors for Yeast, in Methods in Enzymology, Eds. Wu and Grossman, 31987, 
Acad. Press, N.Y., Vol. 153, pp. 516-544; Glover, 1986, DNA Cloning, Vol. 
II, IRL Press, Wash., D.C., Ch. 3; and Bitter, 1987, Heterologous Gene 
Expression in Yeast, Methods in Enzymology, Eds. Berger and Kimmel, Acad. 
Press, N.Y., Vol. 152, pp. 673-684; and The Molecular Biology of the Yeast 
Saccharomyces, 1982, Eds. Strathern, et al., Cold Spring Harbor Press, 
Vols. I and II. A constitutive yeast promoter such as ADH or LEU2 or an 
inducible promoter such as GAL may be used (Cloning in Yeast, Ch. 3, R. 
Rothstein In: DNA Cloning Vol.11, A Practical Approach, Ed. D M Glover, 
1986, IRL Press, Wash., D.C.). Alternatively, vectors may be used which 
promote integration of foreign DNA sequences into the yeast chromosome. 
In cases where plant expression vectors are used, the expression of the 
JAK3 coding sequence may be driven by any of a number of promoters. For 
example, viral promoters such as the 35S RNA and 19S RNA promoters of CaMV 
(Brisson, et al., Nature, 310:511, 1984), or the coat protein promoter to 
TMV (Takamatsu, et al., EMBO J., 6:307, 1987) may be used; alternatively, 
plant promoters such as the small subunit of RUBISCO (Coruzzi, et al., 
EMBO J., 3:1671-1680, 1984; Broglie, et al., Science, 224:838, 1984); or 
heat shock promoters, e.g., soybean hsp17.5-E or hsp17.3-B (Gurley, et 
al., Mol. Cell. Biol., 6:559, 1986) may be used. These constructs can be 
introduced into plant cells using Ti plasmids, Ri plasmids, plant virus 
vectors, direct DNA transformation, microinjection, electroporation, etc. 
For reviews of such techniques see, for example, Weissbach and Weissbach, 
1988, Methods for Plant Molecular Biology, Academic Press, NY, Section 
VIII, pp. 421-463; and Grierson and Corey, 1988, Plant Molecular Biology, 
2d Ed., Blackie, London, Ch. 7-9. 
An alternative expression system which could be used to express is an 
insect system. In one such system, Autographa californica nuclear 
polyhedrosis virus (AcNPV) is used as a vector to express foreign genes. 
The virus grows in Spodoptera frugiperda cells. The JAK3 coding sequence 
may be cloned into non-essential regions (for example the polyhedrin gene) 
of the virus and placed under control of an AcNPV promoter (for example 
the polyhedrin promoter). Successful insertion of the JAK3 coding sequence 
will result in inactivation of the polyhedrin gene and production of 
non-occluded recombinant virus (i.e., virus lacking the proteinaceous coat 
coded for by the polyhedrin gene). These recombinant viruses are then used 
to infect Spodoptera frugiperda cells in which the inserted gene is 
expressed. (e.g., see Smith, et al., J. Viol., 46:584, 1983; Smith, U.S. 
Pat. No. 4,215,051). 
Eukaryotic systems, and preferably mammalian expression systems, allow for 
proper post-translational modifications of expressed mammalian proteins to 
occur. Eukaryotic cells which possess the cellular machinery for proper 
processing of the primary transcript, glycosylation, phosphorylation, and 
advantageously, secretion of the gene product may be used as host cells 
for the expression of JAK3. Mammalian cell lines may be preferable. Such 
host cell lines may include but are not limited to CHO, VERO, BHK, HeLa, 
COS, MDCK, -293, and WI38. 
Mammalian cell systems which utilize recombinant viruses or viral elements 
to direct expression may be engineered. For example, when using adenovirus 
expression vectors, the JAK3 coding sequence may be ligated to an 
adenovirus transcription/translation control complex, e.g., the late 
promoter and tripartite leader sequence. This chimeric gene may then be 
inserted in the adenovirus genome by in vitro or in vivo recombination. 
Insertion in a non-essential region of the viral genome (e.g., region E1 
or E3) will result in a recombinant virus that is viable and capable of 
expressing the protein in infected hosts (e.g., see Logan and Shenk, Proc. 
Natl. Acad. Sci. USA, 81:3655, 1984). Alternatively, the vaccinia virus 
7.5K promoter may be used. (e.g., see, Mackett, et al., Proc. Natl. Acad. 
Sci. USA, 79:7415, 1982; Mackett, et al., J. Virol., 49: 857, 1984; 
Panicali, et al., Proc. Natl. Acad. Sci. USA, 79:4927, 1982). Of 
particular interest are vectors based on bovine papilloma virus which have 
the ability to replicate as extrachromosomal elements (Sarver, et al., 
Mol. Cell. Biol, 1:486, 1981). Shortly after entry of this DNA into mouse 
cells, the plasmid replicates to about 100 to 200 copies per cell. 
Transcription of the inserted cDNA does not require integration of the 
plasmid into the host's chromosome, thereby yielding a high level of 
expression. These vectors can be used for stable expression by including a 
selectable marker in the plasmid, such as, for example, the neo gene. 
Alternatively, the retroviral genome can be modified for use as a vector 
capable of introducing and directing the expression of the JAK3 gene in 
host cells (Cone and Mulligan, Proc. Natl. Acad. Sci. USA, 81:6349, 1984). 
High level expression may also be achieved using inducible promoters, 
including, but not limited to, the metallothionine IIA promoter and heat 
shock promoters. 
For long-term, high-yield production of recombinant proteins, stable 
expression is preferred. Rather than using expression vectors which 
contain viral origins of replication, host cells can be transformed with 
the JAK3 cDNA controlled by appropriate expression control elements (e.g., 
promoter, enhancer, sequences, transcription terminators, polyadenylation 
sites, etc.), and a selectable marker. The selectable marker in the 
recombinant plasmid confers resistance to the selection and allows cells 
to stably integrate the plasmid into their chromosomes and grow to form 
foci which in turn can be cloned and expanded into cell lines. For 
example, following the introduction of foreign DNA, engineered cells may 
be allowed to grow for 1-2 days in an enriched media, and then are 
switched to a selective media. A number of selection systems may be used, 
including but not limited to the herpes simplex virus thymidine kinase 
(Wigler, et al., Cell, 11: 223, 1977), hypoxanthine-guanine 
phosphoribosyltransferase (Szybalska and Szybalski, Proc. Natl. Acad. Sci. 
USA, 48:2026, 1962), and adenine phosphoribosyltransferase (Lowy, et al., 
Cell, 22: 817, 1980) genes can be employed in tk.sup.-, hgprt.sup.- or 
aprt.sup.- cells respectively. Also, antimetabolite resistance can be 
used as the basis of selection for dhfr, which confers resistance to 
methotrexate (Wigler, et al., Natl. Acad. Sci. USA, 77: 3567, 1980; 
O'Hare, et al., Proc. Natl. Acad. Sci. USA, 78: 1527, 1981); gpt, which 
confers resistance to mycophenolic acid (Mulligan and Berg, Proc. Natl. 
Acad. Sci. USA, 78: 2072, 1981; neo, which confers resistance to the 
aminoglycoside G-418 (Colberre-Garapin, et al., J. Mol. Biol., 150:1, 
1981); and hygro, which confers resistance to hygromycin (Santerre, et 
al., Gene, 30:147, 1984) genes. Recently, additional selectable genes have 
been described, namely trpB, which allows cells to utilize indole in place 
of tryptophan; hisD, which allows cells to utilize histinol in place of 
histidine (Hartman and Mulligan, Proc. Natl. Acad. Sci. USA, 85:8047, 
1988); and ODC (ornithine decarboxylase) which confers resistance to the 
ornithine decarboxylase inhibitor, 2-(difluoromethyl)-DL-ornithine, DFMO 
(McConlogue L., 1987, In: Current Communications in Molecular Biology, 
Cold Spring Harbor Laboratory ed.). 
Transformation of a host cell with recombinant DNA may be carried out by 
conventional techniques as are well known to those skilled in the art. 
Where the host is prokaryotic, such as E. coli, competent cells which are 
capable of DNA uptake can be prepared from cells harvested after 
exponential growth phase and subsequently treated by the CaCl.sub.2 method 
using procedures well known in the art. Alternatively, MgCl.sub.2 or RbCl 
can be used. Transformation can also be performed after forming a 
protoplast of the host cell if desired. 
When the host is a eukaryote, such methods of transfection of DNA as 
calcium phosphate co-precipitates, conventional mechanical procedures such 
as microinjection, electroporation, insertion of a plasmid encased in 
liposomes, or virus vectors may be used. Eukaryotic cells can also be 
cotransformed with DNA sequences encoding the JAK3 of the invention, and a 
second foreign DNA molecule encoding a selectable phenotype, such as the 
herpes simplex thyridine kinase gene. Another method is to use a 
eukaryotic viral vector, such as simian virus 40 (SV40) or bovine 
papilloma virus, to transiently infect or transform eukaryotic cells and 
express the protein. (see for example, Eukaryotic Viral Vectors, Cold 
Spring Harbor Laboratory, Gluzman ed., 1982). 
Isolation and purification of microbial expressed polypeptide, or fragments 
thereof, provided by the invention, may be carried out by conventional 
means including preparative chromatography and immunological separations 
involving monoclonal or polyclonal antibodies. 
The invention includes antibodies immunoreactive with or which bind to JAK3 
polypeptide or functional fragments thereof. Antibody which consists 
essentially of pooled monoclonal antibodies with different epitopic 
specificities, as well as distinct monoclonal antibody preparations are 
provided. Monoclonal antibodies are made from antigen containing fragments 
of the protein by methods well known to those skilled in the art (Kohler, 
et al., Nature, 256:495, 1975). The term antibody as used in this 
invention is meant to include intact molecules as well as fragments 
thereof, such as Fab and F(ab').sub.2, which are capable of binding an 
epitopic determinant on JAK3. The antibodies of the invention include 
antibodies which bind to the polypeptide of SEQ ID NO:2 and which bind 
with immunoreactive fragments of SEQ ID NO:2. 
The term "antibody" as used in this invention includes intact molecules as 
well as fragments thereof, such as Fab, F(ab').sub.2, and Fv which are 
capable of binding the epitopic determinant. These antibody fragments 
retain some ability to selectively bind with its antigen or receptor and 
are defined as follows: 
(1) Fab, the fragment which contains a monovalent antigen-binding fragment 
of an antibody molecule can be produced by digestion of whole antibody 
with the enzyme papain to yield an intact light chain and a portion of one 
heavy chain; 
(2) Fab', the fragment of an antibody molecule can be obtained by treating 
whole antibody with pepsin, followed by reduction, to yield an intact 
light chain and a portion of the heavy chain; two Fab' fragments are 
obtained per antibody molecule; 
(3) (Fab').sub.2, the fragment of the antibody that can be obtained by 
treating whole antibody with the enzyme pepsin without subsequent 
reduction; F(ab').sub.2 is a dimer of two Fab' fragments held together by 
two disulfide bonds; 
(4) Fv, defined as a genetically engineered fragment containing the 
variable genetically fused single chain molecule. 
Methods of making these fragments are known in the art. (See for example, 
Harlow and Lane, Antibodies: A Laboratory Manual, Cold Spring Harbor 
Laboratory, New York (1988), incorporated herein by reference). 
As used in this invention, the term "epitope" means any antigenic 
determinant on an antigen to which the paratope of an antibody binds. 
Epitopic determinants usually consist of chemically active surface 
groupings of molecules such as amino acids or sugar side chains and 
usually have specific three dimensional structural characteristics, as 
well as specific charge characteristics. 
Antibodies which bind to the JAK3 polypeptide of the invention can be 
prepared using an intact polypeptide or fragments containing small 
peptides of interest as the immunizing antigen. The polypeptide such as 
SEQ ID NO:2 used to immunize an animal can be derived from translated cDNA 
or chemical synthesis which can be conjugated to a carrier protein, if 
desired. Such commonly used carriers which are chemically coupled to the 
peptide include keyhole limpet hemocyanin (KLH), thyroglobulin, bovine 
serum albumin (BSA), and tetanus toxoid. The coupled peptide is then used 
to immunize the animal (e.g., a mouse, a rat, or a rabbit). 
If desired, polyclonal or monoclonal antibodies can be further purified, 
for example, by binding to and elution from a matrix to which the 
polypeptide or a peptide to which the antibodies were raised is bound. 
Those of skill in the art will know of various techniques common in the 
immunology arts for purification and/or concentration of polyclonal 
antibodies, as well as monoclonal antibodies (See for example, Coligan, et 
al., Unit 9, Current Protocols in Immunology, Wiley Interscience, 1991, 
incorporated by reference). 
The invention also provides a method for detecting a cell proliferative 
disorder associated with JAK3 in a subject, comprising contacting a target 
cellular component containing JAK3, with a reagent which detects JAK3. The 
target cell component can be nucleic acid, such as DNA or RNA, or it can 
be protein. When the component is nucleic acid, the reagent is a nucleic 
acid probe or PCR primer. When the cell component is protein, the reagent 
is an antibody probe. The probes can be detectably labeled, for example, 
with a radioisotope, a fluorescent compound, a bioluminescent compound, a 
chemiluminescent compound, a metal chelator, or an enzyme. Those of 
ordinary skill in the art will know of other suitable labels for binding 
to the antibody, or will be able to ascertain such, using routine 
experimentation. 
For purposes of the invention, an antibody or nucleic acid probe specific 
for JAK3 may be used to detect the presence of JAK3 polypeptide (using 
antibody) or polynucleotide (using nucleic acid probe) in biological 
fluids or tissues. Oligonucleotide primers based on any coding sequence 
region in the JAK3 sequence are useful for amplifying DNA, for example by 
PCR. Any specimen containing a detectable amount of polynucleotide or 
antigen can be used. A preferred sample of this invention is blood or a 
tissue of liver, lung, kidney, spleen and intestine. Preferably the 
subject is human. When the cell proliferative disorder associated with 
JAK3 is a hematopoietic cell disorder, it may include leukemia, 
myelodysplasia, polyethemia vera, thrombocytosis and aplastic anemia, for 
example. 
Monoclonal antibodies used in the method of the invention are suited for 
use, for example, in immunoassays in which they can be utilized in liquid 
phase or bound to a solid phase carrier. In addition, the monoclonal 
antibodies in these immunoassays can be detectably labeled in various 
ways. Examples of types of immunoassays which can utilize monoclonal 
antibodies of the invention are competitive and non-competitive 
immunoassays in either a direct or indirect format. Examples of such 
immunoassays are the radioimmunoassay (RIA) and the sandwich 
(immunometric) assay. Detection of the antigens using the monoclonal 
antibodies of the invention can be done utilizing immunoassays which are 
run in either the forward, reverse, or simultaneous modes, including 
immunohistochemical assays on physiological samples. Those of skill in the 
art will know, or can readily discern, other immunoassays formats without 
undue experimentation. 
The term "immunometric assay" or "sandwich immunoassay", includes 
simultaneous sandwich, forward sandwich and reverse sandwich immunoassays. 
These terms are well understood by those skilled in the art. Those of 
skill will also appreciate that antibodies according to the present 
invention will be useful in other variations and forms of assays which are 
presently known or which may be developed in the future. These are 
intended to be included within the scope of the present invention. 
Monoclonal antibodies can be bound to many different carriers and used to 
detect the presence of JAK3 polypeptide. Examples of well-known carriers 
include glass, polystyrene, polypropylene, polyethylene, dextran, nylon, 
amylases, natural and modified celluloses, polyacrylamides, agaroses and 
magnetite. The nature of the carrier can be either soluble or insoluble 
for purposes of the invention. Those skilled in the art will know of other 
suitable carriers for binding monoclonal antibodies, or will be able to 
ascertain such using routine experimentation. 
In performing the assays it may be desirable to include certain "blockers" 
in the incubation medium (usually added with the labeled soluble 
antibody). The "blockers" are added to assure that non-specific proteins, 
proteases, or anti-heterophilic immunoglobulins to anti-JAK3 
immunoglobulins present in the experimental sample do not crosslink or 
destroy the antibodies on the solid phase support, or the radiolabeled 
indicator antibody, to yield false positive or false negative results. The 
selection of "blockers" therefore may add substantially to the specificity 
of the assays described in the present invention. 
It has been found that a number of nonrelevant (i.e., nonspecific) 
antibodies of the same class or subclass (isotype) as those used in the 
assays (e.g., IgG1, IgG2a, IgM, etc.) can be used as "blockers". The 
concentration of the "blockers" (normally 1-100 .mu.g/.mu.l) may be 
important, in order to maintain the proper sensitivity yet inhibit any 
unwanted interference by mutually occurring cross reactive proteins in the 
specimen. 
In using a monoclonal antibody for the in vivo detection of antigen, the 
detectably labeled monoclonal antibody is given in a dose which is 
diagnostically effective. The term "diagnostically effective" means that 
the amount of detectably labeled monoclonal antibody is administered in 
sufficient quantity to enable detection of the site having the JAK3 
antigen for which the monoclonal antibodies are specific. 
The concentration of detectably labeled monoclonal antibody which is 
administered should be sufficient such that the binding to those cells 
having JAK3 is detectable compared to the background. Further, it is 
desirable that the detectably labeled monoclonal antibody be rapidly 
cleared from the circulatory system in order to give the best 
target-to-background signal ratio. 
As a rule, the dosage of detectably labeled monoclonal antibody for in vivo 
diagnosis will vary depending on such factors as age, sex, and extent of 
disease of the individual. The dosage of monoclonal antibody can vary from 
about 0.001 mg/m.sup.2 to about 500 mg/m.sup.2, preferably 0.1 mg/m.sup.2 
to about 200 mg/m.sup.2, most preferably about 0.1 mg/m.sup.2 to about 10 
mg/m.sup.2. Such dosages may vary, for example, depending on whether 
multiple injections are given, tumor burden, and other factors known to 
those of skill in the art. 
For in vivo diagnostic imaging, the type of detection instrument available 
is a major factor in selecting a given radioisotope. The radioisotope 
chosen must have a type of decay which is detectable for a given type of 
instrument. Still another important factor in selecting a radioisotope for 
in vivo diagnosis is that the half-life of the radioisotope be long enough 
so that it is still detectable at the time of maximum uptake by the 
target, but short enough so that deleterious radiation with respect to the 
host is minimized. Ideally, a radioisotope used for in vivo imaging will 
lack a particle emission, but produce a large number of photons in the 
140-250 keV range, which may be readily detected by conventional gamma 
cameras. 
For in vivo diagnosis, radioisotopes may be bound to immunoglobulin either 
directly or indirectly by using an intermediate functional group. 
Intermediate functional groups which often are used to bind radioisotopes 
which exist as metallic ions to immunoglobulins are the bifunctional 
chelating agents such as diethylenetriaminepentacetic acid (DTPA) and 
ethylenediaminetetraacetic acid (EDTA) and similar molecules. Typical 
examples of metallic ions which can be bound to the monoclonal antibodies 
of the invention are .sup.111 In, .sup.97 Ru, .sup.67 Ga, .sup.68 Ga, 
.sup.72 As, .sup.89 Zr, and .sup.201 Tl. 
A monoclonal antibody useful in the method of the invention can also be 
labeled with a paramagnetic isotope for purposes of in vivo diagnosis, as 
in magnetic resonance imaging (MRI) or electron spin resonance (ESR). In 
general, any conventional method for visualizing diagnostic imaging can be 
utilized. Usually gamma and positron emitting radioisotopes are used for 
camera imaging and paramagnetic isotopes for MRI. Elements which are 
particularly useful in such techniques include .sup.157 Gd, .sup.55 Mn, 
.sup.162 Dy, .sup.52 Cr, and .sup.56 Fe. 
The present invention also provides a method for treating a subject with a 
cell proliferative disorder associated with JAK3 comprising administering 
to a subject with the disorder a therapeutically effective amount of 
reagent which modulates JAK3. In hematopoietic cancers, for example, the 
JAK3 nucleotide sequence may be under-expressed as compared to expression 
in a normal cell, therefore, it is possible to design appropriate 
therapeutic or diagnostic techniques directed to this sequence. Thus, 
where a cell-proliferative disorder is associated with the expression of 
JAK3 associated with malignancy, nucleic acid sequences that modulate JAK3 
expression at the transcriptional or translational level can be used. In 
cases when a cell proliferative disorder or abnormal cell phenotype is 
associated with the under expression of JAK3, for example, nucleic acid 
sequences encoding JAK3 (sense) could be administered to the subject with 
the disorder. 
The term "cell-proliferative disorder" denotes malignant as well as 
non-malignant cell populations which often appear to differ from the 
surrounding tissue both morphologically and genotypically. Such disorders 
may be associated, for example, with absence of expression of JAK3. 
Essentially, any disorder which is etiologically linked to expression of 
JAK3 could be considered susceptible to treatment with a reagent of the 
invention which modulates JAK3 expression. 
The term "modulate" envisions the suppression of JAK3 gene expression when 
JAK3 is over-expressed. When JAK3 is over-expressed, an antisense 
polynucleotide for JAK3 can be introduced into the cell. Alternatively, 
when a cell proliferative disorder is associated with under-expression of 
JAK3 polypeptide, a sense polynucleotide sequence (the DNA coding strand) 
encoding JAK3 polypeptide can be introduced into the cell. The term 
"therapeutically effective" amount refers to that amount of reagent 
includes that amount which modulates JAK3 expression o kinase activity 
such that the symptoms of the disorder are reduced. 
The present invention also provides gene therapy for the treatment of cell 
proliferative disorders which are mediated by JAK3. Such therapy would 
achieve its therapeutic effect by introduction of the appropriate JAK3 
polynucleotide which contains a JAK3 gene (sense), into cells of subjects 
having the proliferative disorder. Delivery of sense JAK3 polynucleotide 
constructs can be achieved using a recombinant expression vector such as a 
chimeric virus or a colloidal dispersion system. An expression vector 
including the JAK3 polynucleotide sequence could be introduced to the 
subject's cells ex vivo after removing, for example, stem cells from a 
subject's bone marrow. The cells are then reintroduced into the subject, 
(e.g., into subject's bone marrow). 
Various viral vectors which can be utilized for gene therapy as taught 
herein include adenovirus, herpes virus, vaccinia, or, preferably, an RNA 
virus such as a retrovirus. Preferably, the retroviral vector is a 
derivative of a murine or avian retrovirus. Examples of retroviral vectors 
in which a single foreign gene can be inserted include, but are not 
limited to: Moloney murine leukemia virus (MoMuLV), Harvey murine sarcoma 
virus (HaMuSV), murine mammary tumor virus (MuMTV), and Rous Sarcoma Virus 
(RSV), and gibbon ape leukemia virus (GaLV), which provides a broader host 
range than many of the murine viruses. A number of additional retroviral 
vectors can incorporate multiple genes. All of these vectors can transfer 
or incorporate a gene for a selectable marker so that transduced cells can 
be identified and generated. By inserting a JAK3 sequence (including 
promoter region) of interest into the viral vector, along with another 
gene which encodes the ligand for a receptor on a specific target cell, 
for example, the vector is now target specific. Preferred targeting is 
accomplished by using an antibody to target the retroviral vector. Those 
of skill in the art will know of, or can readily ascertain without undue 
experimentation, specific polynucleotide sequences which can be inserted 
into the retroviral genome to allow target specific delivery of the 
retroviral vector containing the JAK3 sense or antisense polynucleotide. 
Since recombinant retroviruses are defective, they require assistance in 
order to produce infectious vector particles. This assistance can be 
provided, for example, by using helper cell lines that contain plasmids 
encoding all of the structural genes of the retrovirus under the control 
of regulatory sequences within the LTR. These plasmids are missing a 
nucleotide sequence which enables the packaging mechanism to recognize an 
RNA transcript for encapsidation. Helper cell lines which have deletions 
of the packaging signal include but are not limited to .PSI.2, 17 and 
2, for example. These cell lines produce empty virions, since no genome 
is packaged. If a retroviral vector is introduced into such cells in which 
the packaging signal is intact, but the structural genes are replaced by 
other genes of interest, the vector can be packaged and vector virion 
produced. 
Another targeted delivery system for JAK3 polynucleotide is a colloidal 
dispersion system. Colloidal dispersion systems include macromolecule 
complexes, nanocapsules, microspheres, beads, and lipid-based systems 
including oil-in-water emulsions, micelles, mixed micelles, and liposomes. 
The preferred colloidal system of this invention is a liposome. Liposomes 
are artificial membrane vesicles which are useful as delivery vehicles in 
vitro and in vivo. It has been shown that large unilamellar vesicles 
(LUV), which range in size from 0.2-4.0 um can encapsulate a substantial 
percentage of an aqueous buffer containing large macromolecules. RNA, DNA 
and intact virions can be encapsulated within the aqueous interior and be 
delivered to cells in a biologically active form (Fraley, et al., Trends 
Biochem. Sci., 6:77, 1981). In addition to mammalian cells, liposomes have 
been used for delivery of polynucleotides in plant, yeast and bacterial 
cells. In order for a liposome to be an efficient gene transfer vehicle, 
the following characteristics should be present: (1) encapsulation of the 
genes of interest at high efficiency while not compromising their 
biological activity; (2) preferential and substantial binding to a target 
cell in comparison to non-target cells; (3) delivery of the aqueous 
contents of the vesicle to the target cell cytoplasm at high efficiency; 
and (4) accurate and effective expression of genetic information (Mannino, 
et al., Biotechniques, 6:682, 1988). 
The composition of the liposome is usually a combination of phospholipids, 
particularly high-phase-transition-temperature phospholipids, usually in 
combination with steroids, especially cholesterol. Other phospholipids or 
other lipids may also be used. The physical characteristics of liposomes 
depend on pH, ionic strength, and the presence of divalent cations. 
Examples of lipids useful in liposome production include phosphatidyl 
compounds, such as phosphatidylglycerol, phosphatidylcholine, 
phosphatidylserine, phosphatidylethanolamine, sphingolipids, cerebrosides, 
and gangliosides. Particularly useful are diacylphosphatidylglycerols, 
where the lipid moiety contains from 14-18 carbon atoms, particularly from 
16-18 carbon atoms, and is saturated. Illustrative phospholipids include 
egg phosphatidylcholine, dipalmitoylphosphatidylcholine and 
distearoylphosphatidylcholine. 
The targeting of liposomes has been classified based on anatomical and 
mechanistic factors. Anatomical classification is based on the level of 
selectivity, for example, organ-specific, cell-specific, and 
organelle-specific. Mechanistic targeting can be distinguished based upon 
whether it is passive or active. Passive targeting utilizes the natural 
tendency of liposomes to distribute to cells of the reticulo-endothelial 
system (RES) in organs which contain sinusoidal capillaries. Active 
targeting, on the other hand, involves alteration of the liposome by 
coupling the liposome to a specific ligand such as a monoclonal antibody, 
sugar, glycolipid, or protein, or by changing the composition or size of 
the liposome in order to achieve targeting to organs and cell types other 
than the naturally occurring sites of localization. 
The surface of the targeted delivery system may be modified in a variety of 
ways. In the case of a liposomal targeted delivery system, lipid groups 
can be incorporated into the lipid bilayer of the liposome in order to 
maintain the targeting ligand in stable association with the liposomal 
bilayer. Various linking groups can be used for joining the lipid chains 
to the targeting ligand. 
In general, the compounds bound to the surface of the targeted delivery 
system will be ligands and receptors which will allow the targeted 
delivery system to find and "home in" on the desired cells. A ligand may 
be any compound of interest which will bind to another compound, such as a 
receptor. 
In general, surface membrane proteins which bind to specific effector 
molecules are referred to as receptors. In the present invention, 
antibodies are preferred receptors. Antibodies can be used to target 
liposomes to specific cell-surface ligands. For example, certain antigens 
expressed specifically on tumor cells, referred to as tumor-associated 
antigens (TAAs), may be exploited for the purpose of targeting JAK3 
antibody-containing liposomes directly to the malignant tumor. Since the 
JAK3 gene product may be indiscriminate with respect to cell type in its 
action, a targeted delivery system offers a significant improvement over 
randomly injecting non-specific liposomes. Preferably, the target tissue 
is human brain, colon, lung, and renal cancers. A number of procedures can 
be used to covalently attach either polyclonal or monoclonal antibodies to 
a liposome bilayer. Antibody-targeted liposomes can include monoclonal or 
polyclonal antibodies or fragments thereof such as Fab, or F(ab').sub.2, 
as long as they bind efficiently to an antigenic epitope on the target 
cells. Liposomes may also be targeted to cells expressing receptors for 
hormones or other serum factors. 
For use in the diagnostic research and therapeutic applications suggested 
above, kits are also provided by the invention. The invention provides a 
diagnostic kit useful for the detection of a target cellular component 
indicative of a cell proliferative disorder associated with JAK3 
comprising carrier means being compartmentalized to receive in close 
confinement therein one or more containers comprising a first container 
containing a probe for detection of JAK3 nucleic acid. Such a kit may 
comprise a carrier means being compartmentalized to receive in close 
confinement one or more container means such as vials, tubes, and the 
like, each of the container means comprising one of the separate elements 
to be used in the method. 
For example, one of the container means may comprise a probe which is or 
can be detectably labelled. Such probe may be an antibody or nucleotide 
specific for a target protein or a target nucleic acid, respectively, 
wherein the target is indicative, or correlates with, the presence of JAK3 
of the invention. Where the kit utilizes nucleic acid hybridization to 
detect the target nucleic acid, the kit may also have containers 
containing nucleotide(s) for amplification of the target nucleic acid 
sequence and/or a container comprising a reporter-means, such as a 
biotin-binding protein, such as avidin or streptavidin, bound to a 
reporter molecule, such as an enzymatic, florescent, or radionucleotide 
label. 
A number of embodiments of the present invention have been described. 
Nevertheless, it will be understood that various modifications may be made 
without departing from the spirit and scope of the invention. Accordingly, 
it is to be understood that the invention is not to be limited by the 
specific illustrated embodiment, but only by the scope of the appended 
claims. 
EXAMPLE 1 
Cloning and Sequencing of JAK3 
In order to clone the cDNA for a new member of the JAK family of 
non-receptor protein tyrosine kinases, degenerate oligonucleotides 
corresponding to parts of the highly conserved tyrosine kinase domain were 
used to amplify first strand cDNA from oligo (dT) primed, reverse 
transcribed, CD34+ total RNA from normal human bone marrow. 
1. Bone Marrow Fractions. Iliac crest bone marrow was aspirated from 
consenting adult volunteers under an IRB approved protocol. Mononuclear 
cells were separated by Ficoll-Hypaque (Pharmacia, Piscataway, N.J.) 
density gradient separation. Cell subsets were purified by immunomagnetic 
separation (Strauss, L. C., et al., The American Journal of Pediatric 
Hematology/Oncology, 13:217, 1991, Civin, C. I., et al, Bone Marrow 
Purging and Processing, 1990). Positive selection of the CD34+ fraction 
was done by incubation of the mononuclear fraction with 0.5 ug CD34 
(HPCA-1, Becton Dickinson, San Jose, Calif.) antibody per 10.sup.6 cells 
for 30 minutes at 4.degree. C. Cells were washed twice in RPMI 1640 
(Sigma, St. Louis, Mo.) then resuspended in RPMI-1640 containing 1% human 
serum albumin at 5.times.10.sup.7 cells/ml and incubated with sheep 
anti-mouse IgG1 conjugated immunomagnetic microspheres for 30 minutes at 
4.degree. C. The CD34+ bound cells were released from the microspheres by 
treatment with chymopapain (Chymodiactin TM, Boots USA, Lincolnshire Ill.; 
final concentration 200 U/ml, 15 min., RT). The microspheres were removed 
from the free (CD34+ enriched) cells using a magnetic particle 
concentrator (Dynal, Great Neck, N.Y.). CD34+ cells were further purified 
to obtain CD34+/Lin- cells by negative selection as described by Gore 
(Gore, S. D. et al., Blood, 8:1681, 1991). 
2. Isolation of RNA. Poly A+ RNA was isolated from human hematopoietic cell 
lines using the Mini Ribosep mRNA isolation kit (Becton Dickinson, San 
Jose, Calif.). Total RNA from bone marrow cells and hematopoietic cell 
lines was extracted using the guanidium thiocyanate method (Chomczynski, 
P. and Sacchi, N., Anal Biochem., 162:156, 1987). 
3. Cloning of JAK3. Total RNA isolated from CD34+ cells (see above) was 
reverse transcribed with Superscript Moloney murine-leukemia-virus reverse 
transcriptase (BRL, Gaithersburg, Md.) using oligo d(T) (Boehringer 
Mannheim, Germany) for printing. PCR amplification was carried out using 
degenerate oligonucleotides based on the highly conserved sequence motifs 
VHRDLA (5' GTNCA(T,C)(T,C)(C,A) GNGA(T,C)(TN GC3') AND DVWSYG (5' 
CCC(G,A)TAN(G,C)(A,T) CCA NAC (G,A)TC3') from the PTK catalytic domain 
(Wilks, A. F., et al., Gene, 85:67, 1989, Wilks, A. F., Methods in 
Enzymology, 200: 533, 1991). To facilitate subcloning of the amplified PCR 
products Not 1 and Sal 1 sites were included as part of the PCR primers. 
The resultant 226 bp bands were isolated after electrophoresis in agarose 
gels and cloned into the Not 1/Sal 1 sites of pBluescript II KS- 
(Stratagene, La Jolla, Calif.). After sequencing, products containing 
known tyrosine kinase motifs were compared to reported sequences using the 
NCBI BlastN program (Altschul, S. F., et al., Journal of Molecular 
Biology, 215:403, 1990). The fragment did not match any other sequences in 
the databases but was most closely related to members of the JAK family of 
tyrosine kinases at 65-70% nucleic acid identity. 
The conditions for RT-PCR and thermal RACE were carried out as described by 
Frohman (Frohman, M. A., Methods in Enzymology, 218:340, 1993). KG1a poly 
A+RNA was used as the substrate for RACE. The 5' and 3' ends of JAK3 were 
also amplified from normal human bone marrow cDNA isolated from a 
.lambda.gt10 human bone marrow library (Clonetech, Palo Alto, Calif.) 
using primers specific for JAK3 with primers specific for the arms of 
.lambda.gt10 under the same PCR conditions used for RACE. 
4. Sequencing of JAK3. To correct for PCR errors, multiple overlapping 
partial clones of JAK3 isolated from KG1a and bone marrow cells were 
sequenced and compared using the dideoxy DNA sequencing method (USB, 
Cleveland, Ohio) (Sanger, F., et al., Proc. Natl. Acad. Sci. USA, 74:5463, 
1977). To verify some regions, RT PCR amplified fragments from normal 
human bone marrow and exon containing portions of normal human JAK3 P1 
genomic clones (see below) were also sequenced. 
5. RNAse Protection Assays. Efforts to obtain the full-length JAK3 clone by 
screening of several libraries proved unsuccessful. Therefore, RNAse 
protection assay were developed utilizing the initial PCR amplified kinase 
domain fragment to screen for leukemic derived cell lines expressing JAK3. 
RNAse protection assays were carried out using the MAXIscript T3 in vitro 
Transcription Kit (Ambion, Austin, Tex.). Briefly, an anti-sense RNA probe 
was synthesized by runoff transcription using Bacteriophage T3 RNA 
polymerase on a pBluescript II KS- (Stratagene, La Jolla, Calif.) template 
linearized downstream of the JAK3 207 nucleotide PTK domain fragment. The 
resulting .sup.32 P.alpha.-UTP labelled 249 base RNA probe was hybridized 
with approximately 5 .mu.g of total RNA from hematopoietic cell lines and 
RNA from approximately equal numbers of cells from normal human bone 
marrow sub-fractions. RNA-RNA hybrids were treated with RNAse A and T, 
denatured and separated on an 8 M-Urea, 6% acrylamide gel and exposed to 
film (Kodak X-OMAT) (Melton, D. A., et al., Nucleic Acids Research, 
12:7035, 1984). As an internal standard, a .beta.-actin probe was also 
included with each hybridization reaction. 
6. JAK3 nucleotide and predicted amino acid sequence: Of the cell lines 
tested in this initial screening, JAK3 was most highly expressed by the 
myeloblastic cell line KG1a. Thermal RACE and PCR was employed to cone the 
full-length cDNA of JAK3 from KG1a and normal human bone marrow cells 
(Frohman, M. A., Therman RACE, Methods in Enzymology, 218:340, 1993). 
Using several rounds of RACE we isolated 3,807 bp of JAK3 cDNA, a region 
which covers the entire predicted coding region for JAK3, 167 bases of 5' 
untranslated and 394 bases of 3' untranslated sequence. FIG. 1 shows the 
nucleotide and predicted amino acid sequence of JAK3. The cDNA includes an 
open reading frame of 3,246 bases that predicts a protein of 1082 amino 
acids with a molecular weight of 121 kD. The putative initiating 
methionine shows the strongest homology with the Kozak consensus sequence 
(Kozak, M., Nucleic Acids Research, 15:8125,1987). At the 3' end an in 
frame stop codon defines the C terminus of the JAK3 protein at position 
3242. 
EXAMPLE 2 
Sequence Comparison Between JAK3 and Other JAK Family Member 
1. Amino acid comparison between JAK3 and other JAK family members: The 
initial identification of JAK3 as the fourth member of the JAK family was 
based on a database search using the 207 bp PCR fragment. The comparison 
of full-length-JAK3 with the other JAK family members is shown in FIG. 2. 
Sequences of JAK family members were aligned using the Pileup program (GCG 
Company, Madison, Wis.). The numbering system begins with the initiating 
methionine of the JAK family members. The numbering system does not take 
into account the insertion of gaps and, therefore, should be only regarded 
as a relative measure of location. The fifth line in the figure shows a 
consensus sequence derived if three out of four JAK family members have 
the identical amino acid in that position. Full-length JAK3 has 48% 
identity and 67% similarity with JAK2 (murine), 41% identity and 61% 
similarity with JAK1, and 40% identity and 60% similarity with TYK-2. In 
addition, recently, small fragments of tyrosine kinases by PCR approaches 
from a human breast cancer cell line (TK5) and rat brain (Ptk-2) have been 
isolated (Cance, W. G., et al., Int. J. Cancer, 54:571, 1993, Sanchez, M. 
P., et al., Proc. Natl. Acad. Sci. USA, 91:1819, 1994). Both of these TK's 
show 93% identity with JAK3 in this short region, while rat Jak3 shares 
99% identity in this region. How JAK3 relates to these PTKs must await the 
isolation of their full coding regions. 
2. Amino acid comparison between JAK3 and rat JAK3: FIG. 3 shows the 
comparison of JAK3 with the recently reported rat Jak3. The sequences of 
human JAK3 and rat Jak3 were aligned using the Pileup program (GCG 
Company, Madison, Wis.). The amino acids of each member are numbered 
beginning with the initiating methionine. The comparison shows 77% 
identity and 84% similarity making it likely that these genes are 
homologies (Takahashi, T. and Shirasawa, T., FEBS Letters, 342:124, 1994). 
EXAMPLE 3 
Characterization of JAK3 Expression 
1. RNAse protection analysis of JAK3 expression in leukemic derived cell 
lines: To investigate the hematopoietic expression of JAK3, the RNAse 
protection assay was used utilizing the 206 bp PCR kinase domain fragment 
of JAK3 to screen leukemic derived cell lines (see above). Briefly, a 32P 
.alpha.-UTP labelled anti-sense RNA probe to the kinase domain of JAK3 was 
hybridized with 5 ugs total RNA from hematopoietic cell lines. A 
.beta.-actin probe was also included with each reaction as an internal 
standard, with the exception of the bone marrow and ML-1 populations, 
which were separately assayed for JAK3 and actin. As shown in FIG. 4a, a 
protected band migrating at the expected size is seen in a number of 
lanes. Positive signals were discernable for the Molt-16, Molt-3, KG1, 
KG1a, PB697, Nalm-16, Nalm-6, and TF-1 cell lines. These positive cell 
lines represent various forms of leukemia; the Molt lines were derived 
from T-ALL, the KG1 lines from AML, PB697 and the Nalm lines are B lineage 
ALL, and TF-1 was established from an erythroleukemia. No signals were 
seen from RNA derived from the ML-1, HL60, K562, or Daudi cell lines 
representing additional AML, APL, CML, and Burkitt's leukemia lines, 
respectively. 
2. Northern blot analysis of JAK3 expression in leukemic derived cell 
lines: Although the coding region of the cDNAs for the JAK family are 
.about.3400 bp, the 5' and 3' untranslated regions and polyadenylation 
result in transcripts ranging from 4.4 kbp for JAK1, 4.8 kbp for JAK2, 5.4 
kbp for TYK-2 and 4.0 kbp for rat Jak3. To investigate the size of JAK3 
message, a Northern blot with poly A+ RNA isolated from a number of 
hematopoietic cell lines was probed with a 1.8 kbp JAK3 fragment. 5 .mu.g 
of poly A+ RNA samples from hematopoietic cell lines were incubated at 
55.degree. C. for 15 minutes with 50% formamide, 6.5% formaldehyde, and 
1.times. MOPS. Following the addition of formaldehyde loading buffer and 
ethidium bromide, RNA samples were electrophoresed in a 1.2% agarose gel 
containing 1.times. MOPS and 11% formaldehyde. 
Following electrophoresis, gels were transferred by capillary action to 
nitrocellulose 47. Sambrook J., Fritsch E. F., Maniatis T.: Molecular 
Cloning. A Laboratory Manual. 1989). As is evident from FIG. 4b (upper 
half), JAK3 is not a very highly expressed message. Even after an exposure 
of 17 days at -80.degree. C. with two intensifying screens, signals were 
barely visible in the lanes containing RNA from the HEL, REH, KG1, and 
KG1a cell lines (HEL represents an erythroleukemia, REH is derived from a 
B-ALL, and the KG1 and KG1a cell lines are myeloblastic). RNA markers give 
an estimate of 5.8 kbp for the JAK3 transcript in these cells. 
3. Northern blot analysis of JAK3 expression in non-hematopoietic tissues: 
To assess the expression of JAK3 in non-hematopoietic normal adult 
tissues, a Northern blot containing 2 .mu.g of poly A+ RNA from human 
heart, brain, placenta, lung, liver, skeletal muscle, kidney, and pancreas 
was screened. (Clonetech, Palo Alto, Calif.). Northern blots were 
prehybridized for 2 hrs in 50% formamide, 5.times. SSPE, 10.times. 
Denhardt's, 2% SDS, and 100 ug/ml denatured salmon sperm DNA (Clonetech). 
Blots were hybridized with a randomly primed .sup.32 P-dCTP labeled probe 
corresponding to a 1.8 kbp fragment of JAK3 cDNA (Feinberg, A. P. and 
Vogelstein, B., Anal. Biochem., 132:6, 1983). The blots were exposed to 
film (Kodak x-OMAT) for 1'/days at -80.degree. C. between two intensifying 
screens. 
When the same JAK3 fragment was also used to probe a Northern blot 
containing RNA from non-hematopoietic human tissues (FIG. 4c), signals are 
seen from placenta, lung, liver, kidney, and pancreas, all with a similar 
message size of 5.8 kbp with possibly an additional less distinct band at 
-7.5 kbp. Unlike rat JAK3, which is expressed in rat heart and brain, no 
signals were seen from the RNA representing heart, brain, or skeletal 
muscle. 
4. RNAse Protection of JAK3 expression in normal bone marrow fractions: 
Although the initial JAK3 fragment was generated by PCR amplification of 
CD34+ enriched bone marrow RNA, it remained a possibility that JAK3 
expression was restricted to contaminating CD34- cells. To determine which 
populations of normal bone marrow express JAK3, fractions representing 
whole BM, CD34+, CD34- (i.e. depleted of CD34+ cells), CD34+/lin-, as well 
as peripheral blood were isolated. RNA was then extracted and used to 
perform the RNAse protection assay. The same probe used in FIG. 4a was 
hybridized with approximately 1-5 .mu.gs of RNA from normal total bone 
marrow, bone marrow subfractions, and from peripheral blood. As an 
internal standard, a .beta.-actin probe was also included with each 
reaction as a standard for the amount and quality of RNA loaded in each 
sample. The presence of a band that migrates between the JAK3 and actin 
bands in all lanes, including the no RNA and tRNA control lanes, is a 
result of incomplete digestion of the probe. All of the sample lanes give 
a protected JAK3 band migrating at the expected size. However, probably 
because of the limited amounts of RNA obtained from several fractions, the 
actin bands indicate a variation in total RNA loaded for each sample. 
To determine the relative expression of JAK3 in these different 
populations, the bands were quantified by phosphorimager scanning and 
normalized relative to the actin signal. FIG. 5b shows the phosphorimage 
analysis of bone marrow fractions. Following exposure to film, the gel 
shown in FIG. 5b was exposed to a phosphor-image screen (Molecular 
Dynamics, Sunnyvale, Calif.). Bands were quantified using the 
ImageQuantify program and normalized relative to the actin signals. FIG. 
5b shows the strongest relative signals result from the CD34+ RNA and the 
even more stem cell enriched CD34+/lin- RNA sample. Thus, JAK3 is most 
highly expressed in this primitive population of cells and may play a role 
in transducing the signal of receptor functioning in the proliferative 
and/or developmental pathways of these cells. JAK3 is also expressed in 
the CD34- and peripheral blood fractions and is thus likely to be involved 
with a subset of receptors involved in differentiated cell signalling, in 
analogy to JAK1, JAK2, and TYK-2. 
EXAMPLE 4 
Chromosomal Localization of the JAK3 Gene 
1. Somatic cell hybrid analysis. To determine the chromosomal localization 
of the JAK3 gene, a human/rodent somatic cell hybrid mapping panel, NIGMS 
#2, which included human, mouse and hamster genomic DNA controls was 
screened by PCR (Drwinga, H., et al., Genomics, 16:311, 1993, Dubois, B. 
and Naylor, S., Genomics, 16:315, 1993). In this panel, most of the 
somatic cell hybrid samples contained DNA from a single specific human 
chromosome in a rodent background. To preclude cDNA contamination 
problems, a primer pair was selected that resulted in a PCR product from 
genomic DNA that was larger than the produce resulting from cDNA due to 
the presence of intronic sequence. The plus strand oligo 
5'AGCCGCCTCCTTCTCT3' (SEQ ID NO:3) and minus strand oligo 
5'CGGCAGCAGCTTAGCTAGG3' (SEQ ID NO:4) amplify an approximate 410 base pair 
product from human genomic DNA and a 156 base pair product from JAK3 cDNA. 
For PCR, 100 ngs of genomic DNA from each hybrid cell line were used as the 
target for amplification. PCR amplification was performed using the 
following parameters: (94.degree. C., 1'.fwdarw.55.degree. C., 
1'.fwdarw.72.degree. C., 2').times.30.fwdarw.72.degree. C., 15'. The final 
concentrations of reagents were 0.2 mM dNTP, 50 mM KCL, 3.0 mM Mg, 0.1 U 
Taq Polymerase/ml, and 2.5 mM each primer. The results from the PCR 
amplification were confirmed by Southern transfer and hybridization with a 
.sup.32 P .gamma.-ATP kinase labelled oligo internal to the primers used 
for amplification. 
Using the primer pair results in a PCR product only with DNA from human 
cells and not from mouse or hamster DNA. These oligonucleotides were then 
used on DNA samples from the library representing each of the human 
chromosomes. The amplified DNA was electrophoresed and after transfer to 
nitrocellulose was hybridized to a radiolabelled oligonucleotide internal 
to the other oligonucleotides used for the PCR. Only the DNA from a cell 
line containing human chromosome 19 gave a significant signal. 
2. Fluorescence in situ hybridization: TYK-2 has also been mapped to 
chromosome 19 (JAK1 and JAK2 have been mapped to 1p31.3 and 9p24, 
respectively).sup.55,56 Several pairs of tyrosine kinases (e.g. 
PDGFR.beta. and c-fms, PDGF.alpha. and c-kit, FLT3 and FLT1) have been 
shown to be closely linked, leading to the hypothesis that these receptor 
tyrosine kinases evolved by a trans duplication followed by a cis 
duplication..sup.57-60 In order to confirm the location of the gene on 
chromosome 19, to sublocalize the gene to a specific band, and to 
investigate the possibility that JAK3 and TYK-2 were linked, FISH 
experiment was carried out. 
First clone containing approximately 80 kbp of the JAK3 gene was isolated 
by PCR screening of a P1 library using the same oligonucleotides used 
above. Briefly, P1 genomic clones of JAK3 were obtained by PCR screening 
of the Du Pont Merck Pharmaceutical Company Human Foreskin Fibroblast P1 
Library #1 (DMPC-HFF#1)--(Genome Systems, St. Louis, Mo.). The clones were 
designated DMPC-HFF#1-1441, DMPC-HFF#1-1442, DMPC-HFF#1-1443 and 
identified using the same primer pairs and PCR conditions used for the 
screening of the human/rodent somatic cell hybrid mapping panel (see 
above). Partial sequencing of these P1 clones has confirmed that they 
represent genomic JAK3 DNA. 
The P1 vector containing the approximate 80 kbp genomic clone 1441 of JAK3 
was nick-translated with biotin-14 dATP (BRL, Gaithersburg, Md.), with 30% 
incorporation determined by tritium tracer incorporation. Slides with 
chromosome spreads were made from normal male lymphocytes cultured with 
BrdU (Bhatt, B., et al., Nucleic Acids Res., 16:3951, 1988). Fluorescence 
in situ hybridization was performed as described by Lichter, et. al., (53. 
Lichter, P., Tang, C., Call, K., Hermanson, G., Evans, G., Housman, D., 
Ward, D.: High resolution mapping of human chromosome 11 by in situ 
hybridization with cosmid clones. Science 247:64, 1990) with 
modifications. Probe mix (2.times.SSCP, 60% formamide, 10% dextran 
sulfate, 4 ng/ul biotinylated probe, 300 ng/ul Cot-1 DNA (to suppress 
repeated sequences) and 150 ng/ul salmon sperm DNA was denatured at 
70.degree. C. for 5 minutes, preannealed at 37.degree. C. for 40 minutes, 
placed on slides and hybridized at 37.degree. C. overnight. Slides were 
washed in 70% formamide/2.times.SSC at 43.degree. C. for 20 minutes, and 2 
changes of 2.times.SSC at 37.degree. C. for 5 minutes each. Biotinylated 
probe was detected with FITC-avidin and amplified with biotinylated 
anti-avidin, using reagents from an in situ hybridization kit (Oncor Inc., 
Gaithersburg, Md.), following the manufacturer's instructions. 
Analysis of 36 metaphase cells showed 20 cells (56%) had at least one pair 
of signals (involving both chromatids of a single chromosome). These 20 
metaphases were photographed on color slide film (Kodak Ekttachrome 400HC) 
and 33 paired signals were seen, with all but one located on the proximal 
short arm of an F-group (chr. 19 or 20) chromosome, an example of which is 
shown in FIG. 6a. To determine the specific chromosome and band location 
of the signals, the hybridized slides were G-banded by FPG (fluorescence 
plus Giemsa), photographed, and aligned with the color slides to determine 
the subband location. FIG. 6b shows the position of the paired FISH 
signals on the G-banded chromosomes. All 33 signals were analyzable after 
banding and all were on chromosome 19, with most on bands p12-13.1 (FIG. 
6c). Thus JAK3 may be located near TYK-2, which has been localized to 
19p13.2. FIG. 6c is the ideogram of human chromosome 19, showing 
localization of JAK3 to 19p12-13.1. Each dot represents a paired signal 
seen on metaphase chromosomes. Signals clearly located on a single band 
are diagrammed to the right of the ideogram; those which could not be 
sublocalized to a single band are assigned to regions diagrammed to the 
left (brackets). 
The foregoing is meant to illustrate, but not to limit, the scope of the 
invention. Indeed, those of ordinary skill in the art can readily envision 
and produce further embodiments, based on the teachings herein, without 
undue experimentation. 
__________________________________________________________________________ 
# SEQUENCE LISTING 
- (1) GENERAL INFORMATION: 
- (iii) NUMBER OF SEQUENCES: 12 
- (2) INFORMATION FOR SEQ ID NO:1: 
- (i) SEQUENCE CHARACTERISTICS: 
#pairs (A) LENGTH: 3807 base 
(B) TYPE: nucleic acid 
(C) STRANDEDNESS: single 
(D) TOPOLOGY: linear 
- (ii) MOLECULE TYPE: DNA 
- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:1: 
- AAACAGTTAA TACATATTTT TTATGTTACG TGTATTCTGT ACAACAAAGT AA - #GCTAGACA 
60 
- AAAGAAAATG TTTTCTCCTT CCTGTGTGGG ACTTTCCTCT CGCTGCCTCC CG - #GCTCTGCC 
120 
#GCA CCT 176CCAGGG TCCCTGCCCG CTAGGCAAGT TGCACTC ATG 
#Ala Pro Met 
#1 
- CCA AGT GAA GAG ACG CCC CTG ATC CCT CAG CG - #T TCA TGC AGC CTC TTG 
224 
Pro Ser Glu Glu Thr Pro Leu Ile Pro Gln Ar - #g Ser Cys Ser Leu Leu 
# 15 
- TCC ACG GAG GCT GGT GCC CTG CAT GTG CTG CT - #G CCC GCT CGG GGC CCC 
272 
Ser Thr Glu Ala Gly Ala Leu His Val Leu Le - #u Pro Ala Arg Gly Pro 
# 35 
- GGG CCC CCC CAG CGC CTA TCT TTC TCC TTT GG - #G GAC CAC TTG GCT GAG 
320 
Gly Pro Pro Gln Arg Leu Ser Phe Ser Phe Gl - #y Asp His Leu Ala Glu 
# 50 
- GAC CTG TGC GTG CAG GCT GCC AAG GCC AGC GC - #G ATC CTG CCT GTG TAC 
368 
Asp Leu Cys Val Gln Ala Ala Lys Ala Ser Al - #a Ile Leu Pro Val Tyr 
# 65 
- CAC TCC CTC TTT GCT CTG GCC ACG GAG GAC CT - #G TCC TGC TGG TTC CCC 
416 
His Ser Leu Phe Ala Leu Ala Thr Glu Asp Le - #u Ser Cys Trp Phe Pro 
# 80 
- CGA GCC ACA TCT TCT CCG TGG AGG ATG CCA GC - #A CCC CAA GTC CTG CTG 
464 
Arg Ala Thr Ser Ser Pro Trp Arg Met Pro Al - #a Pro Gln Val Leu Leu 
# 95 
- TAC AGG ATT CGC TTT TAC TTC CCC AAT TGG TT - #T GGG CTG GAG AAG TGC 
512 
Tyr Arg Ile Arg Phe Tyr Phe Pro Asn Trp Ph - #e Gly Leu Glu Lys Cys 
100 1 - #05 1 - #10 1 - 
#15 
- CAC CGC TTC GGG CTA CGC AAG GAT TTG GCC AG - #T GCT ATC CTT GAC CTG 
560 
His Arg Phe Gly Leu Arg Lys Asp Leu Ala Se - #r Ala Ile Leu Asp Leu 
# 130 
- CCA GTC CTG GAG CAC CTC TTT GCC CAG CAC CG - #C AGT GAC CTG GTG AGT 
608 
Pro Val Leu Glu His Leu Phe Ala Gln His Ar - #g Ser Asp Leu Val Ser 
# 145 
- GGG CGC CTC CCC CGT GGC CTC AGT CTC AAG GA - #G CAG GGT GAG TGT CTC 
656 
Gly Arg Leu Pro Arg Gly Leu Ser Leu Lys Gl - #u Gln Gly Glu Cys Leu 
# 160 
- AGC CTG GCC GTG TTG GAC CTG GCC CGG ATG GC - #G CGA GAG CAG GCC CAG 
704 
Ser Leu Ala Val Leu Asp Leu Ala Arg Met Al - #a Arg Glu Gln Ala Gln 
# 175 
- CGG CGG GGA GAG CTG CTG AAG ACT GTC AGC TA - #C AAG GCC TGC CTA CCC 
752 
Arg Arg Gly Glu Leu Leu Lys Thr Val Ser Ty - #r Lys Ala Cys Leu Pro 
180 1 - #85 1 - #90 1 - 
#95 
- CCA AGC CTG CGC GAC CTG ATC CAG GGC CTG AG - #C TTC GTG ACG GGG AGG 
800 
Pro Ser Leu Arg Asp Leu Ile Gln Gly Leu Se - #r Phe Val Thr Gly Arg 
# 210 
- CGT ATT CGG AGG ACG GTG GAG AGC CCC CTG CG - #C CGG GTG GCC GCC TGC 
848 
Arg Ile Arg Arg Thr Val Glu Ser Pro Leu Ar - #g Arg Val Ala Ala Cys 
# 225 
- CAG GCA GAC CGG CAC TCG CTC ATG GCC AAG TA - #C ATC ATG GAC CTG GAG 
896 
Gln Ala Asp Arg His Ser Leu Met Ala Lys Ty - #r Ile Met Asp Leu Glu 
# 240 
- CGG CTG GAT CCA GCC GGG GCC GCC GAG ACC TT - #C CAC GTG GGC CTC CCT 
944 
Arg Leu Asp Pro Ala Gly Ala Ala Glu Thr Ph - #e His Val Gly Leu Pro 
# 255 
- GGG GCC CTT GGT GGC CAC GAC GGG CTG GGG CT - #C GTC CGC GTG GCT GGT 
992 
Gly Ala Leu Gly Gly His Asp Gly Leu Gly Le - #u Val Arg Val Ala Gly 
260 2 - #65 2 - #70 2 - 
#75 
- GAC GGC GGC ATC GCC TGG ACC CAG GGA GAA CA - #G GAG GTC CTC CAG CCC 
1040 
Asp Gly Gly Ile Ala Trp Thr Gln Gly Glu Gl - #n Glu Val Leu Gln Pro 
# 290 
- TTC TGC GAC TTT CCA GAA ATC GTA GAC ATT AG - #C ATC AAG CAG GCC CCG 
1088 
Phe Cys Asp Phe Pro Glu Ile Val Asp Ile Se - #r Ile Lys Gln Ala Pro 
# 305 
- CGC GTT GGC CCG GCC GGA GAG CAC CGC CTG GT - #C ACT GTT ACC AGG ACA 
1136 
Arg Val Gly Pro Ala Gly Glu His Arg Leu Va - #l Thr Val Thr Arg Thr 
# 320 
- GAC AAC CAG ATT TTA GAG GCC GAG TTC CCA GG - #G CTG CCC GAG GCT CTG 
1184 
Asp Asn Gln Ile Leu Glu Ala Glu Phe Pro Gl - #y Leu Pro Glu Ala Leu 
# 335 
- TCG TTC GTG GCG CTC GTG GAC GGC TAC TTC CG - #G CTG ACC ACG GAC TCC 
1232 
Ser Phe Val Ala Leu Val Asp Gly Tyr Phe Ar - #g Leu Thr Thr Asp Ser 
340 3 - #45 3 - #50 3 - 
#55 
- CAG CAC TTC TTC TGC AAG GAG GTG GAC CCG AG - #G CTG CTG GAG GAA GTG 
1280 
Gln His Phe Phe Cys Lys Glu Val Asp Pro Ar - #g Leu Leu Glu Glu Val 
# 370 
- GCC GAG CAG TGC CAC GGC CCC ATC ACT CTG GA - #C TTT GCC ATC AAC AAG 
1328 
Ala Glu Gln Cys His Gly Pro Ile Thr Leu As - #p Phe Ala Ile Asn Lys 
# 385 
- CTC AAG ACT GGG GGC TCA CGT CCT GGC TCC TA - #T GTT CTC CGC CGC ATC 
1376 
Leu Lys Thr Gly Gly Ser Arg Pro Gly Ser Ty - #r Val Leu Arg Arg Ile 
# 400 
- CCC CAG GAC TTT GAC AGC TTC CTC CTC ACT GT - #C TGT GTC CAG AAC CCC 
1424 
Pro Gln Asp Phe Asp Ser Phe Leu Leu Thr Va - #l Cys Val Gln Asn Pro 
# 415 
- CTT GGT CCT GAT TAT AAG GGC TGC CTC ATC CG - #G CGC AGC CCC ACA GGA 
1472 
Leu Gly Pro Asp Tyr Lys Gly Cys Leu Ile Ar - #g Arg Ser Pro Thr Gly 
420 4 - #25 4 - #30 4 - 
#35 
- ACC TTC CTT CTG GTT GGC CTC AGC CGA CCC CA - #C AGC AGT CTT CGA GAG 
1520 
Thr Phe Leu Leu Val Gly Leu Ser Arg Pro Hi - #s Ser Ser Leu Arg Glu 
# 450 
- CTC CTG GCA ACC TGC TGG GAT GGG GGG CTG CA - #C GTA GAT GGG GTG GCA 
1568 
Leu Leu Ala Thr Cys Trp Asp Gly Gly Leu Hi - #s Val Asp Gly Val Ala 
# 465 
- GTG ACC CTC ACT TCC TGC TGT ATC CCC AGA CC - #C AAA GAA AAG TCC AAC 
1616 
Val Thr Leu Thr Ser Cys Cys Ile Pro Arg Pr - #o Lys Glu Lys Ser Asn 
# 480 
- CTG ATT GTG GTC CAG AGA GGT CAC AGC CCA CC - #C ACA TCA TCC TTG GTT 
1664 
Leu Ile Val Val Gln Arg Gly His Ser Pro Pr - #o Thr Ser Ser Leu Val 
# 495 
- CAG CCC CAA TCC CAA TAC CAG CTG AGT CAG AT - #G ACA TTT CAC AAG ATC 
1712 
Gln Pro Gln Ser Gln Tyr Gln Leu Ser Gln Me - #t Thr Phe His Lys Ile 
500 5 - #05 5 - #10 5 - 
#15 
- CCT GCT GAC AGC CTG GAG TGG CAT GAG AAC CT - #G GGC CAT GGG TCC TTC 
1760 
Pro Ala Asp Ser Leu Glu Trp His Glu Asn Le - #u Gly His Gly Ser Phe 
# 530 
- ACC AAG ATT TAC CGG GGC TGT CGC CAT GAG GT - #G GTG GAT GGG GAG GCC 
1808 
Thr Lys Ile Tyr Arg Gly Cys Arg His Glu Va - #l Val Asp Gly Glu Ala 
# 545 
- CGA AAG ACA GAG GTG CTG CTG AAG GTC ATG GA - #T GCC AAG CAC AAG AAC 
1856 
Arg Lys Thr Glu Val Leu Leu Lys Val Met As - #p Ala Lys His Lys Asn 
# 560 
- TGC ATG GAG TCA TTC CTG GAA GCA GCG AGC TT - #G ATG AGC CAA GTG TCG 
1904 
Cys Met Glu Ser Phe Leu Glu Ala Ala Ser Le - #u Met Ser Gln Val Ser 
# 575 
- TAC CGG CAT CTC GTG CTG CTC CAC GGC GTG TG - #C ATG GCT GGA GAC AGC 
1952 
Tyr Arg His Leu Val Leu Leu His Gly Val Cy - #s Met Ala Gly Asp Ser 
580 5 - #85 5 - #90 5 - 
#95 
- ACC ATG GTC GAG GAA TTT GTA CAC CTG GGG GC - #C ATA GAC ATG TAT CTG 
2000 
Thr Met Val Glu Glu Phe Val His Leu Gly Al - #a Ile Asp Met Tyr Leu 
# 610 
- CGA AAA CGT GGC CAC CTG GTG CCA GCC AGC TG - #G AAG CTG CAG GTG GTC 
2048 
Arg Lys Arg Gly His Leu Val Pro Ala Ser Tr - #p Lys Leu Gln Val Val 
# 625 
- AAA CAG CTG GCC TAC GCC CTC AAC TAT CTG GA - #G GAC AAA GGC CTG TCC 
2096 
Lys Gln Leu Ala Tyr Ala Leu Asn Tyr Leu Gl - #u Asp Lys Gly Leu Ser 
# 640 
- CAT GGC AAT GTC TCT GCC CGG AAG GTG CTC CT - #G GCT CGG GAG GGG GCT 
2144 
His Gly Asn Val Ser Ala Arg Lys Val Leu Le - #u Ala Arg Glu Gly Ala 
# 655 
- GAT GGG AGC CCG CCC TTC ATC AAG CTG AGT GA - #C CCT GGG GTC AGC CCC 
2192 
Asp Gly Ser Pro Pro Phe Ile Lys Leu Ser As - #p Pro Gly Val Ser Pro 
660 6 - #65 6 - #70 6 - 
#75 
- GCT GTG TTA AGC CTG GAG ATG CTC ACC GAC AG - #G ATC CCC TGG GTG GCC 
2240 
Ala Val Leu Ser Leu Glu Met Leu Thr Asp Ar - #g Ile Pro Trp Val Ala 
# 690 
- CCC GAG TGT CTC CGG GAG GCG CAG ACA CTT AG - #C TTG GAA GCT GAC AAG 
2288 
Pro Glu Cys Leu Arg Glu Ala Gln Thr Leu Se - #r Leu Glu Ala Asp Lys 
# 705 
- TGG GGC TTC GGC GCC ACG GTC TGG GAA GTG TT - #T AGT GGC GTC ACC ATG 
2336 
Trp Gly Phe Gly Ala Thr Val Trp Glu Val Ph - #e Ser Gly Val Thr Met 
# 720 
- CCC ATC AGT GCC CTA GAT CCT GCT AAG AAA CT - #C CAA TTT TAT GAG GAC 
2384 
Pro Ile Ser Ala Leu Asp Pro Ala Lys Lys Le - #u Gln Phe Tyr Glu Asp 
# 735 
- CGG CAG CAG CTG TCG GCC CCC AAG TGG ACA GA - #G CTG GCC CTG CTG ATT 
2432 
Arg Gln Gln Leu Ser Ala Pro Lys Trp Thr Gl - #u Leu Ala Leu Leu Ile 
740 7 - #45 7 - #50 7 - 
#55 
- CAA CAG TGC ATG GCC TAT GAG CCG GTC CAG AG - #G CCC TCC TTA CGA GCC 
2480 
Gln Gln Cys Met Ala Tyr Glu Pro Val Gln Ar - #g Pro Ser Leu Arg Ala 
# 770 
- GTC ATT CGT GAC CTC AAT AGT CTC ATC TCT TC - #A GAC TAT GAG CTC CTC 
2528 
Val Ile Arg Asp Leu Asn Ser Leu Ile Ser Se - #r Asp Tyr Glu Leu Leu 
# 785 
- TCA GAC CAC ACC TGG TGC CCT GGC ACT CGT GA - #T GGG CTG TGG AAT GGT 
2576 
Ser Asp His Thr Trp Cys Pro Gly Thr Arg As - #p Gly Leu Trp Asn Gly 
# 800 
- GCC CAG CTC TAT GCC TGC CAA GAC CCC ACG AT - #C TTC GAG GAG AGA CAC 
2624 
Ala Gln Leu Tyr Ala Cys Gln Asp Pro Thr Il - #e Phe Glu Glu Arg His 
# 815 
- CTC AAG TAC ATC TCA CAG CTG GGC AAG GGC TT - #C TTT GGC AGC GTG GAG 
2672 
Leu Lys Tyr Ile Ser Gln Leu Gly Lys Gly Ph - #e Phe Gly Ser Val Glu 
820 8 - #25 8 - #30 8 - 
#35 
- CTG TGC CGC TAT GAC CCG CTA GGC GAC AAT AC - #A GGT GCC CTG GTG GCC 
2720 
Leu Cys Arg Tyr Asp Pro Leu Gly Asp Asn Th - #r Gly Ala Leu Val Ala 
# 850 
- GTG AAA CAG CTG CAG CAC AGC GGG CCA GAC CA - #G CAG AGG GAC TTT CAG 
2768 
Val Lys Gln Leu Gln His Ser Gly Pro Asp Gl - #n Gln Arg Asp Phe Gln 
# 865 
- CGG GAG ATT CAG ATC CTC AAA GCA CAG CAC AG - #T GAT TTC ATT GTC AAG 
2816 
Arg Glu Ile Gln Ile Leu Lys Ala Gln His Se - #r Asp Phe Ile Val Lys 
# 880 
- TAT CGT GGT GTC AGC TAT GGC CCG GGC CGC CA - #G AGC CCT GCG CTG GTC 
2864 
Tyr Arg Gly Val Ser Tyr Gly Pro Gly Arg Gl - #n Ser Pro Ala Leu Val 
# 895 
- ATG GAG TAC CTG CCC AGC GGC TGC TTG CGC GA - #C TTC CTG CAG CGG CAC 
2912 
Met Glu Tyr Leu Pro Ser Gly Cys Leu Arg As - #p Phe Leu Gln Arg His 
900 9 - #05 9 - #10 9 - 
#15 
- CGG GGC CTC GAT GCC AGC CGC CTC CTT CTC TA - #T TCC TCG CAG ATC TGC 
2960 
Arg Gly Leu Asp Ala Ser Arg Leu Leu Leu Ty - #r Ser Ser Gln Ile Cys 
# 930 
- AAG GGC ATG GAG TAC CTG GGC TCC CGC CGC TG - #C GTG CAC CGC GAC CTG 
3008 
Lys Gly Met Glu Tyr Leu Gly Ser Arg Arg Cy - #s Val His Arg Asp Leu 
# 945 
- GCC GCC CGA AAC ATC CTC GTG GAG AGC GAG GC - #A CAC GTC AAG ATC GCT 
3056 
Ala Ala Arg Asn Ile Leu Val Glu Ser Glu Al - #a His Val Lys Ile Ala 
# 960 
- GAC TTC GGC CTA GCT AAG CTG CTG CCG CTT GA - #C AAA GAC TAC TAC GTG 
3104 
Asp Phe Gly Leu Ala Lys Leu Leu Pro Leu As - #p Lys Asp Tyr Tyr Val 
# 975 
- GTC CGC GAG CCA GGC CAG AGC CCC ATT TTC TG - #G TAT GCC CCC GAA TCC 
3152 
Val Arg Glu Pro Gly Gln Ser Pro Ile Phe Tr - #p Tyr Ala Pro Glu Ser 
980 9 - #85 9 - #90 9 - 
#95 
- CTC TCG GAC AAC ATC TTC TCT CGC CAG TCA GA - #C GTC TGG AGC TTC GGG 
3200 
Leu Ser Asp Asn Ile Phe Ser Arg Gln Ser As - #p Val Trp Ser Phe Gly 
# 10105 
- GTC GTC CTG TAC GAG CTC TTC ACC TAC TGC GA - #C AAA AGC TGC AGC CCC 
3248 
Val Val Leu Tyr Glu Leu Phe Thr Tyr Cys As - #p Lys Ser Cys Ser Pro 
# 10250 
- TCG GCC GAG TTC CTG CGG ATG ATG GGA TGT GA - #G CGG GAT GTC CCC CGC 
3296 
Ser Ala Glu Phe Leu Arg Met Met Gly Cys Gl - #u Arg Asp Val Pro Arg 
# 10405 
- CTC TGC CGC CTC TTG GAA CTG CTG GAG GAG GG - #C CAG AGG CTG CCG GCG 
3344 
Leu Cys Arg Leu Leu Glu Leu Leu Glu Glu Gl - #y Gln Arg Leu Pro Ala 
# 10550 
- CCT CCT TGC TGC CCT GC TGAGGTGAGT TGCTACAGTG GCT - #GGAGAGA 
3391 
Pro Pro Cys Cys Pro 
1060 
- CGACATCTGC CTGCCTGCTG AGTGAGTTGC TACAGTGGCT GAGAGACGAC AT - #CTGCTCCA 
3451 
- TGGCTGGTGG CCGACAGTAA TCTCACGCCG GACCTGCCGC AGCCCCTGCC CC - #AGACCTCT 
3511 
- CACCATCACC GCCACCACCG TGCAGCTGCC ACCAACCCTG CACGCTACTG CT - #GCCTCAGT 
3571 
- GGCTGTACCC AACAAGACCT GCTGACCCTC TGTCCCTACT GATTCCTCCT TG - #GGTGCAGC 
3631 
- CTCAGAGTGC CTGAGGCCCA GAGGGTCTGG TCTGGTGAGC TCCTGAGGCC AC - #ACAGCACC 
3691 
- ATAAAGTCTC GCATCTACAG GCCTTTGATT ACCTCCTGGG ATGGGTGCTC AC - #TATCTACC 
3751 
- CCAGACCAAC GCCACCTGCA GCCTGTGGAG TCAACTGCAG AATAAATCAC AC - #CCTA 
3807 
- (2) INFORMATION FOR SEQ ID NO:2: 
- (i) SEQUENCE CHARACTERISTICS: 
#acids (A) LENGTH: 1064 amino 
(B) TYPE: amino acid 
(D) TOPOLOGY: linear 
- (ii) MOLECULE TYPE: protein 
- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:2: 
- Met Ala Pro Pro Ser Glu Glu Thr Pro Leu Il - #e Pro Gln Arg Ser Cys 
# 15 
- Ser Leu Leu Ser Thr Glu Ala Gly Ala Leu Hi - #s Val Leu Leu Pro Ala 
# 30 
- Arg Gly Pro Gly Pro Pro Gln Arg Leu Ser Ph - #e Ser Phe Gly Asp His 
# 45 
- Leu Ala Glu Asp Leu Cys Val Gln Ala Ala Ly - #s Ala Ser Ala Ile Leu 
# 60 
- Pro Val Tyr His Ser Leu Phe Ala Leu Ala Th - #r Glu Asp Leu Ser Cys 
# 80 
- Trp Phe Pro Arg Ala Thr Ser Ser Pro Trp Ar - #g Met Pro Ala Pro Gln 
# 95 
- Val Leu Leu Tyr Arg Ile Arg Phe Tyr Phe Pr - #o Asn Trp Phe Gly Leu 
# 110 
- Glu Lys Cys His Arg Phe Gly Leu Arg Lys As - #p Leu Ala Ser Ala Ile 
# 125 
- Leu Asp Leu Pro Val Leu Glu His Leu Phe Al - #a Gln His Arg Ser Asp 
# 140 
- Leu Val Ser Gly Arg Leu Pro Arg Gly Leu Se - #r Leu Lys Glu Gln Gly 
145 1 - #50 1 - #55 1 - 
#60 
- Glu Cys Leu Ser Leu Ala Val Leu Asp Leu Al - #a Arg Met Ala Arg Glu 
# 175 
- Gln Ala Gln Arg Arg Gly Glu Leu Leu Lys Th - #r Val Ser Tyr Lys Ala 
# 190 
- Cys Leu Pro Pro Ser Leu Arg Asp Leu Ile Gl - #n Gly Leu Ser Phe Val 
# 205 
- Thr Gly Arg Arg Ile Arg Arg Thr Val Glu Se - #r Pro Leu Arg Arg Val 
# 220 
- Ala Ala Cys Gln Ala Asp Arg His Ser Leu Me - #t Ala Lys Tyr Ile Met 
225 2 - #30 2 - #35 2 - 
#40 
- Asp Leu Glu Arg Leu Asp Pro Ala Gly Ala Al - #a Glu Thr Phe His Val 
# 255 
- Gly Leu Pro Gly Ala Leu Gly Gly His Asp Gl - #y Leu Gly Leu Val Arg 
# 270 
- Val Ala Gly Asp Gly Gly Ile Ala Trp Thr Gl - #n Gly Glu Gln Glu Val 
# 285 
- Leu Gln Pro Phe Cys Asp Phe Pro Glu Ile Va - #l Asp Ile Ser Ile Lys 
# 300 
- Gln Ala Pro Arg Val Gly Pro Ala Gly Glu Hi - #s Arg Leu Val Thr Val 
305 3 - #10 3 - #15 3 - 
#20 
- Thr Arg Thr Asp Asn Gln Ile Leu Glu Ala Gl - #u Phe Pro Gly Leu Pro 
# 335 
- Glu Ala Leu Ser Phe Val Ala Leu Val Asp Gl - #y Tyr Phe Arg Leu Thr 
# 350 
- Thr Asp Ser Gln His Phe Phe Cys Lys Glu Va - #l Asp Pro Arg Leu Leu 
# 365 
- Glu Glu Val Ala Glu Gln Cys His Gly Pro Il - #e Thr Leu Asp Phe Ala 
# 380 
- Ile Asn Lys Leu Lys Thr Gly Gly Ser Arg Pr - #o Gly Ser Tyr Val Leu 
385 3 - #90 3 - #95 4 - 
#00 
- Arg Arg Ile Pro Gln Asp Phe Asp Ser Phe Le - #u Leu Thr Val Cys Val 
# 415 
- Gln Asn Pro Leu Gly Pro Asp Tyr Lys Gly Cy - #s Leu Ile Arg Arg Ser 
# 430 
- Pro Thr Gly Thr Phe Leu Leu Val Gly Leu Se - #r Arg Pro His Ser Ser 
# 445 
- Leu Arg Glu Leu Leu Ala Thr Cys Trp Asp Gl - #y Gly Leu His Val Asp 
# 460 
- Gly Val Ala Val Thr Leu Thr Ser Cys Cys Il - #e Pro Arg Pro Lys Glu 
465 4 - #70 4 - #75 4 - 
#80 
- Lys Ser Asn Leu Ile Val Val Gln Arg Gly Hi - #s Ser Pro Pro Thr Ser 
# 495 
- Ser Leu Val Gln Pro Gln Ser Gln Tyr Gln Le - #u Ser Gln Met Thr Phe 
# 510 
- His Lys Ile Pro Ala Asp Ser Leu Glu Trp Hi - #s Glu Asn Leu Gly His 
# 525 
- Gly Ser Phe Thr Lys Ile Tyr Arg Gly Cys Ar - #g His Glu Val Val Asp 
# 540 
- Gly Glu Ala Arg Lys Thr Glu Val Leu Leu Ly - #s Val Met Asp Ala Lys 
545 5 - #50 5 - #55 5 - 
#60 
- His Lys Asn Cys Met Glu Ser Phe Leu Glu Al - #a Ala Ser Leu Met Ser 
# 575 
- Gln Val Ser Tyr Arg His Leu Val Leu Leu Hi - #s Gly Val Cys Met Ala 
# 590 
- Gly Asp Ser Thr Met Val Glu Glu Phe Val Hi - #s Leu Gly Ala Ile Asp 
# 605 
- Met Tyr Leu Arg Lys Arg Gly His Leu Val Pr - #o Ala Ser Trp Lys Leu 
# 620 
- Gln Val Val Lys Gln Leu Ala Tyr Ala Leu As - #n Tyr Leu Glu Asp Lys 
625 6 - #30 6 - #35 6 - 
#40 
- Gly Leu Ser His Gly Asn Val Ser Ala Arg Ly - #s Val Leu Leu Ala Arg 
# 655 
- Glu Gly Ala Asp Gly Ser Pro Pro Phe Ile Ly - #s Leu Ser Asp Pro Gly 
# 670 
- Val Ser Pro Ala Val Leu Ser Leu Glu Met Le - #u Thr Asp Arg Ile Pro 
# 685 
- Trp Val Ala Pro Glu Cys Leu Arg Glu Ala Gl - #n Thr Leu Ser Leu Glu 
# 700 
- Ala Asp Lys Trp Gly Phe Gly Ala Thr Val Tr - #p Glu Val Phe Ser Gly 
705 7 - #10 7 - #15 7 - 
#20 
- Val Thr Met Pro Ile Ser Ala Leu Asp Pro Al - #a Lys Lys Leu Gln Phe 
# 735 
- Tyr Glu Asp Arg Gln Gln Leu Ser Ala Pro Ly - #s Trp Thr Glu Leu Ala 
# 750 
- Leu Leu Ile Gln Gln Cys Met Ala Tyr Glu Pr - #o Val Gln Arg Pro Ser 
# 765 
- Leu Arg Ala Val Ile Arg Asp Leu Asn Ser Le - #u Ile Ser Ser Asp Tyr 
# 780 
- Glu Leu Leu Ser Asp His Thr Trp Cys Pro Gl - #y Thr Arg Asp Gly Leu 
785 7 - #90 7 - #95 8 - 
#00 
- Trp Asn Gly Ala Gln Leu Tyr Ala Cys Gln As - #p Pro Thr Ile Phe Glu 
# 815 
- Glu Arg His Leu Lys Tyr Ile Ser Gln Leu Gl - #y Lys Gly Phe Phe Gly 
# 830 
- Ser Val Glu Leu Cys Arg Tyr Asp Pro Leu Gl - #y Asp Asn Thr Gly Ala 
# 845 
- Leu Val Ala Val Lys Gln Leu Gln His Ser Gl - #y Pro Asp Gln Gln Arg 
# 860 
- Asp Phe Gln Arg Glu Ile Gln Ile Leu Lys Al - #a Gln His Ser Asp Phe 
865 8 - #70 8 - #75 8 - 
#80 
- Ile Val Lys Tyr Arg Gly Val Ser Tyr Gly Pr - #o Gly Arg Gln Ser Pro 
# 895 
- Ala Leu Val Met Glu Tyr Leu Pro Ser Gly Cy - #s Leu Arg Asp Phe Leu 
# 910 
- Gln Arg His Arg Gly Leu Asp Ala Ser Arg Le - #u Leu Leu Tyr Ser Ser 
# 925 
- Gln Ile Cys Lys Gly Met Glu Tyr Leu Gly Se - #r Arg Arg Cys Val His 
# 940 
- Arg Asp Leu Ala Ala Arg Asn Ile Leu Val Gl - #u Ser Glu Ala His Val 
945 9 - #50 9 - #55 9 - 
#60 
- Lys Ile Ala Asp Phe Gly Leu Ala Lys Leu Le - #u Pro Leu Asp Lys Asp 
# 975 
- Tyr Tyr Val Val Arg Glu Pro Gly Gln Ser Pr - #o Ile Phe Trp Tyr Ala 
# 990 
- Pro Glu Ser Leu Ser Asp Asn Ile Phe Ser Ar - #g Gln Ser Asp Val Trp 
# 10050 
- Ser Phe Gly Val Val Leu Tyr Glu Leu Phe Th - #r Tyr Cys Asp Lys Ser 
# 10205 
- Cys Ser Pro Ser Ala Glu Phe Leu Arg Met Me - #t Gly Cys Glu Arg Asp 
# 10401030 - # 1035 
- Val Pro Arg Leu Cys Arg Leu Leu Glu Leu Le - #u Glu Glu Gly Gln Arg 
# 10550 
- Leu Pro Ala Pro Pro Cys Cys Pro 
1060 
- (2) INFORMATION FOR SEQ ID NO:3: 
- (i) SEQUENCE CHARACTERISTICS: 
#pairs (A) LENGTH: 16 base 
(B) TYPE: nucleic acid 
(C) STRANDEDNESS: single 
(D) TOPOLOGY: linear 
- (ii) MOLECULE TYPE: DNA 
- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:3: 
# 16 
- (2) INFORMATION FOR SEQ ID NO:4: 
- (i) SEQUENCE CHARACTERISTICS: 
#pairs (A) LENGTH: 19 base 
(B) TYPE: nucleic acid 
(C) STRANDEDNESS: single 
(D) TOPOLOGY: linear 
- (ii) MOLECULE TYPE: DNA 
- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:4: 
# 19 AGG 
- (2) INFORMATION FOR SEQ ID NO:5: 
- (i) SEQUENCE CHARACTERISTICS: 
#acids (A) LENGTH: 1082 amino 
(B) TYPE: amino acid 
(C) STRANDEDNESS: Not R - #elevant 
(D) TOPOLOGY: linear 
- (ii) MOLECULE TYPE: protein 
- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:5: 
- Met Ala Pro Pro Ser Glu Glu Thr - # Pro Leu Ile Pro Gln Arg Ser 
Cys 
# 15 
- Ser Leu Leu Ser Thr Glu Ala Gly - # Ala Leu His Val Leu Leu Pro 
Ala 
# 30 
- Arg Gly Pro Gly Pro Pro Gln Arg - # Leu Ser Phe Ser Phe Gly Asp 
His 
# 45 
- Leu Ala Glu Asp Leu Cys Val Gln - # Ala Ala Lys Ala Ser Ala Ile 
Leu 
# 60 
- Pro Val Tyr His Ser Leu Phe Ala - # Leu Ala Thr Glu Asp Leu Ser 
Cys 
# 80 
- Trp Phe Pro Arg Ala Thr Ser Ser - # Pro Trp Arg Met Pro Ala Pro 
Gln 
# 95 
- Val Leu Leu Tyr Arg Ile Arg Phe - # Tyr Phe Pro Asn Trp Phe Gly 
Leu 
# 110 
- Glu Lys Cys His Arg Phe Gly Leu - # Arg Lys Asp Leu Ala Ser Ala 
Ile 
# 125 
- Leu Asp Leu Pro Val Leu Glu His - # Leu Phe Ala Gln His Arg Ser 
Asp 
# 140 
- Leu Val Ser Gly Arg Leu Pro Arg - # Gly Leu Ser Leu Lys Glu Gln 
Gly 
# 160 
- Glu Cys Leu Ser Leu Ala Val Leu - # Asp Leu Ala Arg Met Ala Arg 
Glu 
# 175 
- Gln Ala Gln Arg Arg Gly Glu Leu - # Leu Lys Thr Val Ser Tyr Lys 
Ala 
# 190 
- Cys Leu Pro Pro Ser Leu Arg Asp - # Leu Ile Gln Gly Leu Ser Phe 
Val 
# 205 
- Thr Gly Arg Arg Ile Arg Arg Thr - # Val Glu Ser Pro Leu Arg Arg 
Val 
# 220 
- Ala Ala Cys Gln Ala Asp Arg His - # Ser Leu Met Ala Lys Tyr Ile 
Met 
# 240 
- Asp Leu Glu Arg Leu Asp Pro Ala - # Gly Ala Ala Glu Thr Phe His 
Val 
# 255 
- Gly Leu Pro Gly Ala Leu Gly Gly - # His Asp Gly Leu Gly Leu Val 
Arg 
# 270 
- Val Ala Gly Asp Gly Gly Ile Ala - # Trp Thr Gln Gly Glu Gln Glu 
Val 
# 285 
- Leu Gln Pro Phe Cys Asp Phe Pro - # Glu Ile Val Asp Ile Ser Ile 
Lys 
# 300 
- Gln Ala Pro Arg Val Gly Pro Ala - # Gly Glu His Arg Leu Val Thr 
Val 
# 320 
- Thr Arg Thr Asp Asn Gln Ile Leu - # Glu Ala Glu Phe Pro Gly Leu 
Pro 
# 335 
- Glu Ala Leu Ser Phe Val Ala Leu - # Val Asp Gly Tyr Phe Arg Leu 
Thr 
# 350 
- Thr Asp Ser Gln His Phe Phe Cys - # Lys Glu Val Asp Pro Arg Leu 
Leu 
# 365 
- Glu Glu Val Ala Glu Gln Cys His - # Gly Pro Ile Thr Leu Asp Phe 
Ala 
# 380 
- Ile Asn Lys Leu Lys Thr Gly Gly - # Ser Arg Pro Gly Ser Tyr Val 
Leu 
# 400 
- Arg Arg Ile Pro Gln Asp Phe Asp - # Ser Phe Leu Leu Thr Val Cys 
Val 
# 415 
- Gln Asn Pro Leu Gly Pro Asp Tyr - # Lys Gly Cys Leu Ile Arg Arg 
Ser 
# 430 
- Pro Thr Gly Thr Phe Leu Leu Val - # Gly Leu Ser Arg Pro His Ser 
Ser 
# 445 
- Leu Arg Glu Leu Leu Ala Thr Cys - # Trp Asp Gly Gly Leu His Val 
Asp 
# 460 
- Gly Val Ala Val Thr Leu Thr Ser - # Cys Cys Ile Pro Arg Pro Lys 
Glu 
# 480 
- Lys Ser Asn Leu Ile Val Val Gln - # Arg Gly His Ser Pro Pro Thr 
Ser 
# 495 
- Ser Leu Val Gln Pro Gln Ser Gln - # Tyr Gln Leu Ser Gln Met Thr 
Phe 
# 510 
- His Lys Ile Pro Ala Asp Ser Leu - # Glu Trp His Glu Asn Leu Gly 
His 
# 525 
- Gly Ser Phe Thr Lys Ile Tyr Arg - # Gly Cys Arg His Glu Val Val 
Asp 
# 540 
- Gly Glu Ala Arg Lys Thr Glu Val - # Leu Leu Lys Val Met Asp Ala 
Lys 
# 560 
- His Lys Asn Cys Met Glu Ser Phe - # Leu Glu Ala Ala Ser Leu Met 
Ser 
# 575 
- Gln Val Ser Tyr Arg His Leu Val - # Leu Leu His Gly Val Cys Met 
Ala 
# 590 
- Gly Asp Ser Thr Met Val Glu Glu - # Phe Val His Leu Gly Ala Ile 
Asp 
# 605 
- Met Tyr Leu Arg Lys Arg Gly His - # Leu Val Pro Ala Ser Trp Lys 
Leu 
# 620 
- Gln Val Val Lys Gln Leu Ala Tyr - # Ala Leu Asn Tyr Leu Glu Asp 
Lys 
# 640 
- Gly Leu Ser His Gly Asn Val Ser - # Ala Arg Lys Val Leu Leu Ala 
Arg 
# 655 
- Glu Gly Ala Asp Gly Ser Pro Pro - # Phe Ile Lys Leu Ser Asp Pro 
Gly 
# 670 
- Val Ser Pro Ala Val Leu Ser Leu - # Glu Met Leu Thr Asp Arg Ile 
Pro 
# 685 
- Trp Val Ala Pro Glu Cys Leu Arg - # Glu Ala Gln Thr Leu Ser Leu 
Glu 
# 700 
- Ala Asp Lys Trp Gly Phe Gly Ala - # Thr Val Trp Glu Val Phe Ser 
Gly 
# 720 
- Val Thr Met Pro Ile Ser Ala Leu - # Asp Pro Ala Lys Lys Leu Gln 
Phe 
# 735 
- Tyr Glu Asp Arg Gln Gln Leu Ser - # Ala Pro Lys Trp Thr Glu Leu 
Ala 
# 750 
- Leu Leu Ile Gln Gln Cys Met Ala - # Tyr Glu Pro Val Gln Arg Pro 
Ser 
# 765 
- Leu Arg Ala Val Ile Arg Asp Leu - # Asn Ser Leu Ile Ser Ser Asp 
Tyr 
# 780 
- Glu Leu Leu Ser Asp His Thr Trp - # Cys Pro Gly Thr Arg Asp Gly 
Leu 
# 800 
- Trp Asn Gly Ala Gln Leu Tyr Ala - # Cys Gln Asp Pro Thr Ile Phe 
Glu 
# 815 
- Glu Arg His Leu Lys Tyr Ile Ser - # Gln Leu Gly Lys Gly Phe Phe 
Gly 
# 830 
- Ser Val Glu Leu Cys Arg Tyr Asp - # Pro Leu Gly Asp Asn Thr Gly 
Ala 
# 845 
- Leu Val Ala Val Lys Gln Leu Gln - # His Ser Gly Pro Asp Gln Gln 
Arg 
# 860 
- Asp Phe Gln Arg Glu Ile Gln Ile - # Leu Lys Ala Gln His Ser Asp 
Phe 
# 880 
- Ile Val Lys Tyr Arg Gly Val Ser - # Tyr Gly Pro Gly Arg Gln Ser 
Pro 
# 895 
- Ala Leu Val Met Glu Tyr Leu Pro - # Ser Gly Cys Leu Arg Asp Phe 
Leu 
# 910 
- Gln Arg His Arg Gly Leu Asp Ala - # Ser Arg Leu Leu Leu Tyr Ser 
Ser 
# 925 
- Gln Ile Cys Lys Gly Met Glu Tyr - # Leu Gly Ser Arg Arg Cys Val 
His 
# 940 
- Arg Asp Leu Ala Ala Arg Asn Ile - # Leu Val Glu Ser Glu Ala His 
Val 
# 960 
- Lys Ile Ala Asp Phe Gly Leu Ala - # Lys Leu Leu Pro Leu Asp Lys 
Asp 
# 975 
- Tyr Tyr Val Val Arg Glu Pro Gly - # Gln Ser Pro Ile Phe Trp Tyr 
Ala 
# 990 
- Pro Glu Ser Leu Ser Asp Asn Ile - # Phe Ser Arg Gln Ser Asp Val 
Trp 
# 10050 
- Ser Phe Gly Val Val Leu Tyr Glu - # Leu Phe Thr Tyr Cys Asp Lys 
Ser 
# 10205 
- Cys Ser Pro Ser Ala Glu Phe Leu - # Arg Met Met Gly Cys Glu Arg 
Asp 
# 10405 
- Val Pro Arg Leu Cys Arg Leu Leu - # Glu Leu Leu Glu Glu Gly Gln 
Arg 
# 10550 
- Leu Pro Ala Pro Pro Cys Cys Pro - # Ala Glu Val Ser Cys Tyr Ser 
Gly 
# 10700 - # 1065 
- Trp Arg Asp Asp Ile Cys Leu Pro - # Ala Glu 
# 1080 
- (2) INFORMATION FOR SEQ ID NO:6: 
- (i) SEQUENCE CHARACTERISTICS: 
#acids (A) LENGTH: 1129 amino 
(B) TYPE: amino acid 
(C) STRANDEDNESS: Not R - #elevant 
(D) TOPOLOGY: linear 
- (ii) MOLECULE TYPE: protein 
- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:6: 
- Met Gly Met Ala Cys Leu Thr Met - # Thr Glu Met Glu Ala Thr Ser 
Thr 
# 15 
- Ser Pro Val His Gln Asn Gly Asp - # Ile Pro Gly Ser Ala Asn Ser 
Val 
# 30 
- Lys Gln Ile Glu Pro Val Leu Gln - # Val Tyr Leu Tyr His Ser Leu 
Gly 
# 45 
- Gln Ala Glu Gly Glu Tyr Leu Lys - # Phe Pro Ser Gly Glu Tyr Val 
Ala 
# 60 
- Glu Glu Ile Cys Val Ala Ala Ser - # Lys Ala Cys Gly Ile Thr Pro 
Val 
# 80 
- Tyr His Asn Met Phe Ala Leu Met - # Ser Glu Thr Glu Arg Ile Trp 
Tyr 
# 95 
- Pro Pro Asn His Val Phe His Ile - # Asp Glu Ser Thr Arg His Asp 
Ile 
# 110 
- Leu Tyr Arg Ile Arg Phe Tyr Phe - # Pro His Trp Tyr Cys Ser Gly 
Ser 
# 125 
- Ser Arg Thr Tyr Arg Tyr Gly Val - # Ser Arg Gly Ala Glu Ala Pro 
Leu 
# 140 
- Leu Asp Asp Phe Val Met Ser Tyr - # Leu Phe Val Gln Trp Arg His 
Asp 
# 160 
- Phe Val His Gly Trp Ile Lys Val - # Pro Val Thr His Glu Thr Gln 
Glu 
# 175 
- Glu Cys Leu Gly Met Ala Val Leu - # Asp Met Met Arg Ile Ala Lys 
Glu 
# 190 
- Lys Asp Gln Thr Pro Leu Ala Val - # Tyr Asn Ser Val Ser Tyr Lys 
Thr 
# 205 
- Phe Leu Pro Lys Cys Val Arg Ala - # Lys Ile Gln Asp Tyr His Ile 
Leu 
# 220 
- Thr Arg Lys Arg Ile Arg Tyr Arg - # Phe Arg Arg Phe Ile Gln Gln 
Phe 
# 240 
- Ser Gln Cys Lys Ala Thr Ala Arg - # Asn Leu Lys Leu Lys Tyr Leu 
Ile 
# 255 
- Asn Leu Glu Thr Leu Gln Ser Ala - # Phe Tyr Thr Glu Gln Phe Glu 
Val 
# 270 
- Lys Glu Ser Ala Arg Gly Pro Ser - # Gly Glu Glu Ile Phe Ala Thr 
Ile 
# 285 
- Ile Ile Thr Gly Asn Gly Gly Ile - # Gln Trp Ser Arg Gly Lys His 
Lys 
# 300 
- Glu Ser Glu Thr Leu Thr Glu Gln - # Asp Val Gln Leu Tyr Cys Asp 
Phe 
# 320 
- Pro Asp Ile Ile Asp Val Ser Ile - # Lys Gln Ala Asn Gln Glu Cys 
Ser 
# 335 
- Asn Glu Ser Arg Ile Val Thr Val - # His Lys Gln Asp Gly Lys Val 
Leu 
# 350 
- Glu Ile Glu Leu Ser Ser Leu Lys - # Glu Ala Leu Ser Phe Val Ser 
Leu 
# 365 
- Ile Asp Gly Tyr Tyr Arg Leu Thr - # Ala Asp Ala His His Tyr Leu 
Cys 
# 380 
- Lys Glu Val Ala Pro Pro Ala Val - # Leu Glu Asn Ile His Ser Asn 
Cys 
# 400 
- His Gly Pro Ile Ser Met Asp Phe - # Ala Ile Ser Lys Leu Lys Lys 
Ala 
# 415 
- Gly Asn Gln Thr Gly Leu Tyr Val - # Leu Arg Cys Ser Pro Lys Asp 
Phe 
# 430 
- Asn Lys Tyr Phe Leu Thr Phe Ala - # Val Glu Arg Glu Asn Val Ile 
Glu 
# 445 
- Tyr Lys His Cys Leu Ile Thr Lys - # Asn Glu Asn Gly Glu Tyr Asn 
Leu 
# 460 
- Ser Gly Thr Asn Arg Asn Phe Ser - # Asn Leu Lys Asp Leu Leu Asn 
Cys 
# 480 
- Tyr Gln Met Glu Thr Val Arg Ser - # Asp Ser Ile Ile Phe Gln Phe 
Thr 
# 495 
- Lys Cys Cys Pro Pro Lys Pro Lys - # Asp Lys Ser Asn Leu Leu Val 
Phe 
# 510 
- Arg Thr Asn Gly Ile Ser Asp Val - # Gln Ile Ser Pro Thr Leu Gln 
Arg 
# 525 
- His Asn Asn Val Asn Gln Met Val - # Phe His Lys Ile Arg Asn Glu 
Asp 
# 540 
- Leu Ile Phe Asn Glu Ser Leu Gly - # Gln Gly Thr Phe Thr Lys Ile 
Phe 
# 560 
- Lys Gly Val Arg Arg Glu Val Gly - # Asp Tyr Gly Gln Leu His Lys 
Thr 
# 575 
- Glu Val Leu Leu Lys Val Leu Asp - # Lys Ala His Arg Asn Tyr Ser 
Glu 
# 590 
- Ser Phe Phe Glu Ala Ala Ser Met - # Met Ser Gln Leu Ser His Lys 
His 
# 605 
- Leu Val Leu Asn Tyr Gly Val Cys - # Val Cys Gly Glu Glu Asn Ile 
Leu 
# 620 
- Val Gln Glu Phe Val Lys Phe Gly - # Ser Leu Asp Thr Tyr Leu Lys 
Lys 
# 640 
- Asn Lys Asn Ser Ile Asn Ile Leu - # Trp Lys Leu Gly Val Ala Lys 
Gln 
# 655 
- Leu Ala Trp Ala Met His Phe Leu - # Glu Glu Lys Ser Leu Ile His 
Gly 
# 670 
- Asn Val Cys Ala Lys Asn Ile Leu - # Leu Ile Arg Glu Glu Asp Arg 
Arg 
# 685 
- Thr Gly Asn Pro Pro Phe Ile Lys - # Leu Ser Asp Pro Gly Ile Ser 
Ile 
# 700 
- Thr Val Leu Pro Lys Asp Ile Leu - # Gln Glu Arg Ile Pro Trp Val 
Pro 
# 720 
- Pro Glu Cys Ile Glu Asn Pro Lys - # Asn Leu Asn Leu Ala Thr Asp 
Lys 
# 735 
- Trp Ser Phe Gly Thr Thr Leu Trp - # Glu Ile Cys Ser Gly Gly Asp 
Lys 
# 750 
- Pro Leu Ser Ala Leu Asp Ser Gln - # Arg Lys Leu Gln Phe Tyr Glu 
Asp 
# 765 
- Lys His Gln Leu Pro Ala Pro Lys - # Trp Thr Glu Leu Ala Asn Leu 
Ile 
# 780 
- Asn Asn Cys Met Asp Tyr Glu Pro - # Asp Phe Arg Pro Ala Phe Arg 
Ala 
# 800 
- Val Ile Arg Asp Leu Asn Ser Leu - # Phe Thr Pro Asp Tyr Glu Leu 
Leu 
# 815 
- Thr Glu Asn Asp Met Leu Pro Asn - # Met Arg Ile Gly Ala Leu Gly 
Phe 
# 830 
- Ser Gly Ala Phe Glu Asp Arg Asp - # Pro Thr Gln Phe Glu Glu Arg 
His 
# 845 
- Leu Lys Phe Leu Gln Gln Leu Gly - # Lys Gly Asn Phe Gly Ser Val 
Glu 
# 860 
- Met Cys Arg Tyr Asp Pro Leu Gln - # Asp Asn Thr Gly Glu Val Val 
Ala 
# 880 
- Val Lys Lys Leu Gln His Ser Thr - # Glu Glu His Leu Arg Asp Phe 
Glu 
# 895 
- Arg Glu Ile Glu Ile Leu Lys Ser - # Leu Gln His Asp Asn Ile Val 
Lys 
# 910 
- Tyr Lys Gly Val Cys Tyr Ser Ala - # Gly Arg Arg Asn Leu Arg Leu 
Ile 
# 925 
- Met Glu Tyr Leu Pro Tyr Gly Ser - # Leu Arg Asp Tyr Leu Gln Lys 
His 
# 940 
- Lys Glu Arg Ile Asp His Lys Lys - # Leu Leu Gln Tyr Thr Ser Gln 
Ile 
# 960 
- Cys Lys Gly Met Glu Tyr Leu Gly - # Thr Lys Arg Tyr Ile His Arg 
Asp 
# 975 
- Leu Ala Thr Arg Asn Ile Leu Val - # Glu Asn Glu Asn Arg Val Lys 
Ile 
# 990 
- Gly Asp Phe Gly Leu Thr Lys Val - # Leu Pro Gln Asp Lys Glu Tyr 
Tyr 
# 10050 
- Lys Val Lys Glu Pro Gly Glu Ser - # Pro Ile Phe Trp Tyr Ala Pro 
Gln 
# 10205 
- Ser Leu Thr Glu Ser Lys Phe Ser - # Val Ala Ser Asp Val Trp Ser 
Phe 
# 10405 
- Gly Val Val Leu Tyr Glu Leu Phe - # Thr Tyr Ile Glu Lys Ser Lys 
Ser 
# 10550 
- Pro Pro Val Glu Phe Met Arg Met - # Ile Gly Asn Asp Lys Gln Gly 
Gln 
# 10700 - # 1065 
- Met Ile Val Phe His Leu Ile Glu - # Leu Leu Lys Ser Asn Gly Arg 
Leu 
# 10850 
- Pro Arg Pro Glu Gly Cys Pro Asp - # Glu Ile Tyr Val Ile Met Thr 
Glu 
# 11005 
- Cys Trp Asn Asn Asn Val Ser Gln - # Arg Pro Ser Phe Arg Asp Leu 
Ser 
# 11205 
- Phe Gly Trp Ile Lys Cys Gly Thr - # Val 
# 1125 
- (2) INFORMATION FOR SEQ ID NO:7: 
- (i) SEQUENCE CHARACTERISTICS: 
#acids (A) LENGTH: 1154 amino 
(B) TYPE: amino acid 
(C) STRANDEDNESS: Not R - #elevant 
(D) TOPOLOGY: linear 
- (ii) MOLECULE TYPE: protein 
- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:7: 
- Met Gln Tyr Leu Asn Ile Lys Glu - # Asp Cys Asn Ala Met Ala Phe 
Cys 
# 15 
- Ala Lys Met Arg Ser Ser Lys Lys - # Thr Glu Val Asn Leu Glu Ala 
Pro 
# 30 
- Glu Pro Gly Val Glu Val Ile Phe - # Tyr Leu Ser Asp Arg Glu Pro 
Leu 
# 45 
- Arg Leu Gly Ser Gly Glu Tyr Thr - # Ala Glu Glu Leu Cys Ile Arg 
Ala 
# 60 
- Ala Gln Ala Cys Arg Ile Ser Pro - # Leu Cys His Asn Leu Phe Ala 
Leu 
# 80 
- Tyr Asp Glu Asn Thr Lys Leu Trp - # Tyr Ala Pro Asn Arg Thr Ile 
Thr 
# 95 
- Val Asp Asp Lys Met Ser Leu Arg - # Leu His Tyr Arg Met Arg Phe 
Tyr 
# 110 
- Phe Thr Asn Trp His Gly Thr Asn - # Asp Asn Glu Gln Ser Val Trp 
Arg 
# 125 
- His Ser Pro Lys Lys Gln Lys Asn - # Gly Tyr Glu Lys Lys Lys Ile 
Pro 
# 140 
- Asp Ala Thr Pro Leu Leu Asp Ala - # Ser Ser Leu Glu Tyr Leu Phe 
Ala 
# 160 
- Gln Gly Gln Tyr Asp Leu Val Lys - # Cys Leu Ala Pro Ile Arg Asp 
Pro 
# 175 
- Lys Thr Glu Gln Asp Gly His Asp - # Ile Glu Asn Glu Cys Leu Gly 
Met 
# 190 
- Ala Val Leu Ala Ile Ser His Tyr - # Ala Met Met Lys Lys Met Gln 
Leu 
# 205 
- Pro Glu Leu Pro Lys Asp Ile Ser - # Tyr Lys Arg Tyr Ile Pro Glu 
Thr 
# 220 
- Leu Asn Lys Ser Ile Arg Gln Arg - # Asn Leu Leu Thr Arg Met Arg 
Ile 
# 240 
- Asn Asn Val Phe Lys Asp Phe Leu - # Lys Glu Phe Asn Asn Lys Thr 
Ile 
# 255 
- Cys Asp Ser Ser Val Ser Thr His - # Asp Leu Lys Val Lys Tyr Leu 
Ala 
# 270 
- Thr Leu Glu Thr Leu Thr Lys His - # Tyr Gly Ala Glu Ile Phe Glu 
Thr 
# 285 
- Ser Met Leu Leu Ile Ser Ser Glu - # Asn Glu Met Asn Trp Phe His 
Ser 
# 300 
- Asn Asp Gly Gly Asn Val Leu Tyr - # Tyr Glu Val Met Val Thr Gly 
Asn 
# 320 
- Leu Gly Ile Gln Trp Arg His Lys - # Pro Asn Val Val Ser Val Glu 
Lys 
# 335 
- Glu Lys Asn Lys Leu Lys Arg Lys - # Lys Leu Glu Asn Lys Asp Lys 
Lys 
# 350 
- Asp Glu Glu Lys Asn Lys Ile Arg - # Glu Glu Trp Asn Asn Phe Ser 
Phe 
# 365 
- Phe Pro Glu Ile Thr His Ile Val - # Ile Lys Glu Ser Val Val Ser 
Ile 
# 380 
- Asn Lys Gln Asp Asn Lys Lys Met - # Glu Leu Lys Leu Ser Ser His 
Glu 
# 400 
- Glu Ala Leu Ser Phe Val Ser Leu - # Val Asp Gly Tyr Phe Arg Leu 
Thr 
# 415 
- Ala Asp Ala His His Tyr Leu Cys - # Thr Asp Val Ala Pro Pro Leu 
Ile 
# 430 
- Val His Asn Ile Gln Asn Gly Cys - # His Gly Pro Ile Cys Thr Glu 
Tyr 
# 445 
- Ala Ile Asn Lys Leu Arg Gln Glu - # Gly Ser Glu Glu Gly Met Tyr 
Val 
# 460 
- Leu Arg Trp Ser Cys Thr Asp Phe - # Asp Asn Ile Leu Met Thr Val 
Thr 
# 480 
- Cys Phe Glu Lys Ser Glu Gln Val - # Gln Gly Ala Gln Lys Gln Phe 
Lys 
# 495 
- Asn Phe Gln Ile Glu Val Gln Lys - # Gly Arg Tyr Ser Leu His Gly 
Ser 
# 510 
- Asp Arg Ser Phe Pro Ser Leu Gly - # Asp Leu Met Ser His Leu Lys 
Lys 
# 525 
- Gln Ile Leu Arg Thr Asp Asn Ile - # Ser Phe Met Leu Lys Arg Cys 
Cys 
# 540 
- Gln Pro Lys Pro Arg Glu Ile Ser - # Asn Leu Leu Val Ala Thr Lys 
Lys 
# 560 
- Ala Gln Glu Trp Gln Pro Val Tyr - # Pro Met Ser Gln Leu Ser Phe 
Asp 
# 575 
- Arg Ile Leu Lys Lys Asp Leu Val - # Gln Gly Glu His Leu Gly Arg 
Gly 
# 590 
- Thr Arg Thr His Ile Tyr Ser Gly - # Thr Leu Met Asp Tyr Lys Asp 
Asp 
# 605 
- Glu Gly Thr Ser Glu Glu Lys Lys - # Ile Lys Val Ile Leu Lys Val 
Leu 
# 620 
- Asp Pro Ser His Arg Asp Ile Ser - # Leu Ala Phe Phe Glu Ala Ala 
Ser 
# 640 
- Met Met Arg Gln Val Ser His Lys - # His Ile Val Tyr Leu Tyr Gly 
Val 
# 655 
- Cys Val Arg Asp Val Glu Asn Ile - # Met Val Glu Glu Phe Val Glu 
Gly 
# 670 
- Gly Pro Leu Asp Leu Phe Met His - # Arg Lys Ser Asp Val Leu Thr 
Thr 
# 685 
- Pro Trp Lys Phe Lys Val Ala Lys - # Gln Leu Ala Ser Ala Leu Ser 
Tyr 
# 700 
- Leu Glu Asp Lys Asp Leu Val His - # Gly Asn Val Cys Thr Lys Asn 
Leu 
# 720 
- Leu Leu Ala Arg Glu Gly Ile Asp - # Ser Glu Cys Gly Pro Phe Ile 
Lys 
# 735 
- Leu Ser Asp Pro Gly Ile Pro Ile - # Thr Val Leu Ser Arg Gln Glu 
Cys 
# 750 
- Ile Glu Arg Ile Pro Trp Ile Ala - # Pro Glu Cys Val Glu Asp Ser 
Lys 
# 765 
- Asn Leu Ser Val Ala Ala Asp Lys - # Trp Ser Phe Gly Thr Thr Leu 
Trp 
# 780 
- Glu Ile Cys Tyr Asn Gly Glu Ile - # Pro Leu Lys Asp Lys Thr Leu 
Ile 
# 800 
- Glu Lys Glu Arg Phe Tyr Glu Ser - # Arg Cys Arg Pro Val Thr Pro 
Ser 
# 815 
- Cys Lys Glu Leu Ala Asp Leu Met - # Thr Arg Cys Met Asn Tyr Asp 
Pro 
# 830 
- Asn Gln Arg Pro Phe Phe Arg Ala - # Ile Met Arg Asp Ile Asn Lys 
Leu 
# 845 
- Glu Glu Gln Asn Pro Asp Ile Val - # Ser Arg Lys Lys Asn Gln Pro 
Thr 
# 860 
- Glu Val Asp Pro Thr His Phe Glu - # Lys Arg Phe Leu Lys Arg Ile 
Arg 
# 880 
- Asp Leu Gly Glu Gly His Phe Gly - # Lys Val Glu Leu Cys Arg Tyr 
Asp 
# 895 
- Pro Glu Asp Asn Thr Gly Glu Gln - # Val Ala Val Lys Ser Leu Lys 
Pro 
# 910 
- Glu Ser Gly Gly Asn His Ile Ala - # Asp Leu Lys Lys Glu Ile Glu 
Ile 
# 925 
- Leu Arg Asn Leu Tyr His Glu Asn - # Ile Val Lys Tyr Lys Gly Ile 
Cys 
# 940 
- Thr Glu Asp Gly Gly Asn Gly Ile - # Lys Leu Ile Met Glu Phe Leu 
Pro 
# 960 
- Ser Gly Ser Leu Lys Glu Tyr Leu - # Pro Lys Asn Lys Asn Lys Ile 
Asn 
# 975 
- Leu Lys Gln Gln Leu Lys Tyr Ala - # Val Gln Ile Cys Lys Gly Met 
Asp 
# 990 
- Tyr Leu Gly Ser Arg Gln Tyr Val - # His Arg Asp Leu Ala Ala Arg 
Asn 
# 10050 
- Val Leu Val Glu Ser Glu His Gln - # Val Lys Ile Gly Asp Phe Gly 
Leu 
# 10205 
- Thr Lys Ala Ile Glu Thr Asp Lys - # Glu Tyr Tyr Thr Val Lys Asp 
Asp 
# 10405 
- Arg Asp Ser Pro Val Phe Trp Tyr - # Ala Pro Glu Cys Leu Met Gln 
Ser 
# 10550 
- Lys Phe Tyr Ile Ala Ser Asp Val - # Trp Ser Phe Gly Val Thr Leu 
His 
# 10700 - # 1065 
- Glu Leu Leu Thr Tyr Cys Asp Ser - # Asp Ser Ser Pro Met Ala Leu 
Phe 
# 10850 
- Leu Lys Met Ile Gly Pro Thr His - # Gly Gln Met Thr Val Thr Arg 
Leu 
# 11005 
- Val Asn Thr Leu Lys Glu Gly Lys - # Arg Leu Pro Cys Pro Pro Asn 
Cys 
# 11205 
- Pro Asp Glu Val Tyr Gln Leu Met - # Arg Lys Cys Trp Glu Phe Gln 
Pro 
# 11350 
- Ser Asn Arg Thr Ser Phe Gln Asn - # Leu Ile Glu Gly Phe Glu Ala 
Leu 
# 11500 - # 1145 
- Leu Lys 
- (2) INFORMATION FOR SEQ ID NO:8: 
- (i) SEQUENCE CHARACTERISTICS: 
#acids (A) LENGTH: 1187 amino 
(B) TYPE: amino acid 
(C) STRANDEDNESS: Not R - #elevant 
(D) TOPOLOGY: linear 
- (ii) MOLECULE TYPE: protein 
- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:8: 
- Met Pro Leu Arg His Trp Gly Met - # Ala Arg Gly Ser Lys Pro Val 
Gly 
# 15 
- Asp Gly Ala Gln Pro Met Ala Ala - # Met Gly Gly Leu Lys Val Leu 
Leu 
# 30 
- His Trp Ala Gly Pro Gly Gly Gly - # Glu Pro Trp Val Thr Phe Ser 
Glu 
# 45 
- Ser Ser Leu Thr Ala Glu Glu Val - # Cys Ile His Ile Ala His Lys 
Val 
# 60 
- Gly Ile Thr Pro Pro Cys Phe Asn - # Leu Phe Ala Leu Phe Asp Ala 
Gln 
# 80 
- Ala Gln Val Trp Leu Pro Pro Asn - # His Ile Leu Glu Ile Pro Arg 
Asp 
# 95 
- Ala Ser Leu Met Leu Tyr Phe Arg - # Ile Arg Phe Tyr Phe Arg Asn 
Trp 
# 110 
- His Gly Met Asn Pro Arg Glu Pro - # Ala Val Tyr Arg Cys Gly Pro 
Pro 
# 125 
- Gly Thr Glu Ala Ser Ser Asp Gln - # Thr Ala Gln Gly Met Gln Leu 
Leu 
# 140 
- Asp Pro Ala Ser Phe Glu Tyr Leu - # Phe Glu Gln Gly Lys His Glu 
Phe 
# 160 
- Val Asn Asp Val Ala Ser Leu Trp - # Glu Leu Ser Thr Glu Glu Glu 
Ile 
# 175 
- His His Phe Lys Asn Glu Ser Leu - # Gly Met Ala Phe Leu His Leu 
Cys 
# 190 
- His Leu Ala Leu Arg His Gly Ile - # Pro Leu Glu Glu Val Ala Lys 
Lys 
# 205 
- Thr Ser Phe Lys Asp Cys Ile Pro - # Arg Ser Phe Arg Arg His Ile 
Arg 
# 220 
- Gln His Ser Ala Leu Thr Arg Leu - # Arg Leu Arg Asn Val Phe Arg 
Arg 
# 240 
- Phe Leu Arg Asp Phe Gln Pro Gly - # Arg Leu Ser Gln Gln Met Val 
Met 
# 255 
- Val Lys Tyr Leu Ala Thr Leu Glu - # Arg Leu Ala Pro Arg Phe Gly 
Thr 
# 270 
- Glu Arg Val Pro Val Cys His Leu - # Arg Leu Leu Ala Gln Ala Glu 
Gly 
# 285 
- Glu Pro Cys Tyr Ile Arg Asp Ser - # Gly Val Ala Pro Thr Asp Pro 
Gly 
# 300 
- Pro Glu Ser Ala Ala Gly Pro Pro - # Thr His Glu Val Leu Val Thr 
Gly 
# 320 
- Thr Gly Gly Ile Gln Trp Trp Pro - # Val Glu Glu Glu Val Asn Lys 
Glu 
# 335 
- Glu Gly Ser Ser Gly Ser Ser Gly - # Arg Asn Pro Gln Ala Ser Leu 
Phe 
# 350 
- Gly Lys Lys Ala Lys Ala His Lys - # Ala Phe Gly Gln Pro Ala Asp 
Arg 
# 365 
- Pro Arg Glu Pro Leu Trp Ala Tyr - # Phe Cys Asp Phe Arg Asp Ile 
Thr 
# 380 
- His Val Val Leu Lys Glu His Cys - # Val Ser Ile His Arg Gln Asp 
Asn 
# 400 
- Lys Cys Leu Glu Leu Ser Leu Pro - # Ser Arg Ala Ala Ala Leu Ser 
Phe 
# 415 
- Val Ser Leu Val Asp Gly Tyr Phe - # Arg Leu Thr Ala Asp Ser Ser 
His 
# 430 
- Tyr Leu Cys His Glu Val Ala Pro - # Pro Arg Leu Val Met Ser Ile 
Arg 
# 445 
- Asp Gly Ile His Gly Pro Leu Leu - # Glu Pro Phe Val Gln Ala Lys 
Leu 
# 460 
- Arg Pro Glu Asp Gly Leu Tyr Leu - # Ile His Trp Ser Thr Ser His 
Pro 
# 480 
- Tyr Arg Leu Ile Leu Thr Val Ala - # Gln Arg Ser Gln Ala Pro Asp 
Gly 
# 495 
- Met Gln Ser Leu Arg Leu Arg Lys - # Phe Pro Ile Glu Gln Gln Asp 
Gly 
# 510 
- Ala Phe Val Leu Glu Gly Trp Gly - # Arg Ser Phe Pro Ser Val Arg 
Glu 
# 525 
- Leu Gly Ala Ala Leu Gln Gly Cys - # Leu Leu Arg Ala Gly Asp Asp 
Cys 
# 540 
- Phe Ser Leu Arg Arg Cys Cys Leu - # Pro Gln Pro Gly Glu Thr Ser 
Asn 
# 560 
- Leu Ile Ile Met Arg Gly Ala Arg - # Ala Ser Pro Arg Thr Leu Asn 
Leu 
# 575 
- Ser Gln Leu Ser Phe His Arg Val - # Asp Gln Lys Glu Ile Thr Gln 
Leu 
# 590 
- Ser His Leu Gly Gln Gly Thr Arg - # Thr Asn Val Tyr Glu Gly Arg 
Leu 
# 605 
- Arg Val Glu Gly Ser Gly Asp Pro - # Glu Glu Gly Lys Met Asp Asp 
Glu 
# 620 
- Asp Pro Leu Val Pro Gly Arg Asp - # Arg Gly Gln Glu Leu Arg Val 
Val 
# 640 
- Leu Lys Val Leu Asp Pro Ser His - # His Asp Ile Ala Leu Ala Phe 
Tyr 
# 655 
- Glu Thr Ala Ser Leu Met Ser Gln - # Val Ser His Thr His Leu Ala 
Phe 
# 670 
- Val His Gly Val Cys Val Arg Gly - # Pro Glu Asn Ser Met Val Thr 
Glu 
# 685 
- Tyr Val Glu His Gly Pro Leu Asp - # Val Trp Leu Arg Arg Glu Arg 
Gly 
# 700 
- His Val Pro Met Ala Trp Lys Met - # Val Val Ala Gln Gln Leu Ala 
Ser 
# 720 
- Ala Leu Ser Tyr Leu Glu Asn Lys - # Asn Leu Val His Gly Asn Val 
Cys 
# 735 
- Gly Arg Asn Ile Leu Leu Ala Arg - # Leu Gly Leu Ala Glu Gly Thr 
Ser 
# 750 
- Pro Phe Ile Lys Leu Ser Asp Pro - # Gly Val Gly Leu Gly Ala Leu 
Ser 
# 765 
- Arg Glu Glu Arg Val Glu Arg Ile - # Pro Trp Leu Ala Pro Glu Cys 
Leu 
# 780 
- Pro Gly Gly Ala Asn Ser Leu Ser - # Thr Ala Met Asp Lys Trp Gly 
Phe 
# 800 
- Gly Ala Thr Leu Leu Glu Ile Cys - # Phe Asp Gly Glu Ala Pro Leu 
Gln 
# 815 
- Ser Arg Ser Pro Ser Glu Lys Glu - # His Phe Tyr Gln Arg Gln His 
Arg 
# 830 
- Leu Pro Glu Pro Ser Cys Pro Gln - # Leu Ala Thr Leu Thr Ser Gln 
Cys 
# 845 
- Leu Thr Tyr Glu Pro Thr Gln Arg - # Pro Ser Phe Arg Thr Ile Leu 
Arg 
# 860 
- Asp Leu Thr Arg Val Gln Pro His - # Asn Leu Ala Asp Val Leu Thr 
Val 
# 880 
- Asn Arg Asp Ser Pro Ala Val Gly - # Pro Thr Thr Phe His Lys Arg 
Tyr 
# 895 
- Leu Lys Lys Ile Arg Asp Leu Gly - # Glu Gly His Phe Gly Lys Val 
Ser 
# 910 
- Leu Tyr Cys Tyr Asp Pro Thr Asn - # Asp Gly Thr Gly Glu Met Val 
Ala 
# 925 
- Val Lys Ala Leu Lys Ala Asp Cys - # Gly Pro Gln His Arg Ser Gly 
Trp 
# 940 
- Lys Gln Glu Ile Asp Ile Leu Arg - # Thr Leu Tyr His Glu His Ile 
Ile 
# 960 
- Lys Tyr Lys Gly Cys Cys Glu Asp - # Gln Gly Glu Lys Ser Leu Gln 
Leu 
# 975 
- Val Met Glu Tyr Val Pro Leu Gly - # Ser Leu Arg Asp Tyr Leu Pro 
Arg 
# 990 
- His Ser Ile Gly Leu Ala Gln Leu - # Leu Leu Phe Ala Gln Gln Ile 
Cys 
# 10050 
- Glu Gly Met Ala Tyr Leu His Ala - # His Asp Tyr Ile His Arg Asp 
Leu 
# 10205 
- Ala Ala Arg Asn Val Leu Leu Asp - # Asn Asp Arg Leu Val Lys Ile 
Gly 
# 10405 
- Asp Phe Gly Leu Ala Lys Ala Val - # Pro Glu Gly His Glu Tyr Tyr 
Arg 
# 10550 
- Val Arg Glu Asp Gly Asp Ser Pro - # Val Phe Trp Tyr Ala Pro Glu 
Cys 
# 10700 - # 1065 
- Leu Lys Glu Tyr Lys Phe Tyr Tyr - # Ala Ser Asp Val Trp Ser Phe 
Gly 
# 10850 
- Val Thr Leu Tyr Glu Leu Leu Thr - # His Cys Asp Ser Ser Gln Ser 
Pro 
# 11005 
- Pro Thr Lys Phe Leu Glu Leu Ile - # Gly Ile Ala Gln Gly Gln Met 
Thr 
# 11205 
- Val Leu Arg Leu Thr Glu Leu Leu - # Glu Arg Gly Glu Arg Leu Pro 
Arg 
# 11350 
- Pro Asp Lys Cys Pro Cys Glu Val - # Tyr His Leu Met Lys Asn Cys 
Trp 
# 11500 - # 1145 
- Glu Thr Glu Ala Ser Phe Arg Pro - # Thr Phe Glu Asn Leu Ile Pro 
Ile 
# 11650 
- Leu Lys Thr Val His Glu Lys Tyr - # Gln Gly Gln Ala Pro Ser Val 
Phe 
# 11805 
- Ser Val Cys 
1185 
- (2) INFORMATION FOR SEQ ID NO:9: 
- (i) SEQUENCE CHARACTERISTICS: 
#acids (A) LENGTH: 498 amino 
(B) TYPE: amino acid 
(C) STRANDEDNESS: Not R - #elevant 
(D) TOPOLOGY: linear 
- (ii) MOLECULE TYPE: protein 
- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:9: 
- Met Pro Leu Gly Ala Glu Glu Cys - # Ala Ala Ile Pro His Asn Leu 
Phe 
# 15 
- Ala Leu Trp Pro Pro Asn Leu Tyr - # Arg Ile Arg Phe Tyr Phe Asn 
Trp 
# 30 
- Gly Leu Leu Asp Glu Tyr Leu Phe - # Gln Asp Val Glu Gln Glu Cys 
Leu 
# 45 
- Gly Met Ala Val Leu Ala Glu Ser - # Tyr Lys Pro Arg Ile Leu Thr 
Arg 
# 60 
- Arg Ile Arg Phe Phe Leu Phe Leu - # Lys Lys Tyr Leu Leu Glu Leu 
Glu 
# 80 
- Phe Val Val Val Thr Gly Gly Gly - # Ile Gln Trp Glu Phe Cys Asp 
Phe 
# 95 
- Pro Ile Ile Lys Val Gln Asp Asn - # Lys Leu Glu Leu Ser Glu Ala 
Leu 
# 110 
- Ser Phe Val Ser Leu Val Asp Gly - # Tyr Phe Arg Leu Thr Ala Asp 
His 
# 125 
- Tyr Leu Cys Val Ala Pro Pro Ile - # Cys His Gly Pro Ile Phe Ala 
Ile 
# 140 
- Lys Leu Gly Tyr Val Leu Arg Trp - # Ser Asp Phe Leu Thr Val Val 
Lys 
# 160 
- Ile Gly Leu Gly Arg Phe Ser Leu - # Arg Phe Cys Cys Pro Pro Ser 
Asn 
# 175 
- Leu Leu Val Gln Ser Gln Phe His - # Ile Leu Glu Leu Gly Gly Thr 
Thr 
# 190 
- Ile Tyr Gly Asp Val Leu Lys Val - # Leu Asp His Phe Glu Ala Ala 
Ser 
# 205 
- Met Ser Gln Val Ser His His Leu - # Val Gly Val Cys Val Glu Asn 
Val 
# 220 
- Glu Phe Val Gly Leu Asp Arg Trp - # Lys Val Ala Lys Gln Leu Ala 
Ala 
# 240 
- Leu Tyr Leu Glu Asp Leu Leu His - # Gly Asn Val Cys Asn Ile Leu 
Leu 
# 255 
- Ala Arg Glu Gly Pro Phe Ile Lys - # Leu Ser Asp Pro Gly Val Leu 
Ser 
# 270 
- Glu Arg Ile Pro Trp Ala Pro Glu - # Cys Asn Leu Ser Ala Asp Lys 
Trp 
# 285 
- Phe Gly Thr Leu Trp Glu Cys Gly - # Pro Leu Lys Phe Tyr Glu Leu 
Pro 
# 300 
- Glu Leu Ala Leu Cys Met Tyr Glu - # Pro Gln Arg Pro Phe Arg Ala 
Arg 
# 320 
- Asp Leu Asn Leu Pro Asp Pro Thr - # Phe Glu Arg Leu Lys Ile Leu 
Gly 
# 335 
- Gly Phe Gly Val Glu Leu Cys Arg - # Tyr Asp Pro Asp Asn Thr Gly 
Glu 
# 350 
- Val Ala Val Lys Leu Ser Gly His - # Asp Glu Ile Ile Leu Leu His 
Ile 
# 365 
- Val Lys Tyr Lys Gly Cys Gly Leu - # Met Glu Tyr Leu Pro Gly Ser 
Leu 
# 380 
- Arg Asp Tyr Leu Ile Leu Leu Leu - # Tyr Leu Leu Gln Ile Cys Lys 
Gly 
# 400 
- Met Tyr Leu Gly Tyr His Arg Asp - # Leu Ala Ala Arg Asn Leu Val 
Glu 
# 415 
- Glu Val Lys Ile Gly Asp Phe Gly - # Leu Lys Pro Asp Lys Glu Tyr 
Tyr 
# 430 
- Val Glu Gly Ser Pro Phe Trp Tyr - # Ala Pro Glu Leu Leu Phe Ala 
Ser 
# 445 
- Asp Val Trp Ser Phe Gly Val Leu - # Tyr Glu Leu Thr Tyr Cys Asp 
Ser 
# 460 
- Ser Pro Phe Leu Met Ile Gly Gly - # Gln Met Val Arg Leu Glu Leu 
Leu 
# 480 
- Gly Arg Leu Pro Pro Cys Pro Glu - # Val Tyr Leu Met Cys Trp Ser 
Arg 
# 495 
- Phe Leu 
- (2) INFORMATION FOR SEQ ID NO:10: 
- (i) SEQUENCE CHARACTERISTICS: 
#acids (A) LENGTH: 1082 amino 
(B) TYPE: amino acid 
(C) STRANDEDNESS: Not R - #elevant 
(D) TOPOLOGY: linear 
- (ii) MOLECULE TYPE: protein 
- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:10: 
- Met Ala Pro Pro Ser Glu Glu Thr - # Pro Leu Ile Pro Gln Arg Ser 
Cys 
# 15 
- Ser Leu Leu Ser Thr Glu Ala Gly - # Ala Leu His Val Leu Leu Pro 
Ala 
# 30 
- Arg Gly Pro Gly Pro Pro Gln Arg - # Leu Ser Phe Ser Phe Gly Asp 
His 
# 45 
- Leu Ala Glu Asp Leu Cys Val Gln - # Ala Ala Lys Ala Ser Ala Ile 
Leu 
# 60 
- Pro Val Tyr His Ser Leu Phe Ala - # Leu Ala Thr Glu Asp Leu Ser 
Cys 
# 80 
- Trp Phe Pro Arg Ala Thr Ser Ser - # Pro Trp Arg Met Pro Ala Pro 
Gln 
# 95 
- Val Leu Leu Tyr Arg Ile Arg Phe - # Tyr Phe Pro Asn Trp Phe Gly 
Leu 
# 110 
- Glu Lys Cys His Arg Phe Gly Leu - # Arg Lys Asp Leu Ala Ser Ala 
Ile 
# 125 
- Leu Asp Leu Pro Val Leu Glu His - # Leu Phe Ala Gln His Arg Ser 
Asp 
# 140 
- Leu Val Ser Gly Arg Leu Pro Arg - # Gly Leu Ser Leu Lys Glu Gln 
Gly 
# 160 
- Glu Cys Leu Ser Leu Ala Val Leu - # Asp Leu Ala Arg Met Ala Arg 
Glu 
# 175 
- Gln Ala Gln Arg Arg Gly Glu Leu - # Leu Lys Thr Val Ser Tyr Lys 
Ala 
# 190 
- Cys Leu Pro Pro Ser Leu Arg Asp - # Leu Ile Gln Gly Leu Ser Phe 
Val 
# 205 
- Thr Gly Arg Arg Ile Arg Arg Thr - # Val Glu Ser Pro Leu Arg Arg 
Val 
# 220 
- Ala Ala Cys Gln Ala Asp Arg His - # Ser Leu Met Ala Lys Tyr Ile 
Met 
# 240 
- Asp Leu Glu Arg Leu Asp Pro Ala - # Gly Ala Ala Glu Thr Phe His 
Val 
# 255 
- Gly Leu Pro Gly Ala Leu Gly Gly - # His Asp Gly Leu Gly Leu Val 
Arg 
# 270 
- Val Ala Gly Asp Gly Gly Ile Ala - # Trp Thr Gln Gly Glu Gln Glu 
Val 
# 285 
- Leu Gln Pro Phe Cys Asp Phe Pro - # Glu Ile Val Asp Ile Ser Ile 
Lys 
# 300 
- Gln Ala Pro Arg Val Gly Pro Ala - # Gly Glu His Arg Leu Val Thr 
Val 
# 320 
- Thr Arg Thr Asp Asn Gln Ile Leu - # Glu Ala Glu Phe Pro Gly Leu 
Pro 
# 335 
- Glu Ala Leu Ser Phe Val Ala Leu - # Val Asp Gly Tyr Phe Arg Leu 
Thr 
# 350 
- Thr Asp Ser Gln His Phe Phe Cys - # Lys Glu Val Asp Pro Arg Leu 
Leu 
# 365 
- Glu Glu Val Ala Glu Gln Cys His - # Gly Pro Ile Thr Leu Asp Phe 
Ala 
# 380 
- Ile Asn Lys Leu Lys Thr Gly Gly - # Ser Arg Pro Gly Ser Tyr Val 
Leu 
# 400 
- Arg Arg Ile Pro Gln Asp Phe Asp - # Ser Phe Leu Leu Thr Val Cys 
Val 
# 415 
- Gln Asn Pro Leu Gly Pro Asp Tyr - # Lys Gly Cys Leu Ile Arg Arg 
Ser 
# 430 
- Pro Thr Gly Thr Phe Leu Leu Val - # Gly Leu Ser Arg Pro His Ser 
Ser 
# 445 
- Leu Arg Glu Leu Leu Ala Thr Cys - # Trp Asp Gly Gly Leu His Val 
Asp 
# 460 
- Gly Val Ala Val Thr Leu Thr Ser - # Cys Cys Ile Pro Arg Pro Lys 
Glu 
# 480 
- Lys Ser Asn Leu Ile Val Val Gln - # Arg Gly His Ser Pro Pro Thr 
Ser 
# 495 
- Ser Leu Val Gln Pro Gln Ser Gln - # Tyr Gln Leu Ser Gln Met Thr 
Phe 
# 510 
- His Lys Ile Pro Ala Asp Ser Leu - # Glu Trp His Glu Asn Leu Gly 
His 
# 525 
- Gly Ser Phe Thr Lys Ile Tyr Arg - # Gly Cys Arg His Glu Val Val 
Asp 
# 540 
- Gly Glu Ala Arg Lys Thr Glu Val - # Leu Leu Lys Val Met Asp Ala 
Lys 
# 560 
- His Lys Asn Cys Met Glu Ser Phe - # Leu Glu Ala Ala Ser Leu Met 
Ser 
# 575 
- Gln Val Ser Tyr Arg His Leu Val - # Leu Leu His Gly Val Cys Met 
Ala 
# 590 
- Gly Asp Ser Thr Met Val Glu Glu - # Phe Val His Leu Gly Ala Ile 
Asp 
# 605 
- Met Tyr Leu Arg Lys Arg Gly His - # Leu Val Pro Ala Ser Trp Lys 
Leu 
# 620 
- Gln Val Val Lys Gln Leu Ala Tyr - # Ala Leu Asn Tyr Leu Glu Asp 
Lys 
# 640 
- Gly Leu Ser His Gly Asn Val Ser - # Ala Arg Lys Val Leu Leu Ala 
Arg 
# 655 
- Glu Gly Ala Asp Gly Ser Pro Pro - # Phe Ile Lys Leu Ser Asp Pro 
Gly 
# 670 
- Val Ser Pro Ala Val Leu Ser Leu - # Glu Met Leu Thr Asp Arg Ile 
Pro 
# 685 
- Trp Val Ala Pro Glu Cys Leu Arg - # Glu Ala Gln Thr Leu Ser Leu 
Glu 
# 700 
- Ala Asp Lys Trp Gly Phe Gly Ala - # Thr Val Trp Glu Val Phe Ser 
Gly 
# 720 
- Val Thr Met Pro Ile Ser Ala Leu - # Asp Pro Ala Lys Lys Leu Gln 
Phe 
# 735 
- Tyr Glu Asp Arg Gln Gln Leu Ser - # Ala Pro Lys Trp Thr Glu Leu 
Ala 
# 750 
- Leu Leu Ile Gln Gln Cys Met Ala - # Tyr Glu Pro Val Gln Arg Pro 
Ser 
# 765 
- Leu Arg Ala Val Ile Arg Asp Leu - # Asn Ser Leu Ile Ser Ser Asp 
Tyr 
# 780 
- Glu Leu Leu Ser Asp His Thr Trp - # Cys Pro Gly Thr Arg Asp Gly 
Leu 
# 800 
- Trp Asn Gly Ala Gln Leu Tyr Ala - # Cys Gln Asp Pro Thr Ile Phe 
Glu 
# 815 
- Glu Arg His Leu Lys Tyr Ile Ser - # Gln Leu Gly Lys Gly Phe Phe 
Gly 
# 830 
- Ser Val Glu Leu Cys Arg Tyr Asp - # Pro Leu Gly Asp Asn Thr Gly 
Ala 
# 845 
- Leu Val Ala Val Lys Gln Leu Gln - # His Ser Gly Pro Asp Gln Gln 
Arg 
# 860 
- Asp Phe Gln Arg Glu Ile Gln Ile - # Leu Lys Ala Gln His Ser Asp 
Phe 
# 880 
- Ile Val Lys Tyr Arg Gly Val Ser - # Tyr Gly Pro Gly Arg Gln Ser 
Pro 
# 895 
- Ala Leu Val Met Glu Tyr Leu Pro - # Ser Gly Cys Leu Arg Asp Phe 
Leu 
# 910 
- Gln Arg His Arg Gly Leu Asp Ala - # Ser Arg Leu Leu Leu Tyr Ser 
Ser 
# 925 
- Gln Ile Cys Lys Gly Met Glu Tyr - # Leu Gly Ser Arg Arg Cys Val 
His 
# 940 
- Arg Asp Leu Ala Ala Arg Asn Ile - # Leu Val Glu Ser Glu Ala His 
Val 
# 960 
- Lys Ile Ala Asp Phe Gly Leu Ala - # Lys Leu Leu Pro Leu Asp Lys 
Asp 
# 975 
- Tyr Tyr Val Val Arg Glu Pro Gly - # Gln Ser Pro Ile Phe Trp Tyr 
Ala 
# 990 
- Pro Glu Ser Leu Ser Asp Asn Ile - # Phe Ser Arg Gln Ser Asp Val 
Trp 
# 10050 
- Ser Phe Gly Val Val Leu Tyr Glu - # Leu Phe Thr Tyr Cys Asp Lys 
Ser 
# 10205 
- Cys Ser Pro Ser Ala Glu Phe Leu - # Arg Met Met Gly Cys Glu Arg 
Asp 
# 10405 
- Val Pro Arg Leu Cys Arg Leu Leu - # Glu Leu Leu Glu Glu Gly Gln 
Arg 
# 10550 
- Leu Pro Ala Pro Pro Cys Cys Pro - # Ala Glu Val Ser Cys Tyr Ser 
Gly 
# 10700 - # 1065 
- Trp Arg Asp Asp Ile Cys Leu Pro - # Ala Glu 
# 1080 
- (2) INFORMATION FOR SEQ ID NO:11: 
- (i) SEQUENCE CHARACTERISTICS: 
#acids (A) LENGTH: 1100 amino 
(B) TYPE: amino acid 
(C) STRANDEDNESS: Not R - #elevant 
(D) TOPOLOGY: linear 
- (ii) MOLECULE TYPE: protein 
- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:11: 
- Met Ala Pro Pro Ser Glu Glu Thr - # Pro Leu Ile Ser Gln Arg Ser 
Cys 
# 15 
- Ser Leu Ser Ser Ser Glu Ala Gly - # Ala Leu His Val Leu Leu Pro 
Pro 
# 30 
- Arg Gly Pro Gly Pro Pro Gln Arg - # Leu Ser Phe Ser Phe Gly Asp 
Tyr 
# 45 
- Leu Ala Glu Asp Leu Cys Val Arg - # Ala Ala Lys Ala Cys Gly Ile 
Leu 
# 60 
- Pro Val Tyr His Ser Leu Phe Ala - # Leu Ala Thr Glu Asp Leu Ser 
Cys 
# 80 
- Trp Phe Pro Pro Ser His Ile Phe - # Ser Ile Glu Asp Val Asp Thr 
Gln 
# 95 
- Val Leu Val Tyr Arg Leu Arg Phe - # Tyr Phe Pro Gly Trp Phe Gly 
Leu 
# 110 
- Glu Thr Cys His Arg Phe Gly Leu - # His Lys Asp Leu Thr Ser Ala 
Ile 
# 125 
- Leu Asp Val His Val Leu Glu His - # Leu Phe Ala Gln His Arg Ser 
Asp 
# 140 
- Leu Val Ser Gly Arg Leu Pro Val - # Gly Leu Ser Leu Lys Asp Gln 
Gly 
# 160 
- Glu Phe Leu Ser Leu Ala Val Leu - # Asp Leu Ala Gln Met Ala Arg 
Lys 
# 175 
- Gln Ala Gln Arg Pro Gly Glu Leu - # Leu Lys Ser Val Ser Tyr Lys 
Ala 
# 190 
- Cys Leu Pro Pro Ser Leu Arg Asp - # Leu Ile Gln Gly Gln Ser Phe 
Val 
# 205 
- Thr Arg Arg Arg Ile Arg Arg Thr - # Val Val Gln Ala Leu Ala Pro 
Cys 
# 220 
- Ser Ser Leu Pro Ser Arg Pro Tyr - # Ala Leu Met Ala Lys Tyr Ile 
Leu 
# 240 
- Asp Leu Glu Arg Leu His Pro Ala - # Ala Thr Thr Glu Ser Phe Leu 
Val 
# 255 
- Gly Leu Pro Gly Ala Gln Glu Glu - # Pro Gly Cys Leu Arg Val Thr 
Gly 
# 270 
- Asp Asn Gly Ile Ala Trp Ser Ser - # Lys Asp Gln Glu Leu Phe Gln 
Thr 
# 285 
- Phe Cys Asp Phe Pro Glu Ile Val - # Asp Val Ser Ile Lys Gln Ala 
Pro 
# 300 
- Arg Val Gly Pro Ala Gly Glu His - # Arg Leu Val Thr Ile Thr Arg 
Met 
# 320 
- Asp Gly His Ile Leu Glu Ala Glu - # Phe Pro Gly Leu Pro Glu Ala 
Leu 
# 335 
- Ser Phe Val Ala Leu Val Asp Gly - # Tyr Phe Arg Leu Ile Cys Asp 
Ser 
# 350 
- Arg His Phe Phe Cys Lys Glu Val - # Ala Pro Pro Arg Leu Leu Glu 
Glu 
# 365 
- Glu Ala Glu Leu Cys His Gly Pro - # Ile Thr Leu Asp Phe Ala Ile 
His 
# 380 
- Lys Leu Lys Ala Ala Gly Ser Leu - # Pro Gly Ser Tyr Ile Leu Arg 
Arg 
# 400 
- Ser Pro Gln Asp Tyr Asp Ser Phe - # Leu Leu Thr Ala Cys Val Gln 
Thr 
# 415 
- Pro Leu Gly Pro Asp Tyr Lys Gly - # Cys Leu Ile Arg Gln Asp Pro 
Ser 
# 430 
- Gly Ala Phe Ser Leu Val Gly Leu - # Ser Gln Leu His Arg Ser Leu 
Gln 
# 445 
- Glu Leu Leu Thr Ala Cys Trp His - # Ser Gly Leu Gln Val Asp Gly 
Thr 
# 460 
- Ala Leu Asn Leu Thr Ser Cys Cys - # Val Pro Arg Pro Lys Glu Lys 
Ser 
# 480 
- Asn Leu Ile Val Val Arg Arg Gly - # Arg Asn Pro Thr Pro Ala Pro 
Gly 
# 495 
- His Ser Pro Ser Cys Cys Ala Leu - # Thr Lys Leu Ser Phe His Thr 
Ile 
# 510 
- Pro Ala Asp Ser Leu Glu Trp His - # Glu Asn Leu Gly His Gly Ser 
Phe 
# 525 
- Thr Lys Ile Phe His Gly His Arg - # Arg Glu Val Val Asp Gly Glu 
Thr 
# 540 
- His Asp Thr Glu Val Leu Leu Lys - # Val Met Asp Ser Arg His Gln 
Asn 
# 560 
- Cys Met Glu Ser Phe Leu Glu Ala - # Ala Ser Leu Met Ser Gln Val 
Ser 
# 575 
- Tyr Pro His Leu Val Leu Leu His - # Gly Val Cys Met Ala Gly Asp 
Ser 
# 590 
- Ile Met Val Gln Glu Phe Val Tyr - # Leu Gly Ala Ile Asp Thr Tyr 
Leu 
# 605 
- Arg Lys Arg Gly His Leu Val Pro - # Ala Ser Trp Lys Leu Gln Val 
Thr 
# 620 
- Lys Gln Leu Ala Tyr Ala Leu Asn - # Tyr Leu Glu Asp Lys Gly Leu 
Pro 
# 640 
- His Gly Asn Val Ser Ala Arg Lys - # Val Leu Leu Ala Arg Glu Gly 
Val 
# 655 
- Asp Gly Asn Pro Pro Phe Ile Lys - # Leu Ser Asp Pro Gly Val Ser 
Pro 
# 670 
- Thr Val Leu Ser Leu Glu Met Leu - # Thr Asp Arg Ile Pro Trp Val 
Ala 
# 685 
- Pro Glu Cys Leu Gln Glu Ala Gly - # Thr Leu Asn Leu Glu Ala Asp 
Lys 
# 700 
- Trp Gly Phe Gly Ala Thr Thr Trp - # Glu Val Phe Ser Gly Ala Pro 
Met 
# 720 
- His Ile Thr Ser Leu Glu Pro Ala - # Lys Lys Leu Lys Phe Tyr Glu 
Asp 
# 735 
- Arg Gly Gln Leu Pro Ala Leu Lys - # Trp Thr Glu Leu Glu Gly Leu 
Ile 
# 750 
- Ala Gln Cys Met Ala Tyr Asp Pro - # Gly Arg Arg Pro Ser Phe Arg 
Ala 
# 765 
- Ile Leu Arg Asp Leu Asn Gly Leu - # Ile Thr Ser Asp Tyr Glu Leu 
Leu 
# 780 
- Ser Asp Pro Thr Pro Gly Ile Pro - # Asn Pro Arg Asp Glu Leu Cys 
Gly 
# 800 
- Gly Ala Gln Leu Tyr Ala Cys Gln - # Asp Pro Ala Ile Phe Glu Glu 
Arg 
# 815 
- His Leu Lys Tyr Ile Ser Leu Leu - # Gly Lys Gly Asn Phe Gly Ser 
Val 
# 830 
- Glu Leu Cys Arg Tyr Asp Pro Leu - # Gly Asp Asn Thr Gly Pro Leu 
Val 
# 845 
- Ala Val Lys Gln Leu Gln His Ser - # Gly Pro Glu Gln Gln Arg Asp 
Phe 
# 860 
- Gln Arg Glu Ile Gln Ile Leu Lys - # Ala Leu His Cys Asp Phe Ile 
Val 
# 880 
- Lys Tyr Arg Gly Val Ser Tyr Gly - # Pro Gly Arg Gln Glu Leu Arg 
Leu 
# 895 
- Val Met Glu Tyr Leu Pro Ser Gly - # Cys Leu Arg Asp Phe Leu Gln 
Arg 
# 910 
- His Arg Ala Arg Leu His Asn Asp - # Arg Leu Leu Leu Phe Ala Trp 
Gln 
# 925 
- Ile Cys Lys Gly Met Glu Tyr Leu - # Gly Ala Arg Arg Cys Val His 
Arg 
# 940 
- Asp Leu Ala Ala Arg Asn Ile Leu - # Val Glu Ser Glu Ala His Val 
Lys 
# 960 
- Ile Ala Asp Phe Gly Leu Ala Lys - # Leu Leu Pro Leu Gly Lys Asp 
Tyr 
# 975 
- Tyr Val Val Arg Val Pro Gly Gln - # Ser Pro Ile Phe Trp Tyr Ala 
Pro 
# 990 
- Glu Ser Leu Ser Asp Asn Ile Phe - # Ser Arg Gln Ser Asp Val Trp 
Ser 
# 10050 
- Phe Gly Val Val Leu Tyr Glu Leu - # Phe Thr Tyr Ser Asp Lys Ser 
Cys 
# 10205 
- Ser Pro Ser Thr Glu Phe Leu Arg - # Met Ile Gly Pro Glu Arg Glu 
Gly 
# 10405 
- Ser Pro Leu Cys His Leu Leu Glu - # Leu Leu Ala Glu Gly Arg Arg 
Leu 
# 10550 
- Pro Pro Pro Ser Thr Cys Pro Thr - # Glu Val Gln Glu Leu Met Gln 
Leu 
# 10700 - # 1065 
- Cys Trp Ser Pro Asn Pro Gln Asp - # Arg Pro Ala Phe Asp Thr Leu 
Ser 
# 10850 
- Pro Gln Leu Asp Ala Leu Trp Arg - # Gly Ser Pro Gly 
# 11005 
- (2) INFORMATION FOR SEQ ID NO:12: 
- (i) SEQUENCE CHARACTERISTICS: 
#acids (A) LENGTH: 846 amino 
(B) TYPE: amino acid 
(C) STRANDEDNESS: Not R - #elevant 
(D) TOPOLOGY: linear 
- (ii) MOLECULE TYPE: protein 
- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:12: 
- Met Ala Pro Pro Ser Glu Glu Thr - # Pro Leu Ile Gln Arg Ser Cys 
Ser 
# 15 
- Leu Ser Glu Ala Gly Ala Leu His - # Val Leu Leu Pro Arg Gly Pro 
Gly 
# 30 
- Pro Pro Gln Arg Leu Ser Phe Ser - # Phe Gly Asp Leu Ala Glu Asp 
Leu 
# 45 
- Cys Val Ala Ala Lys Ala Ile Leu - # Pro Val Tyr His Ser Leu Phe 
Ala 
# 60 
- Leu Ala Thr Glu Asp Leu Ser Cys - # Trp Phe Pro Gln Val Leu Tyr 
Arg 
# 80 
- Arg Phe Tyr Phe Pro Trp Phe Gly - # Leu Glu Cys His Arg Phe Gly 
Leu 
# 95 
- Lys Asp Leu Ser Ala Ile Leu Asp - # Val Leu Glu His Leu Phe Ala 
Gln 
# 110 
- His Arg Ser Asp Leu Val Ser Gly - # Arg Leu Pro Gly Leu Ser Leu 
Lys 
# 125 
- Gln Gly Glu Leu Ser Leu Ala Val - # Leu Asp Leu Ala Met Ala Arg 
Gln 
# 140 
- Ala Gln Arg Gly Glu Leu Leu Lys - # Val Ser Tyr Lys Ala Cys Leu 
Pro 
# 160 
- Pro Ser Leu Arg Asp Leu Ile Gln - # Gly Ser Phe Val Thr Arg Arg 
Ile 
# 175 
- Arg Arg Thr Val Leu Leu Met Ala - # Lys Tyr Ile Asp Leu Glu Arg 
Leu 
# 190 
- Pro Ala Glu Phe Val Gly Leu Pro - # Gly Ala Gly Arg Val Gly Asp 
Gly 
# 205 
- Ile Ala Trp Gln Glu Gln Phe Cys - # Asp Phe Pro Glu Ile Val Asp 
Ser 
# 220 
- Ile Lys Gln Ala Pro Arg Val Gly - # Pro Ala Gly Glu His Arg Leu 
Val 
# 240 
- Thr Thr Arg Asp Ile Leu Glu Ala - # Glu Phe Pro Gly Leu Pro Glu 
Ala 
# 255 
- Leu Ser Phe Val Ala Leu Val Asp - # Gly Tyr Phe Arg Leu Asp Ser 
His 
# 270 
- Phe Phe Cys Lys Glu Val Pro Arg - # Leu Leu Glu Glu Ala Glu Cys 
His 
# 285 
- Gly Pro Ile Thr Leu Asp Phe Ala - # Ile Lys Leu Lys Gly Ser Pro 
Gly 
# 300 
- Ser Tyr Leu Arg Arg Pro Gln Asp - # Asp Ser Phe Leu Leu Thr Cys 
Val 
# 320 
- Gln Pro Leu Gly Pro Asp Tyr Lys - # Gly Cys Leu Ile Arg Pro Gly 
Phe 
# 335 
- Leu Val Gly Leu Ser His Ser Leu - # Glu Leu Leu Cys Trp Gly Leu 
Val 
# 350 
- Asp Gly Ala Leu Thr Ser Cys Cys - # Pro Arg Pro Lys Glu Lys Ser 
Asn 
# 365 
- Leu Ile Val Val Arg Gly Pro Thr - # Ser Leu Phe His Ile Pro Ala 
Asp 
# 380 
- Ser Leu Glu Trp His Glu Asn Leu - # Gly His Gly Ser Phe Thr Lys 
Ile 
# 400 
- Gly Arg Glu Val Val Asp Gly Glu - # Thr Glu Val Leu Leu Lys Val 
Met 
# 415 
- Asp His Asn Cys Met Glu Ser Phe - # Leu Glu Ala Ala Ser Leu Met 
Ser 
# 430 
- Gln Val Ser Tyr His Leu Val Leu - # Leu His Gly Val Cys Met Ala 
Gly 
# 445 
- Asp Ser Met Val Glu Phe Val Leu - # Gly Ala Ile Asp Tyr Leu Arg 
Lys 
# 460 
- Arg Gly His Leu Val Pro Ala Ser - # Trp Lys Leu Gln Val Lys Gln 
Leu 
# 480 
- Ala Tyr Ala Leu Asn Tyr Leu Glu - # Asp Lys Gly Leu His Gly Asn 
Val 
# 495 
- Ser Ala Arg Lys Val Leu Leu Ala - # Arg Glu Gly Asp Gly Pro Pro 
Phe 
# 510 
- Ile Lys Leu Ser Asp Pro Gly Val - # Ser Pro Val Leu Ser Leu Glu 
Met 
# 525 
- Leu Thr Asp Arg Ile Pro Trp Val - # Ala Pro Glu Cys Leu Glu Ala 
Thr 
# 540 
- Leu Leu Glu Ala Asp Lys Trp Gly - # Phe Gly Ala Thr Trp Glu Val 
Phe 
# 560 
- Ser Gly Met Ile Leu Pro Ala Lys - # Lys Leu Phe Tyr Glu Asp Arg 
Gln 
# 575 
- Leu Ala Lys Trp Thr Glu Leu Leu - # Ile Gln Cys Met Ala Tyr Pro 
Arg 
# 590 
- Pro Ser Arg Ala Arg Asp Leu Asn - # Leu Ile Ser Asp Tyr Glu Leu 
Leu 
# 605 
- Ser Asp Thr Pro Arg Asp Leu Gly - # Ala Gln Leu Tyr Ala Cys Gln 
Asp 
# 620 
- Pro Ile Phe Glu Glu Arg His Leu - # Lys Tyr Ile Ser Leu Gly Lys 
Gly 
# 640 
- Phe Gly Ser Val Glu Leu Cys Arg - # Tyr Asp Pro Leu Gly Asp Asn 
Thr 
# 655 
- Gly Leu Val Ala Val Lys Gln Leu - # Gln His Ser Gly Pro Gln Gln 
Arg 
# 670 
- Asp Phe Gln Arg Glu Ile Gln Ile - # Leu Lys Ala His Asp Phe Ile 
Val 
# 685 
- Lys Tyr Arg Gly Val Ser Tyr Gly - # Pro Gly Arg Gln Leu Val Met 
Glu 
# 700 
- Tyr Leu Pro Ser Gly Cys Leu Arg - # Asp Phe Leu Gln Arg His Arg 
Ala 
# 720 
- Leu Arg Leu Leu Leu Gln Ile Cys - # Lys Gly Met Glu Tyr Leu Gly 
Arg 
# 735 
- Arg Cys Val His Arg Asp Leu Ala - # Ala Arg Asn Ile Leu Val Glu 
Ser 
# 750 
- Glu Ala His Val Lys Ile Ala Asp - # Phe Gly Leu Ala Lys Leu Leu 
Pro 
# 765 
- Leu Lys Asp Tyr Tyr Val Val Arg - # Pro Gly Gln Ser Pro Ile Phe 
Trp 
# 780 
- Tyr Ala Pro Glu Ser Leu Ser Asp - # Asn Ile Phe Ser Arg Gln Ser 
Asp 
# 800 
- Val Trp Ser Phe Gly Val Val Leu - # Tyr Glu Leu Phe Thr Tyr Asp 
Lys 
# 815 
- Ser Cys Ser Pro Ser Glu Phe Leu - # Arg Met Gly Glu Arg Leu Cys 
Leu 
# 830 
- Leu Glu Leu Leu Glu Gly Arg Leu - # Pro Pro Cys Pro Glu Val 
# 845 
__________________________________________________________________________