CA2448116A1 - Molecules for disease detection and treatment - Google Patents

Abstract

The invention provides human molecules for disease detection and treatment (MDDT)and polynucleotides which identify and encode MDDT. The invention also provides expression vectors, host cells, antibodies, agonists, and antagonists. The invention also provides methodsfor diagnosing, treating, or preventing disorders associated with aberrant expression of MDDT.

Description

MOLECULES FOR DISEASE DETECTION AND TREATMENT
TECHNICAL FIELD
This invention relates to nucleic acid and amino acid sequences of molecules for disease detection and treatment and to the use of these sequences in the diagnosis, treatment, and prevention of cell proliferative, autoimmune/inflammatory, developmental, and neurological disorders, and in the assessment of the effects of exogenous compounds on the expression of nucleic acid and amino acid sequences of molecules for disease detection and treatment.
1o BACKGROUND OF THE INVENTION
The human genome is comprised of thousands of genes, many encoding gene products that function in the maintenance and growth of the various cells and tissues in the body. Aberrant expression or mutations in these genes and their products is the cause of, or is associated with, a variety of human diseases such as cancer and other cell proliferative disorders. The identification of these genes and their products is the basis of an ever-expanding effort to find markers for early detection of diseases, and targets for their prevention and treatment.
It is estimated that only 2% of mammalian DNA encodes proteins, and only a small fraction of the genes that encode'proteins are actually expressed in a particular cell at any time. The various types of cells in a multicellular organism differ dramatically both in structure and function, and the identity of a particular cell is conferred by its unique pattern of gene expression. In addition, different cell types express overlapping but distinctive sets of genes throughout development. Cell growth and proliferation, cell differentiation, the immune response, apoptosis, and other processes that contribute to organismal development and survival are governed by regulation of gene expression. Appropriate gene regulation also ensures that cells function efficiently by expressing only those genes whose functions are required at a given time. Factors that influence gene expression include extracellular signals that mediate cell-cell communication and coordinate the activities of different cell types. Gene expression is regulated at the level of DNA and RNA transcription, and at the level of mRNA
translation.
Cancer represents a type of cell proliferative disorder that affects nearly every tissue in the body. A wide variety of molecules, either aberrantly expressed or mutated, can be the cause of, or involved with, various cancers because tissue growth involves complex and ordered patterns of cell proliferation, cell differentiation, and apoptosis. Cell proliferation must be regulated to maintain both the number of cells and their spatial organization. This regulation depends upon the appropriate expression of proteins which control cell cycle progression in response to extracellular signals such as growth factors and other mitogens, and intracellular cues such as DNA damage or nutrient starvation.
Molecules which directly or indirectly modulate cell cycle progression fall into several categories, including growth factors and their receptors, second messenger and signal transduction proteins, oncogene products, tumor-suppresser proteins, and mitosis-promoting factors.
Aberrant expression or mutations in genes and their products may cause, or increase susceptibility to, a variety of human diseases such as cancer and other cell proliferative disorders. The identification of these genes and their products is the basis of an ever-expanding effort to find markers for early detection of diseases and targets for their prevention and treatment. For example, cancer represents a type of cell proliferative disorder that affects nearly every tissue in the body. The development of cancer, or oncogenesis, is often correlated with the conversion of a normal gene into a cancer-causing gene, or oncogene, through abnormal expression or mutation.
Oncoproteins, the products of oncogenes, include a variety of molecules that influence cell proliferation, such as growth factors, growth factor receptors, intracellular signal transducers, nuclear transcription factors, and cell-cycle control proteins. In contrast, tumor-suppresser genes are involved in inhibiting cell proliferation. Mutations which reduce or abrogate the function of tumor-suppresser genes result in aberrant cell proliferation and cancer. Thus a wide variety of genes and their products have been found that are associated with cell proliferative disorders such as cancer, but many more may exist that are yet to be discovered.
Mammalian peripheral blood comprises cells of the erythroid, myeloid, and lymphoid lineages.
Each lineage is derived from a pluripotent stem cell which, upon exposure to various molecules and other types of cells, differentiate into effector cells which migrate into the blood and other organs.
These cells include red blood cells and platelets (erythroid), macrophages and granulocytes (myeloid), and T and B lymphocytes (lymphoid). Myeloid and lymphoid cells mediate immune responses to pathogens such as bacteria, parasites, and viruses.
Functional interaction of the cell types involved in immune responses involves transfer of signals via soluble messenger molecules known as cytokines. Both hematopoietic cells and non-hematopoietic cells produce cytokines which stimulate the activation, differentiation and proliferation of T cells, B cells, macrophages, and granulocytes during an active immune response. Cytokines bind to specific receptors expressed on cellular membranes and transduce a signal through the cell.
Depending on the type of cytokine and the cell to which it binds, this signal initiates activation, differentiation, growth, and/or apoptosis.
T cells, which respond to and produce a variety of cytokines, are divided into two major groups, CD4+ T helper (Th) cells, and CD8+ cytotoxic T lymphocytes (CTL).
Tmmune responses are primarily regulated by CD4+ Th cells which fall into two subclasses based on the kinds of cytokines they secrete. Th1 cells secrete primarily IL-2 and IFN-y; regulate the responses of CTLs, B cells, and macrophages; and orchestrate the removal of intracellular pathogens. In contrast, Th2 cells secrete primarily IL-4 and IL-10 and promote the development of certain antibody responses such as IgGl, IgA, and IgE. In addition, Th2 cells remove extracellular pathogens, which include various bacteria and parasites. (See, e.g., Morel and Oriss (1998) Crit. Rev.
T_m_m__unol. 18:275-303.) Studies have shown that the Th1 cytokine response predominates in organ-speciftc autoimmune disorders such as insulin-dependent diabetes mellitus (IDDM), multiple sclerosis (MS), rheumatoid arthritis (RA), and Crolm's disease. A Th1 response also predominates in acute allograft rejection, eradication of tumors, and unexplained recurrent abortions. Th2 responses predominate in allergy and other atopic disorders, transplantation tolerance, chronic graft versus host disease (GVFiD), and systemic autoimmune disease such as systemic lupus erythmatosus (Romangnani et al. .(1997) Int.
Arch. Allergy Tmmunol.
113:153-156). Genes affected by these molecules may reasonably be expected to be markers of immune cell development, function, and activity.
Tumor necrosis factor (TNF) a is a pleiotropic cytokine that mediates immune regulation and inflammatory responses. TNF-a-related cytokines generate.partially overlapping cellular responses, including differentiation, proliferation, nuclear factor-xB (NF-xB) activation, and cell death, by triggering the aggregation of receptor monomers (Smith, C.A. et al. (1994) Cell 76:959-962). The cellular responses triggered by TNF-a are initiated through its interaction with distinct cell surface receptors (TNFRs). Treatment of confluent cultures of vascular smooth muscle cells (SMCs) with TNF-a suppresses the incorporation of [3H]proliue into both collagenase-digestible proteins (CDP) and noncollagenous proteins (NCP). Such suppression by TNF-a is not observed in confluent bovine aortic endothelial cells and human fibroblastic IIVVIR-90 cells. TNF-a decreases the relative proportion of collagen types IV and V suggesting that TNF-a modulates collagen synthesis by SMCs depending on their cell density and therefore may modify formation of atherosclerotic lesions (Hiraga, S. et al.
(2000) Life Sci. 66:235-244). Primary human endothelial cell lines such as human umbilical vein endothelial cells (IiUVECs) have been used as an experimental model for investigating in vitro the role of the endothelium in human vascular biology. Activation of the vascular endothelium is considered to be a central event in a wide range of both physiological and pathophysiological processes, such as vascular tone regulation, coagulation and thrombosis, atherosclerosis, and inflammation.
DNA-based arrays can provide an efficient, high-throughput method to examine gene expression and genetic variability. For example, SNPs, or single nucleotide polymorphisms, are the most common type of human genetic variation. DNA based arrays can dramatically accelerate the discovery of SNPs in hundreds and even thousands of genes. Likewise, such arrays can be used for SNP genotyping in which DNA samples from individuals or populations are assayed for the presence of selected SNPs. These approaches will ultimately lead to the systematic identification of all genetic variations in the human genome and the correlation of certain genetic variations with disease susceptibility, responsiveness to drug treatments, and other medically relevant information. (See, for example, Wang, D.G. et al. (1998) Science 280:1077-1082.) DNA-based array technology is especially important for the rapid analysis of global gene expression patterns. For example, genetic predisposition, disease, or therapeutic treatment may directly or indirectly affect the expression of a large number of genes in a given tissue. In this case, it is useful to develop a profile, or transcript image, of all the genes that are expressed and the levels at which they are expressed in that particular tissue. A profile generated from an individual or population affected with a certain disease or undergoing a particular therapy may be compared with a profile generated from a control individual or population. Such analysis does not require knowledge of gene function, as the expression profiles. can be subjected to mathematical analyses which simply treat each gene as a marker. Furthermore, gene expression profiles may help dissect biological pathways by identifying all the genes expressed, for example, at a certain developmental stage, in a particular tissue, or in response to disease or treatment. (See, for example, Larder, E.S. et al.
(1996) Science 274:536-539.) Dendritic cells (DC) are antigen presenting cells (APC) that play a key role in the primary 20' immune response because of their unique ability to present antigens to naive T-cells. In addition, DC
differentiate into separate subsets of mature immune cells that sustain and regulate immune responses following initial contact with antigen. DC subsets include those that preferentially induce particular T
helper 1 (Th1) or T helper 2 (Th2) responses and those that regulate B cell responses. Moreover, DC
are being used with increasing frequency to manipulate immune responses, either to downregulate aberrant autoimmune response or to enhance vaccination or tumor-specific response.
DC are functionally specialized in correlation with their particular differentiation state. CD34+
myeloid cells found in the bone marrow mature in response to signals into CD14+ CDllc+ monocytes.
An innate or antigen non-specific response takes place initially when monocytes circulate to nonlymphoid tissues and respond to lipopolysaccharide (LPS), a bacterially-derived mitogen, and viruses. Such direct encounters with antigen cause secretion of pro-inflammatory cytokines that attract and regulate natural killer cells, macrophages, and eosinophils in the first line of defense against invading pathogens. Monocytes then mature into DC, which efficiently capture antigen through endocytosis and antigen-receptor uptake. Antigen processing and presentation trigger activation and differentiation into mature DC that express MHC class II molecules on the cell surface and efficiently activate T-cells, initiating antigen-specific T-cell and B-cell responses. In turn, T-cells activate DC
through CD40 ligaud - CD40 interactions, which stimulate expression of the costimulatory molecules CD80 and CD86, the latter most potent in amplifying T-cell responses. DC
interaction via CD40 with T cells also stimulates the production of inflammatory cytokines such as TNF
alpha and IL-1.
Engagement of RANK, a member of the TNF receptor family by its ligand, TRANCE, which is expressed on activated T cells, enhances the survival of DC through inhibition of apoptosis, thereby enhancing T cell activation. The maturation and differentiation of monocytes into mature DC links the antigen non-specific innate immune response to the antigen-specific adaptive immune response.
Certain genes are known to be associated with diseases because of their chromosomal location, such as the genes in the myotonic dystrophy (DM) regions of mouse and human. The mutation underlying DM has been localized to a gene encoding the DM-kinase protein, but another active gene, DMR-N9, is in close proximity to the DM-kinase gene (Jausen, G.
et al. (1992) Nat.
Genet. 1:261-266). DMR-N9 encodes a 650 amino acid protein that contains WD
repeats, motifs found in cell signaling proteins. DMR-N9 is expressed in all neural tissues and in the testis, suggesting a role for DMR-N9 in the manifestation of mental and testicular symptoms in severe cases of DM
(Jansen, G. et al. (1995) Hum. Mol. Genet. 4:843-852).
Other genes are identified based upon their expression patterns or association with disease syndromes. For example, autoautibodies to subcellular organelles are found in patients with systemic rheumatic diseases. A recently identified protein, golgin-67, belongs to a family of Golgi autoantigens having alpha-helical coiled-coil domains (Eystathioy, T. et al. (2000) J.
Autoimmun. 14:179-187). The Stac gene was identified as a brain specific, developmentally regulated gene.
The Stac protein contains an SH3 domain, and is thought to be involved in neuron-specific signal transduction (Suzuki, H. et al. (1996) Biochem. Biophys. Res. Commun. 229:902-909).
Intracellular Si ng aling Cell-cell communication is essential for the growth, development, and survival of multicellular organisms. Cells communicate by sending and receiving molecular signals. An example of a molecular signal is a growth factor, which binds and activates a specific transmembrane receptor on the surface of a target cell. The activated receptor transducer the signal intracellularly, thus initiating a cascade of biochemical reactions that ultimately affect gene transcription and cell cycle progression in the target cell.
Intracellular signaling is the process by which cells respond to extracellular signals (hormones, neurotransmitters, growth and differentiation factors, etc.) through a cascade of biochemical reactions that begins with the binding of a signaling molecule to a cell membrane receptor and ends with the activation of an intracellular target molecule. Intermediate steps in the process involve the activation of various cytoplasmic proteins by phosphorylation via protein kinases, and their deactivation by protein phosphatases, and the eventual translocation of some of these activated proteins to the cell nucleus where the transcription of specific genes is triggered. The intracellular signaling process regulates all types of cell functions including cell proliferation, cell differentiation, and gene transcription, and involves a diversity of molecules including protein kinases and phosphatases, and second messenger molecules such as cyclic nucleotides, calcium-calmodulin, inositol, and various mitogens that regulate protein phosphorylation.
Cells also respond to changing conditions by switching off signals. Many signal transduction proteins are short-lived and rapidly targeted for degradation by covalent ligation to ubiquitin, a highly conserved small protein. Cells also maintain mechanisms to monitor changes in the concentration of denatured or unfolded proteins in membrane-bound extracytoplasmic compartments, including a transmembrane receptor that monitors the concentration of available chaperone molecules in the endoplasmic reticulum and transmits a signal to the cytosol to activate the transcription of nuclear genes encoding chaperones in the endoplasmic reticulum.
Certain proteins in intracellular signaling pathways serve to link or cluster other proteins involved in the signaling cascade. These proteins are referred to as scaffold, anchoring, or adaptor proteins. (For review, see Pawson, T. and J.D. Scott (1997) Science 278:2075-2080.) As many , intracellular signaling proteins such as protein kinases and phosphatases have relatively broad substrate specificities, the adaptors help to organize the component signaling proteins into specific biochemical pathways,. Many of the above signaling molecules are characterized by the presence of particular domains that promote protein-protein interactions. A sampling of these domains is discussed below, along with other important intracellular messengers.
Intracellular Signaling Second Messenger Molecules Protein Phosphorylation Protein kinases and phosphatases play a key role in the intracellular signaling process by controlling the phosphorylation and activation of various signaling proteins.
The high energy phosphate for this reaction is generally transferred from the adenosine triphosphate molecule (ATP to a particular protein by a protein kinase and removed from that protein by a protein phosphatase. Protein kinases are roughly divided into two groups: those that phosphorylate serine or threonine residues (serine/threonine kinases, STK) and those that phosphorylate tyrosine residues (protein tyrosine kinases, PTK). A few protein kinases have dual specificity for serine/threonine and tyrosine residues.
Almost all kinases contain a conserved 250-300 amino acid catalytic domain containing specific residues and sequence motifs characteristic of the kinase family (Hardie, G.
and S. Hanks (1995) The Protein Kinase Facts Books, Vol I:7-20, Academic Press, San Diego, CA).
STKs include the second messenger dependent protein kinases such as the cyclic-AMP
dependent protein kinases (PKA), involved in mediating hormone-induced cellular responses;
calcium-calmodulin (CaM) dependent protein kinases, involved in regulation of smooth muscle contraction, glycogen breakdown, and neurotransmission; and the mitogen-activated protein kinases (MAP kinases) which mediate signal transduction from the cell surface to the nucleus via , phosphorylation cascades. Altered PKA expression is implicated in a variety of disorders and diseases including cancer, thyroid disorders, diabetes, atherosclerosis, and cardiovascular disease (Isselbacher, K.J. et al. (1994) Harrison's Principles of Internal Medicine, McGraw-Hill, New York, 1o NY, pp. 416-431, 1887).
PTKs are divided into transmembrane, receptor PTKs and nontransmembrane, non-receptor PTKs. Transmembrane PTKs are receptors for most growth factors. Non-receptor PTKs lack transmembrane regions and, instead, form complexes with the intracellular regions of cell surface receptors. Receptors that function through non-receptor PTKs include those for cytokines and hormones (growth hormone and prolactin) and antigen-specific receptors on T
and B lymphocytes.
Many of these PTKs were first identified as the products of mutant oncogenes in cancer cells in which their activation was no longer subject to normal cellular controls. In fact, about one third.of the ..
known oncogenes encode PTKs, and it is well known that.cellular transformation (oncogenesis) is often accompanied by increased tyrosine phosphorylation activity (Charbonneau H. and N.K. Tonks (1992) Annu. Rev. Cell Biol. 8:463-493).
An additional family of protein kinases previously thought to exist only in prokaryotes is the histidine protein kinase family (HPK). HPKs bear little homology with mammalian STKs or PTKs but have distinctive sequence motifs of their own (Davie, J.R. et al. (1995) J.
Biol. Chem.
270:19861-19867). A histidine residue in the N-terminal half of the molecule (region I) is an autophosphorylation site. Three additional motifs located in the C-terminal half of the molecule include an invariant asparagine residue in region II and two glycine-rich loops characteristic of nucleotide binding domains in regions III and 1V. Recently a branched chain alpha-ketoacid dehydrogenase kinase has been found with characteristics of HPK in rat (Davie et al., supra).
Protein phosphatases regulate the effects of protein kinases by removing phosphate groups from molecules previously activated by kinases. The two principal categories of protein phosphatases are the protein (serine/threonine) phosphatases (PPs) and the protein tyrosine phosphatases (PTPs).
PPs dephosphorylate phosphoserine/threonine residues and are important regulators of many cAMP-mediated hormone responses (Cohere, P. (1989) Annu. Rev. Biochem. 58:453-508). PTPs reverse the effects of protein tyrosine kinases and play a significant role in cell cycle and cell signaling processes (Charbonneau and Tonks, supra). As previously noted, many PTKs are encoded by oncogenes, and oncogenesis is often accompanied by increased tyrosine phosphorylation activity. It is therefore possible that PTPs may prevent or reverse cell transformation and the growth of various cancers by controlling the levels of tyrosine phosphorylation in cells. This hypothesis is supported by studies showing that overexpression of PTPs can suppress transformation in cells, and that specific inhibition of PTPs can enhance cell transformation (Charbonneau and Tonks, su ra).
Phospholipid and Iuositol-phosphate Si~alina Inosito] phospholipids (phosphoinositides) are involved in an intracellular signaling pathway that begins with binding of a signaling molecule to a G-protein linked receptor in the plasma membrane.
This leads to the phosphorylation of phosphatidylinositol (PI) residues on the inner side of the plasma membrane to the biphosphate state (PIPZ) by inositol kinases. Simultaneously, the G-protein linked receptor binding stimulates. a trimeric G-protein which in turn activates a phosphoinositide-specific phospholipase C-(3. Phospholipase C-(3 then cleaves PIPZ into two products, inositol triphosphate (IP3) and diacylglycerol. These two products act as mediators for separate signaling events. 1P3 diffuses through the plasma membrane to induce calcium release from the endoplasmic reticulum (ER), while .
diacylglycerol remains in the membrane and helps activate protein kinase C, a serine-threonine kinase that phosphorylates selected proteins in the target cell. The calcium response initiated by IP3 is terminated by the dephosphorylation of IP3 by specific inositol phosphatases.
Cellular responses that are mediated by this pathway are glycogen breakdown in the liver in response to. vasopressin, smooth muscle contraction in response to acetylcholine, and thrombin-induced platelet aggregation.
Inositol-phosphate signaling controls tubby, a membrane bound transcriptional regulator that serves as an intracellular messenger of Gccq coupled receptors (Santagata et al. (2001) Science 292:2041-2050). Members of the tubby family contain a C-terminal tubby domain of about 260 amino acids that binds to double-stranded DNA and an N-terminal transcriptional activation domain. Tubby, binds to phosphatidylinositol 4,5-bisphosphate, which localizes tubby to the plasma membrane.
Activation of the G-protein aQ leads to activation of phospholipase C-(3 and hydrolysis of phosphoinositide. Loss of phosphatidylinositol 4,5-bisphosphate causes tubby to dissociate from the plasma membrane and to translocate to the nucleus where tubby regulates transcription of its target genes. Defects in the tubby gene are associated with obesity, retinal degeneration, and hearing loss (Boggon, T.J. et al. (1999) Science 286:2119-2125).
Cyclic Nucleotide Signaling Cyclic nucleotides (CAMP and cGMP) function as intracellular second messengers to transduce a variety of extracellular signals including hormones, light, and neurotransmitters. In particular, cyclic-AMP dependent protein kinases (PKA) are thought to account for all of the effects of cAMP in most mammalian cells, including various hormone-induced cellular responses. Visual excitation and the phototransmission of light signals in the eye is controlled by cyclic-GMP regulated, CaZk-specific channels. Because of the importance of cellular levels of cyclic nucleotides in mediating these various responses, regulating the synthesis and breakdown of cyclic nucleotides is an important matter. Thus adenylyl cyclase, which synthesizes cAMP from AMP, is activated to increase cAMP
levels in muscle by binding of adrenaline to (3-adrenergi.c receptors, while activation of guanylate cyclase and increased cGMP levels in photoreceptors leads to reopening of the Ca2+-specific channels and recovery of the dark state in the eye. There are nine known transmembrane isoforms of mammalian adenylyl cyclase, as well as a soluble form preferentially expressed in testis. Soluble adenylyl cyclase contains a P-loop, or nucleotide binding domain, and may be involved in male fertility (Buck, J. et al. (1999) Proc. Natl. Acad. Sci. USA 96:79-84).
In contrast, hydrolysis of cyclic nucleotides by cAMP and cGMP-specific phosphodiesterases (PDEs) produces the opposite of these and other effects mediated by increased cyclic nucleotide levels. PDEs appear to be particularly important in the regulation of cyclic nucleotides, considering the diversity found in this family of proteins. At least seven families of mammalian PDEs (PDE1-7) have been identified based on substrate specificity and affinity, sensitivity to cofactors, and sensitivity to inhibitory drugs (Beavo, J.A. (1995) Physiol: Rev. 75:725-748). PDE
inhibitors have been found to be particularly useful in treating various clinical disorders. Rolipram, a specific inhibitor of PDE4, has been used in the treatment of depression, and similar inhibitors are undergoing evaluation as anti-inflammatory agents. Theophylline is a nonspecific PDE inhibitor used in the treatment of bronchial asthma and other respiratory diseases (Banner, K.H. and C.P. Page (1995) Eur. Respir. J.
8:996-1000).
Calcium Si~nalin~ Molecules Caz+ is another second messenger molecule that is even more widely used as an intracellular mediator than cAMP. Caz~ can enter the cytosol by two pathways, in response to extracellular signals. One pathway acts primarily in nerve signal transduction where Ca2~
enters a nerve terminal through a voltage-gated Caa* channel. The second is a more ubiquitous pathway in which Ca2~" is released from the ER into the cytosol in response to binding of an extracellular signaling molecule to a receptor. Caa+ directly activates regulatory enzymes, such as protein kinase C, which trigger signal transduction pathways. Ca2+ also binds to specific Ca2k binding proteins (CBPs) such as calmodulin (CaM) which then activate multiple target proteins in the cell including enzymes, membrane transport pumps, and ion channels. CaM interactions are involved in a multitude of cellular processes including, but not limited to, gene regulation, DNA synthesis, cell cycle progression, mitosis, cytokinesis, cytoskeletal organization, muscle contraction, signal trausduction, ion homeostasis, exocytosis, and metabolic regulation (Cello, M.R. et al. (1996) Guidebook to Calcium-binding Proteins, Oxford University Press, Oxford, UK, pp. 15-20). Some Ca2+ binding proteins are characterized by the presence of one or more EF-hand Ca2+ binding motifs, which are comprised of 12 amino acids flanked by a helices (Cello, supra). The regulation of CBPs has implications for the control of a variety of disorders. Calcineurin, a CaM-regulated protein phosphatase, is a target for inhibition by the immunosuppressive agents cyclosporin and FK506. This indicates the importance of calcineurin and CaM in the immune response and immune disorders (Schwaninger M. et al. (1993) J. Biol Chem.
268:23111-23115). The level of CaM is increased several-fold in tumors and tumor-derived cell lines for various types of cancer (Rasmussen, C.D. and A.R. Means (1989) Trends Neurosci. 12:433-438).
The annexins are a family of calcium binding proteins that associate with the cell membrane (Towle, C.A. and B.V. Treadwell (1992) J. Biol. Chem. 267:5416-5423). Annexins reversibly bind to negatively charged phospholipids (phosphatidylcholine and phosphatidylserine) in a calcium dependent manner. Annexins participate in various processes pertaining to signal transduction at the plasma membxane, including membrane-cytoskeletoninteractions, phospholipase inhibition, anticoagulation, and membrane fusion. Annexins contain four to eight repeated segments of about 60 residues. Each repeat folds into five alpha helices wound into a-right-handed superhelix.
G-Protein Signaling Guanine nucleotide binding proteins (G-proteins) are critical mediators of signal transduction between a particular class of extracellular receptors, the G-protein coupled receptors (GPCRs), and intracellular second messengers such as cAMP and Ca2+. G-proteins are linked to the cytosolic side of a GPCR such that activation of the GPCR by ligand binding stimulates binding of the G-protein to GTP, inducing an "active" state in the G-protein. In the active state, the G-protein acts as a signal to trigger other events in the cell such as the increase of cAMP levels or the release of Ca2+ into the, cytosol from the ER, which, in turn, regulate phosphorylation and activation of other intracellular proteins. Recycling of the G-protein to the inactive state involves hydrolysis of the bound GTP to GDP by a GTPase activity in the G-protein. (See Alberts, B. et al. (1994) Molecular Biolog, o Cell Garland Publishing, Inc. New York, NY, pp.734-759.) The superfamily of G-proteins consists of several families which may be grouped as translational factors, heterotrimeric G-proteins involved in transmembrane signaling processes, and low molecular weight (LMW) G-proteins including the proto-oncogene Ras proteins and products of rab, rap, rho, rac, smg2l, smg25, YPT, SEC4, and ARF genes, and tubulins (Kaziro, Y. et al. (1991) Annu. Rev. Biochem. 60:349-400). In all cases, the GTPase activity is regulated through interactions with other proteins.
Heterotrimeric G-proteins are composed of 3 subunits, a, ~3, and y, which in their inactive conformation associate as a trimer at the inner face of the plasma membrane.
Gcc binds GDP or GTP
and contains the GTPase activity. The (3~( complex enhances binding of Ga to a receptor. Gy is necessary for the folding and activity of G(3 (Veer, E.J. et al. (1994) Nature 371:297-300). Multiple homologs of each subunit have been identified in mammalian tissues, and different combinations of subunits have specific functions and tissue specificities (Spiegel, A.M.
(1997) 3. Inner. Metab. Dis.
20:113-121).
The alpha subunits of heterotrimeric G-proteins can be divided into four distinct classes. The a-s class is sensitive to ADP-ribosylation by pertussis toxin which uncouples the receptor:G-protein interaction. This uncoupling blocks signal transduction to receptors that decrease cAMP levels which normally regulate ion channels and activate phospholipases. The inhibitory a-I
class is also susceptible to modification by pertussis toxin which prevents oc-I from lowering CAMP
levels. Two novel classes of a subunits refractory to pertussiS toxin modification are a-q, which activates phospholipase C, and ot-12, which has sequence homology with the Drosophila gene concertina and may contribute to the regulation of embryonic development (Simon, M.I. (1991) Science 252:802-808).
The mammalian G(3 and Gy subunits, each about 340 amino acids long, share more than 80%
homology. The G(3 subunit (also called transducin) contains seven repeating units, each about 43 amino acids long. The activity of both subunits may be regulated by other proteins such as calmodulin and phosducin or the neural protein GAP 43 (Clapham, D. and E. Neer (1993) Nature 365:403-406). , The (3 and 'y subunits are tightly associated. The (3 subunit sequences are highly conserved between 2o species, implying that they perform a fundamentally important role in the organization and function of G-protein. linked systems (Van der Voorn, L. (1992) FEBS Lett. 307:131-134).
They contain seven tandem repeats of the WD-repeat sequence motif, a motif found in many proteins with regulatory functions. WD-repeat proteins contain from four to eight copies of a loosely conserved repeat of approximately 40 amino acids which participates in protein-protein interactions. Mutations and variant expression of (3 transducin proteins are linked with various disorders.
Mutations in LIS 1, a subunit of the human platelet activating factor acetylhydrolase, cause Miller-Dieker lissencephaly. RACK1 binds activated protein kinase C, and RbAp48 binds retinoblastoma protein.
CstF is required for polyadenylation of mammalian pre-mRNA in vitro and associates with subunits of cleavage-stimulating factor. Defects in the regulation of (3-catenin contribute to the neoplastic transformation of human cells. The WD40 repeats of the human F box protein bTrCP mediate binding to ~i-catenin, thus regulating the targeted degradation of (3-catenin by ubiquitin ligase (Neer, su ra; Hart, M. et al. (1999) Curr. Biol. 9:207-210). The y subunit primary structures are more variable than those of the (3 subunits. They are often post-translationally modified by isoprenylation and carboxyl-methylation of a cysteine residue four amino acids from the C-terminus; this appears to be necessary for the interaction of the (3'y subunit with the membrane and with other G-proteins. The (3y subunit has been shown to modulate the activity of isoforms of adenylyl cyclase, phospholipase C, and some ion channels. It is involved in receptor phosphorylation via specific kinases, and has been implicated in the p2lras-dependent activation of the MAP kinase cascade and the recognition of specific receptors by G-proteins (Clapham and Neer, supra).
G-proteins interact with a variety of effectors including adenylyl cyclase (Clapham and Neer, su ra). The signaling pathway mediated by cAMP is mitogenic in hormone-dependent endocrine tissues such as adrenal cortex, thyroid, ovary, pituitary, and testes. Cancers in these tissues have been related to a mutationally activated form of a Goc S known as the gsp (Gs protein) oncogene (Dhanasekaran, N. et al. (1998) Oncogene 17:1383-1394). Another effector is phosducin, a retinal phosphoprotein, which forms a specific complex with retinal G(3 and Gy (G(3~y) and modulates the ability of G(3y to interact with retinal Goc (Clapham and Neer, su ra).
Irregularities in the G-pxotein signaling cascade may result in abnormal activation of leukocytes and lymphocytes, leading to the tissue damage and destruction seen in many inflammatory and autoimmune diseases such as rheumatoid arthritis, biliary cirrhosis, hemolytic anemia, lupus erythematosus, and thyroiditis. Abnormal cell proliferation, including cyclic AMP stimulation of brain, thyroid, adrenal, and gonadal tissue proliferation is regulated by G proteins.
Mutations in Gc~ subunits have been found in growth-hormone-secreting pituitary somatotroph tumors, hyperfunctioning thyroid adenomas, and ovarian and adrenal neoplasms (Meij, J.T.A. (1996) Mol. Cell Biochem. 157:31-38;
Aussel, C. et al. (1988) J. Tmmunol. 140:215-220).
LMW G-proteins are GTPases which regulate cell growth, cell cycle control, protein secretion, and intracellular vesicle interaction. They consist of single polypeptides which, like the alpha subunit of the heterotrimeric G-proteins, are able to bind to and hydrolyze GTP, thus cycling between an inactive and an active state. LMW G-proteins respond to extracellular signals from receptors and activating proteins by transducing mitogenic signals involved in various cell functions. The binding and hydrolysis of GTP regulates the response of LMW G-proteins and acts as an energy source during this process (Bokoch, G.M. and C.J. Der (1993) FASEB J. 7:750-759).
At least sixty members of the LMW G-protein superfamily have been identified and are currently grouped into the ras, rho, arf, sarl, ran, and rab subfamilies.
Activated ras genes were initially found in human cancers, and subsequent studies confirmed that ras function is critical in determining whether cells continue to grow or become differentiated. Ras1 and Ras2 proteins stimulate adenylate cyclase (Kaziro, su ra), affecting a broad array of cellular processes. Stimulation of cell surface receptors activates Ras which, in turn, activates cytoplasmic kinases. These kinases translocate to the nucleus and activate key transcription factors that control gene expression and protein synthesis (Barbacid, M. (1987) Annu. Rev. Biochem. 56:779-827, Treisman, R. (1994) Curr.
Opin. Genet. Dev. 4:96-98). Other members of the LMW G-protein superfamily have roles in signal transduction that vary with the function of the activated genes and the locations of the G-proteins that initiate the activity. Rho G-proteins control signal transduction pathways that link growth factor receptors to actin polymerization, which is necessary for normal cellular growth and division. The rab, arf, and sarl families of proteins control the translocation of vesicles to and from membranes for protein processing, localization, and secretion. Vesicle- and target- specific identifiers (v-SNARES
and t-SNARES) bind to each other and dock the vesicle to the acceptor membrane. The budding process is regulated by the closely related ADP ribosylation factors (ARFs) and SAR proteins, while rab proteins allow assembly of SNARE complexes and may play a role in removal of defective complexes (Rothman, J. and F. Wieland (1996) Science 272:227-234). Ran G-proteins are located in the nucleus of cells and have a key role in nuclear protein import, the control of DNA synthesis, and cell-cycle progression (Hall, A. (1990) Science 249:635-640; Barbacid, M.
(1987) Annu. Rev.
Biochem. 56:779-827; Ktistakis, N. (1998) BioEssays 20:495-504; and Sasaki, T.
and Y. Takai (1998) Biochem. Biophys. Res. Commun. 245:641-645).
Rab proteins have a highly variable amino terminus containing membrane-specific signal information and a prenylated carboxy terminus which determines the target membrane to which the Rab proteins anchor. More than 30 Rab proteins have been identified in a variety of species, and each has a characteristic intracellular location and distinct transport function.
In particular, Rab1 and Rab2 are important in ER-to-Golgi transport; Rab3 transports secretory vesicles to the extracellular membrane; RabS is localized to endosomes and regulates the fusion of early endosomes into late endosomes; Rab6 is specific to the Golgi apparatus and regulates intra-Golgi transport events; Rab7 and Rab9 stimulate the fusion of late endosomes and Golgi vesicles with lysosomes, respectively; and RablO mediates vesicle fusion from the medial Golgi to the traps Golgi. Mutant forms of Rab proteins are able to block protein transport along a given pathway or alter the sizes of entire organelles.
Therefore, Rabs play key regulatory roles in membrane trafficking (Schimtnoller,1.5. and S.R. Pfeffer (1998) J. Biol. Chem. 243:22161-22164).
The function of Rab proteins in vesicular transport requires the cooperation of many other proteins. Specifically, the membrane-targeting process is assisted by a series of escort proteins (Khosravi-Far, R. et al. (1991) Proc. Natl. Acad. Sci. USA 88:6264-6268). In the medial Golgi, it has been shown that GTP bound Rab proteins initiate the binding of VAMP-like proteins of the transport vesicle to syntaxin-like proteins on the acceptox membrane, which subsequently triggers a cascade of protein-binding and membrane-fusion events. After transport, GTPase-activating proteins (GAPS) in the target membrane are responsible for converting the GTP-bound Rab proteins to their GDP bound state. And finally, guanine-nucleotide dissociation inhibitor (GDI) recruits the GDP-bound proteins to their membrane of origin.
The cycling of LMW G-proteins between the GTP bound active form and the GDP
bound inactive form is regulated by a variety of proteins. Guanosine nucleotide exchange factors (GEFs) increase the rate of nucleotide dissociation by several orders of magnitude, thus facilitating release of GDP and loading with GTP. The best characterized is the mammalian homolog of the Drosophila Son-of Sevenless protein. Certain Ras-family proteins are also regulated by guanine nucleotide dissociation inhibitors (GDIs), which inhibit GDP dissociation. The intrinsic rate of GTP hydrolysis of the LMW G-proteins is typically very slow, but it can be stimulated by several orders of magnitude by GAPS (Geyer, M. and A. Wittinghofer (1997) Curr. Opin. Struct. Biol. 7:786-?92). Both GEF and GAP activity may be controlled in response to extracellular stimuli and modulated by accessory proteins such as RalBP1 and POB1. Mutant Ras-family proteins, which bind but cannot hydrolyze GTP, are permanently activated, and cause cell proliferation or cancer, as do GEFs that inappropriately activate LMW G-proteins; such as the human oncogene NET1, a Rho-GEF. (Drives, G.T. et al. (1990) Mol. Cell Biol. 10:1793-1798; Alberts; A.S. and R. Treisman (1998) EMBO J.
14:4075-4085).
A member of the ARF family of G-proteins is centaurin beta 1A, a regulator of membrane traffic and the actin cytoskeleton. The centaurin (3 family of GTPase-activating proteins (GAPS) and Arf guanine nucleotide exchange factors contain pleckstrin homology (PH) domains which are activated by phosphoinositides. PH domains bind phosphoinositides, implicating PH domains in signaling processes. Phosphoinositides have a role in converting Arf GTP to Arf GDP via the centaurin j3 family and a role in Arf activation (Kam, J.L. et al. (2000) J.
Biol. Chem. 275:9653-9663).
The rho GAP family is also implicated in the regulation of actin polymerization at the plasma membrane and in several cellular processes. The gene ARHGAP6 encodes GTPase-activating protein 6 isoform 4. Mutations in ARHGAP6, seen as a deletion of a 500 kb critical region in Xp22.3, causes the syndrome microphthalmia with linear skin defects (MLS). MLS is an X-linked dominant, male-lethal syndrome (Prakash, S.K. et al. (2000) Hum. Mol. Genet. 9:477-488).
A member of the Rho family of G-proteins is CDC42, a regulator of cytoskeletal rearrangements required for cell division. CDC42 is inactivated by a specific GAP (CDC42GAP) that strongly stimulates the GTPase activity of CDC42 while having a much lesser effect on other Rho family members. CDC42GAP also contains an SH3-binding domain that interacts with the SH3 domains of cell signaling proteins such as p85 alpha and c-Src, suggesting that CDC42GAP may serve ~4 as a link between CDC42 and other cell signaling pathways (Barfod, E.T. et al.
(1993) J. Biol. Chem.
268:26059-26062).
The Dbl proteins are a family of GEFs for the Rho and Ras G-proteins (Whitehead, I:P. et al.
(1997) Biochim. Biophys. Acta 1332:F1-F23). All Dbl family members contain a Dbl homology (DH) domain of approximately 180 amino acids, as well as a pleckstrin homology (PH) domain located immediately C-terminal to the DH domain. Most Dbl proteins have oncogenic activity, as demonstrated by the ability to transform various cell lines, consistent with roles as regulators of Rho-mediated oncogenic signaling pathways. The kalirin proteins are neuron-specific members of the Dbl family, which are located to distinct subcellular regions of cultured neurons (Johnson, R.C. (2000) J.
1o Cell Biol. 275:19324-19333).
Other regulators of G-protein signaling (RGS) also exist that act primarily by negatively regulating the G-protein pathway by an unknown mechanism (Druey, I~.M. et al.
(1996) Nature 379:742-746). Some 15 members of the RGS family have been identified. RGS
family members are related structurally through similarities in an approximately 120 amino acid region termed the RGS' domain and functionally by their ability to inhibit the interleukin (cytokine) induction of MAP kinase in cultured mammalian 293T cells (Druey et al.~ su ra).
The Tmmuno-associated nucleotide (IAN) family of proteins has GTP-binding activity as indicated by the conserved ATP/GTP-binding site P-loop motif The IAN family includes IAN-1, IAN-4, IAP38, and IAG-1. IAN-1 is expressed in the immune system, specifically in T cells and 2o thymocytes. Its expression is induced during thymic events (Poirier, G.M.C.
et al. (1999) J. Tmmunol.
163:4960-4969). IAP38 is expressed in B cells and macrophages and its expression is induced in splenocytes by pathogens. IAG-1, which is a plant molecule, is induced upon bacterial infection (Krucken, J. et al. (1997) Biochem. Biophys. Res. Commun. 230:167-170). IAN-4 is a mitochondrial membrane protein which is preferentially expressed in hematopoietic precursor 32D cells transfected with wild-type versus mutant forms of the bcr/abl oncogene. The bcr/abl oncogene is known to be associated with chronic myelogenous leukemia, a clonal myelo-proliferative disorder, which is due to the translocation between the bcr gene on chromosome 22 and the abl gene on chromosome 9. Bcr is the breakpoint cluster region gene and abl is the cellular homolog of the transforming gene of the Abelson murine leukemia virus. Therefore, the IAN family of proteins appears to play a role in cell survival in immune responses and cellular transformation (Daheron, L. et al.
(2001) Nucleic Acids Res. 29:1308-1316).
Formin-related genes (FRL) comprise a large family of morphoregulatory genes and have been shown to play important roles in morphogenesis, embryogenesis, cell polarity, cell migration, and cytokinesis through their interaction with Rho family small GTPases. Formin_ was first identified in mouse limb deformi (1d) mutants where the distal bones and digits of all limbs are Eased and reduced in size. FRL contains formin homology domains FH1, FI32, and FH3. The FH1 domain has been shown to bind the Src homology 3 (SH3) domain, WWP/WW domains, and profilin.
The FH2 domain is conserved and was shown to be essential for formin function as disruption at the FHZ domain results in the characteristic td phenotype. The FH3 domain is located at the N-terminus of FRL, and is required for associating with Rac, a Rho family GTPase (Yayoshi-Yamamoto, S. et al. (2000) Mol.
Cell. Biol. 20:6872-6881).
Signaling Complex Protein Domains 1o PDZ domains were named for three proteins in which this domain was initially discovered.
These pxoteins include PSD-95 (postsynaptic density 95), Dlg (Drosophila lethal(1)discs large-1), and ZO-1 (zonula occludens-1). These proteins play important roles in neuronal synaptic transmission, tumor suppression, and cell junction formation, respectively. Since the discovery of these proteins, over sixty additional PDZ-containing proteins have been identified in diverse prokaryotic and eukaryotic organisms. This domain has been implicated in receptor and ion channel clustering and in the targeting of multiprotein signaling complexes to specialized functional regions of the cytosolic face of the plasma membrane: (For a review of PDZ domain-containing proteins, see Pouting, C.P. et al.
(1997) Bioessays 1,9:469-479.) A large proportion of PDZ domains are found in the eukaryotic MAGUK (membrane-associated guanylate kinase) protein family, members of which bind to the intracellular domains of receptors and channels. However, PDZ domains are also found in diverse membrane-localized proteins such as protein tyrosine phosphatases, serinelthreonine kinases, G-protein cofactors, and synapse-associated proteins such as syntrophins and neuronal nitric oxide synthase (nNOS). Generally, about one to three PDZ domains are found in a given protein, although up to nine PDZ domains have been identified in a single protein. The glutamate receptor interacting protein (GRIP) contains seven PDZ domains. GRIP is an adaptor that links certain glutamate receptors to other proteins and may be responsible for the clustering of these receptors at excitatory synapses in the brain (Doug, H. et a1. (1997) Nature 386:279-284). The Drosophila scribble (SCR1B) protein contains both multiple PDZ domains and leucine-rich repeats. SCRIB is located at the epithelial septate junction, which is analogous to the vertebrate tight junction, at the boundary of the apical and basolateral cell surface. SCRIB is involved in the distribution of apical proteins and correct placement of adherens junctions to the basolateral cell surface (Bilder, D. and N.
Perrimon (2000) Nature 403:676-680).
The PX domain is an example of a domain specialized for promoting protein-protein interactions. The PX domain is found in sorting nexins and in a variety of other proteins, including the PhoX components of NADPH oxidase and the Cpk class of phosphatidylinositol 3-kinase. Most PX
domains contain a polyproline motif which is characteristic of SH3 domain binding proteins (Pouting, C.P. (1996) Protein Sci. 5:2353-2357). 5H3 domain-mediated interactions involving the PhoX
components of NADPH oxidase play a role in the formation of the NADPH oxidase multi-protein complex (Leto, T.L. et al. (1994) Proc. Natl. Acad. Sci. USA 91:10650-10654;
Wilson, L. et al.
(1997) Inflamm. Res. 46:265-271).
The SH3 domain is defined by homology to a region of the proto-oncogene c-Src, a cytoplasmic protein tyrosine kinase. 5H3 is a small domain of 50 to 60 amino acids that interacts with proline-rich ligands. SH3 domains are found in a variety of eukaxyotic proteins involved in signal transduction, cell polarization, and membrane-cytoskeleton interactions. In some cases, SH3 domain-containing proteins interact directly with receptor tyrosine kinases. For example, the SLAP-130 protein is a substrate of the T-cell receptor (TCR) stimulated protein kinase.
SLAP-130 interacts via its 5H3 domain with the protein SLP-76 to affect the TCR-induced expression of interleukin-2 (Musci, M.A. et al. (1997) J. Biol. Chem. 272:11674-11677). Another recently identified SH3 domain protein is macrophage actin-associated tyrosine-phosphorylated protein (MAYP) which is phosphorylated during the response of macrophages to colony stimulating factor-1 (CSF-1) and is likely to play a role in regulating the CSF-1-induced reorganization of the actin cytoskeleton (Yeung, Y.-G. et al. (1998) J.
Biol. Chem. 273:30638-30642). The structure of the SH3 domain is characterized by two antiparallel beta sheets packed against each other at right angles. This packing forms a hydrophobic pocket lined 2o with residues that are highly conserved between different SH3 domains. This pocket makes critical hydrophobic contacts with proline residues in the ligand (Feng, S. et a1.
(1994) Science 266:1241-1247).
A novel domain, called the WW domain, resembles the SH3 domain. in its ability to bind proline-rich ligands. This domain was originally discovered in dystrophin, a cytoskeletal protein with direct involvement in Duchenne muscular dystrophy (Bork, P. and M. Sudol (1994) Trends Biochem.
Sci. 19:531-533). WW domains have since been discovered in a variety of intracellular signaling molecules involved in development, cell differentiation, and cell proliferation. The structure of the WW domain is composed of beta strands grouped around four conserved aromatic residues, generally tryptophan.
Like SH3, the SH2 domain is defined by homology to a region of c-Src. SH2 domains interact directly with phospho-tyrosine residues, thus providing an immediate mechanism for the regulation and transduction of receptor tyrosine kinase-mediated signaling pathways. For example, as many as ten distinct SH2 domains are capable of binding to phosphorylated tyrosine residues in the activated PDGF
receptor, thereby providing a highly coordinated and finely tuned response to ligand-mediated receptor activation. (Reviewed in Schaffhausen, B. (1995) Biochim. Biophys. Acta.
1242:61-75.) The BLNK
protein is a linker protein involved in B cell activation, that bridges B cell receptor-associated kinases with SH2 domain effectors that link to various signaling pathways (Fu, C. et al. (1998) Immunity 9:93-103).
The pleckstrin homology (PH) domain was originally identified in pleckstrin, the predominant substrate for protein kinase C in platelets. Since its discovery, this domain has been identified in over 90 proteins involved in intracellular signaling or cytoskeletal organization.
Proteins containing the pleckstrin homology domain include a variety of kinases, phospholipase-C
isoforms, guanine nucleotide release factors, and GTPase activating proteins. For example, members of the FGD1 family contain both Rho-guanine nucleotide exchange factor (GEF) and PH domains, as well as a FYVE zinc finger domain. FGD1 is the gene responsible for faciogenital dysplasia, an inherited skeletal dysplasia (Pasteris, N.G. and J.L. Gorski (1999) Genomics 60:57-66). Many PH domain proteins function in association with the plasma membrane, and this association appears to be mediated by the PH domain itself. PH domains share a common structure composed of two antiparallel beta sheets flanked by an amphipathic alpha helix. Variable loops connecting the component beta strands generally occur within a positively charged environment and may function as ligand binding sites (Lemmon, M.A. et al.
(1996) Cell 85:621-624). Ankyrin (ANK) repeats mediate protein-protein interactions associated with diverse intracellular signaling functions. For example, ANI~ repeats are found in proteins involved in cell proliferation such as kinases, kinase inhibitors, tumor suppressors, and cell cycle control proteins.
(See, for example, Kalus, W. et al. (1997) FEBS Lett. 401:127-132; Ferrante, A.W. et al. (1995) Proc.
Natl. Acad. Sci. USA 92:1911-1915.) These proteins generally contain multiple ANK repeats, each composed of about 33 amino acids. Myotrophin is an ANK repeat protein that plays a key role in the development of cardiac hypertrophy, a contributing factor to many heart diseases. Structural studies show that the myotrophin ANK repeats, like other ANK repeats, each form a helix-turn-helix core preceded by a protruding "tip." These tips are of variable sequence and may play a role in protein-protein interactions. The helix-turn-helix region of the ANK repeats stack on top of one another and are stabilized by hydrophobic interactions (Yang, Y. et al. (1998) Structure 6:619-626). Members of the ASB protein family contain a suppressor of cytokine signaling (SOCS) domain as well as multiple ankyrin repeats (Hilton, D.J. et al. (1998) Proc. Natl. Acad. Sci. USA 95:114-119).
The tetratricopeptide repeat (TPR) is a 34 amino acid repeated motif found in organisms from bacteria to humans. TPRs are predicted to form ampipathic helices, and appear to mediate protein-protein interactions. TPR domains are found in CDC16, CDC23, and CDC27, members of the anaphase promoting complex which targets proteins for degradation at the onset of anaphase. Other processes involving TPR proteins include cell cycle control, transcription repression, stress response, and protein kinase inhibition (Lamb, J.R. et al. (1995) Trends Biochem. Sci.
20:257-259).
The armadillo/beta-catenin repeat is a 42 amino acid motif which forms a superhelix of alpha helices when tandemly repeated. The structure of the armadillo repeat region from beta-catenin revealed a shallow groove of positive charge on one face of the superhelix, which is a potential binding surface. The armadillo repeats of beta-catenin, plakoglobin, and p120'~ bind the cytoplasmic domains of cadherins. Beta-catenin/cadherin complexes are targets of regulatory signals that govern cell adhesion and mobility (Huber, A.H. et al. (1997) Cell 90:871-882).
Eight tandem repeats of about 40 residues (WD-40 repeats), each containing a central Trp-Asp motif, make up beta-transducin (G-beta), which is one of the three subunits (alpha, beta, and gamma) of the guanine nucleotide-binding proteins (G proteins). In higher eukaryotes G-beta exists as a small multigene family of highly conserved proteins of about 340 amino acid residues.
Expressionprofilin~
Array technology can provide a simple way to explore the expression of a single polymorphic 15: gene or the expression profile of a large number of xelated or unrelated genes. When the expression of a single gene is examined, arrays are employed to detect the expression of a specific gene or its variants. When an expression profile is examined,' arrays provide a platform for identifying genes that are tissue specific, are affected by a substance being tested in a toxicology assay, are part of a signaling cascade, carry out housekeeping functions; or are specifically related to a particular genetic predisposition, condition, disease, or disorder.
Breast Cancer There are more than 180,000 new cases of breast cancer diagnosed each year, and the mortality rate for breast cancer approaches 10% of all deaths in females between the ages of 45-54 (I~. Gish (1999) AWIS Magazine 28:7-10). However the survival rate based on early diagnosis of localized breast cancer is extremely high (97 %), compared with the advanced stage of the disease in which the tumor has spread beyond the breast (22%). Current procedures for clinical breast examination are lacking in sensitivity and specificity, and efforts are underway to develop comprehensive gene expression profiles for breast cancer that may be used in conjunction with conventional screening methods to improve diagnosis and prognosis of this disease (Perou CM et al.
(2000) Nature 406:747-752).
Breast cancer is a genetic disease commonly caused by mutations in cellular disease.
Mutations in two genes, BRCA1 and BRCA2, are known to greatly predispose a woman to breast cancer and may be passed on from parents to children (Gish, su ra). However, this type of hereditary breast cancer accounts for only about 5% to 9% of breast cancers, while the vast majority of breast cancer is due to noninherited mutations that occur in breast epithelial cells.
A good deal is already known about the expression of specific genes associated with breast cancer. For example, the relationship between expression of epidermal growth factor (EGF) and its receptor, EGFR, to human mammary carcinoma has been particularly well studied.
(See Khazaie et al., su ra, and references cited therein for a review of this area.) Overexpression of EGFR, particularly coupled with down-regulation of the estrogen receptor, is a marker of poor prognosis in breast cancer patients. In addition, EGFR expression in breast tumor metastases is frequently elevated relative to the primary tumor, suggesting that EGFR is involved in tumor progression and metastasis. This is supported by accumulating evidence that EGF has effects on cell functions related to metastatic potential, such as cell motility, chemotaxis, secretion and differentiation. Changes in expression of other members of the erbB receptor family, of which EGFR is one, have also been implicated in breast cancer. The abundance of erbB receptors, such as HER-2/neu, HER-3, and HER-4, and their ligands in breast cancer points to their functional importance in the pathogenesis of the disease, and may therefore provide targets for therapy of the disease (Bacus, SS et al. (1994) Am ° 15 J Clip Pathol 102:513-S24). Other known markers of breast cancer include a human secreted frizzled protein mRNA that is downregulated in breast tumors; the matrix G1a protein which is overexpressed is human breast. carcinoma cells; Drg1 or RTP, a gene whose expression is diminished in colon, breast, and prostate tumors; maspin, a tumor suppressor gene dovvnregulated in invasive breast carcinomas;
and CaNl9, a member of the S 100 protein family, all of which are down regulated in mammary carcinoma cells relative to normal mammary epithelial cells (Zhou Z et al.
(1998) Int J Cancer 78:95-99; Chen, L et al. (1990} Oncogene 5:1391-1395; UJrix W et al (1999) FEBS Lett 455:23-26; Sager, R
et al. (1996) Curr Top Microbiol Tmmunol 213:51-64; and Lee, SW et al. (1992) Proc Natl Acad Sci USA 89:2504-2508).
Cell lines derived from human mammary epithelial cells at various stages of breast cancer provide a useful model to study the process of malignant transformation and tumor progression as it has been shown that these cell lines retain many of the properties of their parental tumors for lengthy culture periods (Wistuba II et al. (1998) Clip Cancer Res 4:2931-2938). Such a model is particularly useful for comparing phenotypic and molecular characteristics of human mammary epithelial cells at various stages of malignant transformation.
Prostate Cancer Prostate cancer is a common malignancy in men over the age of 50, and the incidence increases with age. In the US, there are approximately 132,000 newly diagnosed cases of prostate cancer and more than 33,000 deaths from the disorder each year.
Once cancer cells arise in the prostate, they are stimulated by testosterone to a more rapid growth. Thus, removal of the testes can indirectly reduce both rapid growth and metastasis of the cancer. Over 95 percent of prostatic cancers are adenocarcinomas which originate in the prostatic acini. The remaining 5 percent are divided between squamous cell and transitional cell carcinomas, both of which arise in the prostatic ducts or other parts of the prostate gland.
As with most cancers, prostate cancer develops through a multistage progression ultimately resulting in an aggressive, metastatic phenotype. The initial step in tumor progression involves the hyperproliferation of normal luminal and/or basal epithelial cells that become hyperplastic and evolve into early-stage tumors. The early-stage tumors are localized in the prostate but eventually may metastasize, particularly to the bone, brain or lung. About 80°l0 of these tumors remain responsive to androgen treatment, an important hormone controlling the growth of prostate epithelial cells.
However, in its most advanced state, cancer growth becomes androgen-independent and there is currently no known treatment for this condition.
A primary diagnostic marker fox prostate cancer is prostate specific antigen (PSA). PSA is a tissue-specific serine protease almost exclusively produced by prostatic epithelial cells. The quantity of PSA correlates with the number and volume of the prostatic epithelial cells, and consequently, the levels of PSA are an excellent indicator of abnormal prostate growth. Men with prostate cancer exhibit an early linear increase in PSA levels followed by an exponential increase prior to diagnosis.
However, since PSA levels are also influenced by factors such as inflammation, androgen and other growth factors, some scientists maintain that changes in PSA levels-'are not useful in detecting individual cases of prostate cancer.
Current areas of career research provide additional prospects for markers as well as potential therapeutic targets for prostate cancer. Several growth factors have been shown to play a critical role in tumor development, growth, and progression. The growth factors Epidermal Growth Factor (EGF), Fibroblast Growth Factor (FGF), and Tumor Growth Factor alpha (TGFa) are important in the growth of normal as well as hyperproliferative prostate epithelial cells, particularly at early stages of tumor development and progression, and affect signaling pathways in these cells in various ways (Lin J et al.
(1999) Cancer Res. 59:2891-2897; Putz T et al. (1999) Cancer Res 59:227-233).
The TGF-J3 family of growth factors are generally expressed at increased levels in human cancers and the high expression levels in many cases correlates with advanced stages of malignancy and poor survival (Gold LI (1999) Crit Rev Oncog 10:303-360). Finally, there are human cell lines representing both the androgen-dependent stage of prostate cancer (LNCap) as well as the androgen-independent, hormone refractory stage of the disease (PC3 and DU-145) that have proved useful in studying gene expression patterns associated with the progression of prostate cancer, and the effects of cell treatments on these expressed genes (Chung TD (1999) Prostate 15:199-207).

The discovery of new molecules for disease detection and treatment, and the polynucleotides encoding them, satisfies a need in the art by providing new compositions which are useful in the diagnosis, prevention, and treatment of cell proliferative, autoimmune/inflammatory, developmental, and neurological disorders, and in the assessment of the effects of exogenous compounds on the expression of nucleic acid and amino acid sequences of molecules for disease detection and treatment.
SUMMARY OF THE INVENTION
The invention features purified polypeptides, molecules for disease detection and treatment, referred to collectively as "MDDT" and individually as "MDDT-1," "1VIDDT-2,"
"MDDT-3,"
"MDDT-4," "MDDT-5," "IV1DDT-6," "MDDT-7," "1VIDDT-8," "MDDT-9," "MDDT-10,"
"MDDT-11," "IVmDT-12," "MDDT-13," "MDDT-14," "MDDT-15," "MDDT-16," "MDDT-17," "MDDT-18," "MDDT-19," "MDDT-20," "MDDT-21," "MDDT-22," "MDDT-23," "MDDT-24," "MDDT-25," "MDDT-26," "MDDT-27," "MDDT-28," "MDDT-29," "MDDT-30," "MDDT-31," "MDDT-1.5 32," "MDDT-33," "MDDT-34," "IMDDT-35," "MDDT-36," "MDDT-37," "MDDT-38,"
and "MDDT-39:" In one aspect, the invention provides an isolated polypeptide selected from the group consisting of a) a polypeptide comprising an amino acid sequence selected from the group consisting' of SEQW N0:1-39, b) a polypeptide comprising a naturally occurring amino acid sequence at least 90% identical to an amino. acid sequence selected from the group consisting of SEQ ID N0:1-39, c) a biologically active fragment of a polypeptide having an amino acid sequence selected from the group consisting of SEQ ID N0:1-39~ and d) an immunogenic fragment of a polypeptide having an amino acid sequence selected from the group consisting of SEQ ID N0:1-39. In one alternative, the invention provides an isolated polypeptide comprising the amino acid sequence of SEQ ID N0:1-39.
The invention further provides an isolated polynucleotide encoding a polypeptide selected from the group consisting of a) a polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO:1-39, b) a polypeptide comprising a naturally occurring amino acid sequence at least 90% identical to an amino acid sequence selected from the group consisting of SEQ 117 N0:1-39, c) a biologically active fragment of a polypeptide having an amino acid sequence selected from the group consisting of SEQ ID N0:1-39, and d) an immunogenic fragment of a polypeptide having an amino acid sequence selected from the group consisting of SEQ ID N0:1-39. In one alternative, the polynucleotide encodes a polypeptide selected from the group consisting of SEQ
D7 N0:1-39. In anothex alternative, the polynucleotide is selected from the group consisting of SEQ ID N0:40-78.
Additionally, the invention provides a recombinant polynucleotide comprising a promoter sequence operably linked to a polynucleotide encoding a polypeptide selected from the group consisting of a) a polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID N0:1-39, b) a polypeptide comprising a naturally occurring amino acid sequence at least 90% identical to an amino acid sequence selected from the group consisting of SEQ )D N0:1-39, c) a biologically active fragment of a polypeptide having an amino acid sequence selected from the group consisting of SEQ ID N0:1-39, and d) an immunogenic fragment of a polypeptide having an amino acid sequence selected from the group consisting of SEQ ID N0:1-39. In one alternative, the invention provides a cell transformed with the recombinant polynucleotide. In another alternative, the invention provides a transgenic organism comprising the recombinant polynucleotide.
The invention also provides a method for producing a polypeptide selected from the group consisting of a) a polypeptide comprising an amino acid sequence selected from the group consisting of SEQ B7 N0:1-39, b) a polypeptide comprising a naturally occurring amino acid sequence at least 90% identical to an amino acid sequence selected from the group consisting of SEQ ll~ NO:1-39, c) a biologically active fragment of a polypeptide having an amino acid sequence selected from the group consisting of SEQ ID NO:1-39, and d) an immunogenic fragment of a polypeptide having an amino acid sequence selected from the group co~isisting of SEQ 1D N0:1-39. The method comprises a) culturing a cell under conditions suitable for expression of the polypeptide, wherein said cell is.
transformed with a recombinant polynucleotide comprising a promoter sequence operably linked to a, polynucleotide encoding the polypeptide, and b) recovering the polypeptide so expressed.
Additionally, the invention provides ~an isolated antibody which specifically binds to a polypeptide selected from the group consisting of a) a polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID N0:1-39, b) a polypeptide comprising a naturally occurring amino acid sequence at least 90% identical to an amino acid sequence selected from the group consisting of SEQ ID N0:1-39, c) a biologically active fragment of a polypeptide having an amino acid sequence selected from the group consisting of SEQ ID N0:1-39, and d) an immunogenic fragment of a polypeptide having an amino acid sequence selected from the group consisting of SEQ
ID N0:1-39.
The invention further provides an isolated polynucleotide selected from the group consisting of a) a polynucleotide comprising a polynucleotide sequence selected from the group consisting of SEQ
ID N0:40-78, b) a polynucleotide comprising a naturally occurring polynucleotide sequence at least 90% identical to a polynucleotide sequence selected from the group consisting of SEQ DJ N0:40-78, c) a polynucleotide complementary to the polynucleotide of a), d) a polynucleotide complementary to the polynucleotide of b), and e) an RNA equivalent of a)-d). In one alternative, the polynucleotide comprises at least 60 contiguous nucleotides.
Additionally, the invention provides a method for detecting a target polynucleotide in a sample, said target polynucleotide having a sequence of a polynucleotide selected from the group consisting of a) a polynucleotide comprising a polynucleotide sequence selected from the group consisting of SEQ
ID N0:40-78, b) a polynucleotide comprising a naturally occurring polynucleotide sequence at least 90% identical to a polynucleotide sequence selected from the group consisting of SEQ m N0:40-78, c) a polynucleotide complementary to the polynucleotide of a), d) a polynucleotide complementary to the polynucleotide of b), and e) an RNA equivalent of a)-d). The method comprises a) hybridizing the sample with a probe comprising at least 20 contiguous nucleotides comprising a sequence complementary to said target polynucleotide in the sample, and which probe specifically hybridizes to said target polynucleotide, under conditions whereby a hybridization complex is formed between said probe and said target polynucleotide or fragments thereof, and b) detecting the presence or absence of said hybridization complex, and optionally, if present, the amount thereof. In one alternative, the probe comprises at least 60 contiguous. nucleotides.
The invention further provides a method for detecting a target polynucleotide in a sample, said target polynucleotide having a sequence of a polynucleotide selected from the group consisting of a) a polynucleotide comprising a polynucleotide sequence selected from the group consisting of SEQ >D
N0:40-78, b) a polynucleotide comprising a naturally occurring polynucleotide sequence at least 90%
identical to a polynucleotide sequence selected from the group consisting of SEQ ID N0:40-78~ c) a.
polynucleotide complementary to the polynucleotide of a), d) a polynucleotide complementary to the polynucleotide of b), and. e) an RNA equivalent of a)-d). The method comprises a) amplifying said target polynucleotide or fragment thereof using polymerase chain reaction ampli~xcation, and b) detecting the presence or absence of said amplified target polynucleotide or fragment thereof, and, optionally, if present, the amount thereof.
The invention further provides a composition comprising an effective amount of a polypeptide selected from the group consisting of a) a polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO:1-39, b) a polypeptide comprising a naturally occurring amino acid sequence at least 90% identical to an amino acid sequence selected from the group consisting of SEQ m N0:1-39, c) a biologically active fragment of a polypeptide having an amino acid sequence selected from the group consisting of SEQ ID N0:1-39, and d) an immunogenic fragment of a polypeptide having an amino acid sequence selected from the group consisting of SEQ ID NO:1-39, and a pharmaceutically acceptable excipient. In one embodiment, the composition comprises an amino acid sequence selected from the group consisting of SEQ ID N0:1-39. The invention additionally provides a method of treating a disease or condition associated with decreased expression of functional MDDT, comprising administering to a patient in need of such treatment the composition.
The invention also provides a method for screening a compound for effectiveness as an agonist of a polypeptide selected from the group consisting of a) a polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ~ N0:1-39, b) a polypeptide comprising a naturally occurring amino acid sequence at least 90% identical to an amino acid sequence selected from the group consisting of SEQ ll~ N0:1-39, c) a biologically active fragment of a polypeptide having an amino acid sequence selected from the group consisting of SEQ m N0:1-39, and d) an immunogenic fragment of a polypeptide having an amino acid sequence selected from the group consisting of SEQ ll~ N0:1-39. The method comprises a) exposing a sample comprising the polypeptide to a compound, and b) detecting agonist activity in the sample. In one alternative, the invention provides a composition comprising an agonist compound identified by the method and a pharmaceutically acceptable excipient. In another alternative, the invention provides a method of treating a disease or condition associated with decreased expression of functional MDDT, comprising administering to a patient in need of such treatment the composition.
Additionally, the invention provides a method for screening a compound for effectiveness as an antagonist of a polypeptide selected from the group consisting of a) a polypeptide comprising an amino acid sequence selected from the group consisting of SEQ m N0:1-39, b) a polypeptide comprising a naturally occurring amino acid sequence at least 90% identical to au amino acid sequence selected from the group consisting of SEQ m NO:1-39, c) a biologically active fragment of a polypeptide.having an amino acid sequence selected from the group consisting of SEQ ID N0:1-39;
and d) an immunogenic fragment of a polypeptide having an amino acid sequence selected from the group consisting of SEQ m N0:1-39. The method comprises a) exposing a sample comprising the polypeptide to a compound, and b) detecting antagonist activity in the sample.
In one alternative, the invention provides a composition comprising an antagonist compound identified by the method and a pharmaceutically acceptable excipient. In another alternative, the invention provides a method of treating a disease or condition associated with overexpression of functional MDDT, comprising administering to a patient in need of such treatment the composition.
The invention further provides a method of screening for a compound that specifically binds to a polypeptide selected from the group consisting of a) a polypeptide comprising an amino acid sequence selected from the group consisting of SEQ m N0:1-39, b) a polypeptide comprising a naturally occurring amino acid sequence at least 90% identical to an amino acid sequence~selected from the group consisting of SEQ 1D N0:1-39, c) a biologically active fragment of a polypeptide having an amino acid sequence selected from the group consisting of SEQ ll~
N0:1-39, and d) an immunogenic fragment of a polypeptide having an amino acid sequence selected from the group consisting of SEQ m N0:1-39. The method comprises a) combining the polypeptide with at least one test compound under suitable conditions, and b) detecting binding of the polypeptide to the test compound, thereby identifying a compound that specifically binds to the polypeptide.
The invention further provides a method of screening for a compound that modulates the activity of a polypeptide selected from the group consisting of a) a polypeptide comprising an amino acid sequence selected from the group consisting of SEQ JD NO:1-39, b) a polypeptide comprising a naturally occurring amino acid sequence at least 90% identical to an amino acid sequence selected from the group consisting of SEQ m N0:1-39, c) a biologically active fragment of a polypeptide having an amino acid sequence selected from the group consisting of SEQ JD
N0:1-39, and d) an immunogenic fragment of a polypeptide having an amino acid sequence selected from the group consisting of SEQ m N0:1-39. The method comprises a) combining the polypeptide with at least one test compound under conditions permissive for the activity of the polypeptide, b) assessing the activity of the polypeptide in the presence of the test compound, and c) comparing the activity of the polypeptide in the presence of the test compound with the activity of the polypeptide in the absence of the test compound, wherein a change in the activity of the polypeptide in the presence of the test compound is indicative of a compound that modulates the activity of the polypeptide.
The invention further provides a method for screening a compound for effectiveness in altering expression of a target polynucleotide, wherein said target polynucleotide comprises a polynucleotide sequence selected from the group consisting of SEQ m N0:40-78, the method comprising a) exposing a sample comprising the target polynucleotide to a compound, b) detecting altered expression of the target polynucleotide, and c) comparing the expression of the target polynucleotide in the presence of varying amounts of the compound and in the absence of the compound.
The invention further provides a method for assessing toxicity of a test compound, said method comprising a) treating a biological sample containing nucleic acids with the test compound; b) hybridizing the nucleic acids of the treated biological sample with a probe comprising at least 20 contiguous nucleotides of a polynucleotade selected from the group consisting of i) a polynucleotide comprising a polynucleotide sequence selected from the group consisting of SEQ
m N0:40-78, ii) a polynucleotide comprising a naturally occurring polynucleotide sequence at least 90% identical to a polynucleotide sequence selected from the group consisting of SEQ m N0:40-78, iii) a polynucleotide having a sealuence complementary to i), iv) a polynucleotide complementary to the polynucleotide of ii), and v) an 1ZNA equivalent of i)-iv). Hybridization occurs under conditions whereby a specific hybridization complex is formed between said probe and a target polynucleotide in the biological sample, said target polynucleotide selected from the group consisting of i) a polynucleotide comprising a polynucleotide sequence selected from the group consisting of SEQ B7 N0:40-78, ii) a polynucleotide comprising a naturally occurring polynucleotide sequence at least 90% identical to a polynucleotide sequence selected from the group consisting of SEQ m N0:40-78, iii) a polynucleotide complementary to the polynucleotide of i), iv) a polynucleotide complementary to the polynucleotide of ii), and v) an RNA equivalent of i)-iv). Alternatively, the target polynucleotide comprises a fragment of a polynucleotide sequence selected from the group consisting of i)-v) above; c) quantifying the amount of hybridization complex; and d) comparing the amount of hybridization complex in the treated biological sample with the amount of hybridization complex in an untreated biological sample, wherein a difference in the amount of hybridization complex in the treated biological sample is indicative of toxicity of the test compound.
1o BRIEF DESCRIPTION OF THE TABLES
Table 1 summarizes the nomenclature for the full length polynucleotide and polypeptide sequences of the present invention.
Table 2 shows the GenBank identification number and annotation of the nearest GenBank homolog for polypeptides of the invention. The probability scores for the matches between each polypeptide and its homolog(s) are also shown.
Table 3 shows structural features of polypeptide sequences of the invention, including predicted motifs and domains, along with the; methods, algorithms, and searchable databases used for analysis of the polypeptades.
Table 4 lists the cDNA andlor genomic DNA fragments which were used to assemble polynucleotide sequences of the invention, along with selected fragments of the polynucleotide sequences.
Table 5 shows the representative cDNA library for polynucleotides of the invention.
Table 6 provides an appendix which describes the tissues and vectors used for construction of the cDNA libraries shown in Table 5.
Table 7 shows the tools, programs, and algorithms used to analyze the polynucleotides and polypeptides of the invention, along with applicable descriptions, references, and threshold parameters.
Table 8 shows single nucleotide polymorphisms found in polynucleotide sequences of the invention, along with allele frequencies in different human populations.
3o DESCRIPTION OF THE INVENTION
Before the present proteins, nucleotide sequences, and methods are described, it is understood that this invention is not limited to the particular machines, materials and methods described, as these may vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to limit the scope of the present invention which will be limited only by the appended claims.
It must be noted that as used herein and in the appended claims, the singular forms "a," "an,"
and "the" include plural reference unless the context clearly dictates otherwise. Thus, for example, a reference to "a host cell" includes a plurality of such host cells, and a reference to "an antibody" is a reference to one or more antibodies and equivalents thereof known to those skilled in the art, and so forth.
Unless defined otherwise, all technical and scientific terms used herein have the same meanings as commonly understood by one of ordinary skill in the art to which this invention belongs.
Although any machines, materials, and methods similar or equivalent to those described herein can be used to practice or test the present invention, the preferred machines, materials and methods are now described. All publications mentioned herein are cited for the purpose of describing and disclosing the cell lines, protocols, reagents and vectors which are reported in the publications and which might be used in connection with the invention. Nothing herein is to be construed as an admission that the invention is not entitled to antedate such disclosure by virtue of prior invention.
DEFINITIONS
"MDDT" refers to the amino acid sequences of substantially purified MDDT
obtained from any species, particularly a mammalian species, including bovine, ovine, porcine, murine, equine, and human, and from any source, whether natural, synthetic, semi-synthetic, or recombinant. ..
The term "agonist" refers to a molecule which intensifies or mimics the biological activity of 1VVIDDT. Agonists may include proteins, nucleic acids, carbohydrates, small molecules, or any other compound or composition which modulates the activity of MDDT either by directly interacting with MDDT or by acting on components of the biological pathway in which MDDT
participates.
An "allelic variant" is an alternative form of the gene encoding MDDT. Allelic variants may result from at least one mutation in the nucleic acid sequence and may result in altered mRNAs or in polypeptides whose structure or function may or may not be altered. A gene may have none, one, or many allelic variants of its naturally occurring form. Common mutational changes which give rise to allelic variants are generally ascribed to natural deletions, additions, or substitutions of nucleotides.
Each of these types of changes may occur alone, or in combination with the others, one or more times in a given sequence.
"Altered" nucleic acid sequences encoding MDDT include those sequences with deletions, insertions, or substitutions of different nucleotides, resulting in a polypeptide the same as MDDT or a polypeptide with at least one functional characteristic of MDDT. Included within this definition are polymorphisms which may or may not be readily detectable using a particular oligonucleotide probe of the polynucleotide encoding MDDT, and improper or unexpected hybridization to allelic variants, with a Locus other than the normal chromosomal locus for the polynucleotide sequence encoding MDDT.
The encoded protein may also be "altered," and may contain deletions, insertions, or substitutions of amino acid residues which produce a silent change and result in a functionally equivalent MDDT.
Deliberate amino acid substitutions may be made on the basis of similarity in polarity, charge, solubility, hydrophobicity, hydrophilicity, and/or the amphipathic nature of the residues, as long as the biological or immunological activity of N~DT is retained. For example, negatively charged amino acids may include aspartic acid and glutamic acid, and positively charged amino acids may include lysine and arginine. Amino acids with uncharged polar side chains having similar hydrophilicity values may include: asparagine and glutamine; and serine and threonine. Amino acids with uncharged side chains having similar hydrophilicity values ray include: leucine, isoleucine, and valine; glycine and alanine;
and phenylalanine and tyrosine.
The terms "amino acid" and "amino acid sequence" refer to an oligopeptide, peptide, polypeptide, or protein sequence, or a fragment of any of these, and to naturally occurring or synthetic molecules. Where "amino acid sequence" is recited to refer to a sequence of a naturally occurring protein molecule, "amino acid sequence" and like terms are not meant to Limit the amino acid sequence to the complete native amino acid sequence associated with the recited protein molecule.
"Amplification" relates to the production of additional copies of a nucleic acid sequence.
Amplification is generally carried out using polymerase chain reaction (PCR) technologies well known., in the art. .
The term "antagonist" refers to .a molecule which inhibits or attenuates the biological activity of MDDT. Antagonists may include proteins such as antibodies, nucleic acids, carbohydrates, small molecules, or any other compound or composition which modulates the activity of MDDT either by directly interacting with MDDT or by acting on components of the biological pathway in which MDDT participates.
The term "antibody" refers to intact immunoglobulin molecules as well as to fragments thereof, such as Fab, F(ab')2, and Fv fragments, which are capable of binding an epitopic determinant.
Antibodies that bind MDDT polypeptides can be prepared using intact polypeptides or using fragments containing small peptides of interest as the immunizing antigen. The polypeptide or oligopeptide used to immunize an attimal (e.g., a mouse, a rat, or a rabbit) can be derived from the translation of RNA, or synthesized chemically, and can be conjugated to a carrier protein if desired. Commonly used carriers that are chemically coupled to peptides include bovine serum albumin, thyroglobulin, and keyhole limpet hemocyanin (KLITj. The coupled peptide is then used to immunize the animal.
The term "antigenic determinant" refers to that region of a molecule (i.e., an epitope) that makes contact with a particular antibody. When a protein or a fragment of a protein is used to ?m_m__unize a host animal, numerous xegions of the protein may induce the production of antibodies which bind specifically to antigenic determinants (particular regions or three-dimensional structures on the protein). An antigenic determinant map compete with the intact antigen (i.e., the immunogen used to elicit the immune response) for binding to an antibody.
The term "aptamer" refers to a nucleic acid or oligonucleotide molecule that binds to a specific molecular target. Aptamers are derived from an in vitro evolutionary process (e.g., SELEX
(Systematic Evolution of Ligands by EXponential Enrichment), described in U.S.
Patent No.
5,270,163), which selects for target-specific aptamer sequences from large combinatorial libraries.
Aptamer compositions may be double-stranded or single-stranded, and may include deoxyribonucleotides, ribonucleotides, nucleotide derivatives, or other nucleotide-like molecules. The nucleotide components of an aptamer may have modified sugar groups (e.g., the 2'-OH group of a ribonucleotide may be replaced by 2'-F or 2'-NH2), which may improve a desired property, e.g., resistance to nucleases or longer lifetime in blood. Aptamers may be conjugated to other molecules, e.g., a high molecular weight carrier to slow clearance of the aptamer from the circulatory system.
Aptamers may be specifically cross-linked to their cognate ligands, e.g., by photo-activation of a cross-linker. (See, e.g., Brody, E.N. and L. Gold (2000) J. Biotechnol. 74:5-13.) The term "intramer" refers to an aptamer which is expressed in vivo. For example, a vaccinia virus-based RNA expression system has been used to express specific RNA
aptamers at high levels in the cytoplasm of leukocytes (Blind, M. et al. (1999) Proc. Natl. Acad. Sci.
USA 96:3606-3610).
The term "spiegelmer" refers to an aptamer which includes L-DNA, L-RNA, or other left handed nucleotide derivatives or nucleotide-like molecules. Aptamers containing left-handed nucleotides are resistant to degradation by naturally occurring enzymes, which normally act on substrates containing right-handed nucleotides.
The term "antisense" refers to any composition capable of base-pairing with the "sense"
(coding) strand of a specific nucleic acid sequence. Antisense compositions may include DNA; RNA;
peptide nucleic acid (PNA); oligonucleotides having modified backbone linkages such as phosphorothioates, methylphosphonates, or benzylphosphonates; oligonucleotides having modified sugar groups such as 2'-methoxyethyl sugars or 2'-methoxyethoxy sugars; or oligonucleotides having modified bases such as 5-methyl cytosine, 2'-deoxyuracil, or 7-deaza-2'-deoxyguanosine. Antisense molecules may be produced by any method including chemical synthesis or transcription. Once introduced into a cell, the complementary antisense molecule base-pairs with a naturally occurring nucleic acid sequence produced by the cell to form duplexes which block either transcription or translation. The designation "negative" or "minus" cau refer to the autisense strand, and the designation "positive" or "plus" can refer to the sense strand of a reference DNA molecule.

The term "biologically active" refers to a protein having structural, regulatory, or biochemical functions of a naturally occurring molecule. Likewise, "immunologically active" or "immunogenic"
refers to the capability of the natural, recombinant, or synthetic MDDT, or of any oligopeptide thereof, to induce a specific immune response in appropriate animals or cells and to bind with specific antibodies.
"Complementary" describes the relationship between two single-stranded nucleic acid sequences that anneal by base-pairing. For example, 5'-AGT-3' pairs with its complement, 3'-TCA-S'.
A "composition comprising a given polynucleotide sequence" and a "composition comprising a 1o given amino acid sequence" refer broadly to any composition containing the given polynucleotide or amino acid sequence. The composition may comprise a dry formulation or an aqueous solution.
Compositions comprising polynucleotide sequences encbding MDDT or fragments of MDDT may be employed as hybridization probes. The probes may be stored in freeze-dried form and may be associated with a stabilizing agent such as a carbohydrate. In hybridizations, the probe may be deployed in an aqueous solutian containing salts (e.g., NaCl), detergents (e.g., sodium dodecyl sulfate;
SDS), and other components (e.g., Denhardt's solution, dry milk, salmon sperm DNA, etc.).
. "Consensus sequence" refers to a nucleic acid sequence which has been subjected to repeated DNA sequence analysis to resolve uncalled bases, extended using the XL-PCR kit (Applied .
Biosystems, Foster City CA) in the 5' andlor the 3' direction, and resequenced, or which has been assembled from one or more overlapping cDNA, EST, or genomic DNA fragments using a computer program for fragment assembly, such as the GELVIEW fragment assembly system (GCG, Madison WI) or Phrap (University of Washington, Seattle WA). Some sequences have been both extended and assembled to produce the consensus sequence.
"Conservative amino acid substitutions" are those substitutions that are predicted to least interfere with the properties of the original protein, i.e., the structure and especially the function of the protein is conserved and not significantly changed by such substitutions. The table below shows amino acids which may be substituted for an original amino acid in a protein and which are regarded as conservative amino acid substitutions.
Original Residue Conservative Substitution 3o Ala Gly, Ser Arg His, Lys Asn Asp, Gln, His Asp Asn, Glu Cys Ala, Ser Gln Asn, Glu, His Glu Asp, Glu, His Gly Ala His Asn, Arg, Gln, Glu Ile Leu, Val Leu Ile, Val Lys Arg, Glu, Glu Met Leu, Ile Phe His, Met, Leu, Trp, Tyr Ser Cys, Thr Thr Ser, Val Trp Phe, Tyr Tyr His, Phe, Trp Val Ile, Leu, Thr Conservative amino acid substitutions generally maintain (a) the structure of the polypeptide backbone in the area of the substitution, for example, as a beta sheet or alpha helical conformation, (b) the charge or hydrophobicity of the molecule at the site of the substitution, and/or (c) the bulk of the side chain.
A "deletion" refers to a change in the amino acid or nucleotide sequence that results in the absence of one or more amino acid residues or nucleotides.
~ The term "derivative" refers to a chemically modified polynucleotide or polypeptide.
Chemical modifications of a polynucleotide can include, for example, replacement of hydrogen by an'.
alkyl, acyl, hydroxyl, or amino group. A derivative polynucleotide encodes a polypeptide which retains at least one biological or immunological function of the natural molecule. A
derivative polypeptide is.
one modified by glycosylation, pegylation, or any similar process that retains at least one biological or immunological function of the polypeptide from which it was derived.
A "detectable label" refers to a reporter molecule or enzyme that is capable of generating a measurable signal and is covalently or noncovalently joined to a polynucleotide or polypeptide.
"Differential expression" refers to increased or upregulated; or decreased, downregulated, or absent gene or protein expression, determined by comparing at least two different samples. Such comparisons may be carned out between, for example, a treated and an untreated sample, or a diseased and a normal sample.
"axon shuffling" refers to the recombination of different coding regions (exons). Since an exon may represent a structural or functional domain of the encoded protein, new proteins may be assembled through the novel reassortment of stable substructures, thus allowing acceleration of the evolution of new protein functions.
A "fragment" is a unique portion of MDDT or the polynucleotide encoding MDDT
which is identical in sequence to but shorter in length than the parent sequence. A
fragment may comprise up to the entire length of the defined sequence, minus one nucleotide/amino acid residue. For example, a fragment may comprise from 5 to 1000 contiguous nucleotides or amino acid residues. A fragment used as a probe, primer, antigen, therapeutic molecule, or for other purposes, may be at least 5, 10, 15, 16, 20, 25, 30, 40, 50, 60, 75, 100, 150, 250 or at least 500 contiguous nucleotides or amino acid residues in length. Fragments may be preferentially selected from certain regions of a molecule. For example, a polypeptide fragment may comprise a certain length of contiguous amino acids selected from the first 250 or 500 amino acids (or first 25% or 50%) of a polypeptide as shown in a certain defined sequence. Clearly these lengths are exemplary, and any length that is supported by the specification, including the Sequence Listing, tables, and figures, may be encompassed by the present embodiments.
A fragment of SEQ 117 N0:40-78 comprises a region of unique polynucleotide sequence that specifically identifies SEQ m N0:40-78, for example, as distinct from any other sequence in the genome from which the fragment was obtained. A fragment of SEQ m N0:40-78 is useful, for example, in hybridization and amplification technologies and in analogous methods that distinguish SEQ
m N0:40-78 from related polynucleotide sequences. The precise length of a fragment of SEQ m N0:40-78 and the region of SEQ II7 NO:40-78 to which the fragment corresponds are routinely determinable by one of ordinary skill in the art based on the intended purpose for the fragment.
A fragment of SEQ m N0:1-39 is encoded by a fragment of SEQ m N0:40-78. A
fragment, of SEQ m N0:1-39 comprises a region of unique amino acid sequence that specifically identifies SEQ m N0:1-39. For example, a fragment of SEQ m N0:1-39 is useful as an immunogenic peptide for the development of antibodies that specifically recognize SEQ m N0:1-39.
The precise length of a fragment of SEQ m N0:1-39 and the region of SEQ m NO:l-39 to which the fragment corresponds are routinely determinable by one of ordinary skill in the art based on the intended purpose for the fragment.
A "full length" polynucleotide sequence is one containing at least a translation initiation codon (e.g., methionine) followed by an open reading frame and a translation termination codon. A "full length" polynucleotide sequence encodes a "full length" polypeptide sequence.
"Homology' refers to sequence similarity or, interchangeably, sequence identity, between two or more polynucleotide sequences or two or more polypeptide sequences.
The terms "percent identity" and "% identity," as applied to polynucleotide sequences, refer to the percentage of residue matches between at least two polynucleotide sequences aligned using a standardized algorithm. Such an algorithm may insert, in a standardized and reproducible way, gaps in the sequences being compared in order to optimize alignment between two sequences, and therefore achieve a more meaningful comparison of the two sequences.
Percent identity between polynucleotide sequences may be determined using the default parameters of the CLUSTAL V algorithm as incorporated into the MEGALIGN
version 3.12e sequence alignment program. This program is part of the LASERGENE software package, a suite of molecular biological analysis programs (DNASTAR, Madison WI). CLUSTAL V is described in Higgins, D.G. and P.M. Sharp (1989) CABIOS 5:151-153 and in Higgins, D.G. et al. (1992) CABIOS
8:189-191. For pairwise alignments of polynucleotide sequences, the default parameters are set as follows: Ktuple=2, gap penalty=5, window=4, and "diagonals saved"=4. The "weighted" residue weight table is selected as the default. Percent identity is reported by CLUSTAL V as the "percent similarity" between aligned polynucleotide sequences.
Alternatively, a suite of commonly used and freely available sequence comparison algorithms is provided by the National Center for Biotechnology Information (NCBI) Basic Local Alignment Search Tool (BLAST) (Altschul, S.F. et al. (1990) J. Mol. Biol. 215:403-410), which is available from several sources, including the NCBZ, Bethesda, MD, and on the Internet at http:l/www.ncbi.nhn.nih.govBLAST/. The BLAST software suite includes various sequence analysis programs including "blastn," that is used to align a known polynucleotide sequence with other polynucleotide sequences fxom a variety of databases. Also available is a tool called "BLAST 2 Sequences" that is used for direct pairwise comparison of~two nucleotide sequences. "BLAST 2 Sequences" can be accessed and used interactively at http://www.ncbi.nlm.nih.gov/gorf/bl2.html. The "BLAST 2 Sequences" tool can be used for both blastn and blastp (discussed below). BLAST
programs are commonly used with gap and other parameters set to default settings. For example, to compare two nucleotide sequences, one may use blastn with the "BLAST 2 Sequences" tool Version 2Ø12 (April-21-2000) set at default parameters. Such default parameters may be, fox example:
Matrix: BLOSUM62 Rewat~d for' match: 1 Penalty fof~ mismatch: -2 Open Gap: 5 and Extension Gap: 2 penalties Gap x drop-off. SO
Expect: 10 Word Size: 11 Filter-: on Percent identity may be measured over the length of an entire defined sequence, for example, as defined by a particular SEQ m number, or may be measured over a shorter length, for example, over the length of a fragment taken from a larger, defined sequence, for instance, a fragment of at least 20, at least 30, at least 40, at least 50, at least 70, at least 100, or at least 200 contiguous nucleotides. Such lengths are exemplary only, and it is understood that any fragment length supported by the sequences shown herein, in the tables, figures, or Sequence Listing, may be used to describe a length over which percentage identity may be measured.
Nucleic acid sequences that do not show a high degree of identity may nevertheless encode similar amino acid sequences due to the degeneracy of the genetic code. It is understood that changes in a nucleic acid sequence can be made using this degeneracy to produce multiple nucleic acid sequences that all encode substantially the same pxotein.
The phrases "percent identity" and "% identity," as applied to polypeptide sequences, refer to the percentage of residue matches between at least two polypeptide sequences aligned using a standardized algorithm. Methods of polypeptide sequence alignment are well-known. Some alignment methods take into account conservative amino acid substitutions. Such conservative substitutions, explained in more detail above, generally preserve the charge and hydrophobicity at the site.of substitution, thus preserving the structure (and therefore function) of the polypeptide.
Percent identity between polypeptide sequences may be determined using the default parameters of the CLUSTAL V algorithm as incorporated.into the MEGALIGN
version 3.12e sequence alignment program (described. and referenced above). For pairwise alignments of polypeptide sequences using CLUSTAL V, the default parameters are set as follows: Ktuple=1, gap penalty=3, window=5, and "diagonals saved"=5. The PAM250 matrix is selected as the default residue weight table. As with polynucleotide alignments, the percent identity is reported by CLUSTAL V as the "percent similarity" between aligned polypeptide sequence pairs.
Alternatively the NCBI BLAST software suite may be used. For example, fox a pairwise comparison of two polypeptide sequences, one may use the "BLAST 2 Sequences"
tool Version 2Ø12 (April-21-2000) with blastp set at default parameters. Such default parameters may be, for example:
Matrix: BLOS UM62 Operc Gap: 1l arad Exterzsiotz Gap: 1 pezzalties Gap x drop-off 50 Expect: 10 Wot~d Size: 3 Filter: on Percent identity may be measured over the length of an entire defined polypeptide sequence, for example, as defined by a particular SEQ ID number, or may be measured over a shorter length, for example, over the length of a fragment taken from a larger, defined polypeptide sequence, for instance, a fragment of at least 15, at least 20, at least 30, at least 40, at least 50, at least 70 or at least 150 contiguous residues. Such lengths are exemplary only, and it is understood that any fragment length supported by the sequences shown herein, in the tables, figures or Sequence Listing, may be used to describe a length over which percentage identity may be measured.
"Human artificial chromosomes" (HACs) are linear microchromosomes which may contain DNA sequences of about 6 kb to 10 Mb in size and which contain all of the elements required for chromosome replication, segregation and maintenance.
The term "humanized antibody" refers to an antibody molecule in which the amino acid sequence in the non-antigen binding regions has been altered so that the antibody more closely resembles a human antibody, and still retains its original binding ability.
"Hybridization" refers to the process by which a polynucleotide strand anneals with a complementary strand through base pairing under defined hybridization conditions. Specific hybridization is an indication that two nucleic acid sequences share a high degree of complementarity.
Specific hybridization complexes form under permissive annealing conditions and remain hybridized after the "washing" step(s). The washing steps) is particularly important in determining the stringency of the hybridization process, with more stringent conditions allowing less non-specific binding, i.e., binding between pairs of nucleic acid strands that are not perfectly matched: Permissive conditions for annealing of nucleic acid sequences are routinely determinable by one of ordinary skill in the art and may be consistent among hybxidization.experiments, whereas wash conditions may be varied among experiments to achieve the desired stringency, and therefore hybridization specificity. .
Permissive annealing conditions occur, for eXample, at 68°C in the presence of about 6 x SSC, about 1% (w/v) SDS, and about 100 ~tglml sheared, denatured salmon sperm DNA.
Generally, stringency of hybridization is expressed, in part, with reference to the temperature under which the wash step is carried out. Such wash temperatures are typically selected to be about 5°C to 20°C lower than the thermal melting point (T"~ for the specific sequence at a defined ionic strength and pH. The Tm is the temperature (under defined ionic strength and pH) at which SO% of the target sequence hybridizes to a perfectly matched probe. An equation for calculating Tm and conditions for nucleic acid hybridization are well known and can be found in Sambrook, J. et al. (1989) Molecular Cloning: A Laboratory Manual, 2"d ed., vol. 1-3, Cold Spring Harbor Press, Plainview NY;
specifically see volume 2, chapter 9.
High stringency conditions for hybridization between polynucleotides of the present invention 3o include wash conditions of 68°C in the presence of about 0.2 x SSC
and about 0.1% SDS, for 1 hour.
Alternatively, temperatures of about 65°C, 60°C, 55°C, or 42°C may be used. SSC concentration may be varied from about 0.1 to 2 x SSC, with SD5 being present at about 0.1%.
Typically, blocking reagents are used to block non-specific hybridization. Such blocking reagents include, for instance, sheared and denatured salmon sperm DNA at about 100-200 ~Cg/mI. Organic solvent, such as formamide at a concentration of about 35-50% v/v, may also be used under particular circumstances, such as for RNA:DNA hybridizations. Useful variations on these wash conditions will be readily apparent to those of ordinary skill in the art. Hybridization, particularly under high stringency conditions, may be suggestive of evolutionary similarity between the nucleotides. Such similarity is strongly indicative of a similar role for the nucleotides and their encoded polypeptides.
The term "hybridization complex" refers to a complex formed between two nucleic acid sequences by virtue of the formation of hydrogen bonds between complementary bases. A
hybridization complex may be formed in solution (e.g., Cot or Rot analysis) or formed between one nucleic acid sequence present in solution and another nucleic acid sequence immobilized on a solid 1o support (e.g., paper, membranes, filters, chips, pins or glass slides, or any other appropriate substrate to which cells or their nucleic acids have been fixed).
The words "insertion" and "addition" refer to changes in an amino acid or nucleotide sequence resulting in the addition of one or more amino acid residues or nucleotides, respectively.
"Immune response" can refer to conditions associated with inflammation, trauma, immune disorders, or infectious or genetic disease, etc. These conditions can be characterized by expression of various factors, e.g.; cytokines, chemokines, and other signaling molecules, which may affect .
cellular and systemic defense systems.
An "immunogenic fragment" is a polypeptide or oligopeptide fragment of MDDT
which is capable of eliciting an immune response when introduced into a living organism, for example, a mammal. The term "immunogenic fragment" also includes any polypeptide or oligopeptide fragment of MDDT which is useful in any of the antibody production methods disclosed herein or known in the art.
The term "microarray" refers to an arrangement of a plurality of polynucleotides, polypeptides, or other chemical compounds on a substrate.
The terms "element" and "array element" refer to a polynucleotide, polypeptide, or other chemical compound having a unique and defined position on a microarray.
The term "modulate" refers to a change in the activity of MDDT. For example, modulation may cause an increase or a decrease in protein activity, binding characteristics, or any other biological, functional, or immunological properties of MDDT.
The phrases "nucleic acid" arid "nucleic acid sequence" refer to a nucleotide, oligonucleotide, polynucleotide, or any fragment thereof. These phrases also refer to DNA or RNA of genomic or synthetic origin which may be single-stranded or double-stranded and may represent the sense or the antisense strand, to peptide nucleic acid (PNA), or to any DNA-like or RNA-like material.
"Operably linked" refers to the situation in which a first nucleic acid sequence is placed in a functional relationship with a second nucleic acid sequence. For instance, a promoter is operably linked to a coding sequence if the promoter affects the transcription or expression of the coding sequence. Operably linked DNA sequences may be in close proximity or contiguous and, where necessary to joie two protein coding regions, in the same reading frame.
"Peptide nucleic acid" (PNA) refexs to an antisense molecule or anti-gene agent which comprises au oligonucleotide of at least about 5 nucleotides in length linked to a peptide backbone of amino acid residues ending in lysine. The terminal lysine confers solubility to the composition. PNAs preferentially bind complementary single stranded DNA ox RNA and stop transcript elongation, and may be pegylated to extend their lifespan in the cell.
l0 "Post-translational modification" of an MDDT may involve lipidation, glycosylation, phosphorylation, acetylation, racemization, proteolytic cleavage, and other modifications known in the art. These processes may occux synthetically or biochemically. Biochemical madificativns will vary by cell type depending an the enzymatic milieu of MDDT.
"Probe" refers to nucleic acid sequences encoding MDDT, their complements, or fragments 15 thereof, which are used to detect identical, allelic or related nucleic acid sequences. Probes are isolated oligonucleotides or polynucleotides attached to a detectable label or reporter molecule.
Typical labels include radioactive isotopes, ligands, chemiluminescent agents, and enzymes. "Primers'.' are short Nucleic acids, usually DNA oligonucleotides, which may be annealed to a target polynucleotide by complementary base-pairing. The primer may then be extended along the target 20 DNA strand by a DNA polymerase enzyme. Primer pairs can be used for amplification {and identification) of a ~aucleic acid sequence, e.g., by the polyx~erase chain reaction (PCR).
Probes anal primers as used in the present invention typically comprise at least 1S contiguous nucleotides of a known. sequence. In order to enhance specificity, longer probes and primers may also be employed, such as probes and primers that comprise at least 20, 25, 30, 40, 50, 60, 70, 80, 90, 100, 25 or at least 150 consecutive nucleotides of the disclosed nucleic acid sequences. Probes and pximers may be considerably longer than these examples, and it is understood that any length supported by the specification, including the tables, figures, and Sequence Listing, maybe used.
Methods for preparing and using probes and primers are described in the references, for example Sambrook, J. et al. (1989) Molecular Cloning: A Laboratory Manual, 2nd ed., vol. 1-3, Cold 30 Spring Harbor Press, Plainview NY; Ausubel, F.M. et aI. (1987) Current Protocols in Molecular Biolo , Greene Publ. Assoc. & Wiley-Intersciences, New York NY; Iunis, M. et al. {1990) PCR
Protocols, A Guide to Methods and Applications, Academic Press, San Diego CA.
PCR primer pairs can be derived from a I~aown sequence, for example, by using computer programs intended for that puxpvse such as Primer (Vexsion 0.5, 1991, Whitehead Institute for Biomedical Research, Cambridge MA).
Oligonucleotides for use as primers are selected using software known in the art for such purpose. For example, OLIGO 4.06 software is useful for the selection of PCR
primer pairs of up to 100 nucleotides each, and for the analysis of oligonucleotides and larger polynucleotides of up to 5,000 nucleotides from an input polynucleotide sequence of up to 32 kilobases.
Similar primer selection programs have incorporated additional features for expanded capabilities. For example, the PrimOU
primer selection program (available to the public from the Genome Center at University of Texas South West Medical Center, Dallas TX) is capable of choosing specific primers from megabase sequences and is thus useful fox designing primers on a genome-wide scope. The Primer3 primer selection program (available to the public from the Whitehead Institute/MIT
Center for Genome Research, Cambridge MA) allows the user to input a "mispriming library," in which sequences to avoid as primer binding sites are user-specified. Primer3 is useful, in particular, for the selection of o~igonucleotides for microarrays. (The source code for the latter two primer selection programs may also be obtained from their respective sources and modified to meet the user's specific needs.) The PrimeGen program (available to the public from the UK Human Genome Mapping Project Resource Centre, Cambridge UK) designs primers based on multiple sequence alignments, thereby allowing selecs~on of primers that hybridize to eithex the most conserved or least conserved regions of aligned nucleic acid sequences. Hence, this program is useful for identification of both unique and conserved .
oligonucleotides and polynucleotide fragments. The oligonucleotides and polynucleotide fragments identified by any of the above selection methods are useful in hybridization technologies, for example, as PCR or sequencing primers, microarray elements, or specific probes to identify fully or partially complementary polynucleotides in a sample of nucleic acids. Methods of oligonucleotide selection are not limited to those described above.
A "recombinant nucleic acid" is a sequence that is not naturally occurring or has a sequence that is made by an artificial combination of two or more otherwise separated segments of sequence.
This artificial combination is often accomplished by chemical synthesis or, more commonly, by the artificial manipulation of isolated segments of nucleic acids, e.g., by genetic engineering techniques such as those described in Sambrook, supt-a. The term recombinant includes nucleic acids that have been altered solely by addition, substitution, or deletion of a portion of the nucleic acid. Frequently, a recombinant nucleic acid may include a nucleic acid sequence operably linked to a promoter sequence.
Such a recombinant nucleic acid may be part of a vector that is used, for example, to transform a cell.
Alternatively, such recombinant nucleic acids maybe part of a viral vector, e.g., based on a vaccinia virus, that could be use to vaccinate a mammal wherein the recombinant nucleic acid is expressed, inducing a protective immunological response in the mammal.

A "regulatory element" refers to a nucleic acid sequence usually derived from untranslated regions of a gene and includes enhancers, promoters, introns, and 5' and 3' untranslated regions (UTRs). Regulatory elements interact with host or viral proteins which control transcription, translation, or RNA stability.
"Reporter molecules" are chemical or biochemical moieties used for labeling a nucleic acid, amino acid, or antibody. Reporter molecules include radionuclides; enzymes;
fluorescent, chemiluminescent, or chromogenic agents; substrates; cofactors; inhibitors;
magnetic particles; and other moieties known in the art.
An "RNA equivalent," in reference to a DNA sequence, is composed of the same linear sequence of nucleotides as the reference DNA sequence with the exception that all occurrences of the nitrogenous base thymine are replaced with uracil, and the sugar backbone is composed of ribose instead of deoxyribose.
The term "sample" is used in its broadest sense. A sample suspected of containing MDDT, nucleic acids encoding IV>DDT, or fragments thereof may comprise a bodily fluid; an extractfrom a cell,. chromosome, organelle, or membrane isolated from a cell; a cell;
genomic DNA, RNA, or cDNA, in solution or bound to a substrate; a tissue; a tissue print; etc.
The terms "specific binding" and "specifically binding" refer to that interaction between a protein or peptide and an agonist, an antibody,, an antagonist, a small molecule, or any natural or synthetic binding composition. The interaction is .dependent upon the presence of a particular structure of the protein, e.g., the antigenic determinant or epitope, recognized by the binding molecule: For example, if an antibody is specific for epitope "A," the presence of a polypeptide comprising the epitope A, or the presence of free unlabeled A, in a reaction containing free labeled A and the antibody will reduce the amount of labeled A that binds to the antibody.
The term "substantially purified" refers to nucleic acid or amino acid sequences that are removed from their natural environment and are isolated or separated, and are at least 60% free, preferably at least 75% free, and most preferably at least 90% free from other components with which they are naturally associated.
A "substitution" refers to the replacement of one or more amino acid residues or nucleotides by different amino acid residues or nucleotides, respectively.
"Substrate" refers to any suitable rigid or semi-rigid support including membranes, filters, chips, slides, wafers, fibers, magnetic or nonmagnetic beads, gels, tubing, plates, polymers, microparticles and capillaries. The substrate canhave a variety of surface forms, such as wells, trenches, pins, channels and pores, to which polynucleotides or polypeptides are bound.
A "transcript image" or "expression profile" refers to the collective pattern of gene expression by a particular cell type or tissue under given conditions at a given time.
"Transformation" describes a process by which exogenous DNA is introduced into a recipient cell. Transformation may occur under natural or artificial conditions according to various methods well known in the art, and may rely on any known method for the insertion of foreign nucleic acid sequences into a prokaryotic or eukaryotic host cell. The method for transformation is selected based on the type of host cell being transformed and may include, but is not limited to, bacteriophage or viral infection, electroporation, heat shock, lipofection, and particle bombardment.
The term "transformed cells" includes stably transformed cells iu which the inserted DNA is capable of replication either as an autonomously replicating plasmid or as part of the host chromosome, as well as transiently transformed cells which express the inserted DNA or RNA for limited periods of time.
A "transgenic organism," as used herein, is any organism, including but not limited to animals and plants, in which one or more of the cells of the organism contains heterologous nucleic acid introduced by way of human intervention, such as by transgenic techniques well known in the art. The nucleic acid is introduced into the cell, directly or indirectly by introduction into a precursor of the cell, by way of deliberate genetic manipulation, such as by microinjection or by infection with a recombinant virus. In one alternative, the nucleic .acid can be introduced by infection with a recombinant viral vector, such as a lentiviral vector (Lois, C. et al. (2002) Science 295:868-872). The term genetic manipulation does not include classical cross-breeding, or in vitro fertilization, but rather is directed to the introduction of a recombinant DNA molecule. The transgenic organisms contemplated in accordance with the present invention include bacteria, cyanobacteria, fungi, plants and animals.
The isolated DNA of the present invention can be introduced into the host by methods known in the art, for example infection, transfection, transformation or transconjugation.
Techniques for transferring the DNA of the present invention into such organisms are widely known and provided in references such as Sambrook et al. (1989), supra.
A "variant" of a particular nucleic acid sequence is defined as a nucleic acid sequence having at least 40% sequence identity to the particular nucleic acid sequence over a certain length of one of the nucleic acid sequences using blastn with the "BLAST 2 Sequences" tool Version 2Ø9 (May-07-1999) set at default parameters. Such a pair of nucleic acids may show, for example, at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% or greater sequence identity over a certain defined length. A variant may be described as, for example, an "allelic" (as defined above), "splice," "species," or "polymorphic" variant. A
splice variant may have significant identity to a reference molecule, but will generally have a greater or lesser number of polynucleotides due to alternate splicing of exons during mRNA processing. The corresponding polypeptide may possess additional functional domains or lack domains that are present in the reference molecule. Species variants are polynucleotide sequences that vary from one species to another. The resulting polypeptides will generally have significant amino acid identity relative to each other. A polymorphic variant is a variation in the polynucleotide sequence of a particular gene between individuals of a given species. Polymorphic variants also~may encompass "siugle nucleotide polymorphisms" (SNPs) in which the polynucleotide sequence varies by one nucleotide base. The presence of SNPs may be indicative of, for example, a certain population, a disease state, or a propensity for a disease state.
A "variant" of a particular polypeptide sequence is defined as a polypeptide sequence having at least 40% sequence identity to the particular polypeptide sequence over a certain length of one of the polypeptide sequences using blastp with the "BLAST 2 Sequences" tool Version 2Ø9 (May-07-1999) set at default parameters. Such a pair of polypeptides may show, for example, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%
or greater sequence identity over a certain defined length of one of the polypeptides.
THE INVENTI~N
The invention is based on the.discovery of new human molecules for disease detection and treatment (MDDT), the polynucleotides encoding MDDT, and the use of these compositions for the diagnosis, treatment, or prevention of cell proliferative, autoimmune/inflammatoxy, developmental, and neurological disorders.
Table 1 summarizes the nomenclature for the full length polynucleotide and polypeptide sequences of the invention. Each polynucleotide and its corresponding polypeptide are correlated to a single Incyte project identification number (Incyte Project ID). Each polypeptide sequence is denoted by both a polypeptide sequence identification number (Polypeptide SEQ 1D NO:) and an Incyte polypeptide sequence number (Incyte Polypeptide ID) as shown. Each polynucleotide sequence is denoted by both a polynucleotide sequence identification number (Polynucleotide SEQ 1D NO:) and an Incyte polynucleotide consensus sequence number (Incyte Polynucleotide ll~) as shown. Column 6 shows the Incyte ID numbers of physical, full length clones corresponding to the polypeptide.and polynucleotide sequences of the invention. The full length clones encode polypeptides which have at least 95% sequence identity to the polypeptide sequences shown in column 3.
Table 2 shows sequences with homology to the polypeptides of the invention as identified by BLAST analysis against the GenBank protein (genpept) database. Columns 1 and 2 show the polypeptide sequence identification number (Polypeptide SEQ ID NO:) and the corresponding Incyte polypeptide sequence number (Iucyte Polypeptide )D) for polypeptides of the invention. Column 3 shows the GenBank identification number (GenBank >D NO:) of the nearest GenBank homolog.
Column 4 shows the probability scores for the matches between each polypeptide and its homolog(s).
Column 5 shows the annotation of the GenBank homolog(s) along with relevant citations where applicable, all of which are expressly incorporated by reference herein.
Table 3 shows various structural features of the polypeptides of the invention. Columns 1 and 2 show the polypeptide sequence identification number (SEQ ID NO:) and the corresponding Iucyte polypeptide sequence number (Incyte Polypeptide ID) for each polypeptide of the invention. Column 3 shows the number of amino acid residues in each polypeptide. Column 4 shows potential phosphorylation sites, and column 5 shows potential glycosylation sites, as determined by the MOTIFS
program of the GCG sequence analysis software package (Genetics Computer Group, Madison WI).
Column 6 shows amino acid residues comprising signature sequences, domains, and motifs. Column 7 shows analytical methods for protein structure/function analysis and in some cases, searchable databases to which the analytical methods were applied.
Together, Tables 2 and 3 summarize the properties of polypeptides of the invention, and these properties establish that the claimed polypeptides are molecules for disease detection and treatment.
For example, SEQ ID N0:2 is. 53 % identical, from residue G3 to residue 6172 and A183 to residue 6659, to human mitogen inducible gene mig-2. (GenBank ID g505033) as determined by the Basic .
Local Alignment Search Tool (BLAST)'. (See Table 2.) The BLAST probability score is 1.2 e-197;
which indicates the probability of obtaining the observed polypeptide sequence alignment by chance.
SEQ ID NO:2 also contains a pleckstrin homology (PFD domain as determined by searching for statistically significant matches in the hidden Markov model (I~VVIM)-based PFAM database of conserved protein family domains. (See Table 3.) Data from BLAST analyses provide further corroborative evidence that SEQ ID N0:2 is a cell signaling molecule.
In another example, SEQ ID NO:14 is 91 % identical, from residue M1 to residue V659, to mouse DMR-N9 (GenBank ID g817954) as determined by the Basic Local Alignment Search Tool (BLAST). (See Table 2.) The BLAST probability score is 0.0, which indicates the probability of obtaining the observed polypeptide sequence alignment by chance. SEQ )D NO:14 also contains WD
repeats as determined by searching for statistically significant matches in the hidden Markov model (HMM)-based PFAM database of conserved protein family domains. (See Table 3.) Data from BLM'S and additional BLAST analyses against the PRODOM and DOMO databases provide further corroborative evidence that SEQ ID N0:14 is a protein associated with myotonic dystrophy.
In another example, SEQ ID NO:24 is 41% identical, from residue I97 to residue N378, to sponge longevity gene SDLAGL (GenBank )D g9798556) as determined by the Basic Local Alignment Search Tool (BLAST). (See Table 2.) The BLAST probability score is 9.8e-58, which indicates the probability of obtaining the observed polypeptide sequence alignment by chance. SEQ
ID N0:24 also contains a homeobox domain as determined by searching for statistically significant matches in the hidden Markov model (IllvIM)-based PFAM database of conserved protein family domains. (See Table 3.) Data from additional BLAST analyses provide further corroborative evidence that SEQ ID N0:24 is a longevity assurance gene.
In another example, SEQ m N0:26 is 75% identical, from residue M1 to residue 51273, to a human protein, ORF2, which contains a reverse transcriptase domain (GenBank ID
g339777) as determined by the Basic Local Alignment Search Tool (BLAST). (See Table 2.) The BLAST
probability score is 0.0, which indicates the probability of obtaining the observed polypeptide sequence alignment by chance. SEQ ID N0:26 also contains AP endonuclease family and reverse transcriptase domains as determined by searching for statistically significant matches in the hidden Markov model (I~VIM)-based PFAM database of conserved protein family domains.
(See Table 3.) Data from BLIIVVIPS, and further BLAST analyses provide further corroborative evidence that SEQ
ID N0:26 contains a reverse transcriptase domain.
In another example, SEQ ID NO:33 is 90% identical, from residue M1 to residue N1275, to a predicted polypeptide comprising a reverse transcriptase domain (GenBank ll~
g339771) as determined by the Basic Local Alignment Search Tool (BLAST). (See Table 2.) The BLAST
probability score is 0.0, which indicates the probability of obtaining the observed polypeptide sequence alignment by chance. SEQ m N0:33~ also contains a reverse transcriptase domain and an AP
endonuclease domain' as determined by searching for statistically significant matches in the hidden Markov model (I~VIM)-based PFAM database of conserved protein family domains.
(See Table 3.) Data from additional BLAST analysis provide further corroborative evidence that SEQ )D N0:33 is a reverse transcriptase. SEQ m NO:1, SEQ ID N0:3-13, SEQ ID N0:15-23, SEQ ID
N0:25, SEQ DJ
N0:27-32 and SEQ ID N0:34-39 were analyzed and annotated in a similar manner.
The algorithms and parameters for the analysis of SEQ m N0:1-39 are described in Table 7.
As shown in Table 4, the full length polynucleotide sequences of the present invention were assembled using cDNA sequences or coding (exon) sequences derived from genomic DNA, or any combination of these two types of sequences. Column 1 lists the polynucleotide sequence identification number (Polynucleotide SEQ ID NO:), the corresponding Incyte polynucleotide consensus sequence number (Incyte ll~) for each polynucleotide of the invention, and the length of each polynucleotide sequence in basepairs. Column 2 shows the nucleotide start (5') and stop (3') positions of the cDNA and/or genomic sequences used to assemble the full length polynucleotide sequences of the invention, and of fragments of the polynucleotide sequences which are useful, for example, in hybridization or amplification technologies that identify SEQ ID
N0:40-78 or that distinguish between SEQ D7 N0:40-78 and related polynucleotide sequences.
The polynucleotide fragments described in Column 2 of Table 4 may refer specifically, for example, to Incyte cDNAs derived from tissue-specific cDNA libraries or from pooled cDNA
libraries. Alternatively, the polynucleotide fragments described in column 2 may refer to GenBank cDNAs or ESTs which contributed to the assembly of the full length polynucleotide sequences. In addition, the polynucleotide fragments described in column 2 may identify sequences derived from the ENSEMBL (The Sanger Centre, Cambridge, UK) database (i.e., those sequences including the 1o designation "ENST"). Alternatively, the polynucleotide fragments described in column 2 may be derived from the NCBI RefSeq Nucleotide Sequence Records Database (i.e., those sequences including the designation "NM" or "NT") or the NCBI RefSeq Protein Sequence Records (i.e., those sequences including the designation "NP"). Alternatively, the polynucleotide fragments described in column 2 may refer to assemblages of both cDNA and Genscan-predicted axons brought together by an "axon stitching" algorithm. Fox example, a polynucleotide sequence identified as FL XXXXXX NI NZ YYI'YY_N3 IV4, represents a "stitched" sequence in which XXXXXX is the identification number of the cluster of sequences to which the algorithm was applied, and YYYYf is the number of the prediction generated by the algorithm, and N1,2~3..., if present, represent specific axons that may have been manually edited during aanalysis (See Example V).
Alternatively, the polynucleotide fragments in column 2 may refer to assemblages of axons brought together by an "axon-stretching" algorithm. For example, a polynucleotide sequence identified as FLXXXXXX_gAAAAA~BBBBB_1 N is a "stretched" sequence, with XXXXXX being the Incyte project identification number, gAAAAA being the GenBank identification number of the human genomic sequence to which the "axon-stretching" algorithm was applied, gBBBBB
being the GenBank identification number or NCBI RefSeq identification number of the nearest GenBank protein homolog, and N referring to specific axons (See Example V). In. instances where a RefSeq sequence was used as a protein homolog for the "axon-stretching" algorithm, a RefSeq identifier (denoted by "NM,"
"NP," or "NT") may be used in place of the GenBank identifier (i. e., gBBBBB).
Alternatively, a .prefix identifies component sequences that were hand-edited, predicted from genomic DNA sequences, or derived from a combination of sequence analysis methods. The following Table lists examples of component sequence prefixes and corresponding sequence analysis methods associated with the prefixes (see Example IV and Example V).

Prefix Type of analysis andlor examples of programs GNN, GFG,Exon prediction from genomic sequences using, for example, ENST GENSCAN (Stanford University, CA, USA) or FGENES

(Computer Genomics Group, The Sanger Centre, Cambridge, UK) GBI Hand-edited analysis of genomic sequences.

FL Stitched or stretched genomic sequences (see Example V).

INCY Full length transcript and exon prediction from mapping of EST

sequences to the genome. Genomic location and EST composition data are combined to predict the exons and resulting transcript.

In some cases, Incyte cDNA coverage redundant with the sequence coverage shown in Table 4 was obtained to confirm the final consensus polynucleotide sequence, but the relevant Incyte l0 cDNA identification numbers are not shown.
Table 5 shows the representative cDNA libraries for those full length polynucleotide sequences which were assembled using Incyte cDNA sequences. The representative cDNA library is the Incyte cDNA library which is most frequently represented by the Incyte cDNA sequences which were used to assemble and confirm the above polynucleotide sequences.
The tissues and 15 vectors which were used to construct the cDNA libraries shown in Table 5 are described in Table 6.
Table 8 shows single nucleotide polyinorphisms (SNPs) found in polynucleotide sequences of the invention, along with allele frequencies in different human populations.
Columns 1 and 2 show the polynucleotide sequence identification number (SEQ ID NO:) and the corresponding Incyte project identification number (PID) for polynucleotides of the invention. Column 3 shows the Incyte 20 identification number for the EST in which the SNP was detected (EST >D), and column 4 shows the identification number for the SNP (SNP ID). Column 5 shows the position within the EST sequence at which the SNP is located (EST SNP), and column 6 shows the position of the SNP within the full-length polynucleotide sequence (CB 1 SNP). Column 7 shows the allele found in the EST sequence.
Columns 8 and 9 show the two alleles found at the SNP site. Column 10 shows the amino acid 25 encoded by the codon including the SNP site, based upon the allele found in the EST. Columns 11-14 show the frequency of allele 1 in four different human populations. An entry of n/d (not detected) indicates that the frequency of allele 1 in the population was too low to be detected, while n/a (not available) indicates that the allele frequency was not determined for the population.
The invention also encompasses MDDT variants. A preferred MDDT variant is one which 30 has at least about 80%, or alternatively at least about 90%, or even at least about 95% amino acid sequence identity to the MDDT amino acid sequence, and which contains at least one functional or structural characteristic of MDDT.
The invention also encompasses polynucleotides which encode MDDT. In a particular embodiment, the invention encompasses a polynucleotide sequence comprising a sequence selected from the group consisting of SEQ ID N0:40-78, which encodes MDDT. The polynucleotide sequences of SEQ ll~ N0:40-78, as presented in the Sequence Listing, embrace the equivalent RNA
sequences, wherein occurrences of the nitrogenous base thymine are replaced with uracil, and the sugar backbone is composed of ribose instead of deoxyribose.
The invention also encompasses a variant of a polynucleotide sequence encoding MDDT. In particular, such a variant polynucleotide sequence will have at least about 70%, or alternatively at least about 85%, or even at least about 95% polynucleotide sequence identity to the polynucleotide sequence encoding MDDT. A particular aspect of the invention encompasses a variant of a polynucleotide sequence comprising a sequence selected from the group consisting of SEQ ll~ N0:40-78 which has at least about 70%, or alternatively at least about 85%, or even at least about 95%
polynucleotide sequence identity to a nucleic acid sequence selected from the group consisting of SEQ
ID N0:40-78. Any one of the polynucleotide variants described above can encode an amino acid sequence which contains at least one functional or structural characteristic of MDDT.
In addition, or in the alternative, a polynucleotide variant of the invention is a splice variant of a polynucleotide sequence encoding MDDT. A splice variant may have portions which have significant sequence identity to the polynucleotide sequence encoding MDDT, but will generally have a greater or lesser number of polynucleotides due to additions or deletions of blocks of sequence arising from alternate splicing of axons during mRNA processing. A splice variant may have less than about 70%, or alternatively less than about 60%, or alternatively less than about 50%
polynucleotide sequence identity to the polynucleotide sequence encoding MDDT over its entire length;
however, portions of the splice variant will have at least about 70%, or alternatively at least about 85%, or alternatively at least about 95%, or alternatively 100% polynucleotide sequence identity to portions of the polynucleotide sequence encoding MDDT. For example, a polynucleotide comprising a sequence of SEQ ID N0:78 is a splice variant of a polynucleotide comprising a sequence of SEQ ll~ N0:47. Any one of the splice variants described above can encode an amino acid sequence which contains at least one functional or structural characteristic of MDDT.
It will be appreciated by those skilled in the art that as a result of the degeneracy of the genetic code, a multitude of polynucleotide sequences encoding MDDT, some bearing minimal similarity to the polynucleotide sequences of any known and naturally occurring gene, may be produced. Thus, the invention contemplates each and every possible variation of polynucleotide sequence that could be made by selecting combinations based on possible codon choices. These combinations are made in accordance with the standard triplet genetic code as applied to the polynucleotide sequence of naturally occurring MDDT, and all such variations are to be considered as being specifically disclosed.
Although nucleotide sequences which encode MDDT and its variants are generally capable of hybridizing to the nucleotide sequence of the naturally occurring MDDT under appropriately selected conditions of stringency, it may be advantageous to produce nucleotide sequences encoding MDDT or its derivatives possessing a substantially different codon usage, e.g., inclusion of non-naturally occurring codons. Codons may be selected to increase the rate at which expression of the peptide occurs in a particular prokaryotic or eukaryotic host in accordance with the frequency with which particular codons are utilized by the host. Other reasons for substantially altering the nucleotide sequence encoding MDDT and its derivatives without altering the encoded amino acid sequences include the production of RNA transcripts having more desirable properties, such as a greater half life, than transcripts produced from the naturally occurring sequence.
The invention also encompasses production of DNA sequences which encode MDDT
and MDDT derivatives, or fragments thereof, entirely by synthetic chemistry. After production, the synthetic sequence may be inserted into any of the many available expression vectors and cell systems using reagents well known in the art. Moreover, synthetic chemistry may be used to introduce mutations into a sequence encoding MDDT or any fragment thereof.
Also encompassed by the invention are polynucleotide sequences that are capable of hybridizing to the claimed polynucleotide sequences, and, in particular, to those shown in SEQ ID
N0:40-78 and fragments thereof under various conditions of stringency. (See, e.g., Wahl, G.M. and S.L. Berger (1987) Methods Enzymol. 152:399-407; Kirmnel, A.R. (1987) Methods Enzymol. 152:507-511.) Hybridization conditions, including annealing and wash conditions, are described in "Definitions."
Methods for DNA sequencing are well known in the art and may be used to practice any of the embodiments of the invention. The methods may employ such enzymes as the Klenow fragment of DNA polymerase I, SEQUENASE (US Biochemical, Cleveland OH), Taq polymerase (Applied Biosystems), thermostable T7 polymerase (Amersham Biosciences, Piscataway NJ), or combinations of polymerases and proofreading exonucleases such as those found in the ELONGASE amplification system (Invitrogen, Carlsbad CA). Preferably, sequence pxeparation is automated with machines such as the MICROLAB 2200 liquid transfer system (Hamilton, Reno NV), PTC200 thermal cycler (MJ Research, Watertown MA) and ABI CATALYST 800 thermal cycler (Applied Biosystems).
Sequencing is then carried out using either the ABI 373 or 377 DNA sequencing system (Applied Biosystems), the MEGABACE 1000 DNA sequencing system (Amersham Biosciences), or other systems known in the art. The resulting sequences are analyzed using a variety of algorithms which are well known in the art. (See, e.g., Ausubel, F.M. (1997) Short Protocols in Molecular Biology, John Wiley & Sons, New York NY, unit 7.7; Meyers, R.A. (1995) Molecular Biology and Biotechnology, Wiley VCH, New York NY, pp. 856-853.) The nucleic acid sequences encoding MDDT may be extended utilizing a partial nucleotide sequence and employing various PCR-based methods known in the art to detect upstream sequences, such as promoters and regulatory elements. For example, one method which may be employed, restriction-site PCR, uses universal and nested primers to amplify unknown sequence from genomic DNA within a cloning vector. (See, e.g., Sarkar, G. (1993) PCR Methods Applic.
2:318-322.) Another method, inverse PCR, uses primers that extend in divergent directions to amplify unknown sequence from a circularized template. The template is derived from restriction fragments comprising a known genomic locus and surrounding sequences. (See, e.g., Triglia, T. et al. (1988) Nucleic Acids Res. 16:8186.) A third method, capture PCR, involves PCR amplification of DNA
fragments adjacent to known sequences in human and yeast artificial chromosome DNA. (See, e.g., Lagerstrom, M. et.
al. (1991) PCR Methods Applic..1:111-119.) In this method, multiple restriction enzyme digestions and legations may be used to insert an engineered double-stranded sequence into a region of unknown -sequence before performing PCR. Other methods which may be used to retrieve unlr~nown sequences are known in the art. (See, e.g., Parker, J.D. et al. (1991) Nucleic Acids Res. 19:3055-3060).
Additionally, one may use PCR, nested primers, and PROMOTERFINDER libraries (Clontech, Pala Alto CA) to walk genomic DNA. This procedure avoids the need to screen libraries and is useful in finding intronlexon junctions. For all PCR-based methods, primers may be designed using commercially available software, such as OLIGO 4.06 primer analysis software (National Biosciences, Plymouth MN) or another appropriate program, to be about 22 to 30 nucleotides in length;
to have a GC content of about 50% or more, and to anneal to the template at temperatures of about 68°C to 72°C.
When screening for full length cDNAs, it is preferable to use libraries that have been size-selected to include larger cDNAs. In addition, random-primed libraries, which often include sequences containing the 5' regions of genes, are preferable for situations in which an oligo d(T) library does not yield a full-length cDNA. Genomic libraries may be useful for extension of sequence into 5' non-transcribed regulatory regions.
Capillary electrophoresis systems which are commercially available may be used to analyze the size or confirm the nucleotide sequence of sequencing or PCR products. In particular, capillary sequencing may employ flowable polymers for electrophoretic separation, four different nucleotide-specific, laser-stimulated fluorescent dyes, and a charge coupled device camera for detection of the emitted wavelengths. Output/light intensity may be converted to electrical signal using appropriate software (e.g., GENOTYPER and SEQUENCE NAVIGATOR, Applied Biosystems), and the entire process from loading of samples to computer analysis and electronic data display may be computer controlled. Capillary electrophoresis is especially preferable for sequencing small DNA fragments which may be present in limited amounts in a particular sample.
In another embodiment of the invention, polynucleotide sequences or fragments thereof which encode MDDT may be cloned in recombinant DNA molecules that direct expression of MDDT, or fragments or functional equivalents thereof, in appropriate host cells. Due to the inherent degeneracy of the genetic code, other DNA sequences which encode substantially the same or a functionally equivalent amino acid sequence may be produced and used to express MDDT.
The nucleotide sequences of the present invention can be engineered using methods generally known in the art in order to alter MDDT-encoding sequences for a variety of purposes including, but not limited to, modification of the cloning, processing, and/or expression of the gene product. DNA
shuffling by random fragmentation and PCR reassembly of gene fragments and synthetic oligonucleotides may be used to engineer the nucleotide sequences. For example, oligonucleotide-mediated site-directed mutagenesis may be used to introduce mutations that create new restriction sites, alter glycosylation patterns, change codon preference, produce splice variants, and so forth.
The nucleotides of the present invention may be subjected to DNA shuffling techniques such as MOLECULARBREEDING (Maxygen Inc., Santa Clara CA; described in U.S. Patent No.
5,837,458; Chang, C.-C. et al. (1999) Nat. Biotechnol. 17:793-797; Christians, F.C. et al. (1999) Nat.
Biotechnol. 17:259-264; and Crameri, A. et al. (1996) Nat. Biotechnol. 14:315-319) to alter or improve the biological properties of MDDT, such as its biological or enzymatic activity or its ability to bind to other molecules or compounds. DNA shuffling is a process by which a library of gene variants is produced using PCR-mediated recombination of gene fragments. The library is then subjected to selection or screening procedures that identify those gene variants with the desired properties. These preferred variants may then be pooled and further subjected to recursive rounds of DNA shuffling and selection/screening. Thus, genetic diversity is created through "artificial"
breeding and rapid molecular evolution. For example, fragments of a single gene containing random point mutations may be recombined, screened, and then reshuffled until the desired properties are optimized. Alternatively, fragments of a given gene may be recombined with fragments of homologous genes in the same gene family, either from the same or different species, thereby maximizing the genetic diversity of multiple naturally occurring genes in a directed and controllable manner.
In another embodiment, sequences encoding MDDT may be synthesized, in whole or in part, using chemical methods well known in the art. (See, e.g., Caruthers, M.H. et al. (1980) Nucleic Acids Symp. Ser. 7:215-223; and Horn, T. et al. (1980) Nucleic Acids Symp. Ser.
7:225-232.) Alternatively, MDDT itself or a fragment thereof may be synthesized using chemical methods.
For example, peptide synthesis can be performed using various solution-phase or solid-phase techniques. (See, e.g., Creighton, T. (1984) Proteins, Structures and Molecular Properties, WH
Freeman, New York NY, pp.
55-60; and Roberge, J.Y. et al. (1995) Science 269:202-204.) Automated synthesis maybe achieved using the ABI 431A peptide synthesizer (Applied Biiosystems). Additionally, the amino acid sequence of MDDT, or any part thereof, may be altered during direct synthesis and/or combined with sequences from other proteins, or any part thereof, to produce a variant polypeptide or a polypeptide having a sequence of a naturally occurring polypeptide.
The peptide may be substantially purified by preparative high performance liquid chromatography. (See, e.g., Chiez, R.M. and F.Z. Regnier (1990) Methods Enzymol. 182:392-421.) The composition of the synthetic peptides may be confirmed by amino acid analysis or by sequencing.
(See, e.g., Creighton, supra, pp. 28-53.) In order to express a biologically active MDDT, the nucleotide sequences encoding MDDT or derivatives thereof may be inserted into an appropriate expression vector, i.e., a vector which contains the necessary elements for transcriptional and.translational control of the inserted coding sequence in a suitable host. These elements include regulatory sequences, such as enhancers, constitutive and inducible promoters, and 5' and 3' untranslated regions in the vector and in polynucleotide sequences encoding MDDT. Such elements may vary in their strength and specificity.
Specific initiation signals may also be used to achieve more efficient translation of sequences encoding MDDT. Such signals include the ATG initiation codon and adjacent sequences e.g. the Kozak sequence. In cases where sequences encoding MDDT and its initiation codon and upstream regulatory sequences are inserted into the appropriate expression vector, no additional transcriptional or translational control signals may be needed. However, in cases where only Boding sequence, or a fragment thereof, is inserted, exogenous translational control signals including an in-frame ATG initiation codon should be provided by the vector. Exogenous translational elements and initiation codons may be of various origins, both natural and synthetic. The efficiency of expression may be enhanced by the inclusion of enhancers appropriate for the particular host cell system used. (See, e.g., Scharf, D.
et al. (1994) Results Probl.
Cell Differ. 20:125-162.) .
Methods which are well known to those skilled in the art may be used to construct expression vectors containing sequences encoding MDDT and appropriate transcriptional and translational control elements. These methods include irz vitro recombinant DNA techniques, synthetic techniques, and in vivo genetic recombination. (See, e.g., Sambrook, J. et aI. (1989) Molecular Cloning, A Laboratory Manual, Cold Spring Harbor Press, Plainview NY, ch. 4, 8, and 16-17; Ausubel, F.M. et al. (1995) Current Protocols in Molecular Biology, John Wiley & Sons, New York NY, ch. 9, 13, and 16.) A variety of expression vector/host systems may be utilized to contain and express sequences encoding 1V~DT. These include, but are not limited to, microorganisms such as bacteria transformed with recombinant bacteriophage, plasmid, or cosmid DNA expression vectors;
yeast transformed with yeast expression vectors; insect cell systems infected with viral expression vectors (e.g., baculovirus);
plant cell systems transformed with viral expression vectors (e.g., cauliflower mosaic virus, CaMV, or tobacco mosaic virus, TMV) or with bacterial expression vectors (e.g., Ti or pBR322 plasmids); or animal cell systems. (See, e.g., Sambrook, supra; Ausubel, supt-a; Van Heeke, G. and S.M. Schuster (1989) J. Biol. Chem. 264:5503-5509; Engelhard, E.K. et al. (1994) Proc. Natl.
Acad. Sci. USA
l0 91:3224-3227; Sandig, V. et al. (1996) Hum. Gene Ther. 7:1937-1945;
Takamatsu, N. (1987) EMBO
J. 6:307-311; The McGraw Hill Yearbook of Science and Technology (1992) McGraw Hill, New York NY, pp. 191-196; Logan, J. and T. Shenk (1984) Proc. Natl. Acad. Sci. USA
81:3655-3659; and Harrington, J.J. et al. (1997) Nat. Genet. 15:345-355.) Expression vectors derived from retroviruses, adenoviruses, or herpes- or vaccinia viruses; or.from various bacterial plasmids~ may be used for delivery of nucleotide sequences to the targeted organ, tissue, or cell population. (See, e.g., Di Nicola, M. et al. (1998) Cancer Gen. Ther. 5(6):350-356; Yu, M. et al. (1993) Proc.
Natl. Acad. Sci. USA
90(13):6340-6344; Buller, R.M. et al. (1985) Nature 317(6040):813-815;
McGregor, D.P. et al. (1994) Mol. T_mmunol. 31(3.):219-226; and Verma, I:M. and N. Somia (1997) Nature 389:239-242.) The invention is not limited by the host cell employed.
In bacterial systems a number of cloning and expression vectors may be selected depending upon the use intended for polynucleotide sequences encoding MDDT. For example, routine cloning, subcloning, and propagation of polynucleotide sequences encoding MDDT can be achieved using a multifunctional E. coli vector such as PBLUESCRIPT (Stratagene, La Jolla CA) or PSPORT1 plasmid (Invitrogen). Ligation of sequences encoding MDDT into the vector's multiple cloning site disrupts the lacZ gene, allowing a colorimetric screening procedure for identification of transformed bacteria containing recombinant molecules. In addition, these vectors may be useful for in vitro transcription, dideoxy sequencing, single strand rescue with helper phage, and creation of nested deletions in the cloned sequence. (See, e.g., Van Heeke, G. and S.M. Schuster (1989) J. Biol. Chem.
264:5503-5509.) When large quantities of MDDT are needed, e.g. for the production of antibodies, vectors which direct high level expression of MDDT may be used. For example, vectors containing the strong, inducible SP6 or T7 bacteriophage promoter may be used.
Yeast expression systems may be used for production of MDDT. A number of vectors containing constitutive or inducible promoters, such as alpha factor, alcohol oxidase, and PGH
promoters, may be used in the yeast Saccharomyces cer-evisiae or Piclaia pastoris. In addition, such vectors direct either the secretion or intracellular retention of expressed proteins and enable integration of foreign sequences into the host genome for stable propagation. (See, e.g., Ausubel, 1995, supra;
Bitter, G.A. et al. (1987) Methods Enzymol. 153:516-544; and Scorer, C.A. et al. (1994) Bio/Technology 12:181-184.) Plant systems may also be used for expression of MDDT. Transcription of sequences encoding MDDT may be driven by viral promoters, e.g., the 35S and 19S
promoters of CaMV used alone or in combination with the omega leader sequence from TMV (Takamatsu, N.
(1987) EMBO J.
6:307-311). Alternatively, plant promoters such as the small subunit of RUBISCO or heat shock promoters may be used. (See, e.g., Coruzzi, G. et al. (1984) EMBO J. 3:1671-1680; Brogfie, R. et al.
to (1984) Science 224:838-843; and Winter, J. et al. (1991),Results Probl.
Cell Differ. 17:85-105.) These constructs can be introduced into plant cells by direct DNA transformation or pathogen-mediated transfection. (See, e.g., The McGraw Hill Yearbook of Science and Technology (1992) McGraw Hill, New York NY, pp. 191-196.) In mammalian cells, a number of viral-based expression systems may be utilized. In cases where an adenovirus is used as au expression vector, sequences encoding MDDT
may be ligated into an adenovirus transcription/translation complex consisting of the late promoter 'and tripartite leader sequence. Insertion in~ a non-essential E1 or E3 'region of the viral genome may be used to obtain ,:
infective virus which expresses MDDT in host cells. (See, e.g., Logan, J. and T. Shenk (1984) Proc..
Natl. Acad. Sci. USA 81:3655-3659..) In addition, transcription enhancers, such as the Rous sarcoma 2o virus (RSV) enhancer, may be used to.increase expression in mammalian host cells. SV40 or EBV-based vectors may also be used for high-level protein expression.
Human artificial chromosomes (HACs) may also be employed to deliver larger fragments of DNA than can be contained in and expressed from a plasmid. HACs of about 6 kb to 10 Mb are constructed and delivered via conventional delivery methods (liposomes, polycationic amino polymers, or vesicles) for therapeutic purposes. (See, e.g., Harrington, J.J. et al.
(1997) Nat. Genet. 15:345-355.) For long term production of recombinant proteins in mammalian systems, stable expression of MDDT in cell lines is preferred. For example, sequences encoding MDDT can be transformed into cell lines using expression vectors which may contain viral origins of replication and/or endogenous expression elements and a selectable marker gene on the same or on a separate vector. Following the introduction of the vector, cells may be allowed to grow for about 1 to 2 days in enriched media before being switched to selective media. The purpose of the selectable marker is to confer resistance to a selective agent, and its presence allows growth and recovery of cells which successfully express the introduced sequences. Resistant clones of stably transformed cells may be propagated using tissue culture techniques appropriate to the cell type.
Any number of selection systems may be used to recover transformed cell lines.
These include, but are not limited to, the herpes simplex virus thymidine kinase and adenine phosphoribosyltrausferase genes, for use in tk and apt cells, respectively.
(See, e.g., Wigler, M. et al. (1977) Cell 11:223-232; Lowy, I. et al. (1980) Cell 22:817-823.) Also, autimetabolite, antibiotic, or herbicide resistance can be used as the basis for selection. For eXample, dhfr confers resistance to methotrexate; ~aeo confers resistance to the aminoglycosides neomycin and G-418; and als and pat confer resistance to chlorsulfuron and phosphinotricin acetyltrausferase, respectively. (See, e.g., Wigler, M. et al. (1980) Proc. Natl. Acad. Sci. USA 77:3567-3570; Colbere-Garapin, F. et al. (1981) J. Mol. Biol. 150:1-14.) Additional selectable genes have been described, e.g., trpB and hisD, which alter cellular requirements for metabolites. (See, e.g., Hartmau, S.C. and R.C. Mulligan (1988) Proc.
Natl. Acad. Sci. USA 85:8047-8051.) Visible markers, e.g., anthocyanins, green fluorescent proteins (GFP; Clontech),13 glucuronidase and its substrate 13-glucuronide, or luciferase and its substrate luciferin may be used. These markers can be used not only to identify transformants, but also to . quantify the amount of transient or stable protein expression attributable to a specific vector system.
(See, e:g., Rhodes, C.A. (1995) Methods Mol..Biol. 55:121-131:) Although the presence/absence of marker gene expression suggests that the gene of interest .
is also present, the presence and expression of the gene may .need to be confirmed. For example, if .
the. sequence encoding MDDT is inserted withiu a marker' gene sequence, trausformed cells containing sequences encoding MDDT can be identified by the absence of marker gene function.
Alternatively, a marker gene can be placed in tandem with a sequence encoding MDDT under the control of a single promoter. Expression of the marker gene in response to induction or selection usually indicates expression of the tandem gene as well.
In general, host cells that contain the nucleic acid sequence encoding MDDT
and that express MDDT may be identified by a variety of procedures known to those of skill in the art. These procedures include, but are not limited to, DNA-DNA or DNA-RNA hybridizations, PCR
amplification, and protein bioassay or immunoassay techniques which include membrane, solution, or chip based technologies for the detection and/or quantification of nucleic acid or protein sequences.
T_m_m__unological methods for detecting and measuring the expression of MDDT
using either specific polyclonal or monoclonal antibodies are known in the art. Examples of such techniques include enzyme-linked immunosorbent assays (ELISAs), radioimmunoassays (RIAs}, and fluorescence activated cell sorting (FACS). A two-site, monoclonal based immunoassay utilizing monoclonal antibodies reactive to two non-interfering epitopes on MDDT is preferred, but a competitive binding assay may be employed. These and other assays are well known in the art. (See, e.g., Hampton, R. et al. (I990) Serolo 'cal Methods, a Laboratory Manual, APS
Press, St. Paul MN, Sect. IV; Coligan, J.E. et al. (1997) Current Protocols in Tmrrmunoloay, Greene Pub. Associates and Wiley-Interscience, New York NY; and Pound, J.D. (1998) Immunochemical Protocols, Humana Press, Totowa NJ.) A wide variety of labels and conjugation techniques are known by those skilled in the art and may be used in various nucleic acid and amino acid assays. Means for producing labeled hybridization or PCR probes for detecting sequences related to polynucleotides encoding MDDT
include oligolabeling, nick translation, end-labeling, or PCR amplification using a labeled nucleotide.
Alternatively, the sequences encoding MDDT, or any fragments thereof, may be cloned into a vector for the production of an mRNA probe. Such vectors axe known in the art, are commercially available, and may be used to synthesize RNA probes irt vitt-o by addition of an appropriate RNA polymerase such as T7, T3, or SP6 and labeled nucleotides. These procedures may be conducted using a variety of commercially available kits, such as those provided by Amersham Biosciences, Promega (Madison WI), and US Biochemical. Suitable reporter molecules or labels which may be used for ease of detection include radionuclides, enzymes, fluorescent, chemiluminescent, or chromogenic agents, as well as substrates, cofactors, inhibitors, magnetic particles; and the like.
Host cells transformed with nucleotide, sequences encoding MDDT may be cultured under conditions suitable for the expression and recovery .of the protein from cell culture. The protein produced by a transformed cell may be secreted or retained intracellularly depending on the sequence and/or the vector used. As will be understood by those of skill in the art, expression vectors containing polynucleotides which encode MDDT may be designed to contain signal sequences which direct secretion of MDDT through a prokaryotic or eukaryotic cell membrane.
Iu addition, a host cell strain may be chosen for its ability to modulate expression of the inserted sequences or to process the expressed protein in the desired fashion.
Such modifications of the polypeptide include, but are not limited to, acetylation, carboxylation, glycosylation, phosphorylation, lipidation, and acylation. Post-translational processing which cleaves a "prepro" or "pro" form of the protein may also be used. to specify protein targeting, folding, and/or activity. Different host cells which have specific cellular machinery and characteristic mechanisms for post-translational activities (e.g., CHO, HeLa, MDCK, HE~293, and WI38) are available from the American Type C~.tlture Collection (ATCC, Manassas VA) and may be chosen to ensure the correct modification and processing of the foreign protein.
In another embodiment of the invention, natural, modified, or recombinant nucleic acid sequences encoding MDDT may be ligated to a heterologous sequence resulting in translation of a fusion protein in any of the aforementioned host systems. For example, a chimeric MDDT protein containing a heterologous moiety that can be recognized by a commercially available antibody may facilitate the screening of peptide libraries for inhibitors of MDDT activity.
Heterologous protein and peptide moieties may also facilitate purification of fusion proteins using commercially available affinity matrices. Such moieties include, but are not limited to, glutathione S-transferase (GST), maltose binding protein (MBP), thioredoxin (Trx), calmodulin binding peptide (CBP), 6-His, FLAG, c-myc, and hemagglutinin (HA). GST, MBP, Trx, CBP, and 6-His enable purification of their cognate fusion proteins on immobilized glutathione, maltose, phenylarsine oxide, calmodulin, and metal-chelate resins, respectively. FLAG, c-myc, and hemagglutinin (HA) enable imrnunoaffinity puri~tcation of fusion proteins using commercially available monoclonal and polyclonal antibodies that specifically recognize these epitope tags. A fusion protein may also be engineered to contain a proteolytie cleavage site located between the MDDT encoding sequence and the heterologous protein sequence, so that MDDT may be cleaved away from the heterologous moiety following purification.
Methods for fusion protein expression and purification are discussed in Ausubel (1995, sup~'a, ch. 10}. A variety of commercially available kits may also be used to facilitate expression and purification of fusion proteins.
In a further embodiment of the invention, synthesis of radiolabeled MDDT may be achieved .
in vitro using the TNT rabbit reticulocyte lysate or wheat germ extract system (Promega). These systems couple transcription and translation of protein-coding sequences operably associated with the T7; T3, or SP6 promoters. Translation takes place in the presence of a radiolabeled amino acid precursor, for example, 35S-methionine.
MDDT of the present invention or fragments thereof may be used to screen for compounds that specifically bind to MDDT. At least one and up to a plurality of test compounds may be screened for specific binding to MDDT. Examples of test compounds include antibodies, oligonucleotides, proteins (e.g., Iigands or receptors), or small molecules. In one embodiment, the compound thus identified is closely related to the natural ligand of MDDT, e.g., a ligand or fragment thereof, a natural substrate, a structural or functional mimetic, or a natural binding partner.
(See, e.g., Coligan, J.E. et al.
(1991) Current Protocols in Itnmunolo~y 1(2):Chapter 5.) In another embodiment, the compound thus identified is a natural ligand of a receptor MDDT. (See, e.g., Howard, A.D. et al. (2001) Trends Pharmacol. Sci.22:132-140; Wise, A. et al. (2002) Drug Discovery Today 7:235-246.) In other embodiments, the compound can be closely related to the natural receptor to which MDDT binds, at least a fragment of the receptor, or a fragment of the receptor including all or a portion of the ligand binding site or binding pocket. For example, the compound may be a receptor for MDDT which is capable of propagating a signal, or a decoy receptor for MDDT
which is not capable of propagating a signal (Ashkenazi, A. and V.M. Divit (1999) Curr. Opin. Cell Biol. 11:255-260;
Mantovani, A. et al. (2001) Trends Tmmunol. 22:328-336). The compound can be rationally designed using known techniques. Examples of such techniques include those used to construct the compound etanercept (ENBREL; Tmmunex Corp., Seattle WA), which is efficacious for treating rheumatoid arthritis in humans. Etanercept is an engineered p75 tumor necrosis factor (TNF) receptor dimer linked to the Fc portion of human IgGl (Taylor, P.C. et al. (2001) Curr. Opin.
Trmmunol. 13:611-616).
In one embodiment, screening for compounds which specifically bind to, stimulate, or inhibit MDDT involves producing appropriate cells which express MDDT, either as a secreted protein or on the cell membrane. Preferred cells include cells from mammals, yeast, Dr-osophila, or E. coli. Cells expressing MDDT or cell membrane fractions which contain MDDT are then contacted with a test compound and binding, stimulation, or inhibition of activity of either MDDT or the compound is analyzed.
An assay may simply test binding of a test compound to the polypeptide, wherein binding is detected by a fluorophore, radioisotope, enzyme conjugate, or other detectable label. For example; the assay may.comprise the steps of combining at least one test compound with MDDT, either in solution .
or affixed to a solid support, and detecting the binding of MDDT to the compound. Alternatively, the assay may detect or measure binding of a test compound in the presence of a labeled competitor.
Additionally, the assay may be carried out using cell-free preparations, chemical libraries, ox natural product mixtures, and the test compounds) may be free in solution or affixed to a solid support.
An assay can be used to assess the ability of a compound to bind to its natural ligand and/or to inhibit the binding of its natural ligand to its natural receptors. Examples of such assays include radio labeling assays such as those described in U.S. Patent No. 5,914,236 and U.S.
Patent No. 6,372,724.
In a related embodiment, one or more amino acid substitutions can be introduced into a polypeptide compound (such as a receptor) to improve or alter its ability to bind to its natural ligands. (See, e.g., Matthews, D.J. and J.A. Wells. (1994) Chem. Biol. 1:25-30.) In another related embodiment, one or more amino acid substitutions can be introduced into a polypeptide compound (such as a ligand) to improve or alter its ability to bind to its natural receptors. (See, e.g., Cunningham, B.C. and J.A. Wells (1991) Proc. Natl. Acad. Sci. USA 88:3407-3411; Lowman, H.B. et al. (1991) J.
Biol. Chem.
266:10982-10988.) MDDT of the present invention or fragments thereof may be used to screen for compounds that modulate the activity of MDDT. Such compounds may include agonists, antagonists, or partial or inverse agonists. In one embodiment, an assay is performed under conditions permissive for MDDT
activity, wherein MDDT is combined with at least one test compound, and the activity of MDDT in the presence of a test compound is compared with the activity of MDDT in the absence of the test compound. A change in the activity of MDDT in the presence of the test compound is indicative of a compound that modulates the activity of MDDT. Alternatively, a test compound is combined with an in vitro or cell-free system comprising MDDT under conditions suitable for MDDT activity, and the assay is performed. In either of these assays, a test compound which modulates the activity of MDDT may do so indirectly and need not come in direct contact with the test compound. At least one and up to a plurality of test compounds may be screened.
In another embodiment, polynucleotides encoding MDDT or their mammalian homologs may be "knocked out" in au animal model system using homologous recombination in embryonic stem (ES) cells. Such techniques are well known in the art and are useful for the generation of animal models of human disease. (See, e.g., U.S. Patent No. 5,175,383 and U.S. Patent No.
5,767,337.) For example, mouse ES cells, such as the mouse 129/SvJ cell line, are derived from the early mouse embryo and grown in culture. The ES cells are transformed with a vector containing the gene of interest disrupted by a marker gene, e.g., the neomycin phosphotrausferase gene (neo; Capecchi, M.R. (1989) Science 244:1288-1292). The vector integrates into the corresponding region of the host genome by homologous recombination. Alternatively, homologous recombination takes place using the Cre-loxP
system to knockout a gene of interest in a tissue- or developmental stage-specific manner (Marth, J.D.
(1996) Clip. Invest. 97:1999-2002; Wagner, K.U. et al. (1997) Nucleic Acids Res. 25:4323-4330).
Transformed ES cells are identified and microinjected into mouse cell blastocysts such as those from the C57BIJ/6 mouse strain. The blastocysts are surgically transferred to pseudopregnani: dams; and, the resulting chimeric.progeny are genotyped and bred to produce heterozygous or homozygous ,.
strains. Transgenic animals thus generated may be tested with potential therapeutic or toxic agents.::, Polynucleotides encoding MDDT may also be manipulated in vitro in ES cells derived from human blastocysts. Human ES cells have the potential to differentiate into at least eight separate cell lineages including endoderm, mesoderm, and ectodermal cell types. These cell lineages differentiate into, for example, neural cells, hematopoietic lineages, and cardiomyocytes (Thomson, J.A. et al.
(1998) Science 282:1145-1147).
Polynucleotides encoding MDDT can also be used to create "knockin" humanized animals (pigs} or trausgenic animals (mice or rats) to model human disease. With knockin technology, a region of a polynucleotide encoding MDDT is injected into animal ES cells, and the injected sequence integrates into the animal cell genome. Transformed cells are injected into blastulae, and the blastulae are implanted as described above. Transgenic progeny or inbred lines are studied and treated with potential pharmaceutical agents to obtain information on treatment of a human disease. Alternatively, a mammal inbred to overexpress MDDT, e.g., by secreting MDDT in its milk, may also serve as a convenient source of that protein (Janne, J. et al. (1998) Biotechnol. Annu.
Rev. 4:55-74).
THERAPEUTICS
Chemical and structural similarity, e.g., in the context of sequences and motifs, exists between regions of MDDT and molecules for disease detection and treatment. In addition, examples of tissues expressing MDDT can be found in Table 6 and can also be found in Example XI.
Therefore, MDDT
appears to play a role in cell proliferative, autoimmune/inflammatory, developmental, and neurological disorders. In the treatment of disorders associated with increased MDDT
expression or activity, it is desirable to decrease the expression or activity of MDDT. In the treatment of disorders associated with decreased MDDT expression or activity, it is desirable to increase the expression or activity of MDDT.
Therefore, in one embodiment, MDDT or a fragment or derivative thereof may be administered to a subject to treat or prevent a disorder associated with decreased expression or activity of MDDT. Examples of such disorders include, but are not limited to, a cell proliferative disorder such as actinic keratosis, arteriosclerosis, atherosclerosis, bursitis, cirrhosis, hepatitis, mixed connective tissue disease (MCTD), myelofibrosis, paroxysmal nocturnal hemoglobinuria, polycythemia vera, psoriasis, primary thrombocythemia, and cancers including adenocarcinoma, leukemia, lymphoma, melanoma, myeloma, sarcoma, teratocarcinoma, and, in~particular, cancers of the adrenal. ' gland, bladder, bone, bone marrow, brain, breast, cervix, gall bladder, ganglia, gastrointestinal tract, heart, kidney,.liver, lung, muscle, ovary, pancreas,~parathyroid, penis, prostate, salivary glands, skin, .
spleen; testis, thymus, thyroid, and uterus; an autoimmune/inflarnmatory disorder such as acquired imrnunodeficiency syndrome (AmS), Addison's disease; adult respiratory distress syndrome, allergies ankylosing spondylitis, amyloidosis, anemia, asthma, atherosclerosis;
autoimmune hemolytic anemia, autoimmune thyroiditis, autoimmune polyendocrinopathy-candidiasis-ectoderinal dystrophy (APECED), bronchitis, cholecystitis, contact dermatitis, Crohn's disease, atopic dermatitis, derxnatomyositis, diabetes mellitus, emphysema, episodic lymphopenia with lymphocytotoxins, erythroblastosis fetalis, erythema nodosum, atrophic gastritis, glomerulonephritis, Goodpasture's syndrome, gout, Graves' disease, Hashimoto's thyroiditis, hypereosinophilia, irritable bowel syndrome, multiple sclerosis, myasthenia gravis, myocardial or pericardial inflammation, osteoarthritis, osteoporosis, pancreatitis, polymyositis, psoriasis, Reiter's syndrome, rheumatoid arthritis, scleroderma, Sjogren's syndrome, systemic anaphylaxis, systemic lupus erythematosus, systemic sclerosis, thrombocytopenic purpura, ulcerative colitis, uveitis, Werner syndrome, complications of cancer, . hemodialysis, and extracorporeal circulation, viral, bacterial, fungal, parasitic, protozo~l, and hehninthic infections, and trauma; a developmental disorder such as renal tubular acidosis, anemia, C~shing's syndrome, achondroplastic dwarfism, Duchenne and Becker muscular dystrophy, epilepsy, gonadal dysgenesis, WAGR syndrome (Wilms' tumor, aniridia, genitourinary abnormalities, and mental retardation), Smith-Magenis syndrome, myelodysplastic syndrome, hereditary mucoepithelial dysplasia, hereditary keratodermas, hereditary neuropathies such as Charcot-Marie-Tooth disease and neurofibromatosis, hypothyroidism, hydrocephalus, seizure disorders such as Syndenham's chorea and cerebral palsy, spina bifida, anencephaly, craniorachischisis, congenital glaucoma, cataract, and sensorineural hearing loss; and a neurological disorder such as epilepsy, ischemic cerebrovascular disease, stroke, cerebral neoplasms, Alzheimer's disease, Pick's disease, Huntington's disease, dementia, Parkinson's disease and other extrapyramidal disorders, amyotrophic lateral sclerosis and other motor neuron disorders, progressive neural muscular atrophy, retinitis pigmentosa, hereditary ataxias, multiple sclerosis and other demyelinating diseases, bacterial and viral meningitis, brain abscess, subdural empyema, epidural abscess, suppurative intracranial thrombophlebitis, myelitis and radiculitis, viral central nervous system disease, prion diseases including kuru, Creutzfeldt-Jakob disease, and Gerstmann-Straussler-Scheinker syndrome, fatal familial insomnia, nutritional and metabolic diseases of the nervous system, neurofibromatosis, tuberous sclerosis, cerebelloretinal hemangioblastomatosis, encephalotrigeminal syndrome, mental retardation and other developmental disorders of the central nervous system including Down syndrome, cerebral palsy, neuroskeletal disorders, autonomic nervous system disorders, cranial nerve disorders, spinal cord diseases, muscular dystrophy and other neuromuscular disorders peripheral nervous system disorders, dermatomyositis and polymyositis, inherited, metabolic, endocrine, and toxic myopathies, myasthenia gravis, periodic paralysis, mental disorders including mood, anxiety, and schizophrenuc disorders, seasonal affective disorder (SAD), akathesia, amnesia, catatonia, diabetic neuropathy, tardive dyskinesia, dystonias, paranoid psychoses, postherpetic neuralgia, Tourette's disorder,~progressive supranuclear palsy, 2o corticobasal degeneration, and familial frontotemporal dementia.
In another embodiment, a vector capable of expressing MDDT or a fragment or derivative thereof may be administered to a subject to treat or prevent a disorder associated with decreased expression or activity of MDDT including, but not limited to, those described above.
In a further embodiment, a composition comprising a substantially purified MDDT in conjunction with a suitable pharmaceutical carrier may be administered to a subject to treat or prevent a disorder associated with decreased expression or activity of MDDT including, but not limited to, those provided above.
In still another embodiment, an agonist which modulates the activity of MDDT
may be administered to a subject to treat or prevent a disorder associated with decreased expression or activity of IVIDDT including, but not limited to, those listed above.
In a further embodiment, an antagonist of 1VIDDT may be administered to a subject to treat or prevent a disorder associated with increased expression or activity of MDDT.
Examples of such disorders include, but are not limited to, those cell proliferative, autoimmune/inflammatory, developmental, and neurological disorders described above. In one aspect, an antibody which specifically binds 1VIDDT may be used directly as an antagonist or indirectly as a targeting or delivery mechanism for bringing a pharmaceutical agent to cells or tissues which express MDDT.
In an additional embodiment, a vector expressing the complement of the polynucleotide encoding MDDT may be administered to a subject to treat or prevent a disorder associated with increased expression or activity of MDDT including, but not limited to, those described above.
In other embodiments, any of the proteins, antagonists, antibodies, agonists, complementary sequences, or vectors of the invention may be administered in combination with other appropriate therapeutic agents. Selection of the appropriate agents for use in combination therapy may be made by one of ordinary skill in the art, according to conventional pharmaceutical principles. The to combination of therapeutic agents may act synergistically to effect the treatment or prevention of the various disorders described above. Using this approach, one may be able to achieve therapeutic efficacy with lower dosages of each agent, thus xeducing the potential for adverse side effects.
An antagonist of MDDT may be produced using methods which are generally known in the art. In particular, purified MDDT maybe used to produce antibodies or to screen libraries. of pharmaceutical agents to identify those which specifically bind MDDT.
Antibodies to MDDT may also be generated using methods that are well known in the art. Such antibodies may include, but are not limited to, polyclonal, monoclonal, chimeric, and single chain antibodies, Fab fragments, and fragments produced by a Fab expression library. Neutralizing antibodies (i.e.~
those which inhibit dimer formation) are generally preferred for therapeutic use. Single chain antibodies (e.g., from camels or llamas) may be potent enzyme inhibitors and may have advantages in the design of peptide mimetics, and in the development of immuno-adsorbents and biosensors (Muyldermans, S: (2001) J.
Biotechnol. 74:277-302).
For the production of antibodies, various hosts including goats, rabbits, rats, mice, camels, dromedaries, llamas, humans, and others may be immunized by injection with MDDT or with any fragment or oligopeptide thereof which has immunogenic properties. Depending on the host species, various adjuvants may be used to increase immunological response. Such adjuvants include, but are not limited to, Freund's, mineral gels such as aluminum hydroxide, and surface active substances such as lysolecithin, pluronic polyols, polyanions, peptides, oil emulsions, KLH, and dinitrophenol. Among adjuvants used in humans, BCG (bacilli Calmette-Guerin) and Corynebacterium parvum are especially preferable.
It is preferred that the oligopeptides, peptides, or fragments used to induce antibodies to MDDT have au amino acid sequence consisting of at least about 5 amino acids, and generally will consist of at least about 10 amino acids. It is also preferable that these ofigopeptides, peptides, or fragments are identical to a portion of the amino acid sequence of the natural protein. Short stretches of MDDT amino acids may be fused with those of another protein, such as KLH, and antibodies to the chimeric molecule may be produced.
Monoclonal antibodies to MDDT may be prepared using any technique which provides for the.
production of antibody molecules by continuous cell lines in culture. These include, but are not limited to, the hybridoma technique, the human B-cell hybridoma technique, and the EBV-hybridoma technique. (See, e.g., Kohler, G. et al. (1975) Nature 256:495-497; Kozbor, D.
et al. (1985) J.
T_m_m__unol. Methods 81:31-42; Cote, R.J. et al. (1983) Proc. Natl. Acad. Sci.
USA 80:2026-2030; and Cole, S.P. et al. (1984) Mol. Cell Biol. 62:109-120.) In addition, techniques developed for the production of "chimeric antibodies,"
such as the splicing of mouse antibody genes to human antibody genes to obtain a molecule with appropriate antigen specificity and biological activity, can be used. (See, e.g., Morrison, S.L. et al. (1984) Proc.
Natl. Acad. Sci. USA 81:6851-6855; Neuberger, M.S. et al. (1984) Nature 312:604-608; and Takeda, S. et al. (1985) Nature 314:452-454.) Alternatively, techniques described for the production of single chain antibodies may be adapted, using methods known in the art, to produce MDDT-specific single chain antibodies. Antibodies with related specificity, but of distinct idiotypic composition, may be generated by chain shuffling from random combinatorial immunoglobulin libraries. (See, e.g., Burton, D.R. (1991) Proc. Natl. Acad. Sci. USA 88:10134-10137.).
.. Antibodies may also be produced by inducing in vivo production in the lymphocyte population or by screening immunoglobulin libraries or panels of highly specific binding reagents as disclosed in the literature. (See, e.g., Orlandi, R. et al. (1989) Proc. Natl. Acad. Sci.
USA 86:3833-3837; Winter, G. et al. (1991) Nature 349:293-299.) .
Antibody fragments which contain specific binding sites for MDDT may also be generated.
For example, such fragments include, but are not limited to, F(ab')2 fragments produced by pepsin digestion of the antibody molecule and Fab fragments generated by reducing the disulfide bridges of the F(ab')2 fragments. Alternatively, Fab expression libraries may be constructed to allow rapid and easy identification of monoclonal Fab fragments with the desired specificity.
(See, e.g., Huse, W.D.
et al. (1989) Science 246:1275-1281.) Various immunoassays may be used for screening to identify. antibodies having the desired specificity. Numerous protocols for competitive binding or immunoradiometric assays using either polyclonal or monoclonal antibodies with established specificities are well known in the art. Such immunoassays typically involve the measurement of complex formation between MDDT and its specific antibody. A two-site, monoclonal-based immunoassay utilizing monoclonal antibodies reactive to two non-interfering MDDT epitopes is generally used, but a competitive binding assay may also be employed (Pound, supra).

Various methods such as Scatchard analysis in conjunction with radioimmunoassay techniques may be used to assess the affinity of antibodies for MDDT. Affinity is expressed as an association constant, Ka, which is defined as the molar concentration of MDDT-antibody complex divided by the molar concentrations of free antigen and free antibody under equilibrium conditions. The Ka determined for a preparation of polyclonal antibodies, which are heterogeneous in their affinities for multiple MDDT epitopes, represents the average affinity, or avidity, of the antibodies for MDDT. The Ka determined for a preparation of monoclonal antibodies, which are monospecific for a particular MDDT epitope, represents a true measure of affinity. High-affinity antibody preparations with Ka ranging from about 109 to 1012 L/mole are preferred for use in immunoassays in which the MDDT-antibody complex must withstand rigorous manipulations. Low-affinity antibody preparations with K
ranging from about 106 to 10' L/mole are preferred for use in immunopurification and similar procedures which ultimately require dissociation of MDDT, preferably in active form, from the antibody (Catty, D. (1988) Antibodies, Volume I: A Practical Approach, IRL
Press, Washington DC;
Liddell, J.E. and A. Cryer (1991) A Practical Guide to Monoclonal Antibodies, John Wiley & Sons, New fork NY).
The titer and avidity of polyclonal antibody preparations may be further evaluated to determine the quality and suitability of such preparations for certain downstream applications. For example, a polyclonal antibody preparation containing at least 1-2 rng specific antibody/ml, preferably S-10 mg specific antibody/mh is generally employed in procedures .requiring precipitation of MDDT-antibody complexes. Procedures for evaluating antibody specificity, titer, and avidity, and guidelines for antibody quality and usage in various applications, are generally available.
(See, e.g., Catty, supra, and Coligan et al. supra.) In another embodiment of the invention, the polynucleotides encoding MDDT, or any fragment or complement thereof, may be used for therapeutic purposes. In one aspect, .modifications of gene expression can be achieved by designing complementary sequences or antisense molecules (DNA, RNA, PNA, or modified oligonucleotides) to the coding or regulatory regions of the gene encoding MDDT. Such technology is well known in the art, and antisense oligonucleotides or larger fragments can be designed from various locations along the coding or control regions of sequences encoding MDDT. (See, e.g., Agrawal, S., ed. (1996) Antisense Therapeutics, Humana Press Inc., Totawa 3o NJ.) In therapeutic use, any gene delivery system suitable for introduction of the antisense sequences into appropriate target cells can be used. Antisense sequences can be delivered intracellularly in the form of an expression plasmid which, upon transcription, produces a sequence . complementary to at least a portion of the cellular sequence encoding the target protein. (See, e.g., Slater, J.E. et al. (1998) J. Allergy Clip. Tm_m__unol. 102(3):469-475; and Scanlon, K.J. et al. (1995) 9(13):1288-1296.) Antisense sequences can also be introduced intracellularly through the use of viral vectors, such as retrovirus and adeno-associated virus vectors. (See, e.g., Miller, A.D. (1990) Blood 76:271; Ausubel, supra; Uckert, W. and W. Walther (1994) Pharmacol. Ther.
63(3):323-347.) Other gene delivery mechanisms include liposome-derived systems, artificial viral envelopes, and other systems known in the art. (See, e.g., Rossi, J.J. (1995) Br. Med. Bull.
51(1):217-225; Boado, R.J. et al. (1998) J. Pharm. Sci. 87(11):1308-1315; and Morris, M.C. et al. (1997) Nucleic Acids Res.
25(14):2730-2736.) Iu another embodiment of the invention, polynucleotides encoding MDDT may be used for somatic or germline gene therapy. Gene therapy may be performed to (i) correct a genetic deficiency (e.g., in the cases of severe combined immunodeficiency (SLID)-X1 disease characterized by X-linked inheritance (Cavazzana-Calvo, M. et al. (2000) Science 288:669-672), severe combined immunodeficiency syndrome associated with an inherited adenosine deaminase (ADA) deficiency (Blaese, R.M. et al. (1995) Science 270:475-480; Bordignon, C. et al. (1995) Science 270:470-475), cystic fibrosis (Zabner, J. et al. (1993) Cell 75:207-216; Crystal, R.G. et al. (1995) Hum. Gene Therapy 6:643-666; Crystal, R.G. et al. (1995) Hum. Gene Therapy 6:667-703), thalassamias, familial hypercholesterolemia, and hemophilia resulting from Factor VI1T or Factor 1X
deficiencies (Crystal, R.G. (1995) Science 270:404-410; Verma, LM. and N. Somia (1997)' Nature 389:239-242)), (ii) express a conditionally lethal gene product (e.g., in the case of cancers which result from unregulated 2o cell proliferation), or (iii) express a protein which affords protection against intracellular parasites (e.g., against human retroviruses, such as human immunodeficiency virus (HIV) (Baltimore, D. (1988) Nature 335:395-396; Poeschla, E. et al. (1996) Proc. Natl. Acad. Sci. USA
93:11395-11399), hepatitis B or C virus (HBV, HCV); fungal parasites, such as Candida albicans and Paracoccidioides bf asiliensis; and protozoan parasites such as Plasntodium falciparum and Ttypaftosonta cruzi). In the case where a genetic deficiency in MDDT expression or regulation causes disease, the expression of MDDT from an appropriate population of trausduced cells may alleviate the clinical manifestations caused by the genetic deficiency.
In a further embodiment of the invention, diseases or disorders caused by deficiencies in MDDT are treated by constructing mammalian expression vectors encoding MDDT
and introducing these vectors by mechanical means into MDDT-deficient cells. Mechanical transfer technologies for use with cells in vivo or ex vitro include (i) direct DNA microinjection into individual cells, (ii) ballistic gold particle delivery, (iii) liposome-mediated trausfection, (iv) receptor-mediated gene transfer, and (v) the use of DNA transposons (Morgan, R.A. and W.F. Anderson (1993) Annu.
Rev. Biochem.

62:191-217; Ivics, Z. (1997) Cell 91:501-510; Boulay, T-L, and H. Recipon (1998) Curr. Opin.
Biotechtrol. 9:445-450).
Expression vectors that may be effective for the expression of MDDT include, but are not limited to, the PCDNA 3.1, EP1TAG, PRCCMV2, PREP, PVAX, PCR2-TOPOTA vectors (Invitrogen, Carlsbad CA}, PCMV-SCRIPT, PCMV-TAG, PEGSHIPERV (Stratagene, La Jolla CA), and PTET-OFF, PTET-ON, PTRE2, PTRE2-LUC, PTK-HYG (Clontech, Palo Alto CA).
MDDT
may be expressed using (l) a constitutavely active promoter, (e.g., from cytomegalovirus (CMV), Rous sarcoma virus (RSV), SV40 virus, thymidine kinase (TIC), or ~i-actin genes), (ii) an inducible promoter (e.g., the tetracycline-regulated promoter (Gossen, M. and H. Bujard (1992) Proc. Natl. Aced. Sci.
USA 89:5547-5551; Gossen, M. et al. (1995) Science 268:1766-1769; Rossi, F.M.V. and H.M. Bleu (1998) Curr. Opin. Biotechnol. 9:451-456), commercially available in the T-REX
plasmid (Invitrogen)};
the ecdysone-inducible promoter (available in the plasmids PVGRXR and PIND;
Invitrogen}; the FK.506irapamycin inducible promoter; or the RU486imifepristone znducible promoter (Rossi, F.M.V.
and H.M. Bleu, supra)), or (iii) a tissue-specific promoter or the native promoter of the endogenous gene encoding MDDT from a normal individual.
Commercially available liposome transformation kits (e.g., the PERFECT Lll'ID
TRANSFECTION K1T, available fxom Invitrogen) allow. one with ordinary skill in: the art to deliver polynucleotides to target cells in culture and require minimal effort to optimize experimental parameters. Iu the alternative, transformataon.is performed using the calciurr~ phosphate method (Graham, F.L. and A.J. Eb (1973) Virology 52:456-46'7}, ox by electroporation (Neumann, E. et al.
(1982) EMBO J. 1:841.-845). The introduction of DNA.to primary cells requires modification of these standardized mammalian transfection protocols.
In another embodiment of the invention, diseases or disorders caused by genetic defects with respect to MDDT expression are treated by constructing a retrovirus vector consisting of (l) the polynucleotide encoding MDDT under the control of an independent promoter or the retrovirus long terminal repeat (LTR) promoter, (ii) appropriate RNA packaging signals, and (iii) a Rev-responsive element (RRE) along with additional retrovirus cis-acting RNA sequences and coding sequences required for efficient vector propagation. Retrovirus vectors (e.g., PFB and PFBNEO} are commercially available (Stxatagene) and are based on published data (Riviere, I. et al. (1995) Proc.
3o Natl. Aced. Sci. USA 92:6733-6737), incorporated by reference herein. The vector is propagated in an appropriate vector producing cell line (VPCL} that expresses an envelope gene with a tropism for receptors on the target Bells ox a promiscuous envelope protein such as VSVg (Armentano, D. et aI.
(1987) J. Virol. 61:1647-1650; Bender, M.A. et al. (1987)J. Virol. 61:1639-1646; Adam, M.A. and A.D. Miller (1988) 3. Virol. 62:3802-3806; Dull, T. et al. (1998) J. Virol.
72:8463-8471; Zufferey, R. et al. (1998) J. Virol. 72:9873-9880). U.S. Patent No. 5,910,434 to Rigg ("Method for obtaining retrovirus packaging cell lines producing high transducing efficiency retroviral supernatant") discloses a method for obtaining retrovirus packaging cell lines and is hereby incorporated by reference.
Propagation of retrovirus vectors, transduction of a population of cells (e.g., CD4+ T-cells), and the return of transduced cells to a patient are procedures well known to persons skilled in the art of gene therapy and have been well documented (Ranga, U. et al. (1997) J. Virol.
71:7020-7029; Bauer, G. et al. (1997) Blood 89:2259-2267; Bonyhadi, M.L. (1997) J. Virol. 71:4707-4716;
Ranga, U. et al. (1998) Proc. Natl. Acad. Sci. USA 95:1201-1206; Su, L. (1997) Blood 89:2283-2290).
In the alternative, an adenovirus-based gene therapy delivery system is used to deliver polynucleotides encoding MDDT to cells which have one or more genetic abnormalities with respect to the expression of MDDT. The construction and packaging of adenovirus-based vectors are well known to those with ordinary skill in the art. Replication defective adenovirus .vectors have proven to be versatile for importing genes encoding immunoregulatory proteins into intact islets in the pancreas (Csete, M.E. et al. (1995) Transplantation 27:263-268). Potentially useful adenoviral vectors are described in U.S. Patent No. 5,707,618 to Armentano ("Adenovirus vectors for gene therapy"), hereby incorporated by reference. For adenoviral vectors, see also.Antinozzi, P.A. et al. (1999) . . Annu. Rev. Nutr. 19:511-544 and Verma, LM. and N. Somia (1997) Nature 18:389:239-24~2.,'ooth ' incorporated by reference herein.
In another alternative, a herpes-based, gene therapy delivery system is used to deliver polynucleotides encoding MDDT to target cells which have one or more genetic abnormalities with' respect to the expression of MDDT. The use of hexpes simplex virus (HSV)-based vectors may be especially valuable for introducing MDDT to cells of the central nervous system, for which HSV has a tropism. The construction and packaging of herpes based vectors are well known to those with ordinary skill in the art. A replication-competent herpes simplex virus (HSV) type 1-based vector has been used to deliver a reporter gene to the eyes of primates (Liu, X. et al.
(1999) Exp. Eye Res.
169:385-395). The construction of a HSV-1 virus vector has also been disclosed in detail in U.S.
Patent No. 5,804,413 to DeLuca ("Herpes simplex virus strains for gene transfer"), which is hereby incorporated by reference. U.S. Patent No. 5,804,413 teaches the use of recombinant HSV d92 which consists of a genome containing at least one exogenous gene to be transferred to a cell under the control of the appropriate promoter for purposes including human gene therapy. Also taught by this patent are the construction and use of recombinant HSV strains deleted for ICP4, ICP27 and ICP22. For HSV vectors, see also Goins, W.F. et al. (1999) J. Virol. 73:519-532 and Xu, H. et al.
(1994) Dev. Biol. 163:152-161, hereby incorporated by reference. The manipulation of cloned herpesvirus sequences, the generation of recombinant virus following the transfection of multiple plasmids containing different segments of the large herpesvirus genomes, the growth and propagation of herpesvirus, and the infection of cells with herpesvirus are techniques well known to those of ordinary skill in the art.
In another alternative, an alphavirus (positive, single-stranded RNA virus) vector is used to deliver polynucleotides encoding MDDT to target cells. The biology of the prototypic alphavirus, Semlik_i Forest Virus (SFV), has been studied extensively and gene transfer vectors have been based ' on the SFV genome (Garoff, H. and K.-J. Li (1998) Curr: Opin. Biotechnol.
9:464-469). During alphavirus RNA replication, a subgenomic RNA is generated that normally encodes the viral capsid proteins. This subgenomic RNA replicates to higher levels than the full length genomic RNA, l0 resulting in the overproduction of capsid proteins relative to the viral proteins with enzymatic activity (e.g., protease and polymerase). Similarly, inserting the coding sequence for MDDT into the alphavirus genome in place of the capsid-coding region results in the production of a large number of MDDT-coding RNAs and the synthesis of high levels of MDDT in vector transduced cells. While alphavirus infection is typically associated with cell lysis within a few days, the ability to establish a persistent infection in hamster normal kidney cells (BHI~-21) with a variant of Sindbis virus (SIN) indicates that the lytic replication of alphaviruses can be altered to suit the needs of the gene therapy application (Dryga, S.A. et al. (1997) Virology 228:74-83). The wide host range of alphaviruses wi3l~ :~
allow the introduction of MDDT into a variety of cell types. The specific transduction of a subset of cells in a population may require the sorting of cells prior to transduction.
The methods of manipulating infectious cDNA clones of alphaviruses, performing alphavirus cDNA and RNA
transfections, and performing alphavirus infections, are well known to those with ordinary skill in the art.
Oligonucleotides derived from the transcription initiation site, e.g., between about positions -10 and +10 from the start site, may also be employed to inhibit gene expression.
Similarly, inhibition can be achieved using triple helix base-pairing methodology. Triple helix pairing is useful because it causes inhibition of the ability of the double helix to open sufficiently for the binding of polymerases, transcription factors, or regulatory molecules. Recent therapeutic advances using triplex DNA have been described in the literature. (See, e.g., Gee, J.E. et al. (1994) in Huber, B.E. and B.I. Carr, Molecular and Itnmunolo~ic Approaches, Futura Publishing, Mt. Kisco NY, pp.
163-17?.) A
complementary sequence or antisense molecule may also be designed to block translation of mRNA
by preventing the transcript from binding to ribosomes.
Ribozymes, enzymatic RNA molecules, may also be used to catalyze the specific cleavage of RNA. The mechanism of ribozyme action involves sequence-specific hybridization of the ribozyme molecule to complementary target RNA, followed by endonucleolytic cleavage.
For example, engineered hammerhead motif ribozyme molecules may specifically and efficiently catalyze endonucleolytic cleavage of sequences encoding MDDT.
Specific ribozyme cleavage sites within any potential RNA target are initially identified by scanning the target molecule for ribozyme cleavage sites, including the following sequences: GUA, GUU, and GUC. Once identified, short RNA sequences of between 15 and 20 ribonucleotides, corresponding to the region of the target gene containing the cleavage site, may be evaluated for secondary structural features which may render the oligonucleotide inoperable.
The suitability of candidate targets may also be evaluated by testing accessibility to hybridization with complementary oligonucleotides using ribonuclease protection assays.
Complementary ribonucleic acid molecules and ribozymes of the invention may be prepared by any method known in the art for the synthesis of nucleic acid molecules.
These include techniques for chemically synthesizing ofigonucleotides such as solid phase phosphoramidite chemical synthesis.
Alternatively, RNA molecules may be generated by in vitro and in vivo transcription of DNA
sequences encoding MDDT. Such DNA sequences may be incorporated into a wide variety of vectors with suitable RNA polymerase promoters such as T7 or SP6.
Alternatively, these cDNA
constructs that synthesize complementary RNA, constitutively or inducibly, can be introduced into cell lines, cells, or tissues. ' '' RNA molecules may be modified to increase intracellular stability and half life. Possible .
modifications include, but are not limited to, 'the addition of flanking sequences at the 5' and/or 3' ends of the molecule, or the use. of phosphorothioate or 2' O-methyl rather than phosphodiesterase linkages within the backbone of the molecule. This concept is inherent in the production of PNAs and can be extended in all of these molecules by the inclusion of nontraditional bases such as inosine, queosine, and wybutosine, as well as acetyl-, methyl-, thio-, and similarly modified forms of adenine, cytidine, guanine, thymine, and uridine which are not as easily recognized by endogenous endonucleases.
An additional embodiment of the invention encompasses a method for screening for a compound which is effective in altering expression of a polynucleotide encoding IVIDDT. Compounds which may be effective in altering expression of a specific polynucleotide may include, but are not limited to, oligonucleotides, antisense oligonucleotides, triple helix-forming oligonucleotides, transcription factors and other polypeptide transcriptional regulators, and non-macromolecular chemical entities which are capable of interacting with specific polynucleotide sequences. Effective compounds may alter polynucleotide expression by acting as either inhibitors or promoters of polynucleotide expression. Thus, in the treatment of disorders associated with increased MDDT
expression or activity, a compound which specifically inhibits expression of the polynucleotide encoding MDDT may be therapeutically useful, and in the treatment of disorders associated with decreased MDDT expression or activity, a compound which specifically promotes expression of the polynucleotide encoding MDDT may be therapeutically useful.
At least one, and up to a plurality, of test compounds may be screened for effectiveness in altering expression of a specific polynucleotide. A test compound may be obtained by any method commonly known in the art, including chemical modification of a compound known to be effective in altering polynucleotide expression; selection from an existing, commercially-available or proprietary library of naturally-occurring or non-natural chemical compounds; rational design of a compound based on chemical and/or structural properties of the target polynucleotide;
and selection from a library of chemical compounds created combinatorially or randomly. A sample comprising a 1o polynucleotide encoding MDDT is exposed to at least one test compound thus obtained. The sample may comprise, for example, an intact or permeabifized cell, or an irt vitt~o cell-free or reconstituted biochemical system. Alterations in the expression of a polynucleotide encoding MDDT are assayed by any method commonly known in the art. Typically, the expression of a specific nucleotide is detected by hybridization with a probe having a nucleotide sequence complementary to the sequence of the polynucleotide encoding MDDT. The amount of . hybridization may be quantified, thus forming the basis for a comparison of the expression of.the polynucleotide both with and without exposure to one or more test compounds. Detection of a change in the expression of a polynucleotide exposed to a test compound indicates that: the test compound is effective in altering the expression of the polynucleotide. A screen for a compound effective in altering expression of a specific polynucleotide can be carried out, for example, using a Schizosaccharornyces pombe gene expression system (Atkins, D. et al. (1999) U.S. Patent No. 5,932,435; Arndt, G.M. et al. (2000) Nucleic Acids Res.
28:E15) or a human cell line such as HeLa cell (Clarke, M.L. et al. (2000) Biochem. Biophys. Res.
Commun. 268:8-13). A particular embodiment of the present invention involves screening a combinatorial library of oligonucleotides (such as deoxyribonucleotides, ribonucleotides, peptide nucleic acids, and modified oligonucleotides) for antisense activity against a specific polynucleotide sequence (Bruice, T.W. et al. (1997) U.S. Patent No. 5,686,242; Bruice, T.W. et al.
(2000) U.S. Patent No.
6,022,691).
Many methods for introducing vectors into cells or tissues are available and equally suitable for use in vivo, in vitro, and ex vivo. For ex vivo therapy, vectors may be introduced into stem cells taken from the patient and clonally propagated for autologous transplant back into that same patient.
Delivery by transfection, by liposome injections, or by polycationic amino polymers may be achieved using methods which are well known in the art. (See, e.g., Goldman, C.K. et al. (1997) Nat.
Biotechnol. 15:462-466.) Any of the therapeutic methods described above may be applied to any subject in need of such therapy, including, for example, mammals such as humans, dogs, cats, cows, horses, rabbits, and monkeys.
An additional embodiment of the invention relates to the administration of a composition which generally comprises an active ingredient formulated with a pharmaceutically acceptable excipient.
Excipients may include, for example, sugars, starches, celluloses, gums, and proteins. Various formulations are commonly known and are thoroughly discussed in the latest edition of Remin~ton's Pharmaceutical Sciences (Maack Publishing, Easton PA). Such compositions may consist of MDDT, antibodies to MDDT, and mimetics, agonists, antagonitsts, or inhibitors of MDDT.
The compositions utilized in this invention may be administered by any number of routes 1o including, but not limited to, oral, intravenous, intramuscular, intra-arterial, intramedullary, intrathecal, intraventricular, pulmonary, transdermal, subcutaneous, intraperitoneal, intranasal, enteral, topical, sublingual, or rectal means.
Compositions for pulmonary administration may be prepared in liquid or dry powder form.
These compositions are generally aerosolized immediately prior to .inhalation by the patient. In the case of small molecules (e.g. traditional low molecular weight organic drugs), aerosol delivery of fast-acting formulations is well-known in the art. In the case of macromolecules (e.g. larger peptides and proteins), recent developments in the held of pulmonary delivery via the alveolar region of the lung have enabled the practical delivery of drugs such as insulin. to blood circulation (see, e.g., Patton, J.S.
et al., U.S. Patent No. 5,997,848). Pulmonary delivery has the advantage of administration without needle injection, and obviates the need for potentially toxic penetration enhancers.
Compositions suitable for use in the invention include compositions wherein the active ingredients are contained in an effective amount to achieve the intended purpose. The determination of an effective dose is well within the capability of those skilled in the art.
Specialized forms of compositions may be prepared for direct intracellular delivery of macromolecules comprising MDDT or fragments thereof. For example, liposome preparations containing a cell-impermeable macromolecule may promote cell fusion and intracellular delivery of the macromolecule. Alternatively, MDDT or a fragment thereof may be joined to a short cationic N-terminal portion from the HIV Tat-1 protein. Fusion proteins thus generated have been found to transduce into the cells of all tissues, including the brain, in a mouse model system (Schwarze, S.R. et al. (1999) Science 285:1569-1572).
For any compound, the therapeutically effective dose can be estimated initially either in cell culture assays, e.g., of neoplastic cells, or in animal models such as mice, rats, rabbits, dogs, monkeys, or pigs. An animal model may also be used to determine the appropriate concentration range and route of administration. Such information can then be used to determine useful doses and routes for administration in. humans.
A therapeutically effective dose refers to that amount of active ingredient, for example NIDDT or fragments thereof, antibodies of IVIDDT, and agonists, antagonists or inhibitors of 1VIDDT, which ameliorates the symptoms or condition. Therapeutic efficacy and toxicity may be determined by standard pharmaceutical procedures in cell cultures or with experimental animals, such as by calculating the EDSO (the dose therapeutically effective in 50% of the population) or LDso (the dose lethal to 50% of the population) statistics. The dose ratio of toxic to therapeutic effects is the therapeutic index, which can be expressed as the LDSO/EDSO ratio. Compositions which exhibit large therapeutic indices are preferred. The data obtained from cell culture assays and animal studies are used to formulate a range of dosage for human use. The dosage contained in such compositions is preferably within a range of circulating concentrations that includes the EDso with little or no toxicity.
The dosage varies within this range depending upon the dosage form employed, the sensitivity of the patient, and the route of administration.
The exact dosage will be determined by the practitioner, in light of factors related to the subject requiring treatment. Dosage and administration are adjusted to provide sufficient levels of the active moiety or to maintain the desired effect. Factors which may be taken into account include the severity of the disease state, the general health of the subject, the age, weight, and gender of the ' ~ .
subject, time and frequency of administration, drug combination(s), reaction sensitivities, and response to therapy. Long-acting compositions may be administered every 3 ta~ 4 days, every week, or biweekly depending on the half life and clearance rate of the particular formulation.
Normal dosage amounts may vary from about 0.1 /,cg to 100,000 /.tg, up to a total dose of about 1 gram, depending upon the route of administration. Guidance as to particular dosages and methods of delivery is provided in the literature and generally available to practitioners in the art.
Those skilled in the art will employ different formulations for nucleotides than for proteins or their inhibitors. Similarly, delivery of polynucleotides or polypeptides will be specific to particular cells, conditions, locations, etc.
DIAGNOSTICS
In another embodiment, antibodies which specifically bind MDDT may be used for the diagnosis of disorders characterized by expression of MDDT, or in assays to monitor patients being treated with MDDT or agonists, antagonists, or inhibitors of MDDT. Antibodies useful for diagnostic purposes maybe prepared in the same manner as described above for therapeutics. Diagnostic assays for MDDT include methods which utilize the antibody and a label to detect MDDT in human body fluids or in extracts of cells or tissues. The antibodies may be used with or without modification, and may be labeled by covalent or non-covalent attachment of a reporter molecule. A wide variety of reporter molecules, several of which are described above, are known in the art and may be used.
A variety of protocols for measuring MDDT, including ELISAs, RIAs, and FACS, are known in the art and provide a basis for diagnosing altered or abnormal levels of MDDT expression. Normal or standard values for MDDT expression are established by combining body fluids or cell extracts taken from normal mammalian subjects, for example, human subjects, with antibodies to MDDT under conditions suitable for complex formation. The amount of standard complex formation may be quantitated by various methods, such as photometric means. Quantities of MDDT
expressed in subject, control, and disease samples from biopsied tissues are compared with the standard values.
Deviation between standard and subject values establishes the parameters for diagnosing disease.
In another embodiment of the invention, the polynucleotides encoding MDDT may be used for diagnostic purposes. The polynucleotides which may be used include oligonucleotide sequences, complementary RNA and DNA molecules, and PNAs. The polynucleotides may be used to detect and quantify gene expression in biopsied tissues in which expression of MDDT
may be correlated with . disease. The diagnostic assay maybe used to 'determine absence, presence, and excess expression of . MDDT, and to monitor regulation of MDDT levels during therapeutic intervention.
In one aspect; hybridization with PCR probes which are capable of detecting polynucleotide sequences, including genomic sequences; encoding MDDT or closely related molecules may be used ;
.' . to identify nucleic acid sequences which encode MDDT. The specificity of the probe, whether it is made from a highly specific region, e.g., the 5'regulatory region; or from a less specific region, e.g., a'.
conserved motif, and the stringency of the hybridization or amplification will determine whether the probe identifies only naturally occurring sequences encoding MDDT, allelic variants, or related sequences.
Probes may also be used for the detection of related sequences, and may have at least 50°l0 sequence identity to any of the MDDT encoding sequences. The hybridization probes of the subject invention may be DNA or RNA and may be derived from the sequence of SEQ TD
N0:40-78 or from genomic sequences including promoters, enhancers, and introns of the MDDT
gene.
Means for producing specific hybridization probes for DNAs encoding MDDT
include the cloning of polynucleotide sequences encoding MDDT or MDDT derivatives into vectors for the production of mRNA probes. Such vectors are known in the art, are commercially available, and may be used to synthesize RNA probes in vitro by means of the addition of the appropriate RNA
polymerases and the appropriate labeled nucleotides. Hybridization probes may be labeled by a variety of reporter groups, for example, by radionuclides such as 32P or 355, or by enzymatic labels, such as alkaline phosphatase coupled to the probe via avidin/biotin coupling systems, and the like.
Polynucleotide sequences encoding MDDT may be used for the diagnosis of disorders associated with expression of MDDT. Examples of such disorders include, but are not limited to, a cell proliferative disorder such as actinic keratosis, arteriosclerosis, atherosclerosis, bursitis, cirrhosis, hepatitis, mixed connective tissue disease (MCTD), myelofibrosis, paroxysmal nocturnal hemoglobinuria, polycythemia vera, psoriasis, primary thrombocythemia, and cancers including adenocarcinoma, leukemia, lymphoma, melanoma, myeloma, sarcoma, teratocarcinoma, and, in particular, cancers of the adrenal gland, bladder, bone, bone marrow, brain, breast, cervix, gall bladder, ganglia, gastrointestinal tract, heart, kidney, liver, lung, muscle, ovary, pancreas, parathyroid, penis, prostate, salivary glands, skin, spleen, testis, thymus, thyroid, and uterus;
an autoimrnune/inflammatory disorder such as acquired immunodeficiency syndrome (A1DS), Addison's disease, adult respiratory distress syndrome, allergies, ankylosing spondylitis, amyloidosis, anemia, asthma, atherosclerosis, autoimmune hemolytic anemia, autoimmune thyroiditis, autoimmune polyendocrinopathy-candidiasis-ectodermal dystrophy (APECED), bronchitis, cholecystitis, contact dermatitis, Crohn's disease, atopic dermatitis, dermatomyositis, diabetes mellitus, emphysema, episodic lymphopenia with lymphocytotoxins, erythroblastosis fetalis, erythema nodosum, atrophic gastritis, glomerulonephritis, Goodpasture's syndrome, gout; Graves' disease, Hashimoto's thyroiditis, hypereosinophilia, irritable bowel syndrome, multiple sclerosis, myasthenia gravis, myocardial or pericardial inflammation, osteoarthritis, osteoporosis, pancreatitis, polymyositis, psoriasis, Reiter's syndrome, rheumatoid . arthritis, scleroderma, Sjogren's syndrome, systemic anaphylaxis, systemic lupus erythematosus, systemic sclerosis, thrombocytopenic purpura, ulcerative colitis, uveitis, Werner. syndrome, ~, complications of cancer, hemodialysis, and extracorporeal~circulation, viral, bacterial, fungal, parasitic protozoal, and helrninthic infections, and trauma; a developmental disorder such as renal tubular acidosis, anemia, Cushing's syndrome, achondroplastic dwarfism, Duchenne and Becker muscular dystrophy, epilepsy, gonadal dysgenesis, WAGR syndrome (Wilms' tumor, aniridia, genitourinary abnormalities; and mental retardation), Smith-Magenis syndrome, myelodysplastic syndrome, hereditary mucoepithelial dysplasia, hereditary keratodermas, hereditary neuropathies such as Charcot-Marie-Tooth disease and neurofibromatosis, hypothyroidism, hydrocephalus, seizure disorders such as Syndenham's chorea and cerebral palsy, spina bifida, anencephaly, craniorachischisis, congenital glaucoma, cataract, and sensorineural hearing loss; and a neurological disorder such as epilepsy, ischemic cerebrovascular disease, stroke, cerebral neoplasms, Alzheimer's disease, Pick's disease, Huntington's disease, dementia, Parkinson's disease and other extrapyramidal disorders, amyotrophic lateral sclerosis and other motor neuron disorders, progressive neural muscular atrophy, retinitis pigmentosa, hereditary ataxias, multiple sclerosis and other demyelinating diseases, bacterial and viral meningitis, brain abscess, subdural empyema, epidural abscess, suppurative intxacranial thrombophlebitis, myelitis and radiculitis, viral central nervous system disease, prion diseases including kuru, Creutzfeldt-Jakob disease, and Gerstmann-Straussler-Scheinker syndrome, fatal familial insomnia, nutritional and metabolic diseases of the nervous system, neurofibromatosis, tuberous sclerosis, cerebelloretinal hemangioblastomatosis, encephalotrigeminal syndrome, mental retardation and other developmental disorders of the central nervous system including Down syndrome, cerebral palsy, neuroskeletal disorders, autonomic nervous system disorders, cranial nerve disorders, spinal cord diseases, muscular dystrophy and other neuromuscular disorders, peripheral nervous system disorders, dermatomyositis and polymyositis, inherited, metabolic, endocrine, and toxic myopathies, myasthenia gravis, periodic paralysis, mental disorders including mood, anxiety, and schizophrenic disorders, seasonal affective disorder (SAD), akathesia, amnesia, catatonia, diabetic neuropathy, tardive dyskinesia, dystonias~ paranoid psychoses, postherpetic neuralgia, Tourette's disorder, progressive supranuclear palsy, corticobasal degeneration, and familial frontotemporal dementia. The polynucleotide sequences encoding MDDT may be used in Southern or northern analysis, dot blot, or other membrane-based technologies; in PCR technologies; in dipstick, pin, and multiformat ELISA-like assays; and in microarrays utilizing fluids or tissues from patients to detect altered MDDT expression.
Such qualitative or quantitative methods are well known in the art.
In a particular aspect, the nucleotide sequences encoding MDDT may be useful in assays that detect the presence of associated disorders, particularly those mentioned above. The nucleotide -.
sequences encoding MDDT may be labeled by standard methods and added to a fluid or tissue sample from a patient under conditions suitable for the formation of hybridization complexes. After a suitable incubation period, the sample is washed and the signal is quantified and compared with a standard value. If the amount of signal in the patient sample is significantly altered in comparison to a control sample then the presence of altered levels of nucleotide sequences encoding MDDT in the sample indicates the presence of the associated disorder. Such assays may also be used to evaluate the efficacy of a particular therapeutic treatment regimen in animal studies, in clinical trials, or to monitor the treatment of an individual patient.
In order to provide a basis for the diagnosis of a disorder associated with expression of MDDT, a normal or standard profile for expression is established. This may be accomplished by combining body fluids or cell extracts taken from normal subjects, either animal or human, with a sequence, or a fragment thereof, encoding MDDT, under conditions suitable for hybridization or amplification. Standard hybridization may be quantified by comparing the values obtained from normal subjects with values from an experiment in which a known amount of a substantially purified polynucleotide is used. Standard values obtained in this manner may be compared with values obtained from samples from patients who are symptomatic for a disorder.
Deviation from standard values is used to establish the presence of a disorder.

Once the presence of a disorder is established and a treatment protocol is initiated, hybridization assays may be repeated on a regular basis to determine if the level of expression in the patient begins to approximate that which is observed in the normal subject.
The results obtained from successive assays may be used to show the efficacy of treatment over a period ranging from several days to months.
With respect to cancer, the presence of an abnormal amount of transcript (either under- or overexpressed) in biopsied tissue from an individual may indicate a predisposition for the development of the disease, or may provide a means for detecting the disease prior to the appearance of actual clinical symptoms. A more definitive diagnosis of this type may allow health professionals to employ preventative measures or aggressive treatment earlier thereby preventing the development or further progression of the cancer.
Additional diagnostic uses for oligonucleotides designed from the sequences encoding MDDT
may involve the use of PCR. These oligomers may be chemically synthesized, generated enzymatically, or produced in vitro. Oligomers will preferably contain a fragment of a polynucleotide encoding MDDT, ox a fragment of a polynucleotide complementary to the polynucleotide encoding MDDT, and will.be employed under optimized conditions for identification of a specific gene or condition. Oligomers may also be employed under less stringent conditions for detection or quantification of closely related.DNA or RNA sequences.
In a.particular aspect, oligonucleotide primers derived from the polynucleotide sequences .
encoding MDDT may be used to detect single nucleotide polymorphisms (SNPs).
SNPs are substitutions, insertions and deletions that are a frequent cause of inherited or acquired genetic disease.
in humans. Methods of SNP detection include, but are not limited to, single-stranded conformation polymorphism (SSCP) and fluorescent SSCP (fSSCP) methods. In SSCP, oligonucleotide primers derived from the polynucleotide sequences encoding MDDT are used to amplify DNA using the polymerase chain reaction (PCR). The DNA may be derived, for example, from diseased or normal tissue, biopsy samples, bodily fluids, and the like. SNPs in the DNA cause differences in the secondary and tertiary structures of PCR products in single-stranded form, and these differences are detectable using gel electrophoresis in non-denaturing gels. In fSCCP, the oligonucleotide primers are fluorescently labeled, which allows detection of the amplimers in high-throughput equipment such as DNA sequencing machines. Additionally, sequence database analysis methods, termed in silico SNP
(isSNP), are capable of identifying polymorphisms by comparing the sequence of individual overlapping DNA fragments which assemble into a common consensus sequence.
These computer-based methods filter out sequence variations due to laboratory preparation of DNA and sequencing errors using statistical models and automated analyses of DNA sequence chromatograms. In. the alternative, SNPs may be detected and characterized by mass spectrometry using, for example, the high throughput MASSARRAY system (Sequenom, Inc., San Diego CA).
SNPs may be used to study the genetic basis of human disease. For example, at least 16 common SNPs have been associated with non-insulin-dependent diabetes mellitus.
SNPs are also useful for examining differences in disease outcomes in monogenic disorders, such as cystic fibrosis, sickle cell anemia, or chronic gra~nulomatous disease. For example, variants in the mannose-binding lectin, MBL2, have been shown to be correlated with deleterious pulmonary outcomes in cystic fibrosis. SNPs also have utility in pharmacogenomics, the identification of genetic variants that influence a patient's response to a drug, such as life-threatening toxicity.
For example, a variation in N-acetyl transferase is associated with a high incidence of peripheral neuropathy in response to the anti-tuberculosis drug isoniazid, while a variation in the core promoter of the ALOXS gene results in diminished clinical response to treatment with an anti-asthma drug that targets the 5-lipoXygenase pathway. Analysis of the distribution of SNPs in different populations is useful for investigating genetic drift, mutation, recombination, and selection, as well as for tracing the origins of populations and their migrations. (Taylor, J.G. et al. (2002) Trends Mol. Med. 7.:507-512;
I~wok, P.-Y. and Z. Gu (1999) Mol. Med. Today 5:538-543; Nowotny, P. et al. (2001) Curr. Opin.
Neurobiol. 11:637-641.) Methods which may also be used to quantify the expression of MDDT include radiolabeling or biotinylating nucleotides, coamplification of a control nucleic acid, and interpolating results from standard curves. (See, e.g., Melby, P.C. et al. (1993) J. Tmmunol. Methods 159:235-244; Duplaa, C.
et al. (1993) Anal. Biochem. 212:229-236.) The speed ~of quantitation of multiple samples may be accelerated by running the assay in a high-throughput format where the oligomer or polynucleotide of interest is presented in various dilutions and a spectrophotometric or colorimetric response gives rapid quantitation.
In further embodiments, oligonucleotides or longer fragments derived from any of the polynucleotide sequences described herein may be used as elements on a microarray. The microarray can be used in transcript imaging techniques which monitor the relative expression levels of large numbers of genes simultaneously as described below. The microarray may also be used to identify genetic variants, mutations, and polymorphisms. This information may be used to determine gene function, to understand the genetic basis of a disorder, to diagnose a disorder, to monitor 3o progression/regression of disease as a function of gene expression, and to develop and monitor the activities of therapeutic agents in the treatment of disease. In particular, this information may be used to develop a pharmacogenomic profile of a patient in order to select the most appropriate and effective treatment regimen for that patient. For example, therapeutic agents which are highly effective and display the fewest side effects may be selected for a patient based on his/her pharmacogenomic pxohle.
In another embodiment, MDDT, fragments of MDDT, or antibodies specific for MDDT may be used as elements on a microarray. The microarray may be used to monitor or measure protein protein interactions, drug-target interactions, and gene expression profiles, as described above.
A particular embodiment relates to the use of the polynucleotides of the present invention to generate a transcript image of a tissue or cell type. A transcript image represents the global pattern of gene expression by a particular tissue or cell type. Global gene expression patterns are analyzed by quantifying the number of expressed genes and their relative abundance under given conditions and at a given time. (See Seilhamer et al., "Comparative Gene Transcript Analysis,"
U.S. Patent No.
5,840,484, expressly incorporated by reference herein.) Thus a transcript image may be generated by hybridizing the polynucleotides of the present invention or their complements to the fatality of transcripts or reverse transcripts of a particular tissue or cell type. In one embodiment, the hybridization takes place in high-throughput format, wherein the polynucleotides of the present invention or their complements comprise a subset of a plurality of elements on a microarray. The 25 resultant transcript image would provide a profile of gene activity.
Transcript images may be generated using transcripts isolated from tissues, cell lznes, biopsies, or other biological samples. The transcript.image may thus reflect gene expression in vivo, as in the case of a tissue or biopsy sample, or in. yitro, as in the case of a cell line.
Transcript images which profile the expression of the polynucleotides of the present invention may also be used in conjunction with in vitro model systems and preclinical evaluation of pharmaceuticals, as well as toxicological testing of industrial and naturally-occurring environmental compounds. All compounds induce characteristic gene expression patterns, frequently termed molecular fingerprints or toxicant signatures, which are indicative of mechanisms of action and toxicity (Nuwaysix, E.F. et al. (1999) Mol. Careiuog. 24:153-159; Steiner, S. and N.L.
Anderson (2000) Toxicol. Left. 112-113:467-471, expressly incorporated by reference herein).
If a test compound has a signature similar to that of a compound with known toxicity, it is likely to share those toxic properties.
These fingerprints or signatures are most useful and refined when they contain expression information from a large number of genes and gene families. Ideally, a genome-wide measurement of expression provides the highest quality signature. Even genes whose expression is not altered by any tested compounds are important as well, as the levels of expression of these genes are used to normalize the rest of the expression data. The normalization procedure is useful for comparison of expression data after treatment with different compounds. While the assignment of gene function to elements of a toxicant signature aids in interpretation of toxicity mechanisms, knowledge of gene function is not necessary for the statistical matching of signatures which leads to prediction of toxicity. (See, for example, Press Release 00-02 from the National Institute of Environmental Health Sciences, released February 29, 2000, available at http://www.niehs.nih.govloc/news/toxchip.htm.) Therefore, it is important and desirable in toxicological screening using toxicant signatures to include all expressed gene sequences.
In one embodiment, the toxicity of a test compound is assessed by treating a biological sample containing nucleic acids with the test compound. Nucleic acids that are expressed in the treated biological sample are hybridized with one or more probes specific to the polynucleotides of the present invention, so that transcript levels corresponding to the polynucleotides of the present invention may be quantified. The transcript levels in the treated biological sample are compared with levels in an untreated biological sample. Differences in the transcript levels between the two samples are indicative of a toxic response caused by the test compound in the treated sample.
Another particular embodiment relates to the use of the polypeptide sequences of the present invention to analyze the proteome of a tissue or cell type. The term proteome refers to the global pattern of protein expression in a particular tissue or cell type. Each protein component of a proteome can be subjected individually to further analysis. Proteome expression patterns, or profiles, are analyzed by quantifying the number of expressed proteins and their relative abundance under given conditions and at a given time. A profile of a cell's proteome may thus be generated by separating and analyzing the polypeptides of a particular tissue or cell type. In one embodiment, the separation is achieved using two-dimensional gel electrophoresis, :in which proteins from a sample are separated by isoelectric focusing in the first dimension, and then according to molecular weight by sodium dodecyl .
sulfate slab gel electrophoresis in the second dimension (Steiner and Anderson, su ra). The proteins are visualized in the gel as discrete and uniquely positioned spots, typically by staining the gel with an agent such as Coomassie Blue or silver or fluorescent stains. The optical density of each protein spot is generally proportional to the level of the protein in the sample. The optical densities of equivalently positioned protein spots from different samples, for example, from biological samples either treated or untreated with a test compound or therapeutic agent, are compared to identify any changes in protein spot density related to the treatment. The proteins in the spots are partially sequenced using, for example, standard methods employing chemical or enzymatic cleavage followed by mass spectrometry. The identity of the protein in a spot may be determined by comparing its partial sequence, preferably of at least 5 contiguous amino acid residues, to the polypeptide sequences of the present invention. In some cases, further sequence data may be obtained for definitive protein identification.
A proteomic profile may also be generated using antibodies specific for MDDT
to quantify the levels of MDDT expression. In one embodiment, the antibodies are used as elements on a 7~

microarray, and protein expression levels are quantified by exposing the microarray to the sample and detecting the levels of protein bound to each array element (Lueking, A. et al. (1999) Anal. Biochem.
270:103-111; Mendoze, L.G. et al. (1999) Biotechniques 27:778-788). Detection may be performed by a variety of methods known in the art, for example, by reacting the proteins in the sample with a thiol-or amino-reactive fluorescent compound and detecting the amount of fluorescence bound at each array element.
Toxicant signatures at the proteome level are also useful for toxicological screening, and should be analyzed in parallel with toxicant signatures at the transcript level. There is a poor correlation between transcript and protein abundances for some proteins in some tissues (Anderson, 1o N.L. and J. Seilhamer (1997) Electrophoresis 18:533-537), so proteome toxicant signatures maybe useful in the analysis of compounds which do not significantly affect the transcript image, but which alter the proteomic profile. In addition, the analysis of transcripts in body fluids is difficult, due to rapid degradation of mRNA, so proteomic profiling may be more reliable and informative in such cases.
In another embodiment, the toxicity of a test compound is assessed by treating a biological sample containing proteins with the test compound. Proteins that are expressed in the treated biological sample are separated so that the amount of each protein can be quantified. The amount of each protein is compared to the amount of the corresponding protein in an untreated biological sample.
A difference in the amount of protein between the two samples is indicative of a toxic response to the test compound in the treated sample. Individual proteins are identified by sequencing the amino acid residues of the individual proteins and comparing. these partial sequences to the polypeptades of the present invention.
In another embodiment, the toxicity of a test compound is assessed by treating a biological sample containing proteins with the test compound. Proteins from the biological sample are incubated with antibodies specific to the polypeptades of the present invention. The amount of protein recognized by the antibodies is quantified. The amount of protein in the treated biological sample is compared with the amount in an untreated biological sample. A difference in the amount of protein between the two samples is indicative of a toxic response to the test compound in the treated sample.
Microarrays may be prepared, used, and analyzed using methods known in the art. (See, e.g., Brennan, T.M. et al. (1995) U.S. Patent No. 5,474,796; Schena, M. et al.
(1996) Proc. Natl. Acad.
3o Sci. USA 93:10614-10619; Baldeschweiler et aI. (1995) PCT application W095/251116; Shalon, D. et al. (1995) PCT application W095/35505; Heller, R.A. et al. (1997) Proe. Natl.
Acad. Sci. USA
94:2150-2155; and Heller, M.J. et al. (1997) U.S. Patent No. 5,605,662.) Various types of microarrays are well known and thoroughly described in DNA Microarrays: A
Practical Approach, M. Schena, ed. (1999) Oxford University Press, London, hereby expressly incorporated by reference.

In another embodiment of the invention, nucleic acid sequences encoding MDDT
may be used to generate hybridization probes useful in mapping the naturally occurring genoxnic sequence. Either coding or noncoding sequences may be used, and in some instances, noncoding sequences may be preferable over coding sequences. For example, conservation of a coding sequence among members of a multi-gene family may potentially cause undesired cross hybridization during chromosomal - mapping. The sequences may be mapped to a particular chromosome, to a specific region of a chromosome, or to artificial chromosome constructions, e.g., human artificial chromosomes (HACs), yeast artificial chromosomes (YACs), bacterial artificial chromosomes (BACs), bacterial P1 constructions, or single chromosome cDNA libraries. (See, e.g., Harrington, J.J. et al. (1997) Nat.
1o Genet. 15:345-355; Price, C.M. (1993) Blood Rev. 7:127-134; and Trask, B.J.
(1991) Trends Genet.
7:149-154.) Once mapped, the nucleic acid sequences of the invention may be used to develop genetic linkage maps, far example, which correlate the inheritance of a disease state with the inheritance of a particular chromosome region or restriction fragment length polymorphism (RFLP).
(See, for example, Lender, E.S. and D. Botstein (-1986) Proc. Natl. Aced. Sci.
USA 83:7353-735?.) Fluorescent in.situ hybridization (FISH) maybe correlated with other physical and genetic . , map data. (See, e.g,., Heinz-Ulrich, et al. (1995) in Meyers, supra, pp.
965-968.) Examples of genetic map data can be found in various scientific journals or at the Online Mendelian Inheritance in Man (pMINI) World Wide Web site. Correlation between the location of the gene encoding MDDT on a -.
physical map and a speeihe disorder, ar a predisposition to a specihe disorder, may help define the ~ region of ANA associated with that disorder and thus may further positional cloning efforts.
In situ hybridization of chromosomal preparations and physical mapping techniques, such as linkage analysis using established chromosomal markers, may be used fox extending genetic maps.
Often the placement of a gene on the chroxrzosome of another mammalian species, such as mouse, may reveal associated markers even if the exact chromosomal locus is not known. This information is valuable to investigators searching fox disease genes using positional cloning or other gene discovery techniques. Once the gene or genes responsible for a disease or syndrome have been crudely localized by genetic linkage to a particular genomic region, e.g., ataxia-telangiectasia to 11q22-23, any sequences mapping to that area may represent associated or regulatory genes for further investigation.
{See, e.g., Gatti, R.A. et al. (1988) Nature 336:577-580.) The nucleotide sequence of the instant invention may also be used to detect differences in the chromosomal location due to trauslocation, inversion, etc., among normal, carrier, or affected individuals.
In another embodiment of the invention, MDDT, its catalytic or immunogenic fragments, or oligopeptides thereof can be used for screening libraries of compounds in any of a variety of drug screening techniques. The fragment employed ~ such screening may be free in solution, affixed to a solid support, borne on a cell surface, or located intracellularly. The formation of binding complexes between MDDT and the agent being tested may be measured.
Another technique for drug screening provides for high throughput screening of compounds having suitable binding affinity to the protein of interest. (See, e.g., Geysers, et al. (1984) PCT
application WO84/03564.) In this method, large numbers of different small test compounds are synthesized on a solid substrate. The test compounds are reacted with IV~DT, or fragments thereof, and washed. Bound MDDT is then detected by methods well known in the art.
Purified MDDT can also be coated directly onto plates for use in the aforementioned drug screening techniques.
Alternatively, non-neutralizing antibodies can be used to capture the peptide and immobilize it on a solid support.
In another embodiment, one may use competitive drug screening assays in which neutralizing antibodies capable of binding MDDT specifically compete with a test compound for binding MDDT.
In this manner, antibodies can be used to detect the presence of any peptide which shares one or more antigenic determinants with MDDT.
~ In additional embodiments, the nucleotide sequences which encode MDDT may be used in any molecular biology techniques that have yet ~to be developed, provided the new techniques rely on . properties of nucleotide sequences that are currently known, including, but not linuted to, such v properties as the triplet genetic code and specific base pair interactions.
Without.further elaboration, it is believed that one skilled. in the art can, using the preceding description, utilize the present invention to its fullest extent. The following embodiments are, therefore, to be construed as merely illustrative, and not litnitative of the remainder of the disclosure in any way whatsoever.
The disclosures of all patents, applications and publications, mentioned above and below, in particular U.S. Ser. No. 60/293,723, U.S. Ser. No. 60/295,257, U.S. Ser. No.
60/297,220, U.S. Ser.
No. 60/300,526, U.S. Ser. No. 60/301,874 and U.S. Ser. No. 60/359,413 are expressly incorporated by reference herein.
EXAMPLES
I. Construction of cDNA Libraries Incyte cDNAs were derived from cDNA libraries described in the LIFESEQ GOLD
database (Incyte Genomics, Palo Alto CA). Some tissues were homogenized and lysed in guanidinium isothiocyanate, while others were homogenized and lysed in phenol or in a suitable mixture of denaturants, such as TRIZOL (Invitrogen), a monophasic solution of phenol and guanidine isothiocyanate. The resulting lysates were centrifuged over CsCl cushions or extracted with chloroform. RNA was precipitated from the lysates with either isopropanol or sodium acetate and ethanol, or by other routine methods.
Phenol extraction and precipitation of RNA were repeated as necessary to increase RNA
purity. In some cases, RNA was treated with DNase. For most libraries, poly(A)+ RNA was isolated using oIigo d(T)-coupled paramagnetic particles (Promega), OLIGOTEX
latex particles (QIAGEN, Chatsworth CA), or an OLIGOTEX mRNA purification kit (QIAGEN).
Alternatively, RNA was isolated directly from tissue lysates using other RNA isolation kits, e.g., the POLY(A)PURE mRNA purification kit (Ambion, Austin TX).
In some cases, Stratagene was provided with RNA and constructed the corresponding cDNA
libraries. Otherwise, cDNA was synthesized and cDNA libraries were constructed with the UNTZAP vector system (Stratagene) or SUPERSCRIPT plasmid system (Invitrogen), using the recommended procedures or similar methods known in the art. (See, e.g., Ausubel, 1997, supra, units 5.1-6.6.) Reverse transcription was initiated using oligo d(T) or random primers. Synthetic oligonucleotide adapters were ligated to double stranded cDNA, and the cDNA
was digested with the appropriate restriction enzyme or enzymes. For most libraries, the cDNA was size-selected (300-1000 bp) using SEPHACRYL S 1000, SEPHAROSE CL2B, or SEPHAROSE CIA.B column chromatography (Amersham Biosciences) or preparative agarose gel electrophoresis. cDNAs were ligated into compatible restriction enzyme sites of the polylinker of a suitable plasmid, e.g., .
PBLUESCRIPT plasmid (Stratagene), PSPORT1 plasmid (Invitrogen), PCDNA2.1 plasmid (Invitrogen, Carlsbad CA), PBK-CMV plasmid (Stratagene), PCR2-TOPOTA plasmid (Invitrogen), PCMV-IC1S plasmid (Stratagene), pIGEN (Incyte Genomics, Palo Alto CA), pRARE
(Incyte Genomics), or pINCY (Incyte Genomics), or derivatives thereof. Recombinant plasmids were transformed into competent E. coli cells including XL1-Blue, XL1-BlueMRF, or SOLR from Stratagene or DHSa, DH10B, or ElectroMAX DH10B from Invitrogen.
II. Isolation of cDNA Clones Plasmids obtained as described in Example I were recovered from host cells by in vivo excision using the UNIZAP vector system (Stratagene) or by cell lysis.
Plasmids were purified using at least one of the following: a Magic or WIZARD Minipreps DNA purification system (Promega); an AGTC Miniprep purification kit (Edge Biosystems, Gaithersburg MD); and QIAWELL
8 Plasmid, 3o QIAWELL 8 Plus Plasmid, QIAWELL 8 Ultra Plasmid purification systems or the R.E.A.L. PREP
96 plasmid purification kit from QIAGEN. Following precipitation, plasmids were resuspended in 0.1 ml of distilled water and stored, with or without lyophilization, at 4°C.
Alternatively, plasmid DNA was amplified from host cell lysates using direct link PCR in a high-throughput format (Rao, V.B. (1994) Anal. Biochem. 216:1-14). Host cell Iysis and thermal cycling steps were carried out in a single reaction mixture. Samples were processed and stored in 384-well plates, and the concentration of amplified plasmid DNA was quantified fluorometrically using PICOGREEN dye (Molecular Probes, Eugene OR) and a FLUOROSKAN II fluorescence scanner (Labsystems Oy, Helsinki, Finland).
III. Sequencing and Analysis Incyte cDNA recovered in plasmids as described in Example II were sequenced as follows.
Sequencing reactions were processed using standard methods or high-throughput instrumentation such as the ABI CATALYST 800 (Applied Biosystems) thermal cycler or the PTC-200 thermal cycler (MJ Research) in conjunction with the HYDRA microdispenser (Robbins Scientific) or the MICROLAB 2200 (Hamilton) liquid transfer system. cDNA sequencing reactions were prepared using reagents provided by Amersham Biosciences or supplied in ABI sequencing kits such as the ABI PRISM BIGDYE Terminator cycle sequencing ready reaction kit (Applied Biosystems).
Electrophoretic separation of cDNA sequencing reactions and detection of labeled polynucleotides were carried' out using .the MEGABACE 1000 DNA sequencing system (Amersham Biosciences);
the ABI PRISM 37.3 or 377 sequencing system (Applied Biosystems) in conjunction with standard ABI protocols and base calling software; or other sequence analysis systems known in the art.
t Reading frames within the cDNA sequences were identified using standard methods (reviewed in. .
Ausubel, 1997, su ra; unit 7.7). Some of the.cDNA sequences were selected for extension using the , techniques disclosed in Example VIII. , , The polynucleotide sequences derived from Incyte cDNAs were validated by removing vector, linker, and poly(A) sequences and by masking ambiguous bases, using algorithms and programs based on BLAST, dynamic programming, and dinucleotide nearest neighbor analysis. The Incyte cDNA sequences or translations thereof were then queried against a selection of public databases such as the GenBank primate, rodent, mammalian, vertebrate, and eukaryote databases, and BLOCKS, PR1NTS, DOMO, PRODOM; PROTEOME databases with sequences from Homo sa iens, Rattus norve~icus, Mus musculus, Caenorhabditis elegans, Saccharomyces cerevisiae, Schizosaccharomyces pombe, and Candida albicans (Incyte Genomics, Palo Alto CA); hidden Markov model (HMM)-based protein family databases such as PFAM, INCY, and TIGRFAM
(Haft, D.H. et al. (2001) Nucleic Acids Res. 29:41-43); and HMM-based protein domain databases such as SMART
(Schultz et al. (1998) Proc. Natl. Acad. Sci. USA 95:5857-5864; Letunic, I. et al. (2002) Nucleic Acids Res. 30:242-244). (HNINI is a probabilistic approach which analyzes consensus primary structures of gene families. See, for example, Eddy, S.R. (1996) Curr. Opin.
Struct. Biol. 6:361-365.) The queries were performed using programs based on BLAST, FASTA, BLIMPS, and I~VIER.
The Incyte cDNA sequences were assembled to produce full length polynucleotide sequences.
~3 Alternatively, GenBank cDNAs, GenBank ESTs, stitched sequences, stretched sequences, or Genscan-predicted coding sequences (see Examples 1V and V) were used to extend lncyte cDNA
assemblages to full length. Assembly was performed using programs based on Phred, Phrap, and Consed, and cDNA assemblages were screened for open reading frames using programs based on GeneMark, BLAST, and FASTA. The full length polynucleotide sequences were translated to derive the corresponding full length polypeptide sequences. Alternatively, a polypeptide of the invention may begin at any of the methionine residues of the full length translated polypeptide. Full length polypeptide sequences were subsequently analyzed by querying against databases such as the GenBank protein databases (genpept), SwissProt, the PROTEOME databases, BLOCKS, PRINTS, DOMO, PRODOM, Prosite, hidden Markov model (HIVIM)-based protein family databases such as PFAM, 1NCY, and TIGRFAM; and IhVVIM-based protein domain databases such as SMART.
Full length polynucleotide sequences are also analyzed using MACDNASIS PRO software (Hitachi Software Engineering; South San Francisco CA) and LASERGENE software (DNASTAR).
Polynucleotide and polypeptide sequence alignments are generated using default parameters specified by the CLLTSTAL algorithm as incorporated into the MEGALIGN multisequence alignment program (DNASTAR), which also calculates the percent identity between aligned sequences.
Table 7 summarizes the tools, programs, and algorithms used for the analysis and assembly of:.
Incyte cDNA~ and full .length sequences and provides applicable descriptions, references, and threshold . .
parameters. The first column of Table 7 shows the tools, programs, and algorithms used, the second column provides brief descriptions thereof, the third column presents appropriate references, all of which are incorporated by reference herein in their entirety, and the fourth column presents, where applicable, the scores, probability values, and other parameters used to evaluate the strength of a match between two sequences (the higher the score or the lower the probability value, the greater the identity between two sequences).
The programs described above for the assembly and analysis of full length polynucleotide and polypeptide sequences were also used to identify polynucleotide sequence fragments from SEQ ll~
N0:40-78. Fragments from about 20 to about 4000 nucleotides which are useful in hybridization and amplification technologies are described in Table 4, column 2.
IV. Identification and Editing of Coding Sequences from Genomic DNA
Putative molecules for disease detection and treatment were initially identified by running the Genscan gene identification program against public genomic sequence databases (e.g., gbpri and gbhtg). Genscan is a general-purpose gene identification program which analyzes genomic DNA
sequences from a variety of organisms (See Burge, C. and S. Karlin (1997) J.
Mol. Biol. 268:78-94, and Burge, C. and S. Karlin (1998) Curr. Opin. Struct. Biol. 8:346-354). The program concatenates predicted exons to form an assembled cDNA sequence extending from a methionine to a stop codon.
The output of Genscan is a FASTA database of polynucleotide and polypeptide sequences. The maximum range of sequence for Genscan to analyze at once was set to 30 kb. To determine which of these Genscan predicted cDNA sequences encode molecules for disease detection and treatment, the encoded polypeptides were analyzed by querying against PFAM models for molecules for disease detection and treatment. Potential molecules for disease detection and treatment were also identified by homology to Incyte cDNA sequences that had been annotated as molecules for disease detection and treatment. These selected Genscan-predicted sequences were then compared by BLAST
analysis to the genpept and gbpri public databases. Where necessary, the Genscan-predicted sequences were then edited by comparison to the top BLAST hit from genpept to correct errors in the sequence predicted by Genscan, such as extra or omitted exons. BLAST analysis was also used to find any Incyte cDNA or public cDNA coverage of the Genscan-predicted sequences, thus providing evidence for transcription. When Incyte cDNA coverage was available, this information was used to correct or confirm the Genscan predicted sequence. Full length polynucleotide sequences were obtained by assembling Genscan-predicted coding sequences with Incyte cDNA
sequences and/or public cDNA sequences using the assembly process described in Example III.
Alternatively, full length polynucleotide sequences were derived entirely from edited or unedited Genscan-predicted coding sequences. . . ~ .
V: Assembly of Genomic Sequence Data with cDNA Sequence Data ~~Stitched" Seauences Partial cDNA sequences were extended with exons predicted by the Genscan gene identification program described in Example IV. Partial cDNAs assembled as described i_n Example III were mapped to genomic DNA and parsed into clusters containing related cDNAs and Genscan exon predictions from one or more genomic sequences. Each cluster was analyzed using an algorithm based on graph theory and dynamic programnning to integrate cDNA and genomic information, generating possible splice variants that were subsequently confirmed, edited, or extended to create a full length sequence. Sequence intervals in which the entire length of the interval was present on more than one sequence in the cluster were identified, and intervals thus identified were considered to be equivalent by transitivity. For example, if an interval was present on a cDNA and two genomic sequences, then all three intervals were considered to be equivalent. This process allows unrelated but consecutive genomic sequences to be brought together, bridged by cDNA
sequence. Intervals thus identified were then "stitched" together by the stitching algorithm in the order that they appear along their parent sequences to generate the longest possible sequence, as well as sequence variants.
Linkages between intervals which proceed along one type of parent sequence (cDNA to cDNA or genomic sequence to genomic sequence) were given preference over linkages which change parent type (cDNA to genomic sequence). The resultant stitched sequences were translated and compared by BLAST analysis to the genpept and gbpri public databases. Incorrect exons predicted by Genscan were corrected by comparison to the top BLAST hit from genpept. Sequences were further extended with additional cDNA sequences, or by inspection of genomic DNA, when necessary.
"Stretched" Sequences Partial DNA sequences were extended to full length with an algorithm based on BLAST
analysis. First, partial cDNAs assembled as described in Example III were queried against public databases such as the GenBank primate, rodent, mammalian, vertebrate, and eukaryote databases using the BLAST program. The nearest GenBank protein homolog was then compared by BLAST
analysis to either Incyte cDNA sequences or GenScan exon predicted sequences described in Example IV. A chimeric protein was generated by using the resultant high-scoring segment pairs (HSPs) to map the translated sequences onto the GenBank protein homolog.
Insertions or deletions may occur in the chimeric protein with respect to the original GenBank protein homolog. The ' GenBank protein homolog, the chimeric protein, or both were used as probes to search for homologous genomic sequences from the public human genome databases. Partial DNA
sequences were therefore "stretched." or extended by the addition of homologous genomic sequences. The resultant stretched sequences were examined to determine whether it contained a complete gene.
VI. Chromosomal Mapping of 1VVIDDT~ Encoding Polynucleotides The sequences which were used to assemble SEQ ID N0:40-78 were compared with sequences from the Incyte LIFESEQ database and public domain databases using BLAST and other implementations of the Smith-Waterman algorithm. Sequences from these databases that matched SEQ ID N0:40-78 were assembled into clusters of contiguous and overlapping sequences using assembly algorithms such as Phrap (Table 7). Radiation hybrid and genetic mapping data available from public resources such as the Stanford Human Genome Center (SHGC), Whitehead Institute for Genome Research (WIGR), and Genethon were used to determine if any of the clustered sequences had been previously mapped. Inclusion of a mapped sequence in a cluster resulted in the assignment of all sequences of that cluster, including its particular SEQ ID NO:, to that map location.
Map locations are represented by ranges, or intervals, of human chromosomes.
The map position of an interval, in centiMorgans, is measured relative to the terminus of the chromosome's p-arm. (The centiMorgan (cM) is a unit of measurement based on recombination frequencies between chromosomal markers. On average,1 cM is roughly equivalent to 1 megabase (Mb) of DNA in humans, although this can vary widely due to hot and cold spots of recombination.) The cM distances are based on genetic markers mapped by Genethon which provide boundaries for radiation hybrid markers whose sequences were included in each of the clusters. Human genome maps and other resources available to the public, such as the NCBI "GeneMap'99" World Wide Web site (http://www.ncbi.nlm.nih.gov/genemapn, can be employed to determine if previously identified disease genes map within or in proximity to the intervals indicated above.
VII. Analysis of Polynucleotide Expression Northern analysis is a laboratory technique used to detect the presence of a transcript of a gene and involves the hybridization of a labeled nucleotide sequence to a membrane on which RNAs from a particular cell type or tissue have been bound. (See, e.g., Sambrook, supra, ch. 7; Ausubel (1995) su ra, ch. 4 and 16.) Analogous computer techniques applying BLAST were used to search for identical or related molecules in cDNA databases such as GenBank or LIFESEQ (Incyte Genomics). This analysis is much faster than multiple membrane based hybridizations. In addition, the sensitivity of the computer search can be modified to determine whether any particular match is categorized as exact or similar.
The basis of the search is the product score, which is defined as:
BLAST Score x Percent Identity 5 x rrlinitrzum {length(Seq. 1), length(Seq. 2)}
The product score takes into account both the degree of similarity between two sequences and the length of the sequence match. The product score is a normalized value between 0 and 100, and is calculated as follows: the BLAST score is multiplied by the percent nucleotide identity and the pxoduct is divided by (5 times the length of the shorter of the two sequences). The BLAST score is calculated by assigning a score of +5 for every base that matches in a high-scoring segment pair (HSP), and -4 for every mismatch. Two sequences may share more than one HSP
(separated by gaps). If there is more than one HSP, then the pair with the highest BLAST
score is used to calculate the product score. The product score represents a balance between fractional overlap and quality in a BLAST alignment. For example, a product score of 100 is produced only for 100%
identity over the entire length of the shorter of the two sequences being compared. A product score of 70 is produced either by 100% identity and 70% overlap at one end, or by 88% identity and 100% overlap at the other. A product score of 50 is produced either by 100% identity and 50%
overlap at one end, or 79%
identity and 100% overlap.
Alternatively, polynucleotide sequences encoding MDDT are analyzed with respect to the tissue sources from which they were derived. For example, some full length sequences are assembled, at least in part, with overlapping Incyte cDNA sequences (see Example DI). Each cDNA

sequence is derived from a cDNA library constructed from a human tissue. Each human tissue is classified into one of the following organ/tissue categories: cardiovascular system; connective tissue;
digestive system; embryonic structures; endocrine system; exocrine glands;
genitalia, female; genitalia, male; germ cells; heroic and immune system; liver; musculoskeletal system;
nervous system;
pancreas; respiratory system; sense organs; skin; stomatognathic system;
unclassified/mixed; or urinary tract. The number of libraries in each category is counted and divided by the total number of libraries across all categories. Similarly, each human tissue is classified into one of the following disease%ondition categories: cancer, cell line, developmental, inflammation, neurological, trauma, cardiovascular, pooled, and other, and the number of libraries in each category is counted and divided by the total number of libraries across all categories. The resulting percentages reflect the tissue- and disease-specific expression of cDNA encoding MDDT. cDNA sequences and cDNA
library/tissue information are found in the L1FESEQ GOLD database (Incyte Genomics, Palo Alto CA).
VIII. Extension of MDDT Encoding Polynucleotides Full length polynucleotide sequences were also produced by extension of an appropriate fragment of the full length molecule using oligonucleotide primers designed from this fragment. One primer was synthesized to initiate 5' extension of the known fragment, and the other primer was synthesized to initiate 3' extension of the known fraganent. The initial primers were designed using OLIGO 4.06 software (National Biosciences), or another appropriate program, to be about 22 to 30 nucleotides in length, to have a GC content of about;50% or more, and to anneal to the target 20. sequence at temperatures of about 68 °C to about 72 °C. Any stretch of nucleotides which would result in hairpin structures and primer-primer dimerizations was avoided.
Selected human cDNA libraries were used to extend the sequence. If more than one ' extension was necessary or desired, additional or nested sets of primers were designed.
High fidelity amplification was obtained by PCR using methods well known in the art. PCR
was performed in 96-well plates using the PTC-200 thermal cycler (MJ Research, Inc.). The reaction mix contained DNA template, 200 nrilol of each primer, reaction buffer containing Mg2+, (NHq}ZSO4, and 2-mercaptoethanol, Taq DNA polymerase (Amersham Biosciences), ELONGASE
enzyme (Invitrogen), and Pfu DNA polymerise (Stratagene}, with the following parameters for primer pair PCI A and PCI B: Step l: 94°C, 3 min; Step 2: 94°C, 15 sec; Step 3: 60°C, 1 min; Step 4: 68°C, 2 min; Step 5: Steps 2, 3, and 4 repeated 20 times; Step 6: 68 °C, 5 min;
Step 7: storage at 4 °C. In the alternative, the parameters for primer pair T7 and SK+ were as follows: Step 1: 94°C, 3 min; Step 2:
94°C, 15 sec; Step 3: 57°C, 1 min; Step 4: 68°C, 2 min;
Step 5: Steps 2, 3, and 4 repeated 20 times;
Step 6: 68°C, 5 min; Step 7: storage at 4°C.
The concentration of DNA in each well was determined by dispensing 100 ~,l PICOGREEN

quantitation reagent (0.25% (v/v) PTCOGREEN; Molecular Probes, Eugene OR) dissolved in 1X TE
and 0.5 p1 of undiluted PCR product into each well of an opaque fluorimeter plate (Corning Costar, Acton MA), allowing the DNA to bind to the reagent. The plate was scanned in a Fluoxoskan II
(Labsystems Oy, Helsinki, Finland) to measure the fluorescence of the sample and to quantify the concentration of DNA. A S ,u1 to 10 ,u1 aliquot of the reaction mixture was analyzed by electrophoresis on a 1 % agarose gel to determine which reactions were successful in extending the sequence.
The extended nucleotides were desalted and concentrated, transferred to 384-well plates, digested with CviJI cholera virus endonuclease (Molecular Biology Research, Madison WI), and 2o sonicated or sheared prior to relegation into pUC 18 vector (Amexsham Biosciences). For shotgun sequencing, the digested nucleotides were separated on low concentration (0.6 to 0.8%) agarose gels, fragments were excised, and agar digested with Agar ACE (Promega). Extended clones were relegated using T4 ligase (New England Biolabs, Beverly MA) into pUC 18 vector (Arnersham Biosciences), treated with Pfu DNA polymerase (Stratagene) to, fill-in restriction site overhangs, and transfected into competent E. coli cells. Transformed cells were selected.on antibiotic-containing media, and,individual colonies were picked and cultured overnight at 37 °C in 384-well plates in LB/2x carb liquid media..
The cells were lysed, and DNA was amplified by PCR using Taq DNA polymerase (Amersham,Biosciences) and Pfu DNA polymerase (Stratagene) with the following.parameters:.Step 1: 94°C, 3 min; Step 2: 94°C, 15 sec; Step 3: 60°C, 1 min; Step 4: 72°C, 2 min; Step 5: steps 2, 3, and 4 repeated 29 tunes; Step 6: 72 °C, 5 min; Step 7: storage at 4 °C. DNA was quantified by PICOGREEN reagent (Molecular Probes) as described above. Samples with low DNA
recoveries were reampli~Zed using the same conditions as described above. Samples were diluted with 20%
dimethysulfoxide (1;2, v/v), and sequenced using DYENAMIC energy transfer sequencing primers and the DYENAMIC DIRECT kit (Amersham Biosciences) or the ABI PRISM BIGDYE
Terminator cycle sequencing ready reaction kit (Applied Biosystems).
In like manner, full length polynucleotide sequences are veriPxed using the above procedure or are used to obtain 5' regulatory sequences using the above procedure along with oligonucleotides designed for such extension, and an appropriate genomic library.
3o IX. Identification of Single Nucleotide Polymorphisms in MDDT Encoding Polynucleotides Common DNA sequence variants known as single nucleotide polymorphisms (SNPs) were identified in SEQ ID N0:40-78 using the LIFESEQ database (Tncyte Genomics).
Sequences from the same gene were clustered together and assembled as described in Example IfI, allowing the identification of all sequence variants in the gene. An algorithm consisting of a series of filters was used to distinguish SNPs from other sequence variants. Preliminary filters removed the majority of basecall errors by requiring a minim__um Phred quality score of 15, and removed sequence alignment errors and errors resulting from improper trimming of vector sequences, chimeras, and splice variants.
An automated procedure of advanced chromosome analysis analysed the original chromatogram files in the vicinity of the putative SNP. Clone error filters used statistically generated algorithms to identify errors introduced during laboratory processing, such as those caused by reverse transcriptase, polymerase, or somatic mutation. Clustering error filters used statistically generated algorithms to identify errors resulting from clustering of close homologs or pseudogenes, or due to contamination by non-human sequences. A final set of filters removed duplicates and SNPs found in immunoglobulins or T-cell receptors.
Certain SNPs were selected for further characterization by mass spectrometry using the high throughput MASSARRAY system (Sequenom, Inc.) to analyze allele frequencies at the SNP sites in four.different human populations. The Caucasian population comprised 92 individuals (46 male, 46 female), including 83 from Utah, four French, three Venezualan, and two Amish individuals. The African population comprised 194 individuals (97 male, 97 female), all African Americans. The Hispanic population comprised 324 individuals (162 male, 162 female), all Mexican Hispanic. The .
Asian population comprised 126 individuals (64 male, 62 female) with a reported parantalbreakdown,~
of 43 % Chinese, 31 % Japanese, 13 % Korean, 5 % Vietnamese, and 8 % other Asian. Allele frequencies were first analyzed in the Caucasian population; in some cases those SNPs which showed no allelic variance in this population were not further tested in the other three populations.
X. Labeling and Use of Individual Hybridization Probes Hybridization probes derived from SEQ ID N0:40-78 are employed to screen cDNAs, genomie DNAs, or mRNAs. Although the labeling of oligonucleotides, consisting of about 20 base pairs, is specifically described, essentially the same procedure is used with larger nucleotide fragments. Oligonucleotides are designed using state-of the-art software such as OLIGO 4.06 software (National Biosciences) and labeled by combining 50 pmol of each oligomer, 250 ~Ci of [y-32p~ adenosine triphosphate (Amersham Biosciences), and T4 polynucleotide kinase (DuPont NEN, Boston MA). The labeled oligonucleotides are substantially purified using a superf'me size exclusion dextran bead column (Amersham Biosciences). An aliquot containing 10' counts per minute of the labeled probe is used in a typical membrane-based hybridization analysis of human genomic DNA digested with one of the following endonucleases: Ase I, Bgl II, Eco RI, Pst I, Xba I, or Pvu II (DuPont NEN).
The DNA from each digest is fractionated on a 0.7% agarose gel and transferred to nylon membranes (Nytran Plus, Schleicher & Schuell, Durham NH). Hybridization is carried out for 16 hours at 40 °C. To remove nonspecific signals, blots are sequentially washed at room temperature under conditions of up to, for example, 0.1 x saline sodium citrate and O.S to sodium dodecyl sulfate.
Hybridization patterns are visualized using autoradiography or an alternative imaging means and compared.
XI. Microarrays The linkage or synthesis of array elements upon a microarray can be achieved utilizing photolithography, piezoelectric printing (ink jet printing, See, e.g., Baldeschweiler, su re.), mechanical microspotting technologies, and derivatives thereof. The substrate in each of the aforementioned technologies should be uniform and solid with a non-porous surface (Schena (1999), supra).
Suggested substrates include silicon, silica, glass slides, glass chips, and silicon wafers. Alternatively, a procedure analogous to a dot or slot blot may also be used to arrange and link elements to the surface of a substrate using thermal, UV, chemical, or mechanical bonding procedures.
A typical array may be produced using available methods and machines well known to those of ordinary shill in the art and may contain any appropriate number of elements. (See, e.g., Schena, M. et al.
(1995) Science 270:467-470; Shalon, D. et al. (1996) Genome Res. 6:639-645; Marshall, A. and J. Hodgson (199g) Nat. Biotechnol. 16:27-3l.) Full length cDNAs, Expressed Sequence Tags (ESTs), or fragments or oligomers thexeof may comprise the elements of the microarray. Fragments or oligomers suitable for hybridization can be selected using software well known in the art such as LASERGENE software (DNASTAR). The array elements are hybridized with polynucleotides in a biological sample. The polynucleotides in the biological sample are conjugated to a fluorescent label or other molecular tag for ease of detection.
After hybridization, nonhybridized nucleotides from the biological sample are removed, and a fluorescence scanner is used to detect hybridization at each array element.
Alternatively, laser desorbtion and mass spectrometry may be used for detection of hybridization.
The degree of complementarity and the relative abundance of each polynucleotide which hybridizes to an element on the microarray may be assessed. In one embodiment, microarray preparation and usage is described in detail below.
Tissue or Cell Sample_ Preparation Total RNA is isolated from tissue samples using the guanidinium thiocyanate method and poly(A)+ RNA is puri$ed using the oligo-(dT) cellulose method. Each poly(A)+
RNA sample is reverse transcribed using MMLV reverse-transcriptase, O.OS pg/~Cl oligo-(dT) primer (2lmer), 1X first strand buffer, 0.03 units/p,l RNase inhibitor, 500 ,uM dATP, 500 ~tM dGTP, 500 ~tM dTTP, 40 ,uM
dCTP, 40 ACM dCTP-Cy3 (BDS) or dCTP-Cy5 (Amersham Biosciences). The reverse transcription reaction is performed in a 25 ml volume containing 200 ng poly(A)+ RNA with GEMBRIGHT kits (Incyte). Specific control poly(A)+ RNAs are synthesized by in vitro transcription from non-coding yeast genomic DNA. After incubation at 37° C for 2 hr, each reaction sample (one with Cy3 and another with Cy5 labeling) is treated with 2.5 ml of 0.5M sodium hydroxide and incubated for 20 minutes at 85° C to the stop the reaction and degrade the RNA. Samples are purified using two successive CHROMA SPIN 30 gel filtration spin columns (CLONTECH Laboratories, Inc.
(CLONTECI~, Palo Alto CA) and after combining, both reaction samples are ethanol precipitated using 1 ml of glycogen (1 mg/ml), 60 ml sodium acetate, and 300 ml of 100%
ethanol. The sample is then dried to completion using a SpeedVAC (Savant Instruments Inc., Holbrook NY) and resuspended to in 14 ~Cl 5X SSC/0.2% SDS.
Microarray Preparation Sequences of the present invention are used to generate array elements. Each array element is amplified from bacterial cells containing vectors with cloned cDNA inserts.
PCR amplification uses primers complementary to the vector sequences flanking the cDNA insert. Array elements are amplified in thirty cycles of PCR from an initial quantity of 1-2 ng to a final quantity greater than 5 yg:
Amplified array elements are then purified using SEPHACRYL-400 (Amersham Biosciences).
Purified array elements are immobilized on polymer-coated glass slides. Glass microscope slides (Corning) are cleaned by ultrasound in 0. T % SDS and acetone, with extensive distilled water washes between and after treatments. Glass slides are etched in 4%
hydrofluoric acid (VWR
Scientific Products Corporation (VWR)~ West Chester PA), washed extensively in distilled water, and coated with 0.05% aminopropyl silane (Sigma) in 95% ethanol. Coated slides are cured in a 110°C
oven.
Array elements are applied to the coated glass substrate using a procedure described in U.S.
Patent No. 5,807,522, incorporated herein by reference. 1 ~.l of the array element DNA, at an average concentration of 100 ng/~.1, is loaded into the open capillary printing element by a high-speed robotic apparatus. The apparatus then deposits about 5 n1 of array element sample per slide.
Microarrays are UV-crosslinked using a STRATALINKER UV-crosslinker (Stratagene).
Microarrays are washed at room temperature once in 0.2% SDS and three times in distilled water.
Non-specific binding sites are blocked by incubation of microarrays in 0.2%
casein in phosphate buffered saline (PBS) (Tropix, Inc., Bedford MA) for 30 minutes at 60°
C followed by washes in 0.2%
SDS and distilled water as before.
Hybridization Hybridization reactions contain 9 ~Cl of sample mixture consisting of 0.2 ~.g each of Cy3 and Cy5 labeled cDNA synthesis products in 5X SSC, 0.2% SDS hybridization buffer.
The sample mixture is heated to 65° C for 5 minutes and is aliquoted onto the microarray surface and covered with an 1.8 cm2 coverslip. The arrays are transferred to a waterproof chamber having a cavity just slightly larger than a microscope slide. The chamber is kept at 100% humidity internally by the addition of 140 p1 of 5X SSC in a corner of the chamber. The chamber containing the arrays is incubated for about 6.5 hours at 60° C. The arrays are washed for 10 min at 45° C in a ~trst wash buffer (1X SSC, 0.1 %
SDS), three times for 10 minutes each at 45° C in a second wash buffer (0.1X SSC), and dried.
Detection Reporter-labeled hybridization complexes are detected with a microscope equipped with an Iunova 70 mixed gas 10 W laser (Coherent, Inc., Santa Clara CA) capable of generating spectral lines at 488 nm for excitation of Cy3 and at 632 nm for excitation of CyS. The excitation laser light is focused on the array using a 20X microscope objective (Nikon, Inc., Melville NY). The slide containing the array is placed on a computer-controlled X-Y stage on the microscope and raster-scanned past the objective. The 1.8 cm x 1.8 cm array used in the present example is scanned with a resolution of 20 micrometers.
In two separate scans, a mixed gas multiline laser excites the two fluorophores sequentially.
Emitted light is split, based on wavelength, into two photomultiplier tube detectors (PMT 81477, Hamamatsu Photonics Systems, Bridgewater NJ) corresponding to the two fluorophores. Appropriate;.
filters position~:d between the array and the photomultiplier cubes are used to filter the signals. The emission maxuna of the fluorophores used are 565 nm for Cy3 and 650 nm for CyS. Each array is typically scanned twice, one scan per fluorophore'using the appropriate filters at the laser source, although the apparatus is capable of recording the spectra from both fluorophores simultaneously.
The sensitivity of the scans is typically calibrated using the signal intensity generated by a cDNA control species added to the sample mixture at a known concentration. A
specific location on the array contains a complementary DNA sequence, allowing the intensity of the signal at that location to be correlated with a weight ratio of hybridizing species of 1:100,000. When two samples from different sources (e.g., representing test and control cells), each labeled with a different fluorophore, are hybridized to a single array for the purpose of identifying genes that are differentially expressed, the calibration is done by labeling samples of the calibrating cDNA with the two fluorophores and adding identical amounts of each to the hybridization mixture.
The output of the photomultiplier tube is digitized using a 12-bit RTI-835H
analog-to-digital (A/D) conversion board (Analog Devices, Inc., Norwood MA) installed in an IBM-compatible PC
computer. The digitized data are displayed as an image where the signal intensity is mapped using a linear 20-color transformation to a pseudocolor scale ranging from blue (low signal) to red (high signal). The data is also analyzed quantitatively. Where two different fluorophores are excited and measured simultaneously, the data are first corrected for optical crosstalk (due to overlapping emission spectra) between the fluorophores using each fluorophore's emission spectrum.
A grid is superimposed over the fluorescence signal image such that the signal from each spot is centered in each element of the grid. The fluorescence signal within each element is then integrated to obtain a numerical value corresponding to the average intensity of the signal. The software used for signal analysis is the GEMTOOLS gene expression analysis program (Incyte).
Array elements that exhibited at least about a two-fold change in expression, a signal-to-background ratio of at least 2.5, and an element spot size of at least 40% were identified as differentially expressed using the GEMTOOLS program (Incyte Genomics).
l0 Expression For example, expression of SEQ m N0:40 was upregulated in PBMC cells stimulated with lipopolysaccharide (LPS), a component of the bacterial cell wall which induces an inflammatory response. PBMCs collected from the blood of four healthy donors was stimulated with 1 ~.g/ml LPS
for 4, 24, and 72 hours. The PBMCs contained about 52% lymphocytes (12% B-cells and 40% T-cells), 20% natural killer cells, 25% monocytes, and 3% various cells including dendritic cells.
Stimulated cells were then compared to untreated, time-matched controls.
Similarly, expression of SEQ ID N0:40 vc~as upregulated in vascular tissue stimulated with the inflammatory cytokine TNFa or a combination of the protein kinase C activator, PMA, and ionomycin. Cells isolated from vascular smooth muscle, including human coronary artery smooth muscle cells (CASMC), and vascular endothelium, including HUVECs, were grown to 85% confluence in SmGM-2 or EGM-2, respectively, at 37°C, 5% CO2. Cells were then stimulated with either 10 ng/ml TNFa or 1 ~,M PMA, 1 p.g/ml ionomycin over a defined time course. Upregulation of SEQ m N0:40 in treated cells relative to untreated, time-matched controls was seen within 1-4 hours following treatment.
Expression of SEQ m NO:42 was downregulated in ovarian adenocarcinoma and a breast adenocarcinoma cell line, BT-20, relative to normal ovary and breast, respectively. Ovarian tumor tissue obtained from a 79-year-old female was compared to normal ovary obtained from the same donor. BT-20 is a breast adenocarcinoma line derived in vitro from cells emigrating out of thin slices of a tumor mass isolated form a 74-year-old female. BT-20 cells were compared to primary mammary epithelial cells (HMEC) and a breast mammary gland cell line (MCF-10A) isolated from a 36-year-old woman with fibrocystic disease. The breast cell lines were grown in basal medium in the absence of growth factors and hormones for 24 hours prior to the comparison.
Expression of SEQ D7 N0:45 was upregulated in THP-1 promonocytes stimulated with PMA
and ionomycin. THP-1 is a promonocyte cell line isolated from the peripheral blood of a 1-year-old male with acute monocytic leukemia. Upon stimulation with PMA, TFiP-1 differentiates into a macrophage-like cell that displays many characteristics of peripheral human macrophages. THP-1 cells stimulated in vitf-o with 0.1 ~,M PMA and 1 ~,g/xnl ionomycin for 0.5, 1, 2, 4, and 8 hours were compared to untreated, time-matched control cells. Expression of SEQ )D N0:45 was downregulated in several breast cell cancer lines relative to F3MECs. Experiments on breast cell lines were as described above. Cell lines included BT-20, BT474, BT483, Hs578T, MCF-7, and MD-AMB-468.
Expression of SEQ m N0:48 was upregulated in HLJVECs stimulated with TNFa following pre-treatment with either PMA or a low dose of TNFa. HUVECs were pre-treated with either 100 nM PMA or 0.1 ng/ml TNFa for 24 hours, washed, and then stimulated with TNFa for an additional 1, 4, and 24 hours. HWECs were cultured in llVIDM, 10% fetal calf serum at 37°C, 5% CO2. Treated cells were compared to untreated, time-matched controls.
For example, SEQ m N0:68 is upregulated 3.4 fold in mature DC versus monocytes, suggesting that SEQ m N0:68, encoding SEQ ID N0:29, could be used for example, to understand the process by which monocytes differentiate into immature dendritic cells and eventually allow manipulation of the immune system leading to potential itnmunotherapies for diseases such as cancer, A)DS, and infectious diseases; and enhancing vaccine efficacy.
In another example, SEQ ID N0:78 showed differential expression in inflammatory responses as determined by microarray analysis.' The. expression of SEQ m N0:78 was increased by at least two fold in TFiP-1 human promonocytc line which had been stimulated for 26 hours with 1 ~,M PMA
(phorbol 12-myristate 13-acetate) when compared to untreated THP-1 cells. PMA
is abroad activator of the protein kinase C-dependent pathways. THP-1 is promonocyte line derived from peripheral blood of a 1 year old male with acute monocytic leukemia. The cell line acquires monocytic characteristics upon stimulation with PMA. Monocytes play a critical role in the initiation and maintenance of inflammatory immune responses. Therefore, SEQ 1D N0:78 is useful in diagnostic assays for inflammatory responses.
Further, as determined by microarray analysis, SEQ m N0:78 showed differential expression in SKBr3 breast carcinoma cell line versus FiMEC primary mammary epithelial cells and MCF10A
breast mammary gland cells. SkBR3 is a breast adenocarcinoma cell line isolated from a malignant pleural effusion of a 43-year-old female. I3MEC, a primary mammary epithelial cell line was derived from normal human mammary tissue (Clonetics, San Diego, CA). MCF10A, a breast mammary gland (luminal ductal characteristics) cell line was isolated from a 36 year old woman with fibrocystic breast disease. The microarray experiments showed that the expression of SEQ m N0:78 was increased by at least two fold in SKBr3 breast adenocarcinoma line relative to cells from the primary mammary epithelial cell line, I~MEC and the breast mammary gland cell line, MCF10A.
Therefore, SEQ 1D
N0:78 is useful as diagnostic markers or as potential therapeutic targets for breast cancer.

In an alternative example, SEQ ID N0:78 showed differential expression in MDAPCa2b prostate adenocarcinoma cell line versus PrEC normal prostate epithelial cells as determuned by microarray analysis. MDAPCa2b is a prostate adenocarcinoma cell line isolated from a metastatic site in the bone of a 63-year-old male.1VIDAPCa2b cell line expresses prostate specific antigen (PSA) and androgen receptor, grows in vitro and in vivo, and is androgen sensitive.
The normal epithelial cell line, PrEC, is a primary prostate epithelial cell line isolated franc a normal donor. The experiment showed that the expression of SEQ ID N0:78 was increased by at least two fold in MDAPCa2b cell line relative to PrECs. Therefore, SEQ 1D N0:78 is useful as a diagnostic marker or as a potential therapeutic target for prostate cancer.
1o XII. Complementary Polynucleotides Sequences complementary to the MPDT-encoding sequences, or any parts thereof, are used to detect, decrease, or inhibit expression of xlaturally occurring MDDT.
Although use of oligonucleotides comprising from about 15 to 30 base pairs is described, essentially the same procedure is used with smaller or with larger sequence fragments.
Appropriate~oligonucleotides are IS designed using OLIGO 4.06 software (National Biosciences) 'and the coding sequence of MDDT. To inhibit transcription, a complementary oligonucleotide is designed.from the most unique S' sequence a~ad used to prevent promoter binding to the coding equence. To inhibit translation, a complementary oligonucleotide is designed to prevent ribos~maZ bidding to the MDDT-encoding transcript..
XIII. Expression of MDDT
20 Expression and purification of MDDT ~is acliieved'using bacterial or virus based expression systems. For expression of lVmDT in bacteria, cDNA is subcloned into an appropriate vector containing an antibiotic resistance gene and an inducible promoter that directs high levels of cDNA
transcription. Examples of such promoters include, but are not limited to, the trp-lac (tic) hybrid promoter and the TS or T7 bacteriophage promoter in conjunction with the lac operator regulatory 25 element. Recombinant vectors are transformed into suitable bacterial hosts, e.g., BL21(DE3).
Antibiotic resistant bacteria express MDDT upon induction with isopropyl beta-D-thiogalactopyxanoside (n'TG). Expression of MDDT in eukaryotic cells is achieved by infecting insect or mammalian cell lines with recombinant Autographica californica nuclear polyhedrosis virus (AcMNPV), commonly known as baculovirus. The nonessential polyhedrin gene of baculovirtzs is 3o replaced with cDNA encoding MDDT by either homologous recombination or bacterial-mediated transposition involving transfer plasn~id intermediates. Viral infectivity is maintained and. the strong polyhedrin promoter drives high levels of cDNA transcription. Recombinant baculovirus is used to infect S,podoptera fxu~ibexda (Sf9) insect cells in most cases, or human hepatocytes, in some cases.
Infection of the latter requires additional genetic modifications to baculovirus. (See Engelhard, E.K. et al. (1994) Proc. Natl. Acad. Sci. USA 91:3224-3227; Sandig, V. et al. (1996) Hum. Gene Ther.

7:1937-1945.) In most expression systems, MDDT is synthesized as a fusion protein with, e.g., glutathione S-transferase (GST) or a peptide epitope tag, such as FLAG or 6-His, permitting rapid, single-step, affinity-based purification of recombinant fusion protein from crude cell lysates. GST, a 26-kilodalton enzyme from Schistosoma iaponicum, enables the purification of fusion proteins on immobilized glutathione under conditions that maintain protein activity and antigenicity (Amersham Biosciences).
Following purification, the GST moiety can be proteolytically cleaved from MDDT at specifically engineered sites. FLAG, an 8-amino acid peptide, enables immunoaffinity purification using commercially available monoclonal and polyclonal anti-FLAG antibodies (Eastman Kodak). 6-His, a stretch of six consecutive histidine residues, enables purification on metal-chelate resins (QIAGEN).
Methods for protein expression and purification are discussed in Ausubel (1995, su ra, ch. 10 and 16).
Purified MDDT obtained by these methods. can be used directly in the assays shown in Examples XVII and XVIZI, where applicable.
XIV. Functional Assays MDDT function is assessed by expressing the sequences encoding MDDT at physiologically elevated levels in mammalian cell culture systems. cDNA is srbcloned into a mammalian. expression vector containing a strong promoter that drives high levels cf cDNA
expression. Vectors of choice include PCMV SPORT plasmid (Invitrogen, Carlsbad CA) and PCR3.1 plasmid (Invitrogen), both of which contain the cytomegalovirus promoter. 5-10 ~g of recombinant vector are transiently transfected into a human cell line, for example, an endothelial or hematopoietic cell line, using either liposome formulations or electroporation. 1-2 /,tg of an additional plasnud containing sequences encoding a marker protein are co-transfected. Expression of a marker protein provides a means to distinguish transfected cells from nontransfected cells and is a reliable predictor of cDNA expression from the recombinant vector. Marker proteins of choice include, e.g., Green Fluorescent Protein (GFP; Clontech), CD64, or a CD64-GFP fusion protein. Flow cytometry (FCM), an automated, laser optics-based technique, is used to identify transfected cells expressing GFP
or CD64-GFP and to evaluate the apoptotic state of the cells and other cellular properties. FCM
detects and quantifies the uptake of fluorescent molecules that diagnose events preceding or coincident with cell death. These events include changes in nuclear DNA content as measured by staining of DNA
with propidium iodide; changes in cell size and granularity as measured by forward light scatter and 90 degree side light scatter; down-regulation of DNA synthesis as measured by decrease in bromodeoxyuridine uptake; alterations in expression of cell surface and intracellular proteins as measured by reactivity with specific antibodies; and alterations in plasma membrane composition as measured by the binding of fiuorescein-conjugated Annexin V protein to the cell surface. Methods in flow cytometry are discussed in Ormerod, M.G. (1994) Flow Cytometry, Oxford, New York NY.
The influence of MDDT on gene expression can be assessed using highly purified populations of cells transfected with sequences encoding MDDT and either CD64 or CD64-GFP.
CD64 and CD64-GFP are expressed on the surface of transfected cells and bind to conserved regions of human immunoglobulin G (IgG). Transfected cells are efficiently separated from nontransfected cells using magnetic beads coated with either human IgG or antibody against CD64 (DYNAL, Lake Success NY). mRNA can be purified from the cells using methods well known by those of skill in the art.
Expression of mRNA encoding MDDT and other genes of interest can be analyzed by northern analysis or micxoarray techniques.
XV. Production of MDDT Specific Antibodies MDDT substantially purified using polyacrylamide gel electrophoresis (PAGE;
see, e.g., Harrington, M.G (1990) Methods Enzymol. 182:488-495), or other purification techniques, is used to immunize animals (e.g., rabbits, mice, etc.) and to produce antibodies using standard protocol's.
Alternatively, the MDDT amino acid sequence is analyzed using LASERGENE
software (DNASTAR) to determine regions of high immunogenicity, and a corresponding oligopeptide is synthesized and used to raise antibodies by means known to those ~of skill in the art. Methods for selection of appropriate. epitopes, such as those near the C-terminus:or. in hydrophilic regions are well described in the art. (See, e.g., Ausubel, 1995, su ra, ch. 11.) ' Typically, oligopeptides of about 15 residues in length are synthesized using an ABI 431A
peptide synthesizer (Applied Biosystems) using FMOC chemistry and coupled to KI,H (Sigma-Aldrich, St. Louis MO) by reaction with N-maleimidobenzoyl-N-hydroxysuccinimide ester (MBS) to increase immunogenicity. (See, e.g., Ausubel, 199, supra.) Rabbits are immunized with the oligopeptide-KLH complex in complete Freund's adjuvant. Resulting antisera are tested for antipeptide and anti-MDDT activity by, for example, binding the peptide or MDDT to a substrate, ' blocking with 1% BSA, reacting with rabbit antisera, washing, and reacting with radio-iodinated goat anti-rabbit IgG.
XVI. Purification of Naturally Occurring MDDT Using Specific Antibodies Naturally occurring or recombinant MDDT is substantially purified by immunoaffrnity chromatography using antibodies specific for MDDT. An immunoaffinity column is constructed by covalently coupling anti-MDDT antibody to an activated chromatographic resin, such as CNBr-activated SEPHAROSE (Amersham Biosciences). After the coupling, the resin is blocked and washed according to the manufacturer's instructions.
Media containing MDDT are passed over the ixnmunoaffmity column, and the column is washed under conditions that allow the preferential absorbance of MDDT (e.g., high ionic strength buffers in the presence of detergent). The column is eluted under conditions that disrupt antibody/1V>DDT binding (e.g., a buffer of pH 2 to pH 3, or a high concentration of a chaotrope, such as urea or thiocyanate ion), and MDDT is collected.
XVII. Identification of Molecules Which Interact with MDDT
MDDT, or biologically active fragments thereof, are labeled with lasl Bolton-Hunter reagent.
(See, e.g., Bolton, A.E. and W.M. Hunter (1973) Biochem. J. 133:529-539.) Candidate molecules previously arrayed in the wells of a multi-well plate are incubated with the labeled MDDT, washed, and any wells with labeled MDDT complex are assayed. Data obtained using different concentrations of MDDT are used to calculate values for the number, affiuity, and association of MDDT with the candidate molecules.
Alternatively, molecules interacting with MDDT are analyzed using the yeast two hybrid system as described in Fields, S. and O. Song (1989) Nature 340:245-246, or using commercially available kits based on the two-hybrid system, such as the MATCHMAKER system (Clontech).
MDDT may also be used in the PATHCALLING process (CuraGen Corp., New Haven CT) which employs the yeast two-hybrid system in a high-throughput manner to determine all interactions.
. ' v between the proteins encoded by two large libraries of genes (Nandabalau, K. et al. (2000) U.S.
Patent No. 6,057,101).' XVIII. Demonstration of MDDT Activity Phorbol ester binding activity of MDDT is measured using au assay based on the fluorescent phorbol ester sapinotoxin-D (SAPD). Binding of SAPD to MDDT is quantified by measuring the resonance energy transfer from MDDT tryptophans to the 2-(N-methylamino)benzoyl fluorophore of the phorbol ester, as described by Slater et al. (((1996) J. Biol. Chem.
271:4627-4631).
MDDT activity is associated with its ability to form protein-protein complexes and is measured by its ability to regulate growth characteristics of NIH3T3 mouse fibroblast cells. A cDNA
encoding MDDT is subcloned into an appropriate eukaryotic expression vector.
This vector is transfected into NIH3T3 cells using methods kuown in the art. Trausfected cells are compared with non-transfected cells for the following quantifiable properties: growth in culture to high density, reduced attachment of cells to the substrate, altered cell morphology, and ability to induce tumors when injected into immunodehcient mice. The activity of MDDT is proportional to the extent of increased growth or frequency of altered cell morphology in NIH3T3 cells transfected with MDDT.
Alternatively, MDDT activity is measured by binding of MDDT to radiolabeled formin polypeptides containiug the proline-rich region that specifically binds to SH3 containing proteins (Char, D.C. et al. (199fr) EMBO J. 15:1045-1054). Samples of MDDT are run on SDS-PAGE
gels, and transferred onto nitrocellulose by electroblotting. The blots are blocked for 1 hr at room temperature in TBST (137 mM NaCl, 2.7 mM KCl, 25 mM Tris (pH 8.0) and 0.1% Tween-20) containing non-fat dry milk. Blots are then incubated with TBST containing the radioactive formin polypeptide for 4 hrs to overnight. After washing the blots four times with TBST, the blots are exposed to autoradiographic film. Radioactivity is quantitated by cutting out the radioactive spots and counting them in a radioisotope counter. The amount of radioactivity recovered is proportional to the activity of MDDT
in the assay.
Alternatively, MDDT protein kinase activity is measured by quantifying the phosphorylation of an appropriate substrate in the presence of gamma-labeled 32P-ATP. MDDT is incubated with the substrate, 3zP-ATP, and an appropriate kinase buffer. The 32P incorporated into the product is separated from free 32P-ATP by electrophoresis, and the incorporated 32P is quantified using a beta radioisotope counter. The amount of incorporated 32P is proportional to the protein kinase activity of MDDT in the assay. A determination of the specific amino acid residue phosphorylated by protein kinase activity is made by phosphoamino acid analysis of the hydrolyzed protein.
Alternatively, an assay for MDDT protein phosphatase activity measures. the hydrolysis of para-nitrophenyl phosphate (PNPP). MDDT is incubated together with PNPP in HEPES buffer pH
7:.5, in the presence of 0.1 % (3-mercaptoethanol at 37 °C for 60 min.
The reaction is stopped by the , addition of 6 ml of 10 N NaOH, and the increase in light absorbance of the reaction mixture at 410 iim resulting from the hydrolysis of PNPP is measured using a spectrophotometer.
The increase in light absorbance is proportional to the activity of MDDT in the assay (Diamond, R.H.
et al. (1994) Mol.
Cell Biol. 14:3752-3762).
Alternatively, adenylyl cylcase activity of MDDT is demonstrated by the ability to convert ATP to cAMP (Mittal, C.K. (1986) Meth. Enzymol. 132:422-428). In this assay MDDT is incubated with the substrate [a-3aP]ATP, following which the excess substrate is separated from the product cyclic [32P] AMP. MDDT activity is determined in 12 x 75 mm disposable culture tubes containing 5 ~l of 0.6 M Tris-HCl, pH 7.5, 5 ~.l of 0.2 M MgCl2, 5 ~,l of 150 mM creative phosphate containing 3 units of creative phosphokinase, 5 p1 of 4.0 mM 1-methyl-3-isobutylxanthine, 5 p1 of 20 mM cAMP, 5 x,120 mM ditlliothreitol, 5 ~1 of 10 mM ATP, 10 ~,1 [a-3zP]ATP (2-4 x 106 cpm), and water in a total volume of 100 ~.1. The reaction mixture is prewarmed to 30°C. The reaction is initiated by adding MDDT to the prewarmed reaction mixture. After 10-15 minutes of incubation at 30 °C, the reaction is terminated by adding 25 ~tl of 30% ice-cold trichloroacetic acid (TCA). Zero-time incubations and reactions incubated in the absence of MDDT are used as negative controls.
Products are separated by ion exchange chromatography, and cyclic [32P] AMP is quantified using a (3-radioisotope counter.
The MDDT activity is proportional to the amount of cyclic [32P] AMP formed in the reaction.

An alternative assay measures MDDT-mediated G-pxotein signaling activity by monitoring the mobilization of Ca2+ as an indicator of the signal transduction pathway stimulation. (See, e.g., Grynkiewicz, G. et al. (1985) J. Biol. Chem. 260:3440; McColl, S. et al.
(1993) 3. Tmmunol.
150:4550-4555; and Aussel su ra). The assay requires preloading neutrophils or T cells with a fluorescent dye such as FURA-2 or BCECF (Universal Imaging Corp, Westchester PA) whose emission characteristics are altered by Ca2+ binding. When the cells are exposed to one or more activating stimuli artificially (e.g., anti-CD3 antibody ligation of the T
cell receptor) or physiologically (e.g., by allogeneic stimulation), Ca2+ flux takes place. This flux can be observed and quantified by assaying the cells in a fluorometer or fluorescent activated cell sorter.
Measurements of Ca2+ flux are compared between cells in their normal state and those transfected with MDDT.
Increased Caz+
mobilization attributable to increased MDDT concentration is proportional to MDDT activity.
Alternatively, GTP binding activity of MDDT is determined in an assay that measures the binding of MDDT to [a-32P]-labeled GTP. Purified MDDT is first blotted onto filters and rinsed in a suitable buffer. The filters are then incubated in buffer containing radiolabeled [a-3aP]-GTP. The filters are washed in buffer to remove unbound GTP and counted in~a radioisotope counter. Non-specific binding is determined in an assay that contains a 100-fold excess of unlabeled GTP. The amount of specific~binding is proportional to the activity of MDDT.
Altexnatively, GTPase activity of MDDT is determined in an assay that measures the .
conversion of [a-3zP]-GTP to [a-32P]-GDP. MDDT is incubated with [a-32P]-GTP
in buffer for an appropriate period of time, and the reaction is terminated.by heating or acid precipitation followed by centrifugation. An aliquot of the supernatant is subjected to polyacrylamide gel electrophoresis (PAGE) to separate GDP and GTP together with unlabeled standards. The GDP spot is cut out and counted in a radioisotope counter. The amount of radioactivity recovered in GDP is proportional to the GTPase activity of MDDT.
Alternatively, MDDT activity is measured by quantifying the amount of a non-hydrolyzable GTP analogue, GTFyS, bound over a 10 minute incubation period. Varying amounts of MDDT are incubated at 30 °C in 50 mM Tris buffer, pH 7.5, containing 1 mM
dithiothreitol, 1 mM EDTA and 1 /,tM [35S]GTFyS. Samples are passed through nitrocellulose filters and washed twice with a buffer consisting of 50 mM Tris-HCl, pH 7.8,1 mM NaN3, 10 mM MgCl2, 1 mM EDTA, 0.5 mM
dithiothreitol, 0.01 mM PMSF, and 200 mM NaCl. The filter-bound counts are measured by liquid scintillation to quantify the amount of bound [35S]GTFyS. MDDT activity may also be measured as the amount of GTP hydrolysed over a 10 minute incubation period at 37 °C. MDDT is incubated in 50mM Tris-HCl buffer, pH 7.8, containing 1mM dithiothreitol, 2mM EDTA, lO,uM
[a-32P]GTP, and 1 ,uM H-rab protein. GTPase activity is initiated by adding MgClz to a final concentration of 10 mM.
Samples are removed at various time points, mixed with an equal volume of ice-cold O.SmM EDTA, and frozen. Aliquots are spotted onto polyethyleneimine-cellulose thin layer chromatography plates, which are developed in 1M LiCl, dried, and autoradiographed. The signal detected is proportional to MDDT activity.
Alternatively, MDDT activity may be demonstrated as the ability to interact with its associated low molecular weight (LMV~ GTPase in an in vitro binding assay. The candidate LMW
GTPases are expressed as fusion proteins with glutathione S-transferase (GST), and purified by affinity chromatography on glutathione-Sepharose. The LMW GTPases are loaded with GDP by l0 incubating 20 mM Tris buffer, pH 8.0, containing 100 mM NaCl, 2 mM EDTA, 5 mM MgClz, 0.2 mM
DTT, 100 ~.M AMP-PNP and 10 ~.M GDP at 30°C for 20 minutes. MDDT is expressed as a FLAG
fusion protein in a baculovirus system. Extracts of these baculovirus cells containing MDDT-FLAG
fusion proteins are precleared with GST beads, then incubated with GST-GTPase fusion proteins.
The complexes formed are precipitated by glutathione-Sepharose and separated by SDS-polyacrylamide gel electrophoresis. The separated proteins are blotted onto nitrocellulose membranes and.probed with commercially available anti-FLAG antibodies: MDDT activity is proportional to the amount of MDDT-FhAG fusion protein. detected in the complex.
Another alternative assay to detect MDDT activity is the use of a yeast two-hybrid system (Zalcman, G. et al. (1996) J. Biol. Chem. 271:30366-30374). Specifically, a plasmid such as pGAD1318 which may contain the coding region of MDDT can be used to transform reporter L40 yeast cells which contain the reporter genes LacZ and HIS3 downstream from the binding sequences for LexA. These yeast cells have been previously transformed with a pLexA-Rab6-GDP (mouse) plasmid or with a plasmid which contains pLexA-lamin C. The pLEXA-lamin C
cells serve as a negative control. The transformed cells are plated on a histidine-free medium and incubated at 30 °C
for 3 days. His+ colonies are subsequently patched on selective plates and assayed for (3-galactosidase activity by a filter assay. MDDT binding with Rab6-GDP is indicated by positive His*llacZ~ activity for the cells transformed with the plasmid containing the mouse Rab6-GDP and negative His+/lacZ+ activity for those transformed with the plasmid containing lamin C.
Alternatively, MDDT activity is measured by binding of NIDDT to a substrate which recognizes WD-40 repeats, such as ElonginB, by coimmunoprecipitation (Kamura, T. et al. (1998) Genes Dev. 12:3872-3881). Briefly, epitope tagged substrate and MDDT are mixed and immunoprecipitated with commercial antibody. against the substrate tag. The reaction solution is run on SDS-PAGE and the presence of MDDT visualized using an antibody to the MDDT
tag. Substrate binding is proportional to MDDT activity.

Alternatively, MDDT activity is measured by its inclusion in coated vesicles.
MDDT can be expressed by transforming a mammalian cell line such as COS7, HeLa, or CHO
with a eukaryotic expression vector encoding MDDT. Eukaryotic expression vectors are commercially available, and the techniques to introduce them into cells are well known to those skilled in the art. A small amount of a second plasmid, which expresses any one of a number of marker genes, such as (3-galactosidase, is co-transformed into the cells in order to allow rapid identification of those cells which have taken up and expressed the foreign DNA. The cells are incubated for 48-72 hours after transformation under conditions appropriate for the cell line to allow expression and accumulation of MDDT and (3-galactosidase.
In the alternative, MDDT activity is measured by its ability to alter vesicle trafficking pathways. Vesicle trafficking in cells transformed with MDDT is examined using fluorescence microscopy. Antibodies specific for vesicle coat proteins or typical vesicle trafficking substrates such as transferrin or the mannose-6-phosphate receptor are commercially available.
Various cellular components such as ER, Golgi bodies, peroxisomes, endosomes, lysosomes, and the plasmalemma are examined. Alterations in the numbers and locations of vesicles in cells transformed with MDDT as compared to. control cells are characteristic of MDDT activity. Transformed cells are collected and cell lysates are assayed for vesicle form_ation.; .A nom-hydrolyzable form of GTP, GTPyS, and an ATP
regenerating system are added to the lysate and the mixture is incubated at 37 °C for 10 minutes.
Under these conditions, over 90% of the vesicles remain coated (Orci, L. et al. (:1989) Cell 56:357-. 368). Transport vesicles are salt-released from the Golgi membranes, loaded under a sucrose gradient, centrifuged, and fractions are collected and analyzed by SDS-PAGE.
Co-localization of MDDT with clathrin or COP coatamer is indicative of MDDT activity in vesicle formation. The contribution of MDDT in vesicle formation canbe confirmed by incubating lysates with antibodies specific for MDDT prior to GTPyS addition. The antibody will bind to MDDT and interfere with its activity, thus preventing vesicle formation.
Various modifications and variations of the described methods and systems of the invention will be apparent to those skilled in the art without departing from the scope and spirit of the invention.
Although the invention has been described in connection with certain embodiments, it should be understood that the invention as claimed should not be unduly limited to such specific embodiments.
Indeed, various modifications of the described modes for carrying out the invention which are obvious to those skilled in molecular biology or related fields are intended to be within the scope of the following claims.

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Table 5 PolynucleotideIncyte ProjectRepresentative Library SEQ ID:

ID NO:

51 1402405CB I~IDNNOT05 I

53 3189084CB BONEUNROl I

61 4159378CB ISLTNOTOl ~

b4 1593038CB1 SI~INDIA01 I

67 5510805CB BRADDIR.O1 68 1577482CB BRAUNOROl I

69 I805054CBI PLACNOBOl 71 7490847CB BRA.IFEE05 75 3580778CB1 THYMDITOl -d -°o ~ .~ -d N ,G O G .d ~ N G1., ~ ~~ .~ ~ U ~
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h h h hh h h hh h h h <110> INCYTE GENOMICS, TNC.
TANG, Y. TOM
YUE, Henry BAUGHN, Mariah R.
DUGGAN, Brendan M.
WARREN, Bridget A.
BANDMAN, Olga RICHARDSON, Thomas W.
BURFORD, Neil SANJANWALA, Bharati BECHA, Shanya D.
YAO, Monique G.
YANG, Junming TRAN, Uyen K.
HAFALTA, April J.A.
GRIFFIN, Jennifer A.
SWARNAKER, Anita ELLIOTT, Vicki S.
RECIPON, Shirley A.
KHAN, Farrah A.
LEE, Ernestine A.
YUE, Huibin LU, Dyung Aina M.
WALIA, Narinder K.
THANGAVELU, Kavitha ARVIZU, Chandra S.
XU, Yuming ISON, Craig H.
HUANG, Jiaqi DING, Li HONCHELL, Cynthia D.
BOROWSKY, Mark L.
EMERLING, Brooke M.
PETERSON, David P.
LU, Yan RAMKUMAR, Jayalaxmi MASON, Patricia M.
ZEBARJADIAN, Yeganeh AZIMZAI, Yalda STUVE, Laura L.
KAMIGAKI, Laura Y.
BARROSO, Ines LEE, Sally KABLE, Amy E.
<120> MOLECULES FOR DISEASE DETECTION AND TREATMENT
<130> PF-0991 PCT
<140> To Be Assigned <141> Herewith <150> 60/293,723; 60/295,257; 60/297,220; 60/300,526; 60/301,874;
60/359,413 <151> 2001-05-25; 2001-06-01; 2001-06-08; 2001-06-21; 2001-06-29;

<160> 78 <170> PERL Program <210> 1 <211> 200 <212> PRT
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 4973222CD1 <400> 1 Met Asn Ser Phe Ser Thr Ser Ala Phe Gly Pro Val Ala Phe Pro Ala Pro Val Pro Pro Gly Glu Asp Ser Lys Asp Val Ala Ala Pro His Arg Gln Pro Leu Thr Ser Ser Glu Arg Ile Asp Lys Gln Ile Arg His Tle Leu Asp Gly Ile Ser Ala Leu Arg Lys Glu Thr Cys Asn Lys Ser Asn Met Cys Glu Ser Ser Lys Glu Ala Leu Ala Glu Asn Asn Leu Asn Leu Pro Lys Met Ala Glu Lys Asp Gly Cys Phe Gln Ser Gly Phe Asn Glu Glu Thr Cys Leu Val Lys Ile Ile Thr Gly Leu Leu Glu Phe Glu Val Tyr Leu Glu Tyr Leu Gln Asn Arg Phe Glu Ser Ser Glu Glu Gln Ala Arg Ala Val Gln Met Ser Thr Lys Val Leu Val Gln Phe Leu Gln Lys Lys Ala Lys Asn Leu Asp Ala Ile Thr Thr Pro Asp Pro Thr Thr Asn Ala Ser Leu Leu Thr Lys Leu Gln Ala Gln Asn Gln Trp Leu Gln Asp Met Thr Thr Arg Leu Ile Leu Arg Ser Phe Lys Glu Phe Leu Gln Ser Ser Leu Arg Ala Leu Arg Gln Met <210> 2 <211> 663 <212> PRT
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 55009060CD1 <400> 2 Met Ala Gly Met Lys Thr Ala Ser Gly Asp Tyr Ile Asp Ser Ser Trp Glu Leu Arg Val Phe Val Gly Glu Glu Asp Pro Glu Ala Glu Ser Val Thr Leu Arg Val Thr Gly Glu Ser His IIe Gly Gly Val Leu Leu Lys Ile Val Glu Gln Ile Asn Arg Lys Gln Asp Trp Ser Asp His Ala Ile Trp Trp Glu Gln Lys Arg Gln Trp Leu Leu Gln Thr His Trp Thr Leu Asp Lys Tyr Gly Ile Leu Ala Asp Ala Arg Leu Phe Phe Gly Pro Gln His Arg Pro Val Ile Leu Arg Leu Pro Asn Arg Arg Ala Leu Arg Leu Arg Ala Ser Phe Ser Gln Pro Leu Phe Gln Ala Val Ala Ala Ile Cys Arg Leu Leu Ser Ile Arg His Pro Glu Glu Leu Ser Leu Leu Arg Ala Pro Glu Lys Lys Glu Lys Lys Lys Lys Glu Lys Glu Pro Glu Glu Glu Leu Tyr Asp Leu Ser Lys Val Val Leu Ala Gly Gly Val Ala Pro Ala Leu Phe Arg Gly Met Pro Ala His Phe Ser Asp Ser Ala Gln Thr Glu Ala Cys Tyr '185 190 195 His Met Leu Ser Arg Pro Gln Pro Pro Pro Asp Pro Leu Leu Leu Gln Arg Leu Pro Arg Pro Ser Ser Leu Ser Asp Lys Thr Gln Leu His Ser Arg Trp Leu Asp Ser Ser Arg Cys Leu Met Gln Gln Gly Ile Lys Ala Gly Asp Ala Leu Trp Leu Arg Phe Lys Tyr Tyr Ser Phe Phe Asp Leu Asp Pro Lys Thr Asp Pro Val Arg Leu Thr Gln Leu Tyr Glu GIn Ala Arg Trp Asp Leu Leu Leu Glu Glu Ile Asp Cys Thr Glu Glu Glu Met Met Val Phe Ala Ala Leu Gln Tyr His Ile Asn Lys Leu Ser Gln Ser Gly Glu Val Gly Glu Pro Ala Gly Thr Asp Pro Gly Leu Asp Asp Leu Asp Val Ala Leu Ser Asn Leu Glu Val Lys Leu Glu Gly Ser Ala Pro Thr Asp Val Leu Asp Ser Leu Thr Thr Ile Pro Glu Leu Lys Asp His Leu Arg Ile Phe Arg Pro Arg Lys Leu Thr Leu Lys Gly Tyr Arg Gln His Trp Val Val Phe Lys Glu Thr Thr Leu Ser Tyr Tyr Lys Ser Gln Asp Glu Ala Pro Gly Asp Pro Ile Gln Gln Leu Asn Leu Lys Gly Cys Glu Val Val Pro Asp Val Asn Val Ser Gly Gln Lys Phe Cys Ile Lys Leu Leu Val Pro Ser Pro Glu Gly Met Ser Glu Ile Tyr Leu Arg Cys GIn Asp Glu Gln Gln Tyr Ala Arg Trp Met Ala Gly Cys Arg Leu Ala Ser Lys Gly Arg Thr Met Ala Asp Ser Ser Tyr Thr Ser Glu Val Gln Ala Ile Leu Ala Phe Leu Ser Leu Gln Arg Thr Gly Ser Gly Gly Pro Gly Asn His Pro His Gly Pro Asp Ala Ser Ala Glu Gly Leu Asn Pro Tyr Gly Leu Val Ala Pro Arg Phe Gln Arg Lys Phe Lys Ala Lys Gln Leu Thr Pro Arg Ile Leu Glu Ala His Gln Asn Val Ala Gln Leu Ser Leu Ala Glu Ala Gln Leu Arg Phe Ile Gln Ala Trp Gln Ser Leu Pro Asp Phe Gly Ile Ser Tyr Val Met Val Arg Phe Lys Gly Ser Arg Lys Asp Glu Ile Leu Gly Ile Ala Asn Asn Arg Leu Ile Arg Ile Asp Leu Ala Val Gly Asp Val Val Lys Thr Trp Arg Phe Ser Asn Met Arg Gln Trp Asn Val Asn Trp Asp Ile Arg Gln Val Ala Ile Glu Phe Asp Glw His Ile Asn Val Ala Phe Ser Cys Val Ser Ala Ser Cys Arg Ile Val His Glu Tyr 620 625 ~ 630 Ile Gly Gly Tyr Ile Phe Leu Ser Thr Arg Glu Arg Ala Arg Gly Glu Glu Leu Asp Glu Asp Leu Phe Leu Gln Leu Thr Gly Gly His Glu Ala Phe <210> 3 <211> 219 <212> PRT
<213> Homo sapiens <220>
<221> misc_feature <223> Incyte ID No: 1985092CD1 <400> 3 Met Ser Glu Gln Glu Ala Gln Ala Pro Gly Gly Arg Gly Leu Pro Pro Asp Met Leu Ala Glu Gln Val Glu Leu Trp Trp Ser Gln Gln Pro Arg Arg Ser Ala Leu Cys Phe Val Val Ala Val Gly Leu Val Ala Gly Cys Gly Ala Gly Gly Val Ala Leu Leu Ser Thr Thr Ser Ser Arg Ser Gly Glu Trp Arg Leu Ala Thr Gly Thr Val Leu Cys Leu Leu Ala Leu Leu Val Leu Val Lys Gln Leu Met Ser Ser Ala Val Gln Asp Met Asn Cys Ile Arg Gln Ala His His Val Ala Leu Leu Arg Ser Gly GIy Gly Ala Asp Ala Leu Val Val Leu Leu Ser Gly Leu Val Leu Leu Val Thr Gly Leu Thr Leu Ala Gly Leu Ala Ala Ala Pro Ala Pro Ala Arg Pro Leu Ala Ala Met Leu Ser Val Gly Ile Ala Leu Ala Ala Leu Gly Ser Leu Leu Leu Leu Gly Leu Leu Leu Tyr Gln Val Gly Val Ser Gly His Cys Pro Sex Ile Cys Met Ala Thr Pro Ser Thr His Ser Gly His Gly Gly His Gly Ser Ile Phe Ser Ile Ser Gly Gln Leu Ser Ala Gly Arg Arg His Glu Thr Thr Ser Ser Ile Ala Ser Leu Ile <210> 4 <211> 318 <212> PRT
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 1553593CD1 <400> 4 Met Asn Thr Arg Asn Arg Val Val Asn Ser Gly Leu Gly Ala Ser Pro Ala Ser Arg Pro Thr Arg Asp Pro Gln Asp Pro Ser Gly Arg Gln Gly Glu Leu Ser Pro Val Glu Asp Gln Arg Glu Gly Leu Glu Ala Ala Pro Lys Gly Pro Ser Arg Glu Ser Val Val His Ala Gly Gln Arg Arg Thr Ser Ala Tyr Thr Leu Ile Ala Pro Asn Ile Asn Arg Arg Asn Glu Ile Gln Arg Ile Ala Glu Gln Glu Leu Ala Asn Leu~Glu Lys Trp Lys Glu Gln Asn Arg Ala Lys Pro Val His Leu Val Pro Arg Arg Leu Gly Gly Ser Gln Ser Glu Thr Glu Val Arg Gln Lys Gln Gln Leu Gln Leu Met Gln Ser Lys Tyr Lys Gln Lys Leu Lys Arg Glu Glu Ser Val Arg Ile Lys Lys Glu Ala Glu Glu 140 '145 150 Ala Glu Leu Gln Lys Met Lys Ala Ile Gln Arg Glu Lys Ser Asn Lys Leu Glu Glu Lys Lys Arg Leu Gln Glu Asn Leu Arg Arg Glu Ala Phe Arg Glu His Gln Gln Tyr Lys Thr Ala Glu Phe Leu Ser Lys Leu Asn Thr Glu Ser Pro Asp Arg Ser Ala Cys Gln Ser Ala Val Cys Gly Pro Gln Ser Ser Thr Trp Lys Leu Pro Ile Leu Pro Arg Asp His Ser Trp Ala Arg Ser Trp Ala Tyr Arg Asp Ser Leu Lys Ala Glu Glu Asn Arg Lys Leu Gln Lys Met Lys Asp Glu Gln 245 ~ 250 255 His Gln Lys Ser Glu Leu Leu Glu Leu Lys Arg Gln Gln Gln Glu Gln Glu Arg Ala Lys Ile His Gln Thr Glu His Arg Arg Val Asn Asn Ala Phe Leu Asp Arg Leu Gln Gly Lys Ser Gln Pro Gly Gly Leu Glu Gln Ser Gly Gly Cys Trp Asn Met Asn Ser Gly Asn Ser Trp Gly Ile <210> 5 <211> 387 <212> PRT
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 1954222CD1 <400> 5 Met Glu Lys Ile Glu Glu Gln Phe Ala Asn Leu His Ile Val Lys Cys Ser Leu Gly Thr Lys Glu Pro Thr Tyr Leu Leu Gly Ile Asp Thr Ser Lys Thr Val Gln Ala Gly Lys Glu Asn Leu Val Ala Val Leu Cys Ser Asn Gly Ser Ile Arg Ile Tyr Asp Lys Glu Arg Leu Asn Val Leu Arg Glu Phe Ser Gly Tyr Pro Gly Leu Leu Asn Gly Val Arg Phe Ala Asn Ser Cys Asp Ser Val Tyr Ser Ala Cys Thr Asp Gly Thr Val Lys Cys Trp Asp Ala Arg Val Ala Arg Glu Lys Pro Val Gln Leu Phe Lys Gly Tyr Pro Ser Asn Ile Phe Ile Ser Phe Asp Ile Asn Cys Asn Asp His Ile Ile Cys Ala Gly Thr Glu Lys Val Asp Asp Asp Ala Leu Leu Val Phe Trp Asp Ala Arg Met Asn Ser Gln Asn Leu Ser Thr Thr Lys Asp Ser Leu Gly Ala Tyr Ser Glu Thr His Ser Asp Asp Val Thr Gln Val Arg Phe His Pro Ser Asn Pro Asn Met Val Val Ser Gly Ser Ser Asp Gly Leu Val Asn Val Phe Asp Ile Asn Ile Asp Asn Glu Glu Asp Ala Leu Val Thr Thr Cys Asn Ser Ile Ser Ser Val Ser Cys Ile Gly Trp Ser Gly Lys Gly Tyr Lys Gln Ile Tyr Cys Met Thr His Asp Glu Gly Phe Tyr Trp Trp Asp Leu Asn His Leu Asp Thr Asp Glu Pro Val Thr Arg Leu Asn Ile Gln Asp Val Arg Glu Val Val Asn Met Lys Glu Asp Ala Leu Asp Tyr Leu Ile Gly Gly Leu Tyr His Glu Lys Thr Asp Thr Leu His Val Ile Gly Gly Thr Asn Lys Gly Arg Ile His Leu Met Asn Cys Ser Met Ser Gly Leu Thr His Val Thr Ser 305 ' 310 315 Leu Gln Gly Gly His Ala Ala Thr Val Arg Ser Phe Cys Trp Asn Val Gln Asp Asp Ser Leu Leu Thr Gly Gly Glu Asp Ala Gln Leu Leu Leu Trp Lys Pro Gly Ala Ile Glu Lys Thr Phe Thr Lys Lys Glu Ser Met Lys Ile Ala Ser Ser Val His Gln Arg Val Arg Val His Ser Asn Asp Ser Tyr Lys Arg Arg Lys Lys Gln <210> 6 <211> 577 <212> PRT
<213> Homo Sapiens <220>
<221> misc feature <223> Incyte ID No: 3159276CD1 <400> 6 Met Met Ser Leu Ser Pro Ile Ser Leu Gly Ala Leu Glu Gly Arg Asn Val Asp Leu Leu Gln Ile Ser Gly Gln Leu Ser Pro Arg Leu Phe Arg Lys Leu Pro Pro Arg Val Cys Val Ser Leu Lys Asn Ile Val Asp Glu Asp Phe Leu Tyr Ala Gly His Ile Phe Leu Gly Phe Ser Lys Cys Gly Arg Tyr Val Leu Ser Tyr Thr Ser Ser Ser Gly Asp Asp Asp Phe Ser Phe Tyr Ile Tyr His Leu Tyr Trp Trp Glu Phe Asn Val His Ser Lys Leu Lys Leu Val Arg Gln Val Arg Leu Phe Gln Asp Glu Glu Ile Tyr Ser Asp Leu Tyr Leu Thr Val Cys Glu Trp Pro Ser Asp Ala Ser Lys Val Ile Val Phe Gly Phe Asn Thr Arg Ser Ala Asn Gly Met Leu Met Asn Met Met Met Met Ser Asp Glu Asn His Arg Asp Ile Tyr Val Ser Thr Val Ala Val Pro Pro Pro Gly Arg Cys Ala Ala Cys Gln Asp Ala Ser Arg Ala His Pro Gly Asp Pro Asn Ala Gln Cys Leu Arg His Gly Phe Met Leu His Thr Lys Tyr Gln Val Val Tyr Pro Phe Pro Thr Phe Gln Pro Ala Phe Gln Leu Lys Lys Asp Gln Val Val Leu Leu Asn Thr Ser Tyr Ser Leu Val Ala Cys Ala Val Ser Val His Ser Ala Gly Asp Arg Ser Phe Cys Gln Ile Leu Tyr Asp His Ser Thr Cys Pro Leu Ala Pro Ala Ser Pro Pro Glu~Pro Gln Ser Pro Glu Leu Pro Pro Ala Leu Pro Ser Phe Cys Pro Glu Ala Ala Pro Ala Arg Ser Ser Gly Ser Pro Glu Pro Ser Pro Ala Ile Ala Lys Ala Lys Glu Phe Val Ala Asp Ile Phe Arg Arg Ala Lys Glu Ala Lys Gly Gly Val Pro Glu Glu Ala Arg Pro Ala Leu Cys Pro Gly Pro Ser Gly Ser Arg Cys Arg Ala His Ser Glu Pro Leu Ala Leu Cys Gly Glu Thr Ala Pro Arg Asp Ser Pro Pro Ala Ser Glu Ala Pro Ala Ser Glu Pro Gly Tyr Val Asn Tyr Thr Lys Leu Tyr Tyr Val Leu Glu Ser Gly Glu Gly Thr Glu Pro Glu Asp Glu Leu Glu Asp Asp Lys Ile Ser Leu Pro Phe Val Val Thr Asp Leu Arg Gly Arg Asn Leu Arg Pro Met Arg Glu Arg Thr Ala Val Gln Gly Gln Tyr Leu Thr Val Glu Gln Leu Thr Leu Asp Phe Glu Tyr Val Ile Asn Glu Val hle Arg His Asp Ala Thr Trp Gly His Gln Phe Cys Ser Phe Ser Asp Tyr Asp Ile Val Ile Leu Glu Val Cys Pro Glu Thr Asn Gln Val Leu Ile Asn Ile Gly Leu Leu Leu Leu Ala Phe Pro Ser Pro Thr Glu Glu Gly Gln Leu Arg Pro Lys Thr Tyr His Thr Ser Leu Lys Val Ala Trp Asp Leu Asn Thr Gly Ile Phe Glu Thr Val Ser Val Gly Asp Leu Thr Glu Val Lys Gly Gln Thr Ser Gly Ser Val Trp Ser Ser Tyr Arg Lys Ser Cys Val Asp Met Val Met Lys Trp Leu Val Pro Glu Ser Ser Gly Arg Tyr Val Asn Arg Met Thr Asn Glu Ala Leu His Lys Gly Cys Ser Leu Lys Val Leu Ala Asp Ser Glu Arg Tyr Thr Trp Ile Val Leu <210> 7 <211> 224 <212> PRT
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 140052CD1 <400> 7 Met Ala Ala Pro Ser Glu Val Ala Ala Ile Ala Pro Gly Glu Gly Asp Gly Gly Gly Gly Gly Phe Gly Ser Trp Leu Asp Gly Arg Leu Glu Ala Leu Gly Val Asp Arg Ala Val Tyr Gly Ala Tyr Ile Leu Gly Ile Leu Gln Glu Glu Glu Glu Glu Glu Lys Leu Asp Ala Leu Gln Gly Ile Leu Ser Ala Phe Leu Glu Glu Asp Ser Leu Leu Asn Ile Cys Lys Glu Ile Val Glu Arg Trp Ser Glu Thr Gln Asn Val 80 ~ 85 90 Val Thr Lys Val Lys Lys Glu Asp Glu Val Gln Ala Ile Ala Thr Leu Ile Glu Lys Gln Ala Gln Ile Val Val Lys Pro Arg Met Val Ser Glu Glu Glu Lys Gln Arg Lys Ala Ala Leu Leu Ala Gln Tyr Ala Asp Val Thr Asp Glu Glu Asp Glu Ala Asp Glu Lys Asp Asp Ser Gly Ala Thr Thr Met Asn Ile Gly Ser Asp Lys Leu Leu Phe Arg Asn Thr Asn Val Glu Asp Val Leu Asn Ala Arg Lys Leu Glu 170 175 . 180 Arg Asp Ser Leu Arg Asp Glu Ser Gln Arg Lys Lys Glu Gln Asp Lys Leu Gln Arg Glu Arg Asp Lys Leu Ala Lys Gln Glu Arg Lys Glu Lys Glu Lys Lys Arg Thr Gln Arg Gly Glu Arg Lys Arg <210> 8 <211> 600 <212> PRT
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 5158048CD1 <400> 8 Met Gly Lys Lys Leu Asp Leu Ser Lys Leu Thr Asp Glu Glu Ala Gln His Val Leu Glu Val Val Gln Arg Asp Phe Asp Leu Arg Arg Lys Glu Glu Glu Arg Leu Glu Ala Leu Lys Gly Lys Ile Lys Lys Glu Ser Ser Lys Arg Glu Leu Leu Ser Asp Thr Ala His Leu Asn Glu Thr His Cys Ala Arg Cys Leu Gln Pro Tyr Gln Leu Leu Val Asn Ser Lys Arg Gln Cys Leu Glu Cys Gly Leu Phe Thr Cys Lys Ser Cys Gly Arg Val His Pro Glu Glu Gln Gly Trp Ile Cys Asp Pro Cys His Leu Ala Arg Val Val Lys Ile Gly Ser Leu Glu Trp Tyr Tyr Glu His Val Lys Ala Arg Phe Lys Arg Phe Gly Ser Ala Lys Val Ile Arg Ser Leu His Gly Arg Leu Gln Gly Gly Ala Gly Pro Glu Leu Ile Ser Glu Glu Arg Ser Gly Asp Ser Asp Gln Thr Asp Glu Asp Gly Glu Pro Gly Ser Glu Ala Gln Ala Gln Ala Gln Pro Phe Gly Ser Lys Lys Lys Arg Leu Leu Ser Val His Asp Phe Asp Phe Glu Gly Asp Ser Asp Asp Ser Thr Gln Pro Gln Gly His Ser Leu His Leu Ser Ser Val Pro Glu Ala Arg Asp Ser Pro Gln Ser Leu Thr Asp Glu Ser Cys Ser Glu Lys Ala Ala Pro His Lys Ala Glu Gly Leu Glu Glu Ala Asp Thr Gly Ala Ser Gly Cys His Ser His Pro Glu Glu Gln Pro Thr Ser Ile Ser Pro Ser Arg His Gly A1a Leu Ala Glu Leu Cys Pro Pro Gly Gly Ser His Arg Met Ala Leu Gly Thr Ala Ala Ala Leu Gly Ser Asn Val Ile Arg Asn Glu Gln Leu Pro Leu Gln Tyr Leu Ala Asp Val Asp Thr Ser Asp Glu Glu Ser Ile Arg Ala His Val Met Ala Ser His His Ser Lys Arg Arg Gly Arg Ala Ser Ser Glu Ser Gln Ile Phe Glu Leu Asn Lys Arg Ile Ser Ala Val Glu Cys Leu Leu Thr Tyr Leu Glu Asn Thr Val Val Pro Pro Leu Ala Lys Gly Leu Gly Ala Gly Val Arg Thr Glu Ala Asp Val Glu Glu Glu Ala Leu Arg Arg Lys Leu Glu Glu Leu Thr Ser Asn Val Ser Asp Gln Glu Thr Ser Ser Glu Glu Glu Glu Ala Lys Asp Glu Lys Ala Glu Pro Asn Arg Asp Lys Ser Val Gly Pro Leu Pro Gln Ala Asp.Pro Glu Val Gly Thr Ala Ala His Gln Thr Asn Arg Gln Glu Lys Ser Pro Gln Asp Pro Gly Asp Pro Val Gln Tyr Asn Arg Thr Thr Asp Glu Glu Leu Ser Glu Leu Glu Asp Arg Val Ala Val Thr Ala Ser Glu Val Gln Gln Ala Glu Ser Glu Val Ser Asp Ile Glu Ser Arg Ile Ala Ala Leu Arg Ala Ala Gly Leu Thr Val Lys Pro Ser Gly Lys Pro Arg Arg Lys Ser Asn Leu Pro Ile Phe Leu Pro Arg Val Ala Gly Lys Leu Gly Lys Arg Pro Glu Asp Pro Asn Ala Asp Pro Ser Ser Glu Ala Lys Ala Met Ala Val Pro Tyr Leu Leu Arg Arg Lys Phe Ser Asn Ser Leu Lys Ser Gln Gly Lys Asp Asp Asp Ser Phe Asp Arg Lys Ser Val Tyr Arg Gly Ser Leu Thr Gln Arg Asn Pro Asn Ala Arg Lys Gly Met Ala Ser His Thr Phe Ala Lys Pro Val Val Ala His Gln Ser <210> 9 <211> 1250 <212> PRT
<213> Homo sapiens <220>
<221> misc feature <223> Incyte ID No: 3127541CD1 <400> 9 Met Glu Gln Leu Ser Asp Glu Glu Ile Asp His Gly Ala Glu Glu Asp Ser Asp Lys Glu Asp Gln Asp Leu Asp Lys Met Phe Gly Ala Trp Leu Gly Glu Leu Asp Lys Leu~Thr Gln Ser Leu Asp Ser Asp Lys Pro Met Glu Pro Val Lys Arg Ser Pro Leu Arg Gln Glu Thr Asn Met Ala Asn Phe Ser Tyr Arg Phe Ser Ile Tyr Asn Leu Asn Glu Ala Leu Asn Gln Gly Glu Thr Val Asp Leu Asp Ala Leu Met Ala Asp Leu Cys Ser Ile Glu Gln Glu Leu Ser Ser Ile Gly Ser Gly Asn Ser Lys Arg Gln Ile Thr Glu Thr Lys Ala Thr Gln Lys Leu Pro Val Ser Arg His Thr Leu Lys His Gly Thr Leu Lys Gly Leu Ser Ser Ser Ser Asn Arg Ile Ala Lys Pro Ser His Ala Ser Tyr Ser Leu Asp Asp Val Thr Ala Gln Leu Glu Gln Ala Ser Leu Ser Met Asp Glu Ala Ala Gln Gln Ser Val Leu Glu Asp Thr Lys Pro Leu Val Thr Asn Gln His Arg Arg Thr Ala Ser Ala Gly Thr Val Ser Asp Ala Glu Val His Ser Ile Ser Asn Ser Ser His Ser Ser Ile Thr Ser Ala Ala Ser Ser Met Asp Ser Leu Asp Ile Asp Lys Val Thr Arg Pro Gln Glu Leu Asp Leu Thr His Gln Gly Gln Pro Ile Thr Glu Glu Glu Gln Ala Ala Lys Leu Lys Ala Glu Lys Ile Arg Val Ala Leu Glu Lys Ile Lys Glu Ala Gln Val Lys Lys Leu Val Ile Arg Val His Met Ser Asp Asp Ser Ser Lys Thr Met Met Val Asp Glu Arg Gln Thr Val Arg Gln Val Leu Asp Asn Leu Met Asp Lys Ser His Cys Gly Tyr Ser Leu Asp Trp Ser Leu Val Glu Thr Val Ser Glu Leu Gln Met Glu Arg Ile Phe Glu Asp His Glu Asn Leu Val Glu Asn Leu Leu Asn Trp Thr Arg Asp Ser Gln Asn Lys Leu Ile Phe Met Glu Arg Ile Glu Lys Tyr Ala Leu Phe Lys Asn Pro Gln Asn Tyr Leu Leu Gly Lys Lys Glu Thr Ala Glu Met Ala Asp Arg Asn Lys Glu Val Leu Leu Glu Glu Cys Phe Cys Gly Ser Ser Val Thr Val Pro Glu Ile Glu Gly Val Leu Trp Leu Lys Asp Asp Gly Lys Lys Ser Trp Lys Lys Arg Tyr Phe Leu Leu Arg Ala Ser Gly Ile Tyr Tyr Val Pro Lys Gly Lys Ala Lys Val Ser Arg Asp Leu Val Cys Phe Leu Gln Leu Asp His Val Asn Val Tyr Tyr Gly Gln Asp Tyr Arg Asn Lys Tyr Lys Ala Pro Thr Asp Tyr Cys Leu Val Leu Lys His Pro Gln Ile Gln Lys Lys Ser Gln Tyr Ile Lys Tyr Leu Cys Cys Asp Asp Val Arg Thr Leu His Gln Trp Val Asn Gly Ile Arg Ile Ala Lys Tyr Gly Lys Gln Leu Tyr Met Asn Tyr Gln Glu Ala Leu Lys Arg Thr Glu Ser Ala Tyr Asp Trp Thr Ser Leu Ser Ser Ser Ser Ile Lys Ser Gly Ser Ser Ser Ser Ser Ile Pro Glu Ser Gln Ser Asn His Ser Asn Gln Ser Asp Ser Gly Val Ser Asp Thr Gln Pro Ala Gly His Val Arg Ser Gln Ser Ile Val Ser Ser Val Phe Ser Glu Ala Trp Lys Arg Gly Thr Gln Leu Glu Glu Ser Ser Lys Ala Arg Met Glu Ser Met Asn Arg Pro Tyr Thr Ser Leu Val Pro Pro Leu Ser Pro Gln Pro Lys Ile Val Thr Pro Tyr Thr Ala Ser Gln Pro Ser Pro Pro Leu Pro Pro Pro Pro Pro Pro Pro Pro Pro Pro Pro Pro Pro Pro Pro Pro Pro Pro Pro Pro Leu Pro Ser Gln Ser Ala Pro Ser Ala Gly Ser Ala Ala Pro Met Phe Val Lys Tyr Ser Thr Ile Thr Arg Leu Gln Asn Ala Ser Gln His Ser Gly Ala Leu Phe Lys Pro Pro Thr Pro Pro Val Met Gln Ser Gln Ser Val Lys Pro Gln Ile Leu Val Pro Pro Asn Gly Val Val Pro Pro Pro Pro Pro Pro Pro Pro Pro Pro Thr 7l0 715 720 Pro Gly Ser Ala Met Ala Gln Leu Lys Pro Ala Pro Cys Ala Pro Ser Leu Pro Gln Phe Ser Ala Pro Pro Pro Pro Leu Lys Ile His Gln Val Gln His Ile Thr Gln Val Ala Pro Pro Thr Pro Pro Pro Pro Pro Pro Ile Pro Ala Pro Leu Pro.Pro Gln Ala Pro Pro Lys Pro Leu Val Thr Ile Pro Ala Pro Thr Ser Thr Lys Thr Val Ala Pro Val Val Thr Gln Ala Ala Pro Pro Thr Pro Thr Pro Pro Val Pro Pro Ala Lys Lys Gln Pro Ala Phe.Pro Ala Ser Tyr Ile Pro Pro Ser Pro Pro Thr Pro Pro Val Pro. Val Pro Pro Pro Thr Leu Pro Lys Gln Gln Ser Phe Cys Ala Lys Pro Pro Pro Ser Pro Leu Ser Pro Val Pro Ser Val Val Lys Gln Ile Ala Ser Gln Phe Pro Pro Pro Pro Thr Pro Pro Ala Met Glu Ser Gln Pro Leu Lys Pro Val Pro Ala Asn Val Ala Pro Gln Ser Pro Pro Ala Val Lys Ala Lys Pro Lys Trp Gln Pro Ser Ser Ile Pro Val Pro Ser Pro Asp Phe Pro Pro Pro Pro Pro Glu Ser Ser Leu Val Phe Pro Pro Pro Pro Pro Ser Pro Val Pro Ala Pro Pro Pro Pro Pro Pro Pro Thr Ala Ser Pro Thr Pro Asp Lys Ser Gly Ser Pro Gly Lys Lys Thr Ser Lys Thr Ser Ser Pro Gly Gly Lys Lys Pro Pro Pro Thr Pro 965 g70 975 Gln Arg Asn Ser Ser Ile Lys Ser Ser Ser Gly Ala Glu His Pro Glu Pro Lys Arg Pro Ser Val Asp Ser Leu Val Ser Lys Phe Thr Pro Pro Ala Glu Ser Gly.Ser Pro Ser Lys Glu Thr Leu Pro Pro Pro Ala Ala Pro Pro Lys Pro Gly Lys Leu Asn Leu Ser Gly Val Asn Leu Pro Gly Val Leu Gln Gln Gly Cys Val Ser Ala Lys Ala Pro Val Leu Ser Gly Arg Gly Lys Asp Ser Val Val Glu Phe Pro Ser Pro Pro Ser Asp Ser Asp Phe Pro Pro Pro Pro Pro Glu Thr Glu Leu Pro Leu Pro Pro Ile Glu Ile Pro Ala Val Phe Ser Gly Asn Thr Ser Pro Lys Val Ala Val Val Asn Pro Gln Pro Gln Gln Trp Ser Lys Met Ser Val Lys Lys Ala Pro Pro Pro Thr Arg Pro Lys Arg Asn Asp Ser Thr Arg Leu Thr Gln Ala Glu Ile Ser Glu Gln Pro Thr Met Ala Thr Val Val Pro Gln Val Pro Thr Ser Pro Lys Ser Ser Leu Ser Val Gln Pro Gly Phe Leu Ala Asp Leu Asn Arg Thr Leu Gln Arg Lys Ser Ile Thr Arg His Gly Ser Leu Ser Ser Arg Met Ser Arg Ala G7:u Pro Thr Ala Thr Met Asp Asp Met Ala Leu Pro Pro Pro Pro Pro Glu Leu Leu Ser Asp Gln Gln Lys Ala Gly Tyr Gly Gly Ser His Ile Ser Gly Tyr Ala Thr Leu Arg Arg Gly Pro Pro Pro Ala Pro Pro Lys Arg Asp Gln Asn Thr Lys Leu Ser Arg Asp Trp <210> 10 <211> 621 <212> PRT
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 8224777CD1 <400> 10 Met Leu Pro Val Asp Gly Glu Glu Arg Lys Ser Glu Gly Ser Asp Thr Val Gly Asp Arg Thr Ser Pro Cys Ala Thr Ser Ser Ala Thr Leu Lys Asp Leu Glu Val Arg Gly Ser Gly Trp Arg Cys Ser Asp Pro Ser Gly Gln Pro Ser Asn Leu Leu Leu Gln Val Gly Leu Gly Ala Pro Leu Pro Ala Glu Thr Ala His Thr His Pro Ser Pro Asn Asp Arg Ser Leu Tyr Leu Ser Pro His Ser Cys Ser Thr Ser Ser Ser Leu His Ala Pro Gln Ser Pro Cys Gln Glu Arg Ala Val Val Leu Asp Ser Thr Ser Val Lys Ile Ser Arg Leu Lys Asn Thr Ile Lys Ser Leu Lys Gln Gln Lys Lys Gln Val Glu His Gln Leu Glu Glu GIu Lys Lys Ala Asn Asn GIu Arg Gln Lys Ala Glu Arg GIu Leu Glu Val Gln Ile Gln Thr Leu Ile Ile Gln Lys Glu Glu Leu Asn Thr Asp Leu Tyr His Met Glu Arg Ser Leu Arg Tyr Phe Glu Glu Glu Ser Lys Asp Leu Ala Val Arg Leu Gln His Ser Leu Gln Cys Lys Gly Glu Leu Glu Arg Ala Leu Ser Ala Val Ile Ala Thr Glu Lys Lys Lys Ala Asn Gln Leu Ser Ser Cys Ser Lys Ala His Thr Glu Trp Glu Leu Glu Gln Ser Leu Gln Asp Gln Ala Leu Leu Lys Ala Gln Leu Thr Gln Leu Lys Glu Ser Phe Gln Gln Leu Gln Leu Glu Arg Asp G1u Cys Ala Glu His Ile Glu Gly Glu Arg Ala 260 ' . 265 270 Arg Trp His Gln Arg Met Ser Lys Met Ser Gln Glu Ile Cys Thr Leu Lys Lys Glu Lys Gln Gln Asp Met Arg Arg Val Glu Glu Leu 290 295 ' 300 Glu Arg Ser Leu Ser Lys Leu Lys Asn Gln Met Ala Glu Pro Leu Pro Pro Glu Pro Pro Ala Val Pro Ser Glu Val Glu Leu Gln His Val Arg Lys Glu Leu Glu Arg Val Ala Gly Glu Leu Gln Ala Gln 335 ' 340 345 Val Lys Asn Asn Gln His Ile Ser Leu Leu Asn Arg Arg Gln Glu Glu Arg Ile Arg Glu Gln Glu Glu Arg Leu Arg Lys Gln Glu Glu Arg Leu Gln Glu Gln His Glu Lys Leu Arg Gln Leu Ala Lys Pro Gln Ser Val Phe Glu Glu Leu Asn Asn Glu Asn Lys Ser Thr Leu Gln Leu Glu Gln Gln Val Lys Glu Leu Gln Glu Lys Leu Gly Glu Val Lys Glu Thr Glu Thr Ser Thr Pro Ser Lys Lys GIy Trp Glu Ala Gly Ser Ser Leu Leu Gly Gly Glu Val Ser Ser Phe Met Asp His Leu Lys Glu Lys Ala Asp Leu Ser Glu Leu Val Lys Lys Gln Glu Leu Arg Phe Ile Gln Tyr Trp Gln Glu Arg Cys His Gln Lys Ile His His Leu Leu Ser Glu Pro Gly Gly Arg Ala Lys Asp Ala Ala Leu Gly Gly Gly His His Gln Ala Gly Ala Gln Gly Gly Asp Glu Gly Glu Ala Ala Gly Ala Ala Ala Asp Gly Ile Ala Ala Tyr Ser Asn Tyr Asn Asn Gly His Arg Lys Phe Leu Ala Ala Ala His Asn Ser Ala Asp Glu Pro Gly Pro Gly Ala Pro Ala Pro Gln Glu Leu Gly Ala Ala Asp Lys His Gly Asp Leu Arg Glu Val Ser Leu Thr Ser Ser A1a Gln G1y Glu Ala Arg Glu Asp Pro Leu Leu Asp Lys Pro Thr Ala Gln Pro Ile Val Gln Asp His Gln Glu His Pro Gly Leu Gly Ser Asn Cys Cys Val Pro Leu Phe Cys Trp Ala Trp Leu Pro Arg Arg Arg Arg <210> 11 <211> 114 <212> PRT
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 587394CD1 <400> 11 Met Phe Met Arg Lys Arg Glu Asn Ser Leu Ala Ser Leu Phe Thr Glu Trp Leu Phe Leu Val Ser Ser Arg Cys Ser Arg Thr Asp Val Arg Phe Gly Ser Ser Glu Ser Thr Lys His Ile Gly Glu Lys Asp Glu Glu Asp Ser Cys Arg Leu Asp Cys Thr Met Ser Pro Ser Arg Thr Gln Gln Arg Ala Ala Arg Pro Arg Gly Glu Gly Ser Leu Lys Gln His Gln Arg Leu His Ser Asn Phe Ser Ser Val Asn Glu Ala Val Phe Ile Val Ser Phe Ser Leu Asn Thr Asn Leu Lys Thr Thr Gly Leu Asn Leu Leu Ser His Leu Ala <210> 12 <211> 527 <212> PRT
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 1402405CD1 <400> 12 Met Gly Pro Leu Ala Leu G1y Ile Leu Lys Leu Glu His Cys Pro Gln Ala Leu Arg Thr Gln A1a Phe Gln Val Leu Leu Gln Pro Leu Ala Cys Val Leu Lys Ala Thr Val Gln Ala Pro Gly Pro Pro Gly Leu Leu Asp Gly Thr Ala Asp Asp Ala Thr Thr Val Asp Thr Leu Leu Ala Ser Lys Ser Ser Cys Ala Gly,Leu Leu Cys Arg Thr Leu Ala His Leu Glu Glu Leu Gln Pro Leu Pro Gln Arg Pro Ser Pro Trp Pro Gln Ala Ser Leu Leu Gly Ala Thr Val Thr Val Leu Arg Leu Cys Asp Gly Ser Ala Ala Pro Ala Ser Ser Val Gly Gly His Leu Cys Gly Thr Leu Ala Gly Cys Val Arg Val Gln Arg Ala Ala Leu Asp Phe Leu Gly Thr Leu Ser Gln Gly Thr Gly Pro Gln Glu Leu Val Thr Gln Ala Leu Ala Val Leu Leu Glu Cys Leu Glu Ser Pro Gly Ser Ser Pro Thr Val Leu Lys Lys Ala Phe Gln Ala Thr Leu Arg Trp Leu Leu Ser Ser Pro Lys Thr Pro Gly Cys Ser Asp Leu Gly Pro Leu Ile Pro Gln Phe Leu Arg Glu Leu Phe Pro Val Leu Gln Lys Arg Leu Cys His Pro Cys Trp Glu Val Arg Asp Ser Ala Leu Glu Phe Leu Thr Gln Leu Ser Arg His Trp Gly Gly Gln Ala Asp Phe Arg Cys Ala Leu Leu Ala Ser Glu Val Pro Gln Leu Ala Leu Gln Leu Leu Gln Asp Pro Glu Ser Tyr Val Arg Ala Ser Ala Val Thr Ala Met Gly Gln Leu Ser Ser Gln Gly Leu His Ala Pro Thr Ser Pro Glu His Ala Glu Ala Arg Gln Ser Leu Phe Leu Glu Leu Leu His Ile Leu Ser Val Asp Ser Glu Gly Phe Pro Arg Arg Ala Val Met Gln Val Phe Thr Glu Trp Leu Arg Asp Gly His Ala Asp Ala Ala Gln Asp Thr Glu Gln Phe Val Ala Thr Val Leu Gln Ala Ala Ser Gln Asp Leu Asp Trp Glu Val Arg Ala Gln Gly Leu Glu Leu Ala Leu Val Phe Leu Gly Gln Thr Leu Gly Pro Pro Arg Thr His Cys Pro Tyr Ala Val Ala Leu Pro Glu Val Ala Pro Ala Gln Pro Leu Thr Glu Ala Leu Arg Ala Leu Cys His Val Gly Leu Phe Asp Phe Ala Phe Cys Ala Leu Phe Asp Cys Asp Arg Pro Val Ala Gln Lys Ser Cys Asp Leu Leu Leu Phe Leu Arg~Asp Lys Ile Ala Ser Tyr Ser Ser Leu Arg Glu Ala Arg Gly Ser Pro Asn Thr Ala Ser Ala Glu Ala Thr Leu Pro Arg Trp Arg Ala Gly Glu Gln Ala Gln Pro Pro Gly Asp Gln Glu Pro Glu Ala Val Leu Ala Met Leu Arg Ser Leu Asp Leu Glu Gly Leu Arg Sex Thr Leu Ala Glu Ser Ser Asp His Val Glu Lys Ser Pro Gln Ser Leu Leu Gln Asp Met Leu Ala Thr Gly Gly Phe Leu Gln Gly Asp Glu Ala Asp Cys Tyr <210> 13 <211> 316 <212> PRT
<213> Homo Sapiens <220>
<221> misc feature <223> Incyte ID No: 1798468CD1 <400> 13 Met Ala Ser Pro Glu His Pro Gly Ser Pro Gly Cys Met Gly Pro Ile Thr Gln Cys Thr Ala Arg Thr Gln Gln Glu Ala Pro Ala Thr Gly Pro Asp Leu Pro His Pro Gly Pro Asp Gly His Leu Asp Thr His Ser Gly Leu Ser Ser Asn Ser Ser Met Thr Thr Arg Glu Leu Gln Gln Tyr Trp Gln Asn Gln Lys Cys Arg Trp Lys His Val Lys Leu Leu Phe Glu Ile Ala Ser Ala Arg Ile Glu Glu Arg Lys Val Ser Lys Phe Val Val Tyr Gln Ile Ile Val Ile Gln Thr Gly Ser Phe Asp Asn Asn Lys Ala Val Leu Glu Arg Arg Tyr Ser Asp Phe Ala Lys Leu Gln Lys Ala Leu Leu Lys Thr Phe Arg Glu Glu Ile Glu Asp Val Glu Phe Pro Arg Lys His Leu Thr Gly Asn Phe Ala Glu Glu Met Ile Cys Glu Arg Arg Arg Ala Leu Gln Glu Tyr Leu Gly Leu Leu Tyr Ala Ile Arg Cys Val Arg Arg Ser Arg Glu Phe Leu Asp Phe Leu Thr Arg Pro Glu Leu Arg Glu Ala Phe Gly Cys Leu Arg Ala Gly Gln Tyr Pro Arg Ala Leu Glu Leu Leu Leu Arg Val Leu Pro Leu Gln Glu Lys Leu Thr Ala His Cys Pro Ala Ala Ala Val Pro Ala Leu Cys Ala Val Leu Leu Cys His Arg Asp Leu Asp Arg Pro Ala Glu Ala Phe Ala Ala Gly Glu Arg Ala Leu Gln Arg Leu Gln Ala Arg Glu Gly His Arg Tyr Tyr Ala Pro Leu Leu Asp Ala Met Val Arg Leu Ala Tyr Ala Leu Gly Lys Asp Phe Val Thr Leu Gln Glu Arg Leu Glu Glu Ser Gln Leu Arg Arg Pro Thr Pro Arg Gly Ile Thr Leu Lys Glu Leu Thr Val Arg Glu Tyr Leu His <210> 14 <211> 659 <222> PRT
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 3189084CD1 <400> 14 Met Gly Asp Cys Ala Glu Ile Lys Ser Gln Phe Arg Thr Arg Glu 1 5 10' 15 Gly Phe Tyr Lys Leu Leu Pro Gly Asp Gly Ala Ala Arg Arg Ser Gly Pro Ala Ser Ala Gln Thr Pro Val Pro Pro Gln Pro Pro Gln Pro Pro Pro Gly Pro Ala Ser Ala Ser Gly Pro Gly Ala Ala Gly Pro Ala Ser Ser Pro Pro Pro Ala Gly Pro Gly Pro Gly Pro Ala Leu Pro Ala Val Arg Leu Ser Leu Val Arg Leu Gly GIu Pro Asp 80 85. 90 Ser Ala Gly Ala Gly Glu Pro Pro Ala Thr Pro Ala Gly Leu Gly Ser Gly Gly Asp Arg Val Cys Phe Asn Leu Gly Arg Glu Leu Tyr Phe Tyr Pro Gly Cys Cys Arg Arg Gly Ser Gln Arg Ser Ile Asp Leu Asn Lys Pro Ile Asp Lys Arg Ile Tyr Lys Gly Thr Gln Pro Thr Cys His Asp Phe Asn Gln Phe Thr Ala Ala Thr Glu Thr Ile Ser Leu Leu Val Gly Phe Ser Ala Gly Gln Val Gln Tyr Leu Asp Leu Ile Lys Lys Asp Thr Ser Lys Leu Phe Asn Glu Glu Arg Leu 185 19'0 195 Ile Asp Lys Thr Lys Val Thr Tyr Leu Lys Trp Leu Pro Glu Ser Glu Ser Leu Phe Leu Ala Ser His Ala Ser Gly His Leu Tyr Leu Tyr Asn Val Ser His Pro Cys Ala Ser Ala Pro Pro Gln Tyr Ser Leu Leu Lys Gln Gly Glu GIy Phe Ser VaI Tyr Ala AIa Lys Ser Lys Ala Pro Arg Asn Pro Leu Ala Lys Trp Ala Val Gly Glu Gly Pro Leu Asn Glu Phe Ala Phe Ser Pro Asp Gly Arg His Leu Ala Cys Val Ser Gln Asp Gly Cys Leu Arg Val Phe His Phe Asp Ser Met Leu Leu Arg Gly Leu Met Lys Ser Tyr Phe Gly Gly Leu Leu Cys Val Cys Trp Ser Pro Asp Gly Arg Tyr Val Val Thr Gly Gly Glu Asp Asp Leu Val Thr Val Trp Ser Phe Thr Glu Gly Arg Val Val Ala Arg Gly His Gly His Lys Ser Trp Val Asn Ala Val Ala Phe Asp Pro Tyr Thr Thr Arg Ala Glu Glu Ala Ala Thr Ala Ala Gly Ala Asp Gly Glu Arg Ser Gly Glu Glu Glu Glu Glu Glu Pro Glu Ala Ala Gly Thr Gly Ser Ala Gly Gly Ala Pro Leu Ser Pro Leu Pro Lys Ala Gly Ser Ile Thr Tyr Arg Phe Gly Ser Ala Gly Gln Asp Thr Gln Phe Cys Leu Trp Asp Leu Thr Glu Asp Val Leu Tyr Pro His Pro Pro Leu Ala Arg Thr Arg Thr Leu Pro Gly Thr Pro Gly Thr Thr Pro Pro Ala Ala Ser Ser Ser Arg Gly Gly Glu Pro Gly Pro Gly Pro Leu Pro Arg Ser Leu Ser Arg Ser Asn Ser Leu Pro His Pro Ala Gly Gly Gly Lys Ala Gly Gly Pro Gly Val Ala Ala Glu Pro Gly Thr Pro Phe Ser Ile Gly Arg Phe Ala Thr Leu Thr Leu Gln Glu Arg Arg Asp Arg Gly Ala Glu Lys Glu His Lys Arg Tyr His Ser Leu Gly Asn Ile Ser Arg Gly Gly Ser Gly Gly Ser Gly Ser Gly Gly Glu Lys Pro Ser Gly Pro Val Pro Arg Ser Arg Leu Asp Pro Ala Lys Val Leu Gly Thr Ala Leu Cys Pro Arg Ile His Glu Val Pro Leu Leu Glu Pro Leu Val Cys Lys Lys Ile Ala Gln Glu Arg Leu Thr Val Leu Leu Phe Leu Glu Asp Cys Ile Ile Thr Ala Cys Gln Glu Gly Leu Ile Cys Thr Trp Ala Arg Pro Gly Lys Ala Phe Thr Asp Glu Glu Thr Glu Ala Gln Thr Gly Glu Gly Ser Trp Pro Arg Ser Pro Ser Lys Ser Val Val Glu Gly Ile Ser Ser Gln Pro Gly Asn Ser Pro Ser Gly Thr Val Val <210> 15 <211> 446 <212> PRT
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 5580384CD1 <400> 15 Met Asn Val Thr Pro Glu Val Lys Ser Arg Gly Met Lys Phe Ala Glu Glu Gln Leu Leu Lys His Gly Trp Thr Gln Gly Lys Gly Leu Gly Arg Lys Glu Asn Gly Ile Thr Gln Ala Leu Arg Val Thr Leu Lys Gln Asp Thr His Gly Val Gly His Asp Pro Ala Lys Glu Phe Thr Asn His Trp Trp Asn Glu Leu Phe Asn Lys Thr Ala Ala Asn Leu Val Val Glu Thr Gly Gln Asp Gly Val Gln Ile Arg Ser Leu Ser Lys Glu Thr Thr Arg Tyr Asn His Pro Lys Pro Asn Leu Leu Tyr Gln Lys Phe Val Lys Met Ala Thr Leu Thr Ser Gly Gly Glu Lys Pro Asn Lys Asp Leu Glu Ser Cys Ser Asp Asp Asp Asn Gln Gly Ser Lys Ser Pro Lys Ile Leu Thr Asp Glu Met Leu Leu Gln Ala Cys Glu Gly Arg Thr Ala His Lys Ala Ala Arg Leu Gly Ile Thr Met Lys Ala Lys Leu Ala Arg Leu Glu Ala Gln Glu Gln Ala Phe Leu Ala Arg Leu Lys Gly Gln Asp Pro Gly Ala Pro Gln Leu Gln Ser Glu Ser Lys Pro Pro Lys Lys Lys Lys Lys Lys Arg Arg Gln Lys Glu Glu Glu Glu Ala Thr Ala Ser Glu Arg Asn Asp Ala Asp Glu Lys His Pro Glu His Ala Glu Gln Asn Ile Arg Lys Ser Lys Lys Lys Lys Arg Arg His Gln Glu Gly Lys Val Ser Asp Glu Arg Glu Gly Thr Thr Lys Gly Asn Glu Lys Glu Asp Ala Ala Gly Thr Ser Gly Leu Gly Glu Leu Asn Ser Arg Glu Gln Thr Asn Gln Ser Leu Arg Lys Gly Lys Lys Lys Lys Arg Trp His His Glu Glu Glu Lys Met Gly Val Leu Glu Glu Gly Gly Lys Gly Lys Glu Ala Ala G1y Ser Val Arg Thr Glu Glu Val Glu Ser Arg Ala Tyr Ala Asp Pro Cys Ser Arg Arg Lys Lys Arg Gln Gln Gln Glu Glu Glu Asp Leu Asn Leu Glu Asp Arg Gly Glu Glu Thr Val Leu Gly Gly Gly Thr Arg Glu Ala Glu Ser Arg Ala Cys Ser Asp Gly Arg Ser Arg Lys Ser Lys Lys Lys Arg Gln Gln His Gln Glu Glu Glu Asp Ile Leu Asp Val Arg Asp Glu Lys Asp Ser Gly Ala Arg Glu Ala Glu Ser Arg Ala His Thr Gly Ser Ser Ser Arg Gly Lys Arg Lys Arg Gln Gln His Pro Lys Lys Glu Arg Ala Gly Val Ser Thr Val Gln Lys Ala Lys Lys Lys Gln Lys Lys Arg Asp <210> 16 <211> 364 <212> PRT
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 5158619CD1 <400> 16 Met Thr Glu Lys Glu Val Leu Glu Ser Pro Lys Pro Ser Phe Pro Ala Glu Thr Arg Gln Ser Gly Leu Gln Arg Leu Lys Gln Leu Leu Arg Lys Gly Ser Thr Gly Thr Lys Glu Met Glu Leu Pro Pro Glu Pro Gln Ala Asn Gly Glu Ala Val Gly Ala Gly Gly Gly Pro Ile Tyr Tyr IIe Tyr Glu GIu Glu GIu Glu Glu Glu GIu Glu Glu Glu Glu Pro Pro Pro Glu Pro Pro Lys Leu Val Asn Asp Lys Pro His Lys Phe Lys Asp His Phe Phe Lys Lys Pro Lys Phe Cys Asp Val Cys Ala Arg Met Ile Val Leu Asn Asn Lys Phe Gly Leu Arg Cys Lys Asn Cys Lys Thr Asn Ile His Glu His Cys Gln Ser Tyr Val Glu Met Gln Arg Cys Phe Gly Lys Ile Pro Pro Gly Phe His Arg Ala Tyr Ser Ser Pro Leu Tyr Ser Asn Gln Gln Tyr Ala Cys Val Lys Asp Leu Ser Ala Ala Asn Arg Asn Asp Pro Val Phe Glu Thr Leu Arg Thr Gly Val Ile Met Ala Asn Lys Glu Arg Lys Lys Gly Gln Ala Asp Lys Lys Asn Pro Val Ala Ala Met Met Glu Glu Glu Pro Glu Ser Ala Arg Pro Glu Glu Gly Lys Pro Gln Asp Gly Asn 215 ~ 220 225 Pro Glu Gly Asp Lys Lys Ala Glu Lys Lys Thr Pro Asp Asp Lys His Lys Gln Pro Gly Phe Gln Gln Ser His Tyr Phe Val AIa Leu Tyr Arg Phe Lys Ala Leu Glu Lys Asp Asp Leu Asp Phe Pro Pro Gly Glu Lys Ile Thr Val Ile Asp Asp Ser Asn Glu Glu Trp Trp Arg Gly Lys Ile Gly Glu Lys Val Gly Phe Phe Pro Pro Asn Phe Ile Ile Arg Val Arg Ala Gly Glu Arg Val His Arg Val Thr Arg Ser Phe Val Gly Asn Arg Glu Ile Gly Gln Ile Thr Leu Lys Lys Asp Gln Ile Val Val Gln Lys Gly Asp Glu Ala Gly Gly Tyr Val Lys Val Tyr Thr Gly Arg Lys Val Gly Leu Phe Pro Thr Asp Phe Leu Glu Glu Ile <210> 17 <211> 91 <212> PRT
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 2792745CD1 <400> 17 Met Glu Phe Cys Ser Phe Ala Gln Thr Gly Val Gln Arg Arg Asp Arg Gly Ser Leu Gln Pro Pro Pro Pro Lys Phe Lys Gln Phe Ser His Leu Ser Leu Leu Ser Ser Trp Asp Tyr Tyr Arg His Pro Pro Ser Arg Pro Asp Asn Phe Trp Ile Phe Val Val Met Gly Leu His His Val Gly Leu Ala Gly Leu Gln Leu Leu Thr Ser Ser Asp Pro Pro Thr Ser Thr Ser Gln Ser Ala Gly Ile Thr Ser Val Asn His Arg <210> 18 <211> 116 <212> PRT
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 2827678CD1 <400> 18 Met Pro Pro Ser Ser Ala Asn Phe Phe Cys Leu Phe Val Phe Val Phe Val Leu Arg Trp Ser Phe Val Leu Val Ala Gln Ala Gly Val Gln Trp Cys Arg Leu Gly Ser Pro Gln Pro Leu Pro Pro Arg Phe Lys Arg Phe Ser Cys Leu Thr Leu Pro Ser Ser Trp Asp Tyr Arg Cys Leu Pro Pro Arg Pro Ala Asn Phe Phe Val Phe Leu Val Glu Thr Gly Phe His His Ile Ala Gln Ala Gly Phe Gln Leu Leu Thr Ser Gly Asp Pro Pro Ala Leu Ser 5er GIn Ser Ala Gly Ile Thr Gly Val Ser His Cys Ala Trp Pro Thr Phe Leu <210> 19 <211> 684 <212> PRT
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 790257CD1 <400> 19 Met Ser Ala Ser Gly Val Leu Ser Phe Thr Gln Gln Gly Trp Glu Gln Val Leu Ala Lys Val Lys Arg Ala Val Val Tyr Leu Asp Ala Ala Cys Ala Glu Ser Leu His Trp Gly Cys Gly Ser Thr Arg Leu Leu Glu Ala Val Gly Gly Pro Asp Cys His Leu Arg Glu Phe Glu Pro Asp Ala Ile Gly Gly Gly Ala Lys Gln Pro Lys Ala Val Phe Val Leu Ser Cys Leu Leu Lys Gly Arg Thr Val Glu Ile Leu Arg Asp Ile Ile Cys Arg Ser His Phe Gln Tyr Cys Val Val Val Thr Thr Val Ser His Ala Val His Leu Thr A'la Asn His Val Pro Ala Ala Ala Ala Ala Glu Met Glu Gly Gln Gln Pro Val Phe Glu Gln Leu Glu Glu Lys Leu Cys Glu Trp Met Gly Asn Met Asn Tyr Thr Ala Glu Val Phe His Val Pro Leu Leu Leu Ala Pro Val Ala Pro His Phe Ala Leu Thr Pro Ala Phe Ala Ser Leu Phe Pro Leu Leu Pro Gln Asp Val His Leu Leu Asn Ser Ala Arg Pro Asp Lys Arg Lys Leu Gly Ser Leu Gly Asp Val Asp Ser Thr Thr Leu Thr Pro Glu Leu Leu Leu Gln Ile Arg Cys Leu Val Ser Gly Leu Ser Ser Leu Cys Glu His Leu Gly Val Arg Glu G1u Cys Phe Ala Val GIy Ser Leu Ser Gln Val Ile Ala'Ala Asp Leu Ala Asn Tyr Ala Pro Ala Lys Asn Arg Lys Lys Thr Ala Ala Gly Arg Ala Ser Val Val Phe Val Asp Arg Thr Leu Asp Leu Thr Gly Ala Val Gly His His Gly Asp Asn Leu Val Glu Lys Ile Ile Ser Ala Leu Pro Gln Leu Pro Gly His Thr Asn Asp Val Met Val Asn Met Ile Ala Leu Thr Ala Leu His Thr Glu Glu Glu Asn Tyr Asn Val Val Ala Pro Gly Cys Leu Ser Gln Ser Ser Asp Thr Thr Ala Lys Ala Leu Trp Glu Ala Leu Leu Asn Thr Lys His Lys Glu Ala Val Met Glu Val Arg Arg His Leu Val Glu Ala Ala Ser Arg Glu Asn Leu Pro Ile Lys Met Ser Met Gly Arg Val Thr Pro Gly Gln Leu Met Ser Tyr Ile Gln Leu Phe Lys Asn Asn Leu Lys Ala Leu Met Asn His Cys Gly Leu Leu Gln Leu Gly Leu Ala Thr Ala Gln Thr Leu Lys His Pro Gln Thr Ala Lys Trp Asp Asn Phe Leu Ala Phe Glu Arg Leu Leu Leu Gln Ser Ile Gly Glu Ser Ala Met Ser Va1 Val Leu Asn Gln Leu Leu Pro Met Ile Lys Pro Val Thr Gln Arg Thr Asn Glu.Asp Tyr Ser Pro Glu Glu Leu Leu Ile Leu Leu Ile Tyr Ile Tyr Ser Val Thr Gly Glu Leu Thr Val Asp Lys Asp Leu Cys Glu Ala Glu Glu Lys Val Lys Lys Ala Leu Ala Gln Val Phe Cys Glu Glu Ser Gly Ser Ser Pro Leu Leu Gln Lys Ile Thr Asp Trp Asp Ser Ser Ile Asn Leu Thr Phe His Lys Ser Lys Ile Ala Val Asp Glu Leu Phe Thr Ser Leu Arg Asp Ile Ala Gly Ala Arg Ser Leu Leu Lys Gln Phe Lys Ser Val Tyr Val Pro Gly Asn His Thr His Gln Ala Ser Tyr Lys Pro Leu Leu Lys Gln Val Val Glu Glu Ile Phe His Pro Glu Arg Pro Asp Ser Val Asp Ile Glu His Met Ser Ser Gly Leu Thr Asp Leu Leu Lys Thr Gly Phe Ser Met Phe Met Lys Val Ser Arg Pro His Pro Ser Asp Tyr Pro Leu Leu Ile Leu Phe Val Val Gly Gly Val Thr Val Ser Glu Val Lys Met Val Lys Asp Leu Val Ala Ser Leu Lys Pro Gly Thr Gln Val Ile Val Leu Ser Thr Arg Leu Leu Lys Pro Leu Asn Ile Pro Glu Leu Leu Phe Ala Thr Asp Arg Leu His Pro Asp Leu Gly Phe <210> 20 <211> 344 <212> PRT
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 2617345CD1 <400> 20 Met Asn Leu Leu Pro Cys Asn Pro His Gly Asn Gly Leu Leu Tyr Ala Gly Phe Asn Gln Asp His Gly Cys Phe Ala Cys Gly Met Glu Asn Gly Phe Arg Val Tyr Asn Thr Asp Pro Leu Lys Glu Lys Glu Lys Gln Glu Phe Leu Glu Gly Gly Val Gly His Val Glu Met Leu Phe Arg Cys Asn Tyr Leu Ala Leu Val Gly Gly Gly Lys Lys Pro Lys Tyr Pro Pro Asn Lys Val Met Ile Trp Asp Asp Leu Lys Lys Lys Thr Val Ile Glu Ile Glu Phe Ser Thr Glu Val Lys Ala Val Lys Leu Arg Arg Asp Arg Ile Val Val Val Leu Asp Ser Met Ile Lys Val Phe Thr Phe Thr His Asn Pro His Gln Leu His Val Phe Glu Thr Cys Tyr Asn Pro Lys Gly Leu Cys Val Leu Cys Pro Asn Ser Asn Asn Ser Leu Leu Ala Phe Pro Gly Thr His Thr Gly His Val Gln Leu Val Asp Leu Ala Ser Thr Glu Lys Pro Pro Val Asp Ile Pro Ala His Glu Gly Val Leu Ser Cys Ile Ala Leu Asn Leu Gln Gly Thr Arg Ile Ala Thr Ala Ser Glu Lys Gly Thr Leu Ile Arg Ile Phe Asp Thr Ser Ser Gly His Leu Ile Gln Glu Leu Arg Arg Gly Ser Gln Ala Ala Asn Ile Tyr Cys Ile Asn Phe Asn Gln Asp Ala Ser Leu Ile Cys Val Ser Ser Asp His Gly Thr Val His Ile Phe Ala Ala Glu Asp Pro Lys Arg Asn Lys Gln Ser Ser Leu Ala Ser Ala Ser Phe Leu Pro Lys Tyr Phe Ser Ser Lys Trp Ser Phe Ser Lys Phe Gln Val Pro Ser Gly Ser Pro Cys Tle Cys Ala Phe Gly Thr Glu Pro Asn Ala Val Ile Ala Ile Cys Ala Asp Gly Ser Tyr Tyr Lys Phe Leu Phe Asn Pro Lys Gly Glu Cys Ile Arg Asp Val Tyr Ala Gln Phe Leu Glu Met Thr Asp Asp Lys Leu <210> 21 <211> 95 <212> PRT
<213> Homo sapiens <220>
<221> misc feature <223> Incyte ID No: 3254666CD1 <400> 21 Met Gly Cys Met Lys Ser Lys Gln Thr Phe Pro Phe Pro Thr Ile Tyr Glu Gly Glu Lys Gln His Glu Ser Glu Glu Pro Phe Met Pro Glu Glu Arg Cys Leu Pro Arg Met Ala Ser Pro Val Asn Val Lys Glu Glu Val Lys Glu Pro Pro Gly Thr Asn Thr Val Ile Leu Glu Tyr Ala His Arg Leu Ser Gln Asp I1e Leu Cys Asp Ala Leu Gln Gln Trp Ala Cys Asn Asn Ile Lys Tyr His Asp Ile Pro Tyr Ile Glu Ser Glu Gly Pro ~ ..
<210> 22 <211> 410 <212> PRT
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 4159378CD1 <400> 22 Met Pro Tyr Ser Thr Asn Lys Glu Leu Ile Leu Gly Ile Met Val Gly Thr Ala Gly Ile Ser Leu Leu Leu Leu Trp Tyr His Lys Val Arg Lys Pro Gly Ile Ala Met Lys Leu Pro Glu Phe Leu Ser Leu Gly Asn Thr Phe Asn Ser Ile Thr Leu Gln Asp Glu Ile His Asp Asp Gln Gly Thr Thr Val Ile Phe Gln Glu Arg Gln Leu Gln Ile Leu Glu Lys Leu Asn Glu Leu Leu Thr Asn Met Glu Glu Leu Lys Glu Glu Ile Arg Phe Leu Lys Glu Ala Ile Pro Lys Leu Glu Glu Tyr Ile Gln Asp Glu Leu Gly Gly Lys Ile Thr Val His Lys Ile Ser Pro Gln His Arg Ala Arg Lys Arg Arg Leu Pro Thr Ile Gln Ser Ser Ala Thr Ser Asn Ser Ser Glu Glu Ala Glu Ser Glu Gly Gly Tyr Ile Thr Ala Asn Thr Asp Thr Glu Glu Gln Ser Phe Pro Val Pro Lys Ala Phe Asn Thr Arg Val Glu Glu Leu Asn Leu Asp Val Leu Leu Gln Lys Val Asp His Leu Arg Met Ser Glu Ser Gly Lys Ser Glu Ser Phe Glu Leu Leu Arg Asp His Lys Glu Lys Phe Arg Asp Glu Ile Glu Phe Met Trp Arg Phe Ala Arg Ala Tyr Gly Asp Met Tyr Glu Leu Ser Thr Asn Thr Gln Glu Lys Lys His Tyr Ala Asn Ile Gly Lys Thr Leu Ser Glu Arg Ala Ile Asn Arg Ala Pro Met Asn Gly His Cys His Leu Trp Tyr Ala Val Leu Cys Gly Tyr Val Ser Glu Phe Glu Gly Leu Gln Asn Lys Ile Asn Tyr Gly His Leu Phe Lys Glu His Leu Asp Ile Ala Ile Lys Leu Leu Pro Glu Glu Pro Phe Leu Tyr Tyr Leu Lys Gly Arg Tyr Cys Tyr Thr Val Ser Lys Leu Ser Trp Ile Glu Lys Lys Met Ala Ala Thr Leu Phe Gly Lys Ile Pro Ser Ser Thr Val Gln Glu Ala Leu His Asn Phe Leu Lys Ala Glu Glu Leu Cys Pro Gly Tyr Ser Asn Pro Asn Tyr Met Tyr Leu Ala Lys Cys Tyr Thr Asp Leu Glu Glu Asn Gln Asn Ala Leu Lys Phe Cys Asn Leu Ala Leu Leu Leu Pro Thr Val Thr Lys Glu Asp Lys Glu Ala Gln Lys Glu Met Gln Lys Ile Met Thr Ser Leu Lys Arg <210> 23 <211> 616 <212> PRT
<213> Homo Sapiens <220>
<221> misc feature <223> Incyte ID No: 4317538CD1 <400> 23 Met Lys Cys Ala Leu Phe Leu Arg Val Lys IIe Leu Gln Arg Val Cys Arg Ser Arg Tyr Ile Val His Ala Thr Cys Asp Ser Thr Ala Ala Met Ser Gly Ile Leu Lys Arg Lys Phe Glu Glu Val Asp Gly Ser Ser Pro Cys Ser Ser Val Arg Glu Ser Asp Asp Glu Val Ser Ser Ser Glu Ser Ala Asp Ser Gly Asp Ser Val Asn Pro Ser Thr Ser Ser His Phe Thr Pro Ser Ser Ile Leu Lys Arg Glu Lys Arg Leu Arg Thr Lys Asn Val His Phe Ser Cys Val Thr Val Tyr Tyr Phe Thr Arg Arg Gln Gly Phe Thr Ser Val Pro Ser G1n Gly Gly 7.10 115 120 Ser Thr Leu Gly Met Ser Ser Arg His Asn Ser Val Arg Gln Tyr Thr Leu Gly Glu Phe Ala Arg Glu Gln Glu Arg Leu His Arg Glu Met Leu Arg Glu His Leu Arg Glu Glu Lys Leu Asn Ser Leu Lys Leu Lys Met Thr Lys Asn Gly Thr Val Glu Ser Glu Glu Ala Ser Thr Leu Thr Leu Asp Asp Ile Ser Asp Asp Asp Ile Asp Leu Asp Asn Thr Glu Val Asp Glu Tyr Phe Phe Leu Gln Pro Leu Pro Thr Lys Lys Arg Arg Ala Leu Leu Arg Ala Ser Gly Val Lys Lys Ile 215. 220 225 Asp Val Glu Glu Lys His Glu Leu Arg Ala Ile Arg Leu Ser Arg Glu Asp Cys Gly Cys Asp Cys Arg Val Phe Cys Asp Pro Asp Thr Cys Thr Cys Ser Leu Ala Gly Ile Lys Cys Gln Val Asp Arg Met Ser Phe Pro Cys Gly Cys Thr Lys Glu Gly Cys Ser Asn Thr Ala Gly Arg Ile Glu Phe Asn Pro Ile Arg Val Arg Thr His Phe Leu His Thr Ile Met Lys Leu Glu Leu Glu Lys Asn Arg Glu Gln Gln Ile Pro Thr Leu Asn Gly Cys His,Ser Glu Ile Ser Ala His Ser Ser Ser Met Gly Pro Val Ala His Ser Val Glu Tyr Ser Ile Ala Asp Ser Phe Glu Ile Glu Thr Glu Pro Gln Ala Ala Val Leu His Leu Gln Ser Ala Glu Glu Leu Asp Cys Gln Gly Glu Glu Glu Glu Glu Glu Glu Asp Gly Ser Ser Phe Cys Gly Gly Val Thr Asp Ser Ser Thr Gln Ser Leu Ala Pro Ser Glu Ser Asp Glu Glu Glu Glu Glu Glu Glu Glu Glu Glu Glu Glu Glu Asp Asp Asp Asp Asp Lys Gly Asp Gly Phe Val Glu Gly Leu Gly Thr His Ala Glu Val Val Pro Leu Pro Ser Val Leu Cys Tyr Ser Asp Gly Thr Ala Val His Glu Ser His Ala Lys Asn Ala Ser Phe Tyr Ala Asn Ser Ser Thr Leu Tyr Tyr Gln Ile Asp Ser His Ile Pro Gly Thr Pro Asn Gln Ile Ser Glu Asn Tyr Ser Glu Arg Asp Thr Val Lys Asn Gly Thr Leu Ser Leu Val Pro Tyr Thr Met Thr Pro Glu Gln Phe Val Asp Tyr Ala Arg Gln Ala Glu Glu Ala Tyr Gly Ala Ser His Tyr Pro Ala Ala Asn Pro Ser Val Ile Val Cys Cys Ser Ser Ser Glu Asn Asp Ser Gly Val Pro Cys Asn Ser Leu Tyr Pro Glu His Arg Ser Asn His Pro Gln Val Glu Ser His Ser Tyr Leu Lys Gly Pro Ser Gln Glu Gly Phe Val Ser Ala Leu Asn Gly Asp Ser His Ile Ser Glu His Pro Ala Glu Asn Ser Leu Ser Leu Ala Glu Lys Ser Ile Leu His Glu Glu Cys Ile Lys Ser Pro Val Val Glu Thr Val Pro Val , <210> 24 <211> 392 <212> PRT
<213> Homo Sapiens <220>
<221> mist feature <223> Incyte ID No: 1881010CD1 <400> 24 Met Ala Thr Ala Ala Gln Gly Pro Leu Ser Leu Leu Trp Gly Trp Leu Trp Ser Glu Arg Phe Trp Leu Pro Glu Asn Val Ser Trp Ala Asp Leu Glu Gly Pro Ala Asp Gly Tyr Gly Tyr Pro Arg Gly Arg His Ile Leu Ser Val Phe Pro Leu Ala Ala Gly Ile Phe Phe Val Arg Leu Leu Phe Glu Arg Phe Ile Ala Lys Pro Cys Ala Leu Cys Ile Gly Ile Glu Asp Ser Gly Pro Tyr Gln Ala Gln Pro Asn Ala Ile Leu Glu Lys Val Phe Ile Ser Ile Thr Lys Tyr Pro Asp Lys Lys Arg Leu Glu Gly Leu Ser Lys Gln Leu Asp Trp Asn Val Arg Lys Ile Gln Cys Trp Phe Arg His Arg Arg Asn Gln Asp Lys Pro Pro Thr Leu Thr Lys Phe Cys Glu Ser Met Trp Arg'Phe Thr Phe Tyr Leu Cys Ile Phe Cys Tyr Gly Ile Arg Phe Leu Trp Ser Ser Pro Trp Phe Trp Asp Ile Arg Gln Cys Trp His Asn Tyr Pro Phe Gln Pro Leu Ser Ser Gly Leu Tyr His Tyr Tyr Ile Met Glu Leu Ala Phe Tyr Trp Ser Leu Met Phe Ser Gln Phe Thr Asp Ile Lys Arg Lys Asp Phe Leu Ile Met Phe Val His His Leu Val Thr Ile Gly Leu Ile Ser Phe Ser Tyr Ile Asn Asn Met Val Arg Val Gly , Thr Leu Ile Met Cys Leu His Asp Val Ser Asp Phe Leu Leu Glu Ala Ala Lys Leu Ala Asn Tyr Ala Lys Tyr Gln Arg Leu Cys Asp Thr Leu Phe Val Ile Phe Sex Ala Val Phe Met Val Thr Arg Leu Gly Ile Tyr Pro Phe Trp Ile Leu Asn Thr Thr Leu Phe Glu Ser Trp Glu Ile Ile Gly Pro Tyr Ala Ser Trp Trp Leu Leu Asn Gly Leu Leu Leu Thr Leu Gln Leu Leu His Val Ile Trp Ser Tyr Leu Ile Ala Arg Ile Ala Leu Lys Ala Leu Ile Arg Gly Lys Val Ser Lys Asp Asp Arg Ser Asp Val Glu Ser Ser Ser Glu Glu Glu Asp Val Thr Thr Cys Thr Lys Ser Pro Cys Asp Ser Ser Ser Ser Asn 365 370 ~ 375 Gly Ala Asn Arg Val Asn Gly His Met Gly Gly Ser Tyr Trp Ala Glu Glu <210> 25 <211> 1125 <212> PRT
<213> Homo sapiens <220>
<221> misc_~eature <223> Incyte ID No: 1593038CD1 <400> 25 Met Ala Ser Glu Glu Ala Ser Leu Arg Ala Leu Glu Ser Leu Met Thr Glu Phe Phe His Asp Cys Thr Thr Asn Glu Arg Lys Arg Glu Ile Glu Glu Leu Leu Asn Asn Phe Ala Gln Gln Ile Gly Ala Trp Arg Phe Cys Leu Tyr Phe Leu Ser Ser Thr Arg Asn Asp Tyr Val Met Met Tyr Ser Leu Thr Val Phe Glu Asn Leu Ile Asn Lys Met Trp Leu Gly Val Pro Ser Gln Asp Lys Met Glu Ile Arg Ser Cys Leu Pro Lys Leu Leu Leu Ala His His Lys Thr Leu Pro Tyr Phe Ile Arg Asn Lys Leu Cys Lys Val Ile Val Asp Ile Gly Arg Gln Asp Trp Pro Met Phe Tyr His Asp Phe Phe Thr Asn Ile Leu Gln Leu Ile Gln Ser Pro Val Thr Thr Pro Leu Gly Leu Ile Met Leu Lys Thr Thr Ser Glu Glu Leu Ala Cys Pro Arg Glu Asp Leu Ser Val Ala Arg Lys Glu Glu Leu Arg Lys Leu Leu Leu Asp Gln Va1 Gln Thr Val Leu Gly Leu Leu Thr Gly Ile Leu Glu Thr Val Trp Asp Lys His Ser Val Thr Ala Ala Thr Pro Pro Pro Ser Pro Thr Ser Gly Glu Ser Gly Asp Leu Leu Ser Asn Leu Leu Gln Ser Pro Ser Ser Ala Lys Leu Leu Asn Gln Pro Ile Pro Ile Leu Asp Val Glu Ser Glu Tyr Ile Cys Ser Leu A1a Leu Glu Cys Leu Ala His Leu Phe Ser Trp Ile Pro Leu Ser Ala Ser Ile Thr Pro Ser Leu Leu Thr Thr Ile Phe His Phe Ala Arg Phe Gly Cys Asp Ile Arg Ala Arg Lys Met Ala Ser Val Asn Gly Ser Ser Gln Asn Cys Val Ser Gly Gln Glu Arg G1y Arg Leu Gly Va1 Leu Ala Met Ser Cys Ile Asn Glu Leu Met Ser Lys Asn Cys Val Pro Met Glu Phe Glu Glu Tyr Leu Leu Arg Met Phe Gln Gln Thr Phe Tyr Leu Leu Gln Lys Ile Thr Lys Asp Asn Asn A1a His Thr Val Lys Ser Arg Leu Glu Glu Leu Asp Glu Ser Tyr Ile Glu Lys Phe Thr Asp Phe Leu Arg Leu Phe Val Ser Val His Leu Arg Arg Ile Glu Ser Tyr Ser Gln Phe Pro Val Val Glu Phe Leu Thr Leu Leu Phe Lys Tyr Thr Phe His Gln Pro Thr His Glu Gly Tyr Phe Ser Cys Leu Asg Ile Trp Thr Leu Fhe Leu Asp Tyr Leu Thr Ser Lys Ile Lys Ser Arg Leu Gly Asp Lys Glu A1a Val Leu Asn Arg Tyr Glu Asp Ala Leu Val Leu Leu Leu Thr Glu Val Leu Asn Arg Ile Gln Phe Arg Tyr Asn Gln Ala Gln Leu G1u Glu Leu Asp Asp Glu Thr Leu Asp Asp Asp Gln Gln Thr Glu Trp Gln Arg Tyr Leu Arg Gln Ser Leu Glu Val Val Ala Lys Val Met Glu Leu Leu Pro Thr His Ala Phe Ser 32!89 Thr Leu Phe Pro Val Leu Gln Asp Asn Leu Glu Val Tyr Leu Gly Leu Gln Gln Phe Ile Val Thr Ser Gly Ser Gly His Arg Leu Asn Ile Thr Ala Glu Asn Asp Cys Arg Arg Leu His Cys Ser Leu Arg Asp Leu Ser Ser Leu Leu Gln Ala Val Gly Arg Leu Ala Glu Tyr Phe Ile Gly Asp Val Phe Ala Ala Arg Phe Asn Asp Ala Leu Thr VaI Val Glu Arg Leu Val Lys Val Thr Leu Tyr Gly Ser Gln Ile Lys Leu Tyr Asn Ile Glu Thr Ala Val Pro Ser Val Leu Lys Pro Asp Leu Ile Asp Val His Ala Gln Ser Leu Ala Ala Leu Gln Ala Tyr Ser His Trp Leu Ala Gln Tyr Cys Ser Glu Val His Arg Gln Asn Thr Gln Gln Phe Val Thr Leu Ile Ser Thr Thr Met Asp Ala Ile Thr Pro Leu Ile Ser Thr Lys Val Gln Asp Lys Leu Leu Leu Ser Ala Cys His Leu Leu Val Ser Leu Ala Thr Thr Val' Arg Pro Val Phe Leu Ile Ser Ile Pro Ala Val Gln Lys Val Phe Asn Arg Ile Thr Asp Ala Ser Ala Leu Arg Leu Val Asp Lys Ala.Gln Val Leu Val Cys Arg Ala Leu Ser Asn Ile Leu Leu Leu Pro Trp Pro Asn Leu Pro Glu Asn Glu Gln Gln Trp Pro Val Arg Ser Ile Asn His Ala Ser Leu Ile Ser Ala Leu Ser Arg Asp Tyr Arg Asn Leu Lys Pro Ser Ala Val Ala Pro Gln Arg Lys Met Pro Leu Asp Asp Thr Lys Leu Ile Ile His Gln Thr Leu Ser Val Leu Glu Asp Ile Val Glu Asn Ile Ser Gly Glu Ser Thr Lys Ser Arg Gln Ile Cys Tyr Gln Ser Leu Gln Glu Ser Val Gln Val Ser Leu Ala Leu Phe Pro AIa Phe Ile His GIn Ser Asp Val Thr Asp Glu Met Leu Ser Phe Phe Leu Thr Leu Phe Arg Gly Leu Arg Val Gln Met Gly Val Pro Phe Thr Glu Gln Ile Ile Gln Thr Phe Leu Asn Met Phe Thr Arg Glu Gln Leu Ala Glu Ser Ile Leu His Glu Gly Ser Thr Gly Cys Arg Val Val Glu Lys Phe Leu Lys Ile Leu Gln Val Val Val 890 ~ 895 900 Gln Glu Pro Gly Gln Val Phe Lys Pro Phe Leu Pro Ser Ile Ile Ala Leu Cys Met Glu Gln Val Tyr Pro Ile Ile Ala Glu Arg Pro Ser Pro Asp Val Lys Ala Glu Leu Phe Glu Leu Leu Phe Arg Thr Leu His His Asn Trp Arg Tyr Phe Phe Lys Ser Thr Val Leu Ala Ser Val Gln Arg Gly Ile Ala Glu Glu Gln Met Glu Asn Glu Pro Gln Phe Ser Ala Ile Met Gln Ala Phe Gly Gln Ser Phe Leu Gln Pro Asp Ile His Leu Phe Lys Gln Asn Leu Phe Tyr Leu Glu Thr Leu Asn Thr Lys Gln Lys Leu Tyr His Lys Lys Ile Phe Arg Thr Ala Met Leu Phe Gln Phe Val Asn Val Leu Leu Gln Val Leu Val His Lys Ser His Asp Leu Leu Gln Glu Glu Ile Gly Ile Ala Ile Tyr Asn Met Ala Ser Val Asp Phe Asp Gly Phe Phe Ala Ala Phe Leu Pro Glu Phe Leu Thr Ser Cys Asp Gly Val Asp Ala Asn Gln Lys Ser Val Leu Gly Arg Asn Phe Lys Met Asp Arg Asp.Leu Pro Ser Phe Thr Gln Asn Val His Arg Leu Val Asn Asp Leu Arg Tyr Tyr Arg Leu Cys Asn Asp Ser Leu Pro Pro Gly Thr Val Lys Leu <210> 26 <211> 1273 <212> PRT
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 7494930CD1 <400> 26 Met Thr Gly Ser Asp Leu His Ile Thr Val Leu Thr Leu His Val Ser Gly Thr Asn Asp Pro Ile Lys Arg His Arg Leu Asp Asn Trp Ile Lys Ser Gln Asp Pro Ser Val Cys Cys Ile Gln Glu Thr His Leu Thr Trp Gln Asp Thr His Gly Ala Gln Val Lys Gly Trp Lys Lys Ala Phe His Ala Asn Gly His Gln Lys Gln Ala Gly Val Ala Ile Leu Ile Ser Asp Lys Thr Asn Cys Lys Ala Thr Ala Val Lys Lys Asp Lys Glu Gly His Tyr Ile Arg Val Lys Gly Leu Val Gln Gln Glu Asn Ile Thr Val Leu Asn Ile Tyr Ala Leu Asn Thr Gly Ala Ser Lys Phe Ile Lys Gln Leu Leu Ile Asp Leu Arg Asn Glu Thr Asp Ser Asn.Thr Ile Val Val Gly Asp Phe Asn Ala Pro Leu Thr Thr Val Asp Arg Ser Ser Arg Gln Lys Val Asn Lys Glu Thr Met Gly Leu Asn Tyr Thr Leu Glu Gln Met Asp Leu Thr Asp Val Tyr Arg Thr Phe His Pro Ala Thr Arg Glu Tyr Thr Phe Tyr Ser Thr Val His Gly Thr Phe Ser Lys Ile Asp His Met Ile Gly His Lys Thr Asn Leu Asn Thr Phe Lys Lys Thr Glu Ile Ile Ser Arg Thr Pro Ser Asp His Ser Val Ile Lys Leu Glu Leu Arg Ile Lys Lys Leu Thr Gln Asn Arg Thr Thr Thr Trp Lys Leu Asn Asn Leu Leu Leu Asn Asp Ser Trp Val Asn Asn Glu Ile Lys Ala Glu Ile Lys Lys Leu Phe Glu Thr Asn Glu Ser Lys Glu Thr Met Tyr Gln Asn Leu Trp Asp Thr Val Thr Ala Val Phe Arg Gly Lys Phe Ile Ala Leu Asn Ala His Gly Arg Lys Gln Glu Arg Ser Lys Ile Asn Thr Leu Thr Ser Gln Leu Lys Glu Leu Glu Lys Gln Lys G1n Ile Asn Ser Lys Ala Ser Arg Arg Gln Glu Ile Thr Lys Ile Arg Ala Glu Leu Lys Glu Thr Glu Thr Gln Lys Thr Leit Gln Lys Ile Asn Glu Thr Lys Ser Trp Phe Phe Glu Lys Asn Lys Ile Asp Arg Pro Leu Ala Arg Phe Ile Gln Thr Lys Arg Asp Lys Asn Gln Thr Asp Thr Ile Lys Asn Asp Lys Val Ala Ile Thr Thr Asp Pro Thr Glu Ile Gln Thr Thr Ile Arg Glu Tyr Tyr Lys His Leu Tyr Ala Lys Lys Leu Asp Asn Leu Glu Glu Met Asp Lys Phe Leu Asp Ser Tyr Thr Leu Pro Arg Leu Asn Gln Glu Glu Thr Glu Asn Leu Asn Arg Thr Ile Met Ser Ser Lys Ile Glu Ser Val Ile Lys Ser Leu Pro Thr Lys Lys Ser Pro Arg Leu Asp Gly Phe Thr Ala Glu Phe Tyr Gln Thr Tyr Lys Glu Leu Ile Leu Ile Ile Leu Lys Gln Phe Gln Lys Ile Glu Lys Asp Gly Thr Leu Pro Asn Ser Phe Tyr Glu Ala Ser Ile Thr Ile Ile Pro Lys Pro Gly Lys Asp Ile Thr Lys Lys Glu Asn Tyr Arg Leu Ile Ser Leu Met Asn Ile Asp Ala Lys Ile Leu Asn Lys Met Leu Ala Asn Gln Ile Gln Gln His Ile Lys Lys Ile Ile His His Asp Gln Val Gly Phe Ile Pro Gly Met Gln Glu Trp Phe Asn Thr His Met Ser Ile Asn Val Met Tyr His Val Asn 575 580 ~ 585 Arg Ile Lys Thr Lys Asn His Leu Ile Ile Ser Ile Asp Ala Glu Lys Ala Phe Asp Lys Ile Gln His Cys Phe Met Ile Lys Thr Leu Ser Lys Ile Gly Gly Gln Gly Thr Tyr Phe Asn Ile Ile Lys Ala Ile Tyr Asp Lys Ser Thr Ala Asn Ile Ile Leu Asn Gly Glu Lys Leu Lys Ala Phe Pro Leu Arg Asn Gly Lys Arg Gln Gly Cys Pro Leu Ser Pro Leu Leu Phe Asn Ile Val Leu Glu Val Leu Ala Arg Ala Ile Arg Gln Glu Lys Glu Ile Lys Gly Ile Gln Ile Gly Lys Glu Glu Val Lys Leu Ser Leu Phe Ala Glu Asp Met Ile Val Tyr Leu Glu Asn Pro Ile Ile Ser Ala Gln Asn Leu Leu Lys Leu Ile Ser Asn Phe Ser Lys Ile Ser Gly Tyr Lys Ile Asn Val Gln Lys Ser Val Ala Leu Leu Cys Ala Asn Ser Asp Gln Ala Glu Asn Gln Ile Lys Asn Ser Thr Pro Phe Thr Ile Ala Ala Lys Lys Asn Lys His Leu Arg Ile Tyr Leu Thr Lys Glu Glu Lys Asp Phe Tyr Lys Glu Asn Tyr Lys Thr Leu Leu Lys Glu Ile Ile His Asp Thr Asn Lys Trp Lys His Ile Pro Cys Ser Trp Ile Gly Arg Ile Asn Ile Val Lys Met Ala Ile Leu Pro Lys Val Ile Tyr Arg Ile Asn Thr Ile Pro Ile Lys Leu Pro Leu Thr Phe Phe Thr Glu Leu Glu Lys Thr Thr Leu Asn Phe Ile Trp Asn Gln Lys Lys Ser Leu Tyr Ser Gln Asp Asn Pro Lys Gln Lys Asn Lys Ala Gly Gly Ile Met Leu Pro Asn Phe Lys Leu His Tyr Lys Ala Thr Val Thr Lys Thr Ala Trp Tyr Trp Tyr Gln Asn Arg Tyr Ile Asp Gln Trp Asn Ser Thr Glu Ala Ser Glu Ile Thr Pro Asp Ile Tyr Asn His Leu Ile Phe Asp Lys Pro Asp Lys Asn Lys Lys Trp Gly Lys Asp Ser Leu Phe Asn Lys Trp Cys Trp Glu Asn Trp Leu Ala Ile Cys Arg Lys Leu Lys Leu Asp Pro Phe Leu Thr Thr Tyr Thr Lys Ile Asn Ser Arg Trp Ile Lys Asp Leu Asn Leu Arg Pro Lys Thr Ile Lys Thr Leu Glu Glu Asn Pro Gly Asn Thr Ile Gln Asp Ile Gly Met Gly Lys Asp Phe Met Thr Lys Thr Pro Lys Ala Ile Ala Thr Lys Ala Lys Ile Asp Lys Trp Asp Leu Ile Lys Leu Lys Ser Phe Cys Thr Ala Lys Glu Thr Ile Ile Arg Val Asn Arg Gln Pro Thr Glu Trp Glu Lys Ile Phe Ala Ile Cys Pro Ser Asp Lys Gly Leu Ile Ser Arg Ile Tyr Lys Glu Leu Lys Gln Ile Tyr Lys Lys Lys Thr Asn Asn Leu Leu Lys Lys Trp Ala Lys Asp Met Asn Arg His Phe Ser Lys Glu Asp Ile Gln Ala Ala Ser Lys His Met Glu Lys Cys Ser Ile Ser Leu Ile Ile Arg Glu Met Gln Ile Lys Thr Thr Met Ile Pro Asn Leu Thr Pro Val Arg Met Ala Ile Ser Lys Glu Ala Lys Asn Ser Gly Cys Leu His Gly Cys Gly Asp Gln Gly Thr Leu Leu His 1130 1135 ~ 1140 Cys Trp~Arg Gln Cys Lys Leu Val Gln Pro Leu Trp Lys Thr Val Trp Arg Phe Leu Lys Glu Leu Lys Val Asp Pro Pro Phe Asp Pro 1160 ' 1165 1170 Ala Ile Pro Leu Leu Gly Ile Tyr Pro Glu Glu Lys Lys Ser Leu Tyr Lys Lys Asp Thr Cys Thr His Met Ser Ile Ala Ala Arg Phe.

Ala Ile Ala Lys Leu Trp Asn Gln Pro Asn Cys Pro Ser Thr Lys Glu Trp Ile Lys Lys Met Trp Gln Met Tyr Thr Leu Glu Tyr Tyr Ala Ala Ile Lys Lys Asp Glu Ile Met Ser Phe Ala Gly Thr Trp Met Met Leu Glu Thr Ile Ile Leu Ser Lys Leu Thr Gln Glu Gln Gln Thr Lys His Cys Met Phe Ser Leu Ile Ser Gly Ser <210> 27 <212> 327 <212> PRT
<213> Homo sapiens <220>
<221> misc_feature <223> Incyte ID No: 7497349CD1 <400> 27 Met Ala Pro Ser Thr Ala Val Ser Val Val Ser Asp Ser Ile Lys Val Gln Pro Leu Leu Ile Ser Ala Asp Asn Lys Val Ile Ile Ile Gln Pro Gln Val Gln Thr Gln Pro Glu Ser Thr Ala Glu Ser Arg Pro Pro Thr Glu Glu Pro Ser Gln Gly Ala G1n Ala Thr Lys Lys Lys Lys Glu Asp Arg Pro Pro Thr Gln Glu Asn Pro Glu Lys Ile Ala Phe Met Val Ala Leu Gly Leu Val Thr Thr Glu His Leu G1u Glu Ile Gln Ser Lys Arg Gln Glu Arg Lys Arg Arg Ser Thr A1a Asn Pro Ala Tyr Ser Gly Leu Leu Glu Thr Glu Arg Lys Arg Leu Ala Ser Asn Tyr Leu Asn Asn Pro Leu Phe Leu Thr Ala Arg Ala Asn Glu Asp Pro Cys Trp Lys Asn Glu Ile Thr His Asp Glu His Cys Ala Ala Cys Lys Arg Gly Ala Asn Leu Gln Pro Cys Gly Thr Cys Pro Gly Ala Tyr His Leu Ser Cys Leu Glu Pro Pro Leu Lys Thr Ala Pro Lys Gly Val Trp Val Cys Pro Arg Cys Gln Gln Lys A1a Leu Lys Lys Asp Glu Gly Val Pro Trp Thr Gly Met Leu Ala Ile Val His Ser Tyr Val Thr His Lys Thr Val Lys Glu Glu Glu Lys Gln Lys Leu Leu Gln Arg Gly Ser Glu Leu Gln Asn Glu His Gln Gln Leu Glu Glu Arg Asp Arg Arg Leu Ala Ser Ala Val Gln Lys Cys Leu Glu Leu Lys Thr Ser Leu Leu Ala Arg Gln Arg Gly Thr Gln Ser Ser Leu Asp Arg Leu Arg Ala Leu Leu Arg Leu Ile Gln Gly Glu Gln Leu Leu Gln Val Thr Met Thr Thr Thr Ser Pro Ala Pro Leu Leu Ala Gly Pro Trp Thr Lys Pro Ser Val Ala Ala Thr His Pro Thr Val Gln His Pro Gln Gly His Asn <210> 28 <211> 79 <212> PRT
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 5510805CD1 <400> 28 Met Leu Ser Ser Lys Ile Ser Val Ile Leu Phe Arg Thr Arg Gln Ser Phe Val Leu Lys Gly Gln Leu Glu Lys Arg Ile Arg Leu Phe Tyr Asp Cys Ser Trp Ser Leu Thr Tyr Pro Tyr Asn Thr Ser Glu Phe Met Lys Gly Trp Arg Ser Cys Leu Met Leu Ile Glu Ala Ala Lys Val Leu Cys Thr Leu His Ile Ile Phe Val Arg Ala Tyr Leu Ile Ala Arg Lys <210> 29 <211> 270 <212> PRT
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 1577482CD1 <400> 29 Met Pro Arg Asp Asn Met Ala Ser Leu Ile Gln Arg Ile Ala Arg Gln Ala Cys Leu Thr Phe Arg Gly Ser Gly Gly Gly Arg Gly Ala Ser Asp Arg Asp Ala Ala Ser Gly Pro Glu Ala Pro Met Gln Pro 35 40 . 45 Gly Phe Pro Glu Asn Leu Ser Lys Leu Lys Ser Leu Leu Thr Gln.

Leu Arg Ala Glu Asp Leu Asn Ile Ala Pro Arg Lys Ala Thr Leu Gln Pro Leu Pro Pro Asn Leu Pro Pro Val Thr Tyr Met His Ile Tyr Glu Thr Asp Gly Phe Ser Leu Gly Val Phe Leu Leu Lys Ser Gly Thr Ser Ile Pro Leu His Asp His Pro Gly Met His Gly Met Leu Lys Val Leu Tyr Gly Thr Val Arg Ile Ser Cys Met Asp Lys Leu Asp Ala Gly Gly Gly.Gln Arg Pro Arg Ala Leu Pro Pro Glu Gln Gln Phe Glu Pro Pro Leu Gln Pro Arg Glu Arg Glu Ala Val Arg Pro Gly Val Leu Arg Ser Arg Ala Glu Tyr Thr Glu Ala Ser Gly Pro Cys Ile Leu Thr Pro His Arg Asp Asn Leu His Gln Ile Asp Ala Val Glu Gly Pro Ala Ala Phe Leu Asp Ile Leu Ala Pro Pro Tyr Asp Pro Asp Asp Gly Arg Asp Cys His Tyr Tyr Arg Val Leu Glu Pro Val Arg Pro Lys Glu Ala Ser Ser Ser Ala Cys Asp Leu Pro Arg Glu Val Trp Leu Leu Glu Thr Pro Gln Ala Asp Asp Phe Trp Cys Glu Gly Glu Pro Tyr Pro Gly Pro Lys Val Phe Pro <210> 30 <211> 692 <212> PRT
<213> Homo sapiens <220>
<221> misc_feature <223> Incyte ID No: 1805054CD1 <400> 30 Met Phe Tyr Phe Arg Gly Cys Gly Arg Trp Val Ala Val Ser Phe Thr Lys Gln Gln Phe Pro Leu Ala Arg Leu Ser Ser~Asp Ser Ala Ala Pro Arg Thr Pro His Phe Asp Val Ile Val Ile Gly Gly Gly His Ala Gly Thr Glu Ala Ala Thr Ala Ala Ala Arg Cys Gly Ser Arg Thr Leu Leu Leu Thr His Arg Val Asp Thr Ile Gly Gln Met Ser Cys Asn Pro Ser Phe Gly Gly Ile Gly Lys Gly His Leu Met Arg Glu Val Asp Ala Leu Asp Gly Leu Cys Ser Arg Ile Cys Asp Gln Ser Gly Val His Tyr Lys Val Leu Asn Arg.Arg Lys Gly Pro Ala Val Trp Gly Leu Arg Ala Gln Ile Asp Arg Lys Leu Tyr Lys Gln Asn Met Gln Lys Glu Ile Leu Asn Thr Pro Leu Leu Thr Val Gln Glu Gly Ala Val Glu Asp Leu Ile Leu Thr Glu Pro Glu Pro Glu His Thr Gly Lys Cys Arg Val Ser Gly Val Val Leu Val Asp Gly Ser Thr Val Tyr Ala Glu Ser Val Ile Leu Thr Thr Gly Thr Phe Leu Arg Gly Met Ile Val Ile Gly Leu Glu Thr His Pro Ala Gly Arg Leu Gly Asp Gln Pro Ser Ile Gly Leu Ala Gln Thr Leu Glu Lys Leu Gly Phe Val Val Gly Arg Leu Lys Thr Gly Thr Pro Pro Arg Ile Ala Lys Glu Ser Ile Asn Phe Ser Ile Leu Asn Lys His Ile Pro Asp Asn Pro Ser Ile Pro Phe Ser Phe Thr Asn Glu Thr Val Trp Ile Lys Pro Glu Asp Gln Leu Pro Cys Tyr Leu Thr His Thr Asn Pro Arg Val Asp Glu Ile Val Leu Lys Asn Leu His Leu Asn Ser His Val Lys Glu Thr Thr Arg Gly Pro Arg Tyr Cys Pro Ser Ile Glu Ser Lys Val Leu Arg Phe Pro Asn Arg Leu His Gln Val Trp Leu Glu Pro Glu Gly Met Asp Ser Asp Leu Ile Tyr Pro Gln Gly Leu Ser Met Thr Leu Pro Ala Glu Leu Gln Glu Lys Met Ile Thr Cys Ile Arg Gly Leu Glu Lys Ala Lys Val Ile Gln Pro Gly Tyr Gly Val Gln Tyr Asp Tyr Leu Asp Pro Arg Gln Ile Thr Pro Ser Leu Glu Thr His Leu Val Gln Arg Leu Phe Phe Ala Gly Gln Ile Asn Gly Thr Thr Gly Tyr Glu Glu A1a Ala Ala Gln Gly Val Ile Ala Gly Ile Asn Ala Ser Leu Arg Val Ser Arg Lys Pro Pro Phe Val Val Ser Arg Thr Glu Gly Tyr Ile Gly Val Leu Ile Asp Asp Leu Thr Thr Leu Gly Thr Ser Glu Pro Tyr Arg Met Phe Thr Ser Arg Val Glu Phe Arg Leu Ser Leu Arg Pro Asp Asn Ala Asp Ser Arg Leu Thr Leu Arg Gly Tyr Lys Asp Ala Gly Cys 485 , 490 495 Val Ser Gln Gln Arg Tyr Glu Arg Ala Cys Trp Met Lys Ser Ser Leu Glu Glu Gly Ile Ser Val Leu Lys Ser Ile Glu Phe Leu Ser Ser Lys Trp Lys Lys Leu Ile Pro Glu Ala Ser Ile Ser.Thr Ser Arg Ser Leu Pro Val Arg Ala Leu Asp Val Leu Lys Tyr Glu Glu Val Asp Met Asp Ser Leu Ala Lys Ala Val Pro Glu Pro Leu Lys Lys Tyr Thr Lys Cys Arg Glu Leu Ala Glu Arg Leu Lys Ile Glu Ala Thr Tyr Glu Ser Val Leu Phe His Gln Leu Gln Glu.Ile Lys Gly Val Gln Gln Asp G1u Ala Leu Gln Leu Pro Lys Asp Leu Asp Tyr Leu Thr Ile Arg Asp Val Ser Leu Ser His Glu Val Arg Glu Lys Leu His Phe Ser Arg Pro Gln Thr Ile Gly Ala Ala Ser Arg Ile Pro Gly Val Thr Pro Ala Ala Ile Ile Asn Leu Leu Arg Phe Val Lys Thr Thr Gln Arg Arg Gln Ser Ala Met Asn Glu Ser Ser Lys Thr Asp Gln Tyr Leu Cys Asp Ala Asp Arg Leu Gln Glu Arg Glu Leu <210> 31 <211> 338 <212> PRT
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 7492708CD1 <400> 31 Met Gly Lys Lys Gln Ser Arg Lys Thr Gly Asn Ser Lys Lys Gln Ser Ala Ser Pro Pro Pro Lys Glu Arg Ser Ser Ser Leu Ala Thr Glu Gln Ser Arg Thr Glu Asn Glu Phe Asp Glu Leu Arg Glu Glu Gly Phe Arg Arg Ser Asn Tyr Ser Glu Leu Gln Glu Glu Ile Gln Thr Lys Gly Lys Glu Val Glu Asn Phe Glu Lys Asn Leu Asp Glu Cys Ile Thr Arg Ile Thr Asn Thr Glu Lys Cys Leu Lys Glu Leu Met Glu Leu Lys Ala Lys Ala Arg Glu Leu Arg Glu Glu Tyr Arg Ser Leu Arg Ser Arg Cys Asp Gln Leu Glu Glu Arg Leu Ser Met Met Glu Asp Glu Met Asn Glu Met Lys Arg G1u Gly Lys Phe Arg Glu Lys Arg Ile Lys Arg Asn Glu Gln Ser Pro Gln Glu Ile Trp Asp Tyr Val Lys Arg Pro Asn Leu Cys Leu Ile Val Val Pro Glu Ser Asp Gly Glu Asn Gly Thr Lys Leu Glu~Asn Thr Leu Arg Asn Ile Phe Gln Glu Asn Phe Pro Asn Leu Ala Arg Gln Al,a Asn Ile Gln Ile Gln Glu Ile Glr~ Arg Thr Pro Gln Arg Tyr Ser Ser Arg Arg Ala Thr Pro Arg Leu Ile Il'e Val Arg Phe Thr Lys Val Glu 215 220 . 225 Met Lys Glu Lys Met Leu Arg Ala Ala Arg Glu Lys.Gly Gln Val Thr His Lys Gly Lys Thr Ile Arg Leu Thr Ala Asp Leu Ser Ala Glu Thr Leu Gln Ala Arg Arg Glu Trp' Gly Pro Ile Phe Asn Ile Leu Lys Glu Lys Asn Phe Gln Pro Arg Ile Ser Tyr Pro Ala Lys Leu Ser Phe Ile Ser Glu Gly Glu Ile Lys Tyr Phe Thr Asp Lys Gln Met Leu Arg Asp Phe Val Thr Thr Arg Pro Ala Leu Lys Glu Leu Leu Lys Glu Ala Leu Asn Met Glu Arg Asn Asn Gln Tyr Gln Pro Leu Gln Asn His Ala Lys Leu <210> 32 <211> 1027 <212> PRT
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 7490847CD1 <400> 32 Met Tyr Glu Gly Lys His Ile His Phe Ser Glu Val Asp Asn Lys Pro Leu Cys Ser Tyr Ser Pro Lys Leu Cys Lys Gln Arg Arg Leu Asn Gly Tyr Ala Phe Cys Ile Arg His Val Leu Glu Asp Lys Thr 35 40 ~ 45 Ala Pro Phe Lys Gln Cys Glu Tyr Val Ala Lys Tyr Asn Ser Gln Arg Cys Thr Asn Pro Ile Pro Lys Ser Glu Asp Arg Arg Tyr Cys Asn Ser His Leu Gln Val Leu Gly Phe Ile Pro Lys Lys Glu Arg Lys Lys Lys Asn Asp Pro Ile Asp Glu Val Lys Val Arg His Gln Met Asp Thr Met Ala Phe Ser Leu Thr Val Pro Thr Leu Ala Leu Lys Met Pro Asn Gly Leu Asp Gly Met Ser Leu Ser Pro Pro Gly Ala Arg Val Pro Leu His Tyr Leu Glu Thr Glu Leu Glu Asp Pro Phe Ala Phe Asn Glu Glu Asp Asp Asp Leu Lys Lys Gljr Ala Thr Val Arg Lys Lys Leu Gln Ser Lys Leu Ala Gln Asn Arg Gln Arg Gln Arg Glu Thr Glu Ile Leu Lys Val Arg Gln Glu.His Phe Ser 185 . 190 195 Pro.Pro Pro Ala Pro Ser Gln Gln:Gln Pro Pro Gln Gln His Ser His Leu Ser Pro Leu Ser Thr Ser Leu Lys Pro Pro Ala Pro Pro Gln Gly Ser Val Cys Lys Ser Pro Gln.Pro Gln Asn Thr Ser Leu Pro Met Gln Gly Val Ala Pro Thr Thr His Thr Ile Ala Gln Ala Arg Gln Leu Ser His Lys Arg Pro Leu Pro Leu Leu Pro Ser Ser Arg Ala Pro Thr Val Asp Pro Pro Arg Thr Asp Arg Ile Leu Met Lys Ala Thr Ala Phe Ser Pro His Phe Ser Cys IIe Ser Arg Leu Gln Arg Leu Val Lys Leu Cys Thr Gln Lys His Gln Leu Asp Thr Asp Leu Phe Pro His Leu Gly Leu Asp Trp Ser Glu Glu Ser Gly Glu Glu Pro Glu Asp Ser Glu Gln Ala Ser Pro Tyr Gln Val Ala Trp Ser Ile Arg Glu Thr Leu Arg Tyr Gln Arg His Ala Ser Asp Asp Asp Asp Ala Glu Ser Arg Ser Ser Gly Val Thr Gln Leu Cys Thr Tyr Phe Gln Gln Lys Tyr Lys His Leu Cys Arg Leu Glu Arg Ala Glu Ser Arg Gln Lys Lys Cys Arg His Thr Phe Arg Lys Ala Leu Leu Gln Ala Ala Ser Lys Glu Pro Glu Cys Thr Gly Gln Leu Ile Gln Glu Leu Arg Arg Ala Ala Cys Ser Arg Thr Ser Ile Ser Arg Thr Lys Leu Arg Glu Val Glu Pro Ala Ala Cys Ser Gly Thr 440 445 ' 450 Val Lys GIy Glu Gln Cys Ala Asn Lys AIa Leu Pro Phe Thr Arg His Cys Phe Gln His Ile Leu Leu Asn His Ser Gln Gln Leu Phe Ser Ser Cys Thr Ala Lys Phe Ala Asp Gly Gln Gln Cys Ser Val Pro Val Phe Asp Ile Thr His Gln Thr Pro Leu Cys Glu Glu His Ala Lys Lys Met Asp Asn Phe Leu Arg Gly Asp Asn Ser Arg Lys Val Gln His Gln Gln Gln Arg Lys Pro Arg Lys Lys Thr Lys Pro Pro Ala Leu Thr Lys Lys His Lys Lys Lys Arg Arg Arg Gly Pro Arg Arg Pro Gln Lys Pro Ile Pro Pro Ala Val Pro Gln Gly Asn Leu Ser Met Pro Ala Ser Val Ser Leu Pro Val Glu Ala Ser His Ile Arg Ser Pro Ser Thr Pro Glu Leu Ser Ala Asp Glu Leu Pro Asp Asp Ile Ala Asn Glu Ile Thr Asp Ile Pro His Asp Leu Glu Leu Asn Gln Glu Asp Phe Ser Asp Val Leu Pro Arg Leu Pro Asp Asp Leu Gln Asp Phe Asp Phe Phe Glu Gly Lys Asn G1y Asp Leu Leu Pro Thr Thr Glu Glu Ala Glu Glu Leu Glu Arg Ala Leu Gln Ala Val Thr Ser Leu Glu Cys Leu Ser Thr Ile Gly Val Leu Ala Gln Ser Asp Gly Val Pro Val Gln Glu Leu Ser Asp Arg Gly Ile Gly Val Phe Ser Thr Gly Thr Gly Ala Ser Gly Ile Gln Ser Leu Ser Arg Glu Val Asn Thr Asp Leu Gly Glu Leu Leu Asn Gly Arg Ile Val His Asp Asn Phe Ser Ser Leu Glu Leu Asp Glu Asn Leu Leu Arg Ser Ala Thr Leu Ser Asn Pro Pro Thr Pro Leu Ala Gly Gln Ile Gln Gly Gln Phe Ser Ala Pro Ala Asn Val Gly Leu Thr Ser Ala Thr Leu Ile Ser Gln Ser Ala Leu Gly Glu Arg Ala Phe Pro Gly Gln Phe His Gly Leu His Asp Gly Ser His Ala Ser Gln Arg Pro His Pro Ala Gln Leu Leu Ser Lys Ala Asp Asp Leu Ile Thr Ser Arg Gln Gln Tyr Ser Ser Asp His Ser His Ser Ser Pro His Gly Ser His Tyr Asp Ser Glu His Val Pro Ser Pro Tyr Ser Asp His Ile Thr Ser Pro His Thr Thr Ser Tyr 5er Gly Asp Asn Met Ala Ala Thr Phe Ser Ala Glu Met Pro Ile Met Ala Gln His Leu Leu Pro Thr Gln Leu Glu Val Pro Leu Gly Gly VaI Val Asn Pro Arg Thr His Trp Gly Asn Leu Pro Val Asn Leu Gly Asp Pro Ser Pro Phe Ser Asn Leu Leu Gly Ala Asp Gly His Leu Leu Ser Thr Ser Leu Ser Thr Pro Pro Thr Thr Ser Asn Ser Glu Thr Thr Gln Pro Ala Phe Ala Thr Val Thr Pro Ser Ser Ser Ser Val Leu Pro Gly Leu Pro Gln Thr Ser Phe Ser Gly Met Gly Pro Ser Ala Glu Leu Met Ala Ser Thr Ser Pro Lys Gln Gln Leu Pro Gln Phe Ser Ala Ala Phe Gly His Gln Leu Ser Ser His Ser Gly Ile Pro Lys Asp Leu Gln Pro Ser His Ser Ser Ile Ala Pro Pro Thr Gly Phe Thr Val Thr Gly Ala Thr Ala Thr Ser Thr Asn Asn Ala Ser Ser Pro Phe Pro Ser Pro Asn . , <210> 33 <211> 1275 <212> PRT
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 7493059CD1 <400> 33 Met Thr Gly Ser Asn Ser His Ile Thr Ile Leu Thr Leu Asn Val Asn Gly Leu Asn Ala Pro Ile Lys Ile His Trp Leu Ala Asn Trp Ile Lys Ser Gln Asp Pro Ser Val Cys Tyr Ile Gln Glu Thr His Phe Thr Cys Arg Asp Thr His Arg Leu Lys Ile Lys Gly Trp Arg Lys Ile Tyr Gln Ala Asn Gly Lys Gln Lys Lys Ala Gly Va1 Ala Ile Leu Val Ser Asp Lys Thr Asp Phe Lys Pro Thr Lys Ile Lys Arg Asp Lys Glu Gly His Cys Ile Met Val Lys Gly Ser Ile Gln Gln Glu GIu Leu Thr Ile Leu Asn Ile Tyr Ala Pro Asn Thr Gly Gly Pro Arg Phe Ile Lys Gln Val Leu Arg Glu Arg Asn Gln Ser Leu Asp Ser His Thr Ile Ile Met Gly Asp Phe Asn Thr Pro Leu Ser Thr Leu Asg Arg Ser Thr Arg Gln Lys Val Asn Lys Asp I1e Gln Glu Leu Asn Ser Ala Leu His Gln Ala Asp Leu Ile Asp Ile Tyr Arg Thr Leu His Pro Gln Ser Thr Glu Tyr Thr Phe Phe Ser Ala Pro His Arg Thr Tyr Ser Lys Ile Asp His Ile Val Gly Ser Lys Ala Leu Leu Ser Lys Cys Lys Arg Thr Glu Ile Ile Thr Asn Cys Leu Ser Asp His Ser Ala Ile Lys Gln Glu Leu Thr Ile Lys Lys Leu Thr Gln Asn His Ser Thr Thr Trp Lys Leu Asn Asn Leu Val Leu Asn Asp Tyr Trp Val His Asn Lys Met Lys Ala Glu Ile Lys Met Phe Phe Glu Thr Asn Glu Asp Lys Asp Thr Thr His Gln Asn Leu Trp Asp Thr Phe Lys Ala Val Cys Arg Gly Lys Phe Ile Ala Leu Asn Ala His Lys Arg Lys Gln Glu Arg Ser Asn Ile Asp Thr Leu Thr Ser Gln Leu Lys Glu Leu Glu Lys Gln Glu Gln Thr Tyr Ser Lys Ala Ser Arg Arg Gln Glu Ile Thr Lys Ile Arg Ala Glu Leu Lys Glu Ile GIu Thr Gln Lys Thr Leu Gln Lys Lys Ile Asn Glu Ser Arg Ser Trp Phe Phe Glu Lys Ile Asn Lys Tle Asp Arg Pro Leu Ala Arg Leu Ile Lys Lys Lys Arg Glu Lys Asn Gln Ile Asp Thr Ile Lys Asn His Lys Gly Asp Ile Thr Thr Gly Pro Thr Glu Ile Gln Thr Thr Ile Arg Glu Tyr Tyr Lys His Leu Tyr Ala Asn Lys Leu Glu Asn Leu Glu Glu Met Asp Lys Phe Leu Asn Thr Tyr Thr Leu Pro Arg Leu Asn Gln Glu Glu Val Glu Ser Leu Asn Arg Pro Ile Thr Gly Ser Glu Ile Glu Ala Ile Ile Ser Ser Leu Pro Thr Gln Lys Ser Pro Gly Pro Asp Gly Phe Thr Ala Glu Phe Tyr Gln Arg Tyr Lys Glu Lys Leu Val Pro Phe Leu Leu Lys Leu Phe Gln Ser Thr Glu Lys Glu Gly Ile Leu Pro Asn Ser Phe Tyr Glu Ala Ser Ile Ile Leu Ile Pro Lys Pro Gly Arg Asp Thr Thr Lys Lys Glu Asn Phe Arg Pro Ile Ala Leu Met Asn Ser Asn Ala Lys Ile Leu Asn Arg Thr Leu Ala Asn Arg Ile Gln Gln His Ile Lys Lys Leu Ile His His Asp Gln Val Gly Phe Ile Pro Glu Met Gln Gly Trp Phe Asn Ile His Lys Ser Ile Asn Val Ile His His Ile Asn Arg Thr Asn Asp Lys Asn His Met Ile Ile Ser Ile Asp Ala Glu Lys Ala Phe Asp Lys Ile Gln Pro Phe Met Leu Lys Thr Leu Ser Lys Leu Gly Ile Asp Gly Thr Tyr Leu Lys Ile Ile Arg Ala Ile Tyr Asp Lys Pro Thr Ala Asn Ile Ile Leu Asn Gly Gln Lys Leu Glu Ala Phe His Leu Thr Thr Gly Ala Arg Gln Gly Cys Pro Leu Leu Leu Leu Leu Phe Asn Val Val Leu Glu Val Leu Ala Arg Ala Ile Arg Gln Glu Lys Glu Ile Lys Gly Val Gln Ile GIy Arg Glu Glu Val Lys Leu Ser Leu Phe Thr Asp Gly Met Ile Val Tyr Leu Gly Asn Pro Ile Val Ser Ala Gln Asn Leu Leu Lys Leu Arg Ser Asn Phe Ser Lys.Val Ser Gly Tyr Lys Ile Asn Val Gln Lys Ser Gln Ala Phe Leu Tyr Thr Asn Asn Arg Gln Thr Glu Ser Gln Ile Met Ser Glu Leu Pro Phe Thr Ile Ala Ser Lys Arg Ile Lys Tyr Leu Gly Ile Gln Leu Thr Arg Asp Val Lys Asp Leu Phe Lys Glu Asn Tyr Lys Pro Leu Leu Asn~Glu Ile Lys Glu Asp Thr Asn Lys Trp Lys Asn Ile,Pro Cys Ser Trp Val Gly Arg Ile Asn Ile Met Lys Met Ala Ile~Leu Pro Lys Val Ile Tyr Arg Phe Asn Gly Ile Pro Ile Lys Leu Pro Met Thr Phe Phe Thr Glu Leu Glu Lys Thr Thr Leu Lys Phe Ile Trp Asn Gln Lys Arg Ala His Ile Ala Lys Thr Ile Leu Ser Gln Lys Asn Lys Ala Gly Gly Ile Met Leu Pro Asp Phe Lys Leu Tyr Tyr Lys Ala Thr Val Thr Lys Thr Ala Trp Tyr Trp Tyr Gln Asn Arg Asp Ile Asp Gln Arg Asn Arg Thr Glu Pro Ser Gln Ile Met Pro His Asn Tyr Asn His Leu 905 ~ 910 915 Ile Phe Asp Lys Pro Asn Lys Asn Lys Lys Trp Gly Lys His Ser Leu Phe Asn Lys Trp Cys Trp Glu Asn Trp Leu Ala Ile Cys Arg Lys Leu Lys Leu Asp Pro Phe Leu Thr Pro Tyr Thr Lys Ile Asn Ser Arg Trp Ile Lys Asp Leu His Val Arg Pro Lys Thr Ile Lys Thr Leu Glu Glu Asn Leu Gly Asn Ile Ile Gln Asp Ile Gly Met Gly Lys Asp Phe Met Ser Lys Thr Pro Lys Ala Met Ala Thr Lys Ala Lys Ile Asp Lys Trp Asp Leu Leu Lys Leu Lys Ser Phe Phe Thr Ala Lys Glu Thr Thr Ile Arg Val Asn Arg Gln Pro Thr Glu Trp~Glu Lys Ile Phe Ala Thr Tyr Ser Ser Asp Arg Gly Leu Ile Ser Arg Tle Tyr Asn Lys Leu Lys Gln Ile Tyr Lys Lys Lys Thr Asn Asn Pro Ile Lys Lys Trp Val Lys Asp Met Asn Arg His Phe Ser Lys Glu Asp 21e Tyr Ala Ala Lys Arg His Met Lys Lys Cys Ser Ser Ser Met Ala Ile Arg Glu Met Gln Ile Lys Thr Ser Met Arg Tyr His Leu Thr Pro Val Arg Met Ala Ile Ile Lys Lys Ser Gly Asn Asn Arg Cys Trp Arg Glu Cys Gly Glu Thr Gly Ile Leu Leu His Cys Trp Trp Asp Cys Lys Leu Val Gln Pro Leu Trp Lys Ser Val Trp Arg Phe Leu Arg Asp.Leu Glu Gln Glu Ile Pro Phe Asp Pro Ala Ile Pro Leu Leu Gly Ile Tyr Pro Lys Asp Tyr Lys Ser Cys Cys Asn Lys Asp Thr Cys Thr Pro.Met Phe Ile Ala Ala Leu Phe Thr Ile Ala Lys Thr Trp Asn Gln Pro Lys Cys Pro Thr Met Val Glu Trp Met Lys Lys Met Trp His Ile Tyr Thr Met Gln Tyr Tyr Ala Ala Ile Lys~Asn Asp Ala Phe Met Ser Phe Val Gly Thr Trp Met Lys Leu Glu Ile Ile Ile Leu Ser Lys Leu Ser Gln Glu Gln Lys Thr Lys His Arg Ile Leu Ser Leu Ile Gly Gly Asn 1265 ' 1270 1275 <210> 34 <211> 635 <212> PRT
<213> Homo Sapiens <220>
<221> misc feature <223> Incyte ID No: 2321130CD1 <400> 34 Met Ala Pro Pro Leu Leu Leu Leu Leu Leu Ala Ser Gly Ala Ala Ala Cys Pro Leu Pro Cys Val Cys Gln Asn Leu Ser Glu Ser Leu Ser Thr Leu Cys Ala His Arg Gly Leu Leu Phe Val Pro Pro Asn Val Asp Arg Arg Thr Val Glu Leu Arg Leu Ala Asp Asn Phe Ile Gln Ala Leu Gly Pro Pro Asp Phe Arg Ser Met Thr Gly Leu Val Asp Leu Thr Leu Ser Arg Asn Ala Ile Thr Arg Ile Gly Ala Arg Ala Phe Gly Asp Leu Glu Ser Leu Arg Ser Leu His Leu Asp Gly Asn Arg Leu Val Glu Leu Gly Thr Gly Ser Leu Arg Gly Pro Val Asn Leu Gln His Leu Ile Leu Ser Gly Asn Gln Leu Gly Arg Ile Ala Pro Gly Ala Phe Asp Asp Phe Leu Glu Ser Leu Glu Asp Leu Asp Leu Ser Tyr Asn Asn Leu Arg Gln Val Pro Trp Ala Gly Ile Gly Ala Met Pro Ala Leu His Thr Leu Asn Leu Asp His Asn Leu Ile Asp Ala Leu Pro Pro Gly Ala Phe Ala Gln Leu Gly Gln Leu Ser Arg Leu Asp Leu Thr Ser Asn Arg Leu Ala Thr Leu Ala Pro Asp Pro Leu Phe Ser Arg Gly Arg Asp Ala Glu Ala Ser Pro Ala Pro Leu Val Leu Ser Phe~Ser Gly Asn Pro Leu His Cys Asn Cys Glu Leu Leu Trp Leu Arg Arg Leu Ala Arg Pro Asp Asp Leu Glu Thr Cys Ala Ser Pro Pro Gly Leu Ala Gly Arg Tyr Phe Trp Ala Val Pro Glu Gly Glu Phe Ser Cys Glu Pro Pro Leu Ile Ala Arg His Thr Gln Arg Leu Trp Val Leu Glu Gly Gln Arg Ala Thr Leu Arg Cys Arg Ala Leu Gly Asp Pro Ala Pro Thr Met His Trp Val Gly Pro Asp Asp Arg Leu Val Gly Asn Ser Ser Arg Ala Arg Ala Phe Pro Asn Gly Thr Leu G1u Ile Gly Val Thr Gly Ala Gly Asp Ala Gly Gly Tyr Thr Cys Ile Ala Thr Asn Pro Ala Gly Glu Ala Thr Ala Arg Val Glu Leu Arg Val Leu Ala Leu Pro His Gly Gly Asn Ser Ser Ala Glu Gly Gly Arg Pro Gly Pro Ser Asp Ile Ala Ala Ser Ala Arg Thr Ala Ala Glu Gly Glu Gly Thr Leu Glu Ser Glu Pro Ala Val Gln Val Thr Glu Val Thr Ala Thr Ser Gly Leu Va1 Ser Trp Gly Pro Gly Arg Pro Ala Asp Pro Val Trp Met Phe Gln Ile Gln Tyr Asn Ser Ser Glu Asp Glu Thr Leu Ile Tyr Arg Ile Val Pro Ala Ser Ser His His Phe Leu Leu Lys His Leu Val Pro Gly Ala Asp Tyr Asp Leu Cys Leu Leu Ala Leu Ser Pro Ala Ala Gly Pro Ser Asp Leu Thr Ala Thr Arg Leu Leu Gly Cys Ala His Phe Ser Thr Leu Pro Ala Ser Pro Leu Cys His Ala Leu Gln Ala His Val Leu Gly.Gly Thr Leu Thr Val Ala Val Gly Gly Val Leu Val Ala Ala Leu Leu Val Phe Thr Val Ala Leu Leu Val Arg Gly Arg Gly Ala Gly Asn Gly Arg Leu Pro Leu Lys Leu Ser His Val Gln Ser Gln Thr Asn Gly Gly Pro Ser Pro Thr Pro Lys Ala His Pro Pro Arg Ser Pro Pro Pro Arg Pro Gln Arg Ser Cys Ser Leu Asp Leu Gly Asp Ala Gly Cys Tyr Gly Tyr Ala Arg Arg Leu Gly Gly Ala Trp Ala Arg Arg Ser.His Ser~Val His Gly Gly Leu Leu Gly Ala Gly Cys Arg Gly Val Gly~Gly Ser Ala Glu.Arg Leu 620 625 ' 630 Glu Glu Ser Val Val <210> 35 <211> 170 <212> PRT
<213> Homo aapi.ens <220>
<222> misc_feature <223> Incyte ID No: 2008365CD1 <400> 35 Met Asn Ala Ala Val Pro Pro Glu Gln Ala His Ser Cys Gly Trp Gly Thr Glu Gly Cys Pro Cys Leu Arg Ser Thr Ala Ile Arg Gln Thr Phe Phe Pro Gly Gly Asp Gln Phe Gln Asn Arg Trp Arg Gly Met Lys Asn Glu Glu His Cys Pro Gly Ser Phe Phe Leu Cys Lys Ile Arg Glu Cys Val Leu Asn Tyr Arg Phe Gln Leu Gln His Pro Gly Phe Gln His Tyr Leu Gln Ser Ser Gly Arg Arg Asp Arg Gly Arg Ser Glu Asp Lys Lys Pro Leu Glu Ala Gly Val Trp Cys Trp Asp Arg Gly Gly Trp Asp Gly Ser Ser Arg Ala Val His Leu Leu Phe Arg Gly Val Ala His Pro Ser Leu Tyr Leu Phe Pro Arg Glu Asp Pro Pro Arg Leu Leu Phe Pro Arg Leu Ser Leu Leu Val Cys Glu Gln Phe Trp Cys Tyr Ser Ala Thr Leu Leu Leu Ala Pro Leu Pro Ala Ser Thr Cys <210> 36 <211> 388 <212> PRT
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 3580778CD1 <400> 36 Met Asn Thr Met Tyr Val Met Met Ala Gln Ile Leu Arg Ser His Leu Ile Lys Ala Thr Val Ile Pro Asn Arg Val Lys Met Leu Pro Tyr Phe Gly Ile Ile Arg Asn Arg Met Met Ser Thr His Lys Ser Lys Lys Lys Ile Arg Glu Tyr Tyr Arg Leu.Leu Asn Val Glu Glu Gly Cys Ser Ala Asp Glu Val Arg Glu Ser Phe His Lys Leu Ala Lys Gln Tyr His Pro Asp Ser Gly Ser Asn Thr Ala Asp Ser Ala Thr Phe Ile Arg Ile Glu Lys Ala Tyr Arg Lys Val Leu Ser His Val Ile Glu Gln Thr Asn Ala Ser Gln Ser Lys Gly Glu Glu Glu Glu Asp Val Glu Lys Phe Lys Tyr Lys Thr Pro Gln His Arg His Tyr Leu Ser Phe Glu Gly Ile Gly Phe Gly Thr Pro Thr Gln Arg Glu Lys His Tyr Arg Gln Phe Arg Ala Asp Arg Ala Ala Glu Gln Val Met Glu Tyr Gln Lys Gln Lys Leu Gln Ser Gln Tyr Phe Pro Asp Ser Val Ile Val Lys Asn Ile Arg Gln Ser Lys Gln Gln Lys Ile Thr Gln Ala Ile Glu Arg Leu Val Glu Asp Leu Ile Gln Glu Ser Met Ala Lys Gly Asp Phe Asp Asn Leu Ser Gly Lys Gly Lys Pro Leu Lys Lys Phe Ser Asp Cys Ser Tyr Ile Asp Pro Met Thr His Asn Leu Asn Arg Ile Leu Ile Asp Asn Gly Tyr Gln Pro Glu Trp Ile Leu Lys Gln Lys Glu Ile Ser Asp Thr Ile Glu Gln Leu Arg Glu Ala Ile Leu Val Ser Arg Lys Lys Leu Gly Asn Pro Met Thr Pro Thr Glu Lys Lys Gln Trp Asn His Val Cys Glu Gln Phe G1n Glu Asn Ile Arg Lys Leu Asn Lys Arg Ile Asn Asp Phe Asn Leu Ile Val Pro Ile Leu Thr Arg G1n Lys Val His Phe Asp Ala Gln Lys G1u Ile Val Arg A1a Gln Lys Ile Tyr Glu Thr Leu Ile Lys Thr Lys Glu Val Thr Asp Arg Asn Pro Asn Asn Leu Asp Gln Gly Glu G1y Glu Lys Thr Pro Glu Ile Lys Lys Gly Phe Leu Asn Trp Met Asn Leu Trp Lys Phe Ile Lys Tle Arg Ser Phe <210> 37 <211> 347 <212> PRT
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 7948785CD1 <400> 37 Met Val Ala Ser Thr Val A1a Asn Gly Pro Arg Ser Pro Trp Asp . 10 15 Ala Ile Ser Leu I1e Ile Met Leu Arg Ile Trp Arg Val Lys Arg Val Ile Asp Ala Tyr Val Leu Pro Val Lys Leu Glu Met Glu Met Val Ile Gln Gln Tyr Glu Lys Al.a Lys Val Ile Gln Asp Glu Gln Leu Glu Arg Leu Thr Gln I1e Cys G1n Glu Gln Gly Phe Glu Ile Arg Gln Leu Arg Ala His Leu Ala Gln Gln Asp Leu Asp Leu Ala A1a Glu Arg Glu Ala Ala Leu Gln Ala Pro His Val Leu Ser Gln Pro Arg Ser Arg Phe Lys Val Leu Glu Ala Gly Thr Trp Asp Glu Glu Thr Ala Ala G1u Ser Val Val Glu Glu Leu Gln Pro Ser Gln G1u Ala Thr Met Lys Asp Asp Met Asn Ser Tyr Ile Ser Gln Tyr Tyr Asn Gly Pro Ser Ser Asp Ser Gly Val Pro Glu Pro Ala Val Cys Met Val Thr Thr Ala Ala Ile Asp Ile His Gln Pro Asn Ile Ser Ser Asp Leu Phe Ser Leu Asp Met Pro Leu Lys Leu Gly Gly Asn Gly Thr Ser A1a Thr Ser Glu Ser Ala Ser Arg Ser Ser Val Thr Arg Ala Gln Ser Asp Ser Ser Gln Thr Leu Gly Ser Ser Met Asp Cys Ser Thr A1a Arg G1u Glu Pro Ser Ser Glu Pro GIy Pro 230 235 ' 240 Ser Pro Pro Pro Leu Pro Ser Gln Gln Gln Val Glu Glu Ala Thr Val Gln Asp Leu Leu Ser Ser Leu Ser Glu Asp Pro Cys Pro Ser Gln Lys Ala Leu Asp Pro Ala Pro Leu Ala Arg Pro Ser Pro Ala Gly Ser Ala Gln Thr Ser Pro Glu Leu Glu His Arg Val Ser Leu Phe Asn Gln Lys Asn Gln Glu Gly Phe Thr Val Phe Gln Ile Arg Pro Val Ile His Phe Gln Pro Thr Val Pro Met Leu Glu Asp Lys Phe Arg Ser Leu Glu Ser Lys Glu Gln Lys Leu His Arg Val Pro Glu Ala <210> 38 <211> 338 <212> PRT
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 7494415CD1 <400> 38 .
Met Arg Lys Asn Gln Pro Gln Lys Thr Glu Asn Ser Lys Lys Gln 1 5 10 ' . 15 Asn Gly Ser Ser Pro Ser Lys Asp His Asp Phe Ser Pro Ala Arg Glu Gln Thr Trp Met Glu Asn Glu Phe Asp Lys Gly Thr Glu Glu Gly Phe Arg Asn Asn Lys Pro Ser Glu Leu Gln Glu His Val Leu Thr Gln Cys Lys Glu Ala Lys Asn Leu Glu Lys Gly Leu Glu Glu Leu Leu Thr Arg Ile Thr Ser Leu Glu Lys Asn Thr Asn Asp Leu Ile Glu Leu Lys Asn Thr Ala Gln Glu Leu His Glu Ala Tyr Thr Ser Ile Asn Ser Pro Ile Asp Gln Ala Glu Glu Arg Ile Ser Glu Ile Glu Asp Gln Leu Asn Lys Ile Lys Cys Glu Gly Lys Phe Arg .

Glu Lys Arg Ile Lys Arg Asn Lys Gln Ser Leu Gln Glu Ile Trp Asp Tyr Val Lys Arg Pro Asn Leu His Leu Ile Gly Val Pro Glu 155 .160 165 Ser Asp Gly Glu Asn Gly Thr Lys Leu Glu Asn Thr Leu Gln Asp Ile Ile GIn Glu Asn Phe Pro Asn Leu Ala Arg Gln AIa Asn Ile Gln Ile Gln Glu Ile Gln Arg Thr Leu Gln Arg Tyr Ser Ser Arg Arg Ala Thr Pro Arg His Ile Ile Val Arg Phe Thr Lys Val Glu Met Lys Glu Lys Met Leu Arg Ala Ala Arg Glu Lys Gly Arg Val Thr Leu Lys Trp Lys Pro Ile Arg Leu Thr Val Asp Leu Ser Ala Gly Thr Leu Leu Ala Arg Arg Glu Trp Gly Pro Ile Phe Asn Ile Leu Lys Glu Lys Asn Phe Gln Ser Arg Ile Ser Tyr Pro Ala Lys Leu Ser Phe Ile Arg Glu Gly Glu Ile Lys Tyr Phe Thr Asp Lys Gln Met Leu Arg Asp Phe Val Thr Thr Arg Pro Ala Leu Lys Glu Leu Leu Lys Glu Ala Leu Asn Met Glu Arg Asn Asn Arg Tyr Gln Leu Leu Gln Asn His Ala Lys Met <210> 39 <211> 520 <212> PRT
<213> Homo sapiens <220>
<221> misc_feature <223> Incyte Ib No: 2234223CD1 <400> 39 Met Gly Lys Lys Leu Asp Leu Ser Lys Leu Thr Asp Glu Glu Ala Gln His Val Leu Glu Val Val Gln Arg Asp Phe Asp Leu Arg Arg Lys Glu Glu Glu Arg Leu Glu Ala Leu Lys Gly Lys Ile Lys Lys Glu Ser Ser Lys Arg Glu Leu Leu Ser Asp Thr Ala His Leu Asn Glu Thr His Cys Ala Arg Cys Leu Gln Pro Tyr Gln Leu Leu Val Asn Ser Lys Arg Gln Cys Leu Glu Cys Gly Leu Phe Thr Cys Lys Ser Cys Gly Arg Val His Pro Glu Glu Gln Gly Trp Ile Cys Asp Pro Cys His Leu Ala Arg Val Val Lys Ile Gly Ser Leu Glu Trp Tyr Tyr Glu His Val Lys Ala Arg Phe Lys Arg Phe Gly Ser Ala Lys Val Ile Arg Ser Leu His Gly Arg Leu Gln Gly Gly Ala Gly Pro Glu Leu Ile Ser Glu Glu Arg Ser Gly Asp Ser Asp Gln Thr Asp Glu Asp Gly Glu Pro Gly Ser Glu Ala Gln Ala Gln Ala Gln Pro Phe Gly Ser Lys Lys Lys Arg Leu Leu Ser Val His Asp Phe Asp Phe Glu Gly Asp Ser Asp Asp Ser Thr Gln Pro Gln Gly His Ser Leu His Leu Ser Ser Val Pro Glu Ala Arg Asp Ser Pro Gln Ser Leu Thr Asp Glu Ser Cys Ser Glu Lys Ala Ala Pro His Lys Ala Glu Gly Leu Glu Glu Ala Asp Thr Gly Ala Ser Gly Cys His Ser His Pro Glu Glu Gln Pro Thr Ser Ile Ser Pro Ser Arg His Gly Ala Leu Ala Glu Leu Cys Pro Pro Gly Gly Ser His Arg Met Ala Leu Gly Thr Ala Ala Ala Leu Gly Ser Asn Val Ile Arg Asn Glu Gln Leu Pro Leu Gln Tyr Leu Ala Asp Val Asp Thr Ser Asp Glu Glu Ser Ile Arg Ala His Val Met Ala Ser His His Ser Lys Arg Arg Gly Arg Ala Ser Ser Glu Ser Gln Gly Leu Gly Ala Gly Ala Arg Thr Glu Ala Asp Val Glu Glu Glu Ala Leu Arg Arg Lys Leu Glu Glu Leu Thr Ser Asn Val Ser Asp Gln Glu Thr Ser Ser Glu Glu Glu Glu Ser Lys Asp Glu Lys Ala Glu Pro Asn Arg Asp Lys Ser Val Gly Pro Leu Pro Gln Ala Asp Pro Glu Val Ser Asp Ile Glu Ser Arg Ile Ala Ala Leu Arg Ala Ala Gly Leu Thr Val Lys Pro Ser Gly Lys Pro Arg Arg Lys Ser Asn Leu Pro Ile Phe Leu Pro Arg Val Ala Gly Lys Leu Gly~Lys Arg Pro Glu Asp. Pro Asn Ala Asp Pro Ser Ser Glu Ala Lys Ala Met Ala Val Pro Tyr Leu Leu Arg Arg Lys Phe Ser Asn Ser Leu Lys Ser Gln Gly Lys Asp Asp Asp Ser Phe Asp Arg Lys Ser Val Tyr Arg Gly Ser Leu Thr Gln Arg Asn Pro Asn Ala Arg Lys Gly Met Ala Ser His Thr Phe Ala Lys Pro Val Val Ala His Gln Ser <210> 40 <211> 733 <212> DNA
<213> Homo Sapiens <220>
<221> misc feature <223> Incyte ID No: 4973222CB1 <400> 40 acgggaacga aagagaagct ctatctcccc tccaggagcc cagctatgaa ctccttctcc 60 acaagcgcct tcggtccagt tgccttccct gccccagtac ccccaggaga agattccaaa 120 gatgtagccg ccccacacag acagccactc acctcttcag aacgaattga caaacaaatt 180 cggcacatcc tcgacggcat ctcagccctg agaaaggaga catgtaacaa gagtaacatg 240 tgtgaaagca gcaaagaggc actggcagaa aacaacctga accttccaaa gatggctgaa 300 aaagatggat gcttccaatc tggattcaat gaggagactt gcctggtgaa aatcatcact 360 ggtcttttgg agtttgaggt atacctagag tacctccaga acagatttga gagtagtgag 420 gaacaagcca gagctgtgca gatgagtaca aaagtcctgg tccagttcct gcagaaaaag 480 gcaaagaatc tagatgcaat aaccacccct gacccaacca caaatgccag cctgctgacg 540 aagctgcagg cacagaacca gtggctgcag gacatgacaa ctcgtctcat tctgcgcagc 600 tttaaggagt tcctgcagtc cagcctgagg gctcttcggc aaatgtagca tgggcacctc 660 agattgttgt tgttaatggg cattccttct tctggtcaga aacctgtcca ctgggcacag 720 aacttatgtt gtt 733 <210> 41 <211> 2502 <212> DNA
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 55009060CB1 <400> 41 ggaagctcca aagagaaagg agggcaggaa gcccacggcc cacaggggtg tagcccgaga 60 cccacctgca gcccccagcc cttgccagga aagcagccgc agccatggcg gggatgaaga 120 cagcctccgg ggactacatc gactcgtcat gggagctgcg ggtgtttgtg ggagaggagg 180 acccagaggc cgagtcggtc accctgcggg tcactgggga gtcgcacatc ggcggggtgc 240 tcctgaagat tgtggagcag atcaatcgca agcaggactg gtcagaccat gctatttggt 300 gggaacagaa gaggcagtgg ctgctgc'aga cccactggac actggacaag tacgggatcc 360 tggccgacgc acgcctcttc tttgggcccc agcaccg'gcc cgtcatcctt cggttgccca 420 accgccgcgc actgcgcctc cgtgccagct tctcccagcc cctcttccag gctgtggctg 480 ccatctgccg cctcctcagc atccggcacc ccgaggagct gtccctgctc cgggctcctg 540 agaagaagga gaagaagaag aaagagaagg agccagagga agagctctat gacttgagca 600 aggttgtctt ggctgggggc gtggcacctg cactgttccg ggggatgcca gctcacttct 660 cggacagcgc ccagactgag gcctgctacc acatgctgag ccggccccag ccgccacccg 720 accccctcct gctccagcgt ctgccacggc ccagctccct gtcagacaag acccagctcc 780 acagcaggtg gctggactcg tcgcggtgtc tcatgcagca gggcatcaag gccggggacg 840 cactctggct gcgcttcaag tactacagct tcttcgattt ggatcccaag acagaccccg 900 tgcggctgac acagctgtat gagcaggccc ggtgggacct gctgctggag gagattgact 960 gcaccgagga ggagatgatg gtgtttgccg ccctgcagta c'cacatcaac aagctgtccc 1020 agagcgggga ggtgggggag ccggctggca cagacccagg gctggacgac ctggatgtgg 1080 ccctgagcaa cctggaggtg aagctggagg ggtcggcgcc cacagatgtg ctggacagcc 1140 tcaccaccat cccagagctc aaggaccatc tccgaatctt tcggccccgg aagctgaccc 1200 tgaagggcta ccgccaacac tgggtggtgt tcaaggagac cacactgtcc tactacaaga 1260 gccaggacga ggcccctggg gaccccattc agcagctcaa cctcaagggc tgtgaggtgg 1320 ttcccgatgt taacgtctcc ggccagaagt tctgcattaa actcctagtg ccctcccctg 1380 agggcatgag tgagatctac ctgcggtgcc aggatgagca gcagtatgcc cgctggatgg 1440 ctggctgccg cctggcctcc aaaggccgca ccatggccga cagcagctac accagcgagg 1500 tgcaggccat cctggccttc ctcagcctgc agcgcacggg cagtgggggc ccgggcaacc 1560 acccccacgg ccctgatgcc tctgccgagg gcctcaaccc ctacggcctc gttgcccccc 1620 gtttccagcg aaagttcaag gccaagcagc tcaccccacg gatcctggaa gcccaccaga 1680 atgtggccca gttgtcgctg gcagaggccc agctgcgctt catccaggcc tggcagtccc 1740 tgcccgactt cggcatctcc tatgtcatgg tcaggttcaa gggcagcagg aaagacgaga 1800 tcctgggcat cgccaacaac cgactgatcc gcatcgactt ggccgtgggc gacgtggtca 1860 agacctggcg tttcagcaac atgcgccagt ggaatgtcaa ctgggacatc cggcaggtgg 1920 ccatcgagtt tgatgaacac atcaatgtgg ccttcagctg cgtgtctgcc agctgccgaa 1980 ttgtacacga gtatatcggg ggctacattt tcctgtcgac gcgggagcgg gcccgtgggg 2040 aggagctgga tgaagacctc ttcctgcagc tcaccggggg ccatgaggcc ttctgagggc 2100 tgtctgattg cccctgccct gctcaccacc ctgtcacagc cactcccaag cccacaccca 2160 caggggctca ctgccccaca cccgctccag gcaggcaccc agctgggcat ttcacctgct 2220 gtcactgact ttgtgcaggc caaggacctg gcagggccag acgctgtacc atcacccagg 2280 ccagggatgg gggtgggggt ccctgagctc atgtggtgcc ccctttcctt gtctgagtgg 2340 ctgaggctga tacccctgac ctatctgcag tcccccagca cacaaggaag accagatgta 2400 gctacaggat gatgaaacat ggtttcaaac gagttctttc ttgttacttt ttaaaatttc 2460 ttttttataa attaatattt tattgttaaa aaaaaaaaaa as 2502 <210> 42 <211> 1591 <212> DNA
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 1985092CB1 <400> 42 ctcctcccga ggaaccagtg gtgacagctg aggccatgtg agtaggatcc tgaatgaggc 60 tttatctctg gctgttcgtc ccatcgtcca ccgtggcacc agctccctca gccagccggg 120 atgggaccag cgactgagag agccagaggc agagaggtga gggtgaccat atcctggact 180 gtgagaggaa tgggactctg ggcctgtagc tgccaagcag gtggcaggtg ctccaggctg 240 tgatctgcac cctctgaccc ctgacattga cctcctaccc tgacccctgc ctgaccaagc 300 catgtctgaa caggaggctc aagccccagg :gggccggggg ctgcccccgg acatgctggc 360 agagcaggtg gagctgtggt ggtcccagca gccgcggcgc tcggcgctct gcttcgtcgt 420 ggccgtgggc ctcgtggcag gctgtggcgc.gggcggcgtg gcactgctgt caaccaccag 480 .
cagccgctca ggtgaatggc ggctagcaac,gggcactgtg ctctgtttgc tggctctgct 540 ggttctggtg aaacagctga tgagctcggc-tgtgcaggac atgaactgca tccgccaggc 600 ccaccatgtg gccctgctgc gcagtggtgg aggggccgac gccctcgtgg tgctgctcag 660 tggcctcgtg ctgctggtca ccggcctgac cctggccggg ctggccgccg cccctgcccc 720 tgctcggccg ctggcc_gcca tgctgtctgt gggcattgct ctggctgcct tgggctcgct 780 tttgctgctg ggcctgctgc tgtatcaagt.gggtgtgagc ggacactgcc cctccatctg 840 tatggccact ccctccaccc acagtggcca tggcggccat ggcagcatct tcagcatctc 900 aggacagttg tctgctggcc ggcgtcacga gaccacatcc agcattgcca gcctcatctg 960 acggagccag agccgtcctt cttctcacag cggcctcagc gtccccagag ccgagccagg 1020 gtgtgagtgc atgtgaacgt tgagtacaca tgagtgcgtg tatgccccca ggctgggtca 1080 gctcttctgt ggattgcatg gcgtgtgatt-aaaagcccat gtgttcccac acatccacat 1140 catgggaagg ttaatgtgtg cctccttgga actgggtgtt ggtgtccatg gaacttcctc 1200 tctgtatctc aggtcagtag gcgcagaaac gcctcatgat gaagattctt gagccccatt 1260 tccaagaccc ctcacatcca atcctgtcct gtaacatcca tcaaggattt ccataggggt 1320 gactggtgcc cacccaagac tgcaccagtg cctgctcatt gaggagagta actgctggcc 1380 aggcagaaag aatatgggct ctgcaatgag acagacctgg aggggactct cccgttgagc 1440 actagcagct ggaggagttg ggagttcatg gctatcatgg ttgtgttaat cgattgtggg 1500 gatgaaatgt cattgtgtat ggaaggcggg gctcatggct gattggcaat aaaatggcgg 1560 ctgccgttgt cattgtctcc aaaaaaaaaa a 1591 <210> 43 <211> 1210 <212> DNA
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 1553593CB1 <400> 43 gaaactcagc tgttaaaacc gctgcctctg cactttggaa tcccatctgg agactegcta 60 agcgtcccag ccgcatccct cccgcagcga cggcggcccg ggacccgcgg gctgtgaacc 120 atgaacaccc gcaatagagt ggtgaactcc gggctcggcg cctcccctgc ctcccgcccg 180 acccgggatc cccaggaccc ttctgggcgg caaggggagc tgagccccgt ggaagaccag 240 agagagggtt tggaggcagc ccctaagggc ccttcgcggg agagcgtcgt gcacgcgggc 300 cagaggcgca caagtgcata caccttgata gcaccaaata taaaccggag aaatgagata 360 caaagaattg cggagcagga gctggccaac ctggagaagt ggaaggagca gaacagagct 420 aaaccggttc acctggtgcc cagacggcta ggtggaagcc agtcagaaac tgaagtcaga 480 cagaaacaac aactccagct gatgcaatct aaatacaagc aaaagctaaa aagagaagaa 540 tctgtaagaa tcaagaagga agctgaagaa gctgaactcc aaaaaatgaa ggcaattcag 600 agagagaaga gcaataaact ggaggagaaa aaaagacttc aagaaaacct tagaagagaa 660 gcatttagag agcatcagca atacaaaacc gctgagttct tgagcaaact gaacacagaa 720 tcgccagaca gaagtgcctg tcaaagtgct gtttgtggcc cacaatcctc aacatggaaa 780 cttcctatcc tgcctaggga tcacagctgg gccagaagct gggcttacag agattctcta 840 aaggcagaag aaaacagaaa attgcaaaag atgaaggatg aacaacatca aaagagtgaa 900 ttactggaac tgaaacggca gcagcaagag caagaaagag ccaaaatcca ccagactgaa 960 cacaggaggg taaataatgc ttttctggac cgactccaag gcaaaagtca accaggtggc 1020 ctcgagcaat ctggaggctg ttggaatatg aatagtggta acagctgggg tatatgagaa 1080 aatattgact cctatctggc cttcatcaac tgacctcgaa aagcctcatg agatgctttt 1140 tcttaatgtg attttgttca gcctcactgt ttttacctta atttcaactg cccacacact 1200.
tgaccgtgca 1210 <210> 44 <211> 3112 <212> DNA
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 1954122CB1 .
<400> .44 cttgaccgct ctaagccgcc gcgcacctgc gttttctccg gtacaggttg aggctactaa 60 ggtatttggc tattataaat aaaatagtga tgaacacgtg tacaggtgtt tgtacggaca 120 taagttgtca tttctcttga gtaaatacct aaaagtgggc ttgctggagt tttactaccc 180 ttggatgctg ctgttgacat ggaaaagatt gaggaacaat ttgctaatct gcacattgtt 240 aaatgttcct taggaaccaa agagcccact taccttcttg gtatagacac atcaaagact 300 gtccaagcag gaaaggaaaa cttggttgct gttttatgtt ctaatggatc aatcagaata 360 tatgataaag aaaggttaaa tgtactacga gaatttagtg gatatcctgg acttcttaat 420 ggagtcagat ttgcaaattc ctgtgacagt gtatattcag catgtactga tggcactgtg 480 aaatgctggg,atgctcgagt agccagagaa aaacctgttc agctcttcaa gggttaccct 540 tccaatattt ttatcagttt tgatattaat tgtaatgatc atattatttg tgctggtaca 600 gaaaaagttg atgatgatgc attgttggtg ttttgggatg caaggatgaa ttctcagaat 660 ttatctacaa ctaaagactc acttggtgca tattcagaga cacatagtga tgatgtcact 720 caagtacgtt tccatcccag caatcccaac atggtagtct caggttcatc tgatggcctg 780 gtaaatgtat ttgatattaa tattgataat gaggaggatg cactggttac aacctgtaac 840 tcaatttcat cagtaagctg tattggttgg tctgggaaag gttataaaca gatttactgc 900 atgacacatg atgaaggatt ttattggtgg gatcttaatc atctggacac tgatgaacca 960 gttacacgtt tgaacatcca ggatgtcaga gaagtagtta acatgaaaga agatgctttg 1020 gactatttga ttggtggcct atatcatgaa aagacagaca cattgcatgt tattggagga 1080 acaaacaaag gaaggattca tttgatgaac tgcagcatgt caggactgac ccatgtgact 1140 agccttcagg gagggcatgc tgctacagtc cgttctttct gttggaatgt gcaagatgat 1200 tctttgttga ctggaggaga agatgcacag ttgttacttt ggaaacctgg agctatagag 1260 aagaccttta caaagaaaga gagtatgaaa atagcatcct ctgtgcacca acgagtacga 1320 gttcatagta atgattctta taaaagaagg aaaaagcagt gataatgcat ttggcacttt 1380 gtttggtagg ttttatagtt tcaaatagtc cttcttgttt actacccatg gtagacatgt 2440 ttaaagcttt atgtaaaaac aagccagtta gcaaacagtc ctggaaaaat atggtgagaa 1500 tggtttaatt ccggtcttca catattctaa aaaattttaa agcctctaag tataaaatca 1560 gtgagttgaa agtaaaattt cttatttaaa gaacccatct gttaatgtag taaatgttgg 1620 ctttaaagaa atagctccga taaggacttt ttagaaggaa atacttggta aaacttaata 1680 aaacaggtgg ttggctgcat tttttaagcc cagtttttta cattttaaat tgtctttatt 1740 gtgtataaaa ccagattaga ctatattctt catctgagga gcatctatat gtttaataca 1800 cctaatgtat tatgctggag tatttaaatg taagtatttt aatgagtaaa tagaattaac 1860 cttttttaca aatgaaatga taggcatcta tgtaaagtga gaaaaatatg tttcgtaaat 1920 atttgcatct ttaatattgt tagtaagtag taaacaggtg ttttgctgat tgaaaggaag 1980 ttatcttggt aaattgagac tcaaagtgaa atacaaaaat gaaaatattt ataataggac 2040 tcgacttgga gaaattgcta ttcttcctgt tgtccaaacc ataaatctga ggataacctg 2100 ggcttttctc tttcctcttc tcctaaccag tttttcacca agtcctgtgc catttcacct 2160 cctaacgtct gtcagctatc cttttgcctt tacctcacca ctattaccct agagaagact 2220 ttcattattt ctcatctgga ttaccagctc gatctctaat ctgcttcagt tcatccttct 2280 tgctacttct aggctaaaca tcaaaaacag atctggtagg ggcggggaaa tgagggggaa 2340 gaaacaaaaa cgtgatggtg cctcatgctg cttaaaatct tcagtacatt gatgttttga 2400 tggcggacta cataagcgtt aaaaattgtg tttttcagaa tctttaaaat ataagacagt 2460 gctatctagt gaataaaaaa attagtttga aagatatctg gagaaatcgc attcataaaa 2520 caattggaag tgaaactatt aaaacaatag ggctttttaa aattaaaaat atttaaaatt 2580 caaaagtaat taatagtgtt ggaagatgta ggtgagaaaa tattcctgaa agtagaactg 2640 aaagagacaa agagaaaaga tgaaagccac agaagataaa tacaggggtc aaaaccagac 2700 taacagtttt agaaagtgaa aaaagttaaa aaagaaatgg gggcagtggg ttattagaaa 2760' taacataaat ggctggtatg gtttgtctgt~gtcctcaccc aaatttcatc tcgaattgta 2820 atccccataa tccccatgtg tctagggaga gacctggtgg gaggtgattg gatcatgggg 2880 gtggtttccc ctacgatgtt ctcctgatag tgggtgagtt ctcacaagat ctgatggttt 2940 tataaagggc tctgcccctt taactcctca ctctttctcc tgaagccttg tgaagaaggt 3000 gctttgcttc ccctttgcgt tcccccatga ttgtaagttt cttgaggcct acctggccat 3060 gctaactctt cagtcaatac actcgtttct tctctgtgtg cgctgcgtct tc 3112 <210> 45 <211> 2398 <212> DNA
<213> Homo Sapiens <220>
<221> misc_feature.
<223> Incyte ID No: 3159276CB1 <400> 45 cgaccacaca tgagctggga gttccagtac ctgggtgggc ctccccattg aaagtgtggg 60 agctccctgg atagggctga gtcttccccc gcagagtgag agttaccgaa gcaggctctg 120 tctcctccat cagactggaa actcccagag ctgggggatg atgtctctct cccccatcag 180 tttgggagct ctggagggca gaaatgtgga tctcttgcag atcagcggac agctctcccc 240 tcgcctcttc cggaagctgc ctccccgggt gtgcgtgtcc ctcaagaaca ttgtggatga 300 ggacttcctc tatgcaggac atatcttcct gggcttttcc aaatgcggcc gctacgtcct 360 ctcctacacc agcagcagtg gggatgacga cttctccttc tacatctacc atctgtactg 420 gtgggagttc aacgttcaca gcaagctcaa gctggtccgg caggttcggc tattccagga 480 cgaggagatc tacagcgacc tgtacctgac cgtatgcgag tggcccagcg acgcctccaa 540 ggtcatcgtc ttcggcttca acacccgctc ggccaacggg atgctcatga acatgatgat 600 gatgagtgac gagaaccacc gtgacatcta cgtcagcacc gtggccgtgc caccgccagg 660 ccgctgtgct gcttgccagg atgccagccg agcccaccca ggagacccga atgcacagtg 720 cctacggcat ggcttcatgc tgcacaccaa gtaccaggtg gtctacccct tccccacctt 780 ccagcccgcc ttccagctca agaaggacca ggtggtgctg ctcaacacca gctactccct 840 ggtggcctgc gccgtctccg tccactcggc aggtgacagg agtttctgcc aaatcctgta 900 tgaccacagc acctgccccc tggcgcctgc cagcccccct gagccccaga gcccagagct 960 gccccctgcc ctccccagct tctgccctga ggcggcccca gcccgttctt ctgggtctcc 1020 tgagccctcg cccgccattg ccaaagccaa ggagtttgtg gctgacatct tccgccgggc 1080 caaagaggcc aagggcgggg tccctgagga agcccggcct gccctgtgcc caggaccctc 1140 tggcagccgc tgccgtgcgc actctg.agcc cctagccctg tgtggagaga cggcaccccg 2200 ggacagcccc cctgcctcgg aggcacctgc ctccgagcct ggctatgtca actacaccaa 1260 gctgtactat gtgctggagt ccggagaggg gacggagccg gaggatgagt tggaggacga 1320 caagatctcc ctgcecttcg tggtgactga tcttcgtggc cgcaacctgc ggcccatgcg 1380 ggagcggact gctgtccagg gccagtacct gacagtggag cagctcacac tagacttcga 1440 atatgttatc aatgaggtca tccgccacga cgctacctgg ggccatcagt tctgttcttt 1500 cagcgactat gacatcgtca ttctggaggt ctgcccagaa accaaccagg tcctcatcaa 1560 cattggcctg ctgctcctgg ccttcccgtc ccccactgag gagggccagc tccgaccaaa 2620 gacctatcac accagcctca aggtggcatg ggacctcaac-acagggatct tcgagacagt 1680 cagtgtaggc gacctgactg aggtcaaagg gcagaccagc ggcagtgtct ggagctccta 1740 ccgcaagagc tgcgtggaca tggtcatgaa gtggetggtg ccggagagca gcggccgcta 180.0 cgtcaacagg atgaccaatg aggcgctgca caaagggtgc tccctgaagg ttctggcgga 1860 cagcgagcga tatacgtgga tcgtgctgtg agggccaggc cgccccggac actgactcca 1920 actacctccg tggcctggga ccggccccct tcctggggtg gcctcttcct ggccggctgg 1980 cccaccgact gatgaccggc actagtgtta gcctgcggaa cggggctggg cagggcagcc 2040 tctgttggcc tgagggtctg gacgcttttt atttatgcct atttaagttg ggaaggggca 2100 gagagagggc gccccctgcc ccaccagcct gagtgccccg ccttcacccc gagctgggca 2160 tgggcctggc ccctcgtgca tttgcccttt tctcggctac agctgtggac gttgccctcg 2220 gggaggtcga atggacccca ttccccctgc cctgcccgcc cccagcctcc ccacccaggc 2280 cggcaacctg gccatcccca ttccgttctt cttcatgtaa taaatgtttt aatttctgaa 2340 cctcaaaaaa aaaaaaaaaa aaaaaaaaag ggcggccgct cgcgatctag aactagtc 2398 <210> 4&
<211> 2127 <212> DNA
<213> Homo Sapiens <220>
<221> mist feature <223> Incyte ID No: 140052CB1 <400> 46 gaccccaaga tggcggcgcc cagcgaagtg gccgcgatag cccctggcga aggcgatggc 60 ggaggcggcg gctttggctc ctggctggac ggacggttgg aggcactggg agtggaccga 120 gccgtttatg gagcctacat cttgggtatc ctgcaggagg aggaggaaga agagaagctg 180 gacgctctgc aggggatcct ctctgctttc ctggaagaag attccctcct taatatctgc 240 aaggagattg tggaacgatg gtcagaaact cagaatgttg tcaccaaagt gaaaaaagaa 300 gatgaagtac aggccattgc caccctaatt gagaagcagg cacaaatcgt agtaaagcca 360 aggatggtgt cagaagagga gaagcagaga aaagctgccc tcctggccca gtatgctgat 420 gtgacagatg aagaggatga agcagatgag aaggatgatt caggtgctac cacaatgaac 480 attggttctg acaaacttct gttccgaaac accaatgtgg aagatgtcct taatgcccga 540 aaactggagc gagactcact tcgggatgaa tcccaaagga agaaggaaca ggacaagctg 600 cagagggaga gagacaaact agccaagcag gagcgcaagg aaaaggaaaa aaaaaggaca 660 cagagagggg agcgaaagcg ataaccttgg cctactctat tctttccccc catgaactta 720 tcaaagggag aactggttgt gtatttaaga gaaatgagag tgcattgcaa agtagtttcc 780 caaggcattt cccctattgt ttacatggtc aaaaggaaaa tcaaaatcac ttctaattac 840 aaaatgtgct gttttggtgg ggtgggcaat cagattatta tagttgatga ctgtaccaaa 900 gatctggaat gggggccacc tttatatttt aatactcaag tcctgtgctc cttttggcca 960 ctttaaaaag tettccctca ccttggataa ataggattca gaaagctaac agttttcatt 1Q20 ctgaaggagc agaaacataa tcaatccctc cctgagataa aatgtgggca tgcacattgc 1080 tttggagtca gtattcagct gtaagtgtag tattgtttta tggctgaaat agaaagctaa 1140 tgaagaggcc tacctggatc cagattttat gttaacatgg cagcaaaact aggaaaatgg 1200 gaagaaatta cttgcggggt gttctagatc tgattttttt tttttttttg aaagtttcag 1260 cctgtgggag gaaaagaggt atcttttaaa atagagatag ttacatattt tcctcatcta 1320 ggctgtgtca ctattgggtg caaggatatc attgttggct agattcattt tgtataatca 1380 tgtatcctct tgtgtgctgg tagagatttt aatcctgatt tttccataaa acatgagtat 1440 taagaaataa ttcctggttt ggagaaactg gagaaatcac ccttttaagg aagaaacact 1500 ggaaatttct gctaacacca agatatttaa gagtgtacat agtaggtgct caacaaattt 1560 attgaatgaa tgagtgaatg gaaaaactgg gagagtcaaa agtgagcaga agctctccat 1620 ttctacttct gtcacaaacc acattaaatt gtaaataagg cccttctcca cttgacttca 1680 ggcagcagat tgtctagaag cctaaggaca gcaatttctc tgacaagaca aagtagatat 1740 tttataccag gggttggcaa actactgccc acgggccgaa tttggcccag tctgtttttg 1800 tatggtgcaa actaaaaatg atttttacat ttttaaagag ttataaaaga aaaaaatatg 1860 tggtctgtga aatctaaaat atttactacc tggcctgttg gaggaaaggt ttgccaatct 1920 ctgttttata ccattaacta tgagattaac aaaaactttt acctttgtgc agaaggttaa 1980 aaaaaaaaaa acatggttaa ggaaaaggag acatgttacc tcttcataca ctcctataac 2040 tgtggcattg caaaaaataa aaataaccac ctttaaaaaa ataaatctta tttaaattgc 2100 aaaaaaaaaa aaaaaaaaag atcggtc 2127 <210> 47 <211> 2407 <212> DNA
<213> Homo sapiens <220>
<221> misc_feature <223> Incyte ID No: 5158048CB1 <400> 47 ggagagccag gcgctaacca gccgctctgc gccccgcgcc ctgcttgccc ccattatcca 60 gccttgcccc ggcgccctga cctgacgccc tggcctgacg ccctgcttcg tcgcctcctt 120 tctctcccag gtgctggacc agggactgag cgtcccccgg agagggtccg gtgtgacccc 180 gacaagaagc agaaatgggg aagaaactgg atctttccaa gctcactgat gaagaggccc 240 agcatgtctt ggaagttgtt caacgagatt ttgacctccg aaggaaagaa gaggaacggc 300 tagaggcgtt gaagggcaag attaagaagg aaagctccaa gagggagctg ctttccgaca 360 ctgcccatct gaacgagacc cactgcgccc gctgcctgca gccctaccag ctgcttgtga 420 atagcaaaag gcagtgcctg gaatgtggcc tcttcacctg caaaagctgt ggccgcgtcc 480 acccggagga gcagggctgg atctgtgacc cctgccatct ggccagagtc gtgaagatcg 540 gctcactgga gtggtactat gagcatgtga aagcccgctt caagaggttc ggaagtgcca 600 aggtcatccg gtccctccac gggcggctgc agggtggagc tgggcctgaa ctgatatctg 660 aagagagaag tggagacagc gaccagacag atgaggatgg agaacctggc tcagaggccc 720 aggcccaggc ccagcccttt ggcagcaaaa aaaagcgcct cctctccgtc cacgacttcg 780 acttcgaggg agactcagat gactccactc agcctcaagg tcactccctg cacctgtcct 840 cagtccctga ggccagggac agcccacagt ccctcacaga tgagtcctgc tcagagaagg 900 cagcccctca caaggctgag ggcctggagg aggctgatac tggggcctct gggtgccact 960 cccatccgga agagcagccg accagcatct caccttccag acacggcgcc ctggctgagc 1020 tctgcccgcc tggaggctcc cacaggatgg ccctggggac tgctgctgca ctcgggtcga 1080 atgtcatcag gaatgagcag ctgcccctgc agtacttggc cgatgtggac acctctgatg 1140 aggaaagcat ccgggctcac gtgatggcct cccaccattc caagcggaga ggccgggcgt 1200 cttctgagag tcagatcttt gagctgaata agcgtatttc agctgtggaa tgcctgctga 1260 cctacctgga gaacacagtt gtgcctccct tggccaaggg tctaggtgct ggagtgcgca 1320 cggaggccga tgtagaggag gaggccctga ggaggaagct ggaggagctg accagcaacg 1380 tcagtgacca ggagacctcg tccgaggagg aggaagccaa ggacgaaaag gcagagccca 1440 acagggacaa atcagttggg cctctccccc aggcggaccc ggaggtgggc acggctgccc 1500 atcaaaccaa cagacaggaa aaaagccccc aggaccctgg ggaccccgtc cagtacaaca 1560 ggaccacaga tgaggagctg tcagagctgg aggacagagt ggcagtgacg gcctcagaag 1620 tccagcaggc agagagcgag gtttcagaca ttgaatccag gattgcagcc ctgagggccg 1680 cagggctcac ggtgaagccc tcgggaaagc cccggaggaa gtcaaacctc ccgatatttc 1740 tccctcgagt ggctgggaaa cttggcaaga gaccagagga cccaaatgca gacccttcaa 1800 gtgaggccaa ggcaatggct gtgccctatc ttctgagaag aaagttcagt aattccctga 1860 aaagtcaagg taaagatgat gattcttttg atcggaaatc agtgtaccga ggctcgctga 1920 cacagagaaa ccccaacgcg aggaaaggaa tggccagcca caccttcgcg aaacctgtgg 1980 tggcccacca gtcctaacgg gacaggacag agagacagag cagccctgca ctgttttccc 2040 tccaccacag ccatcctgtc cctcattggc tctgtgcttt ccactataca cagtcaccgt 2100 cccaatgaga aacaagaagg agcaccctcc acatggactc ccacctgcaa gtggacagcg 2160 acattcagtc ctgcactgct cacctgggtt tactgatgac tcctggctgc cccaccatcc 2220 tctctgatct gtgagaaaca gctaagctgc tgtgacttcc ctttaggaca atgttgtgta 2280 aatctttgaa ggacacaccg aagaccttta tactgtgatc ttttacccct ttcactcttg 2340 gctttcttat gttgctttca tgaatggaat ggaaaaaaga tgactcagtt aaggcaccaa 2400 aaaaaaa 2407 <210> 48 <211> 4549 <212> DNA
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 3127541CB1 <400> 48 cccgcccccg ctcccgcagc cgcccgccag tcagtcagtc agtcagtcag tcagtcagtc 60 agtcagtcac tgagcgcgcg gcgcgggagc tgctggcagt cgctgcgtct ctggcgaggg 120 agcgccgcgc ctggggagga ggcggaggca gcggctggag gagcgcgagc ggcggtttcc 180 ttgcccgggg ccgcgggaag gccgaccgac tgccgcgatg gagcagctat cagatgaaga 240 aattgatcat ggtgctgaag aagacagtga caaggaagat caggacctgg acaaaatgtt 300 tggagcctgg cttggagaac tagacaaact cactcagagt ttggattctg acaagcccat 360 ggaaccagta aaaagatctc ctcttcgcca ggaaacaaac atggccaact tttcttaccg 420 cttctccata tacaacttga atgaagctct gaat.caggga gagactgtgg atctggatgc 480 cttgatggct gatctttgct ctatagagca ggagctcagc agcattggtt caggaaacag 540 taagcgtcaa atcacagaaa cgaaagctac tcagaaattg cctgttagcc gacatacatt 600 gaaacatggc accttgaaag gattatcttc ttcatctaat aggatagcta aaccttccca 660 , tgccagctac tccttggacg acgtcactgc acagttagaa caggcctctt tgagtatgga 720 tgaggctgct cagcaatctg tactagaaga tactaaaccc ttagtaacta atcagcacag 780 aagaaccgcg tcagcaggca cagtgagtga tgctgaagta cactctatta gtaattcctc 840 .
ccattccagc atcacttccg cagcctccag catggactct ttggatattg ataaagtaac 900 acgccctcaa gagctggatt tgacacatca agggcagcca attactgagg aagaacaggc 960 agcaaaattg aaagctgaga agatcagagt tgccctagag aaaattaaag aggcacaagt 1020 gaaaaagctg gtgatcagag tccacatgtc tgatgacagt tctaaaacaa tgatggtgga 1080 tgagaggcag acagtaagac aagtactgga taacctgatg gacaaatccc actgcggtta 1140 tagtttagac tggtcactgg tagaaaccgt ttctgaatta caaatggaga gaatctttga 2200 agaccatgaa aacttggttg aaaatcttct taattggaca agagatagcc aaaacaagct 1260 tatatttatg gagcgtatag aaaaatatgc acttttcaaa aacccacaga attatctttt 1320 ggggaaaaag gaaacagctg agatggcaga tagaaacaaa gaagtcctct tggaggaatg 1380 tttttgtgga agttctgtaa ctgtaccaga aattgaagga gtcctttggt tgaaggatga 1440 tggcaagaag tcctggaaaa agcgttattt tctcttgcga gcatctggta tctactatgt 1500 tcccaaagga aaagcaaagg tctctcggga tctggtgtgc tttctccagc tggatcatgt 1560 caacgtttat tatggccagg actatcggaa caaatacaaa gcacctacag actattgtct 1620 ggtgctgaag catccacaaa tccagaagaa atctcaatat atcaaatacc tttgttgtga 1680 tgatgtgagg acactgcatc agtgggtcaa tgggatccgc attgcaaagt atgggaagca 1740 gctctatatg aactaccaag aagccttgaa gaggacagag tcagcctatg attggacttc 1800 cttatccagc tccagcatta aatcgggatc cagttcttcc agcatcccag agtctcagtc 1860 aaaccactcc aatcagtctg atagcggagt ttctgacacc cagccagcag gacacgtccg 1920 ttcccagagc attgtgagct ccgtattctc tgaagcctgg aaacgaggca ctcagttgga 1980 agagtccagc aaggccagaa tggagtctat gaatcggccc tacacttcac ttgtgccccc 2040 tttatccccg caacctaaga tagtcacccc ctacactgct tcacagcctt caccacctct 2100 acctcctccg ccacccccac ctcctcctcc accaccccct ccaccacccc ctcctccccc 2160 actccccagc cagtctgcac cttctgcagg ctcagcagcc ccaatgttcg tcaagtacag 2220 cacaataaca cggctacaga atgcgtctca gcattcaggg gccctgttta agccgccaac 2280 acccccagtg atgcagtcac agtcagtgaa gcctcagatc ctggtacccc ccaatggagt 2340 tgttccacca ccccctcccc ctcctccacc cccaacccca ggctctgcca tggcccagct 2400 aaagcctgca ccgtgtgccc catcccttcc acagttcagt gccccgcctc ctccactgaa 2460 gatccatcaa gttcagcata ttactcaggt ggctccccca acaccccccc cacctcctcc 2520 tatccctgca cccctccctc cccaagctcc cccaaaaccc cttgtgacca tccccgcacc 2580 aaccagcacc aagactgtgg cacctgttgt gactcaagct gcaccaccca cacctactcc 2640 tccagtgccc ccagcaaaaa agcagccagc tttccctgct tcttacattc caccctctcc 2700 ccctacccct cctgttccag tacccccgcc aacattaccc aagcaacaga gcttctgtgc 2760 aaaaccccct ccctctccac tgtcaccggt gccctcggtc gtgaagcaga tagccagcca 2820 gtttccaccc cctccaactc cccctgccat ggaatctcag cccttaaagc ctgtcccagc 2880 aaatgtagct ccacagtccc ctcctgcagt aaaagcaaag cccaagtggc agcccagctc 2940 catcccagtc ccttctccgg acttccctcc tccccctcct gaaagcagcc tggtgtttcc 3000 tcctccaccc ccatcacctg tcccagcccc accaccgcca cctccaccca cagcttctcc 3060 tacccctgac aaaagtggat ctccaggcaa aaagaccagt aagacgtcca gccctggggg 3120 aaagaaacca cccccaaccc cacagcgcaa ctccagcatt aaatccagca gtggtgcaga 3180 gcaccccgag cccaagagac cctcggtgga cagtctagtc agcaagttta caccgccagc 3240 agaatcaggg tctcccagca aggagaccct accacctcct gcagcacccc ccaagcctgg 3300 aaaactcaat ctttctggag tcaaccttcc tggagttctc caacaagggt gtgtgtcagc 3360 aaaagcccct gttctgagtg ggcgtggaaa ggactccgtg gtggaatttc cttctcctcc 3420 atccgattct gattttccac cccctccacc tgaaacagag cttcctctgc cccccattga 3480 gattccagca gttttctcgg gaaacacctc tccaaaagtg gcagtcgtta atcctcaacc 3540 acaacaatgg tctaaaatgt cagtgaagaa ggcccctcca cccacacgac ccaaacggaa 3600 tgatagcacc cgcctcactc.aagctgagat ttctgagcag ccaacaatgg ccacagttgt 3660 gccacaagtg cccacctcta ccaaatccag ccttagtgtc cagcctggat tcctggctga 3720 cctcaacagg acactgcaac gaaagtccat cactcggcac ggctcactct.cctcccgcat 3780 gtccagagca gaaccaacag ccaccatgga tgatatggca ttgcct~ccac caccccctga 3840 actgctgtct gatcaacaga aggctggtta cgg~ggcagt catatatcag gctatgcaac 3900 gttgcggaga ggaccccctc ctgctccccc caaaagagac cagaacacca agctctccag 3960 agactggtag ccaccatagg actttatttt catgatatct gtaatcactg ctacaatcag 4020 ctacacctga tcatctagta gaattcaggt agtaacagag cctcctggta tgatgttatt 4080 caggtagtag tccagctaat atagtagtac atagtgtagt tagtagtaca cagtagcata 4140 gtaacaacac caagctgtaa cagatagtag tgttataaga tataataaca taataatagt 4200 aatagtgtaa taatagtaat aataagaaag aggacgagcc taggagaagc ttatttacta 4260 atacataatt acccttcgat actctaccta acatcgtgac aaagactagg ggtagttcac 4320 ctgttacata tagatgggcg gttgcagaca agaaggcaat aacaaacccg cgggccatgg 4380 tggcagctag cgtcatcctt gtaaaggaat agtacttaat aatccgctag cgtggagatg 4440 tggaccccca caaaacactt gcctcaatcc acctttagtt ttagcaacca ggtaattagc 4500 actacaacgt agctcgtcag tcgcacaggt aggtaagacc atattaaca 4549 <210> 49 <211> 2598 <212> DNA
<213> Homo sapiens <220>
<221> misc_feature <223> Incyte ID No: 8224777CB1 <400> 49 cccttccgtt aagagaattg cttctagggc ggctcctggc caatccccgt cgcctcgcct 60 gagtcagaga ggacccgttc aaaaaggaag gattctggtt tgatccggcc tctccgtctg 120 ctaggatatg atttagcgca gttgcgtgag ctctttggcc tctgtgttca cgtctgtatg 180 atagaggtgg tattgtttga ctcgtatttg tgaagtttca atgagattga taattgtcga 240 ttttatgttg aatccctagt acatggcctg ctgtcaacac ccaggacacc caggatatgg 300 tctttgctgt ttgattttcc tcatccccag tctcaagggg aagccaggac aatgagaaca 360 gccacttccc atcaggagtc actgcaaggc ccccagggtg ggatggtggg gagataagaa 420 ccgtgagaga agttggcaca aaggagttat gggacaaagg gtccaagata ggcagaaaag 480 aaaatgttgc cagttgatgg ggaagaaagg aagtcagagg gctcagacac tgtgggggac 540 agaacatctc catgtgcaac atcatctgct accctgaaag atctggaggt aagaggctct 600 gggtggaggt gcagtgaccc ttcgggtcaa ccctccaacc tcctcctcca ggtgggactg 660 ggtgcccctc tgccagctga gacagcccac acacacccca gccctaacga tcgttctctc 720 tacctctctc cccactcctg ctccacctcc tcctctctgc atgcacctca gagcccgtgc 780 caagaacgag cagtagtcct ggattcaacg tccgtaaaaa tcagtcgact gaagaacacc 840 atcaaatctt tgaaacaaca gaagaaacaa gtggaacatc agctggaaga agaaaagaaa 900 gcaaacaacg agagacagaa agccgaaagg gagctagagg ttcaaatcca gacattgatc 960 atacagaaag aggaactaaa tacggacctg taccacatgg aacgttctct cagatacttt 7.020 gaagaagagt ccaaggacct ggctgtccgc ctgcaacatt cattgcagtg taaaggagag 1080 ttagagaggg ctctgtctgc tgtcatcgcc acagagaaga agaaggcaaa ccagttgtcc 1140 agctgcagca aagcacatac agagtgggag ttagagcagt ccctacagga ccaggcactg 1200 ctgaaagcgc agctgacaca gttgaaggag tcatttcaac aactccaatt agaaagagat 1260 gagtgtgctg aacatataga aggagagagg gcccggtggc atcagaggat gagtaaaatg 1320 tcgcaggaga tttgcacatt aaagaaagag aagcagcaag atatgcgtcg ggtagaggag 1380 ctggagagga gcttgtccaa actcaaaaac cagatggctg aacccttgcc cccggagccc 1440 ccagcagtgc cctctgaggt ggagctgcag cacgtgagga aggaactaga gagagtggca 1500 ggagagc cc aggcccaggt caaaaacaat cagcacataa gtctcctgaa ccggcgacaa 1560 gaagagagga ttcgggaaca ggaagagagg cttcggaagc aggaggagag gcttcaggag 1620 cagcacgaga agcttcggca gctggccaag ccacagagcg tcttcgagga gctgaacaat 1680 gagaacaaga gcacactgca gttggagcag caagtaaagg agctacagga gaagcttggc 1740 gaggtgaagg agacggaaac ctccacccca tccaagaagg gctgggaggc gggcagcagc 1800 ctcttgggag gggaggtgag cagctttatg gaccacctga aggagaaggc agacctgagt 1860 gagctggtga aaaaacaaga acttcgcttc attcaatact ggcaagagag atgccatcag 1920 , aaaatccatc accttttatc agaaccaggg ggccgtgcca aagatgcagc actggga.gga 1980 ggacaccatc aggctggagc tcagggagga gatgaaggtg aagctgctgg agctgcagca 2040 gatggtattg cggcttacag caactacaac aatgggcaca gaaaattcct ggccgctgcc 2100 cacaactctg ctgatgagcc cggtccagga gccccagccc cccaggagct tggggctgca 2160 gacaagcatg gtgatcttcg tgaggtgagc ctcacctcct ctgcccaagg agaggccagg 2220 .
gaggatcctc tccttgacaa gcctactgca cagccgatcg tgcaggacca ccaggagcac 2280 ccaggcttgg gcagcaactg ctgtgtgcca ttattttgtt gggcttggct gccaagaaga 2340 aggagataaa catcaccatc atcaaadagc tgctcaagaa atttttaaat aagaaaccaa 2400 gttatggggt taatctccta cacaattcat ttacttcctt tgaatgttag actcactcat 2460 gattatttgt gtttctaatt tatagtttaa gtttatttgt aaaaagttaa aagagagtgg 2520 gtgtctgtgg ctctcactga tgttcactct ggctttccag cacactggcg ccgtattagt 2580 gatggagctc gaccgctc 2598 <210> 50 <211> 1353 <212> DNA
<213> Homo Sapiens <220>
<221> misc feature <223> Incyte ID No: 587394CB1 <400> 50 acaccccctg tagtcgccct ttgttcctcc ctctttggtt ctctctctcg ttttttttcc 60 cttttgcttc tcccctttgt ccccctctgt ttgttgttct ccctcttgtg ttccctccgg 120 ttttttctcc ctgtcccccg ccccccgtgc gctccccccc cccccctctt tttttttttt 180 tttttttttc tttttttttt tttttttttt tttttttttt ttttttttta agatttgaag 240 gacttaaaat acccagactt taattcctct aagattatag tcattaatca tgcttttata 300 tcatattatc tcttaacatt atatcatatt atctcttaac atttaatttc taaatataat 360 gttcatgagg aaaagagaaa atagcttggc ttctctcttc actgaatggt tgttcttagt 420 atcttctaga tgttccagaa ctgatgtcag atttggctcg tcagagtcca caaaacatat 480 tggtgagaaa gatgaagagg attcctgccg ccttgactgt acaatgagcc cttccagaac 540 acagcagcgc gcggccaggc cccggggaga gggatcgctc aaacagcacc agaggctgca 600 ttccaacttt tcctccgtca acgaggccgt tttcattgtt agtttctcct taaacacaaa 660 cttaaaaaca actggcttaa atctactatc gcatcttgcg tgatcatgtt tgtgccttac 720 agtgacaggc tgcaaacgat agaaattcaa cccaaactgt cacaagaaaa gaaggaaagg 780 cttgtaacta cttaaacata ttgctaatta aagatgtctg gaaaattgac tcaaaaacaa 840 aaacaaaaac aaaacaacaa caacaaaaaa ttggccgaac acgcgggaac agggaaaacc 900 tgactgaaga atgaggcaca ttaaaactta agggccttgg gtcccggcgc ggtggctcac 960 gcctggaatc ccagcacttt gggaggcaga ggcagggtca tttgaggtca ggagttcgag 1020 accagcctgg gcaaaaacac gcgagcatct cagaggccat tttcccattc cgcgcctggg 1080 ctcaaacaac agctggagca gccccagccc cagccccagt cagcggtccc gggagcagtc 1140 ccgctgactg agggcagacc atgggtccca agagggctcc cggccagccg cgggctccac 1200 cacctcggtg cgtggcgacg gcggccaaga ggggccagcg gcaccccgag tcagccccgc 1260 gccaggagcc ggagaacgca gccctccgcc ctgagccgat tccgagtaag tacgctcata 1320 ctatagtata aaccgatata tcacccttgt ttt 1353 , <210> 51 <211> 2161 <212> DNA
<213> Homo Sapiens <220>
<221> misc_~eature <223> Incyte ID No: 1402405CB1 -<400> 51 agggccccgg gacgcagctg gggggcctgg gtgggcaact gttttcctcg gataacgccc 60 ggggatgtcc cactggcagc ggagactctc tgagacatgg ctgcagtgaa gcggcaccgg 120 ggtgaccctg gaaccctcat ggggaaaggc ctgtcttggg ctgggcgcct ccagtgccgg 180 agccatggca gtctcagcca ggcctcgaat.ggggttttga gggtggggga gtaatttttc 240 tctctggggt ctccctgcag ttctcccgtg ttcagttctt ccgacggcag cctgtgggag 300 acagtggcgc gggctctgag ctgcctgggt cccacccaca tgggacccct ggctttgggg 360 atcctgaagc tcgagcactg tccacaggca ctgaggaccc aggccttcca ggtccttctc 420 cagcccctgg cctgtgtcct gaaggccacg gttcaggccc ccggaccccc aggcttgctg 480 gacgggacgg cagacgatgc cacgacggtg gacacactcc tggcctccaa gtcgtcctgc 540 gccggcctcc tgtgccgcac cctggctcac ctggaggagc tgcagccgct gccccagcgc 600 ccttcaccgt ggccccaggc gtctctactg ggggctacag tgactgtcct gcggctctgt 660 gacggctcgg ctgcccctgc ctccagtgtg gggggccacc tctgtgggac cctggcgggc 720 tgcgtccggg tccagcgagc agccctcgac ttcctgggga cgctgtcaca ggggacaggc 780 ccccaggagc tggtgacgca ggcgcttgct gtcctcctgg agtgcctcga gagccccggc 840 tccagcccca cggttctgaa gaaggccttc caggccacgc tcaggtggct cctgagctca 900 cccaagaccc ccggctgctc tgatctcggc cccctcatcc cgcagttcct cagagagctg 960 ttccctgtgc tgcagaaacg cctgtgccac ccctgctggg aggtgaggga ctccgccctc 1020 gagttcctga cccagctgag caggcactgg ggaggacagg ctgacttcag atgcgcactc 1080 ttggcttcag aggtgcctca gctggccctg cagctcctcc aggaccctga gagttatgtc 1140 cgagcgagtg cagtgaccgc catggggcag ctgtccagcc agggcctgca cgcccccacc 1200 agccctgagc atgcagaggc ccggcagagc ctgttcctgg agctcctgca catcctctcc 1260 gtagactcgg agggcttccc acggcgggcg gtcatgcaag tcttcactga gtggctgcgg 1320 gacggccacg ccgacgcggc ccaggacacg gagcagttcg tggccactgt gctgcaggcg.1380 gcgagccagg acctggactg ggaggtccgc gcccagggcc tggagctggc cctcgtgttc 1440 ctgggccaga ctttggggcc gccgcgtacc cactgcccct atgccgtggc cctacccgag 1500 gtggccccag cccagccact caccgaggca ctgagggctc tctgccacgt ggggctcttt 1560 gacttcgcct tttgtgcctt gtttgactgc gaccgccctg tggcgcagaa gtcttgtgac 1620 ctccttctct tcctgaggga caagattgct tcctacagca gcctgcggga ggccaggggc 1680 agccccaaca ctgcctccgc agaggccacc ctgccgaggt ggcgggcggg tgagcaggcc 1740 cagcccccag gggaccagga gcctgaggct gtgctggcca tgctcaggtc cctagacctg 1800 gagggcctgc ggagcacgct ggccgagagc agcgaccacg tggagaagag tccccagtcc 1860 ctcctgcagg acatgctggc cacgggaggc ttcctgcagg gggacgaggc cgactgctac 1920 tgagcagaac cagactctgc cactggggct caggaccaag ggaggcagca ccatgtcctt 1980 ctgtgggaca ctgccagccc cagggctcca gcccagcccg gtggatcctc tggggaagcc 2040 aggaccagga gagaagcaag gtcaagaaat cccacagttt gatgtattaa agaaatgact 2100 tatttctact caaaataaat ggcattgaag tctttcttta aaaaaaaaaa aaaaaaaaac 2160 c 2161 <210> 52 <211> 1487 <212> DNA
<213> Homo Sapiens <220>
<221> misc feature <223> Incyte ID No: 1798468CB1 <400> 52 gttttcaaac ggggaactta gaaagtggca gcccctcggc ttgtcgccgg agctgagaac 60 caagagctcg aaggggccat atgacactcc tcccggaccc ctggacacac acagccctgg 120 agactggagc cttggagcat ggcaagtcca gagcaccctg ggagccctgg ctgcatggga 180 cccataaccc agtgcacggc aaggacccag caggaagcac cagccactgg ccccgacctc 240 ccgcacccag gacctgacgg gcacttagac acacacagtg gcctgagctc caactccagc 300 atgaccacgc gggagcttca gcagtactgg cagaaccaga aatgccgctg gaagcacgtc 360 aaactgctct ttgagatcgc ttcagctcgc atcgaggaga gaaaagtctc taagtttgtg 420 gtgtaccaaa tcatcgtcat ccagactggg agctttgaca acaacaaggc cgtcctggaa 480 cggcgctatt ccgacttcgc gaagctccag aaagcgctgc tgaagacgtt cagggaggag 540 atcgaagacg tggagtttcc caggaagcac ctgactggga acttcgctga ggagatgatc 600 tgtgagcgtc ggcgcgccct gcaggagtac ctgggcctgc tctacgccat ccgctgcgtg 660 cgccgctccc gggagttcct ggacttcctc acgcggccgg agctgcgcga. ggctttcggc 720 tgcctgcggg ccggccagta cccgcgcgcc ctggagctgc tgctgcgcgt gctgccgctg 780 caggagaagc tcaccgccca ctgccctgcg gccgccgtcc cggccctgtg cgccgtgctg 840 ctgtgccacc gcgacctcga ccgccccgcc gaggccttcg cggccggaga gagggccctg 900 cagcgcctgc aggcccggga gggccatcgc tactatgcgc ctctgctgga cgccatggtc 960 cgcctggcct acgcgctggg caaggacttc gtgactctgc aggagaggct ggaggagagc 1020 cagctccgga ggcccacgcc ccgaggcatc accctgaagg agctcactgt gcgagaatac 1080 ctgcactgag ccggcctggg accccgcagg gacgctggag atttggggtc accatggctc 1140 acagtgggct gtttggggtt cttttttttt atttttcctt ttcttttttg ttatttgaga 1200 cagtcttgct ctgtcaccca gactgaagtg cagtggctca attatgtctc actgcagcct 1260 caaactcctg ggcacaagca atcctcccac ctcagcctcc caagtagctg ggattacagg 1320 tgcacaccac cacaccaggc taatttttaa ttttttagtc gagaccgggt ctctctgttg 1380 gccaggctgg tctctaactt ctgaccttaa atgatcttcc tgctcaggtt cccaaacccg 1440 gggttcccgt gtggccccac cccggctggg gtcccttatg ggaagcc 1487 <210> 53 <211> 2495 <212> DNA
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 3189084CB1 <400> 53 ggcggggccc ggggggcgcc caagatggcg tgcgggcggc gcggagggcg gctcgggccc 60 cggcgccgcc atgggggact gcgcggagat taagtcgcaa ttccgcacgc gcgagggttt 120 ctacaagcta ctcccgggcg acggcgccgc tcgcaggtcg ggtccggctt ccgcccagac 180 tccggtgccg cctcagccac cgcagccccc gcccggccct gcctccgcct ccggtcccgg 240 cgctgcaggc cccgcgtcgt ccccgccgcc cgcaggcccc ggacccgggc ccgccctgcc 300 cgccgtgcgc ctcagcctcg tgcgcctcgg ggagccggac agcgccgggg ccggggagcc 360 gcccgccacg cccgcggggc tgggctcggg gggagaccgc gtctgcttca acttgggccg 420 tgagctctat ttctacccag gctgctgtcg tcgtgggagc caacggtcca ttgacctcaa 480 caagccaatt gacaagcgga tctacaaggg cacccagccc acctgccacg atttcaacca 540 gttcactgct gccaccgaga ccatctcgct gctggtgggc ttctcagcgg gtcaagtgca 600 gtacctggat ctcatcaaaa aggacaccag caagctgttc aatgaggagc ggttgatcga 660 caagaccaag gtgacatatc tgaagtggct gcctgagtcg gagagcctgt tcctggcatc 720 acacgccagt ggccacctgt acctgtacaa cgtcagccac ccctgcgcct cggccccgcc 780 ccagtacagc ctgctgaagc agggcgaggg cttctctgtc tatgctgcca agagcaaggc 840 accccgcaac ccgctggcca agtgggcggt gggtgagggg cccctcaacg agttcgcctt 900 ctcgcccgat ggccggcacc tggcctgtgt gagccaggat ggctgcctgc gcgtcttcca 960 cttcgactcc atgctcctgc gtgggctcat gaagagctac tttgggggcc tgctgtgtgt 1020 gtgctggagc cctgacggcc gctacgtggt gacgggtggc gaagatgacc tggtcaccgt 1080 gtggtccttc accgagggcc gcgtggtggc tcgaggccat ggccacaagt cctgggtcaa 1140 cgctgtggcc tttgacccct acaccacaag ggcagaggag gcggcgacag cagccggtgc 1200 tgatggggag cggagcggcg aagaggagga ggaggagccc gaggctgcgg gcacaggctc 1260 ggccgggggc gccccactct ctccactgcc caaggctggc tccattactt accgctttgg 1320 ctcggcgggc caggacacgc agttctgcct gtgggacctc actgaagacg tgctctaccc 1380 gcacccgccc ctggcccgca cccgcaccct ccctggcaca cctggcacca cgccaccggc 1440 cgccagcagc tcgaggggtg gcgagcctgg cccaggcccc ctgcctcgct cgctgtcccg 1500 ctccaacagt ctcccgcacc cagctggcgg gggcaaggcg ggcggcccgg gtgtggcggc 1560 agagcctggc acaccattca gcattggccg cttcgccacg,ctcacactgc aggagcggcg 1620 ggaccggggg g~cagagaagg agcacaagcg ctaccacagc ctgggcaaca tcagccgggg 1680 tggcagtggc ggcagtggca gtggtgggga gaagcccagc ggccctgttc cccgcagccg 1740 cctggacccc gccaaggtgc tgggcactgc gctgtgcccg cgcatccacg aggtgcccct 1800 gctggagccc cttgtgtgca agaagatcgc ccaggagcgg ctcacagtcc tcetgttcct 1860 ggaggactgc atcatcactg cctgccagga gggcctcatc tgcacctggg cccggccggg 192 0 caaggcgttc acagacgagg agaccgaggc ccagacaggg gaaggaagtt ggcccaggtc 1980 acccagcaag tcagtggtag agggcatctc ctcccaacca ggcaactccc cgagtggcac 2040 agtggtgtga agccatggat atcgggcccc cccaacccca.tgcccccagc ctcctagcca 2100 taaccctccc tgctgacctc acagatcaac gtattaacaa gactaaccat gatggatgga 2160 ctgctccagt ccccccacct gcacaaaatt tgggggcccc ccagactggc ccggacacgg 2220 gcgatgtaat agcccttgtg gcctcagcct tgtcccccac ccactgccaa gtacaatgac 2280 ctcttcctct gaaacatcag tgttaccctc atccctgtcc ccagcatgtg actggtcact 2340 cctggggaga cactccccgc ccctgccaca agagccccag gtctgcagtg tgcccctcag 2400 ttgagtgggc agggccgggg gtggtccagc cctcgcccgg cccccacccc agctgccctt 2460 gctattgtct gtgcttttga agagtgttaa attac 2495 <210> 54 <211> 2227 <212> DNA
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 5580384CB1 <400> 54 ggaagatggc ggcgcacaag tcaggtccgg cacatgtttc cgcggagcgg acccagcaat 60 gacggatgat atcacctctt cttctctggt gagagtctga ggatagagac ttttttctca 120 ccatgaatgt caccccagag gtcaagagtc gtgggatgaa gtttgctgag gagcagctgc 180 taaagcatgg atggactcaa ggcaaaggcc tcggccggaa ggagaatggt atcactcagg 240 ctctcagggt gacactgaag caagacactc atggggtagg acatgaccct gccaaggagt 300 tcacaaacca ctggtggaat gagctcttca acaagactgc ggccaacttg gtagtggaaa 360 ctgggcagga tggagtacag ataaggagcc tttctaagga gaccacccgt tataatcatc 420 ccaagcccaa cttgctgtat cagaagtttg tgaagatggc tacattgact tcaggtggag 480 agaagccaaa caaagacttg gagagctgca gtgatgacga caaccagggg tccaagtccc 540 caaagattct gactgatgag atgctgctcc aagcctgtga ggggcgaaca gcacacaagg 600 ctgcccgtct tgggatcaca atgaaggcca agcttgctcg cctagaggcc caggagcagg 660 ccttcctggc tcgtctcaaa ggccaggacc ctggggcccc tcaactgcag tcagagagca 720 agccccccaa aaaaaaaaaa aagaaaagga ggcagaaaga ggaggaagaa gctacagcat 780 ctgaaaggaa tgatgcagat gagaagcacc cagaacatgc tgagcagaac atcagaaaaa 840 gcaagaagaa gaaaaggcga catcaagaag gaaaggtctc agatgaaaga gagggtacaa 900 ctaaagggaa tgagaaggag gacgctgcag gaacaagtgg gcttggggaa ttgaatagca 960 gagagcaaac caatcagtcc ctcaggaaag ggaagaaaaa gaagaggtgg caccatgaag 1020 aggagaagat gggggtcttg gaggaaggag gaaaaggcaa ggaggctgca ggcagtgtca 1080 ggacagagga ggtagagagc agggcatatg ctgacccatg cagccgaaga aagaagaggc 1140 agcaacagga ggaggaggac ttgaacctag aagatagagg tgaggaaact gttttaggtg 1200 gtggaaccag ggaagcagag agcagagcat gcagtgatgg aagaagcagg aaaagcaaga 1260 agaaaagaca gcagcatcaa gaggaggagg acatcttgga tgtaagggat gagaaggata 1320 gcggggctag ggaagcagag agcagagcac acactggctc aagcagcaga ggtaagagga 1380 agaggcagca gcatcccaag aaggaaagag ctggagtcag cactgtccag aaagccaaaa 1440 agaaacagaa gaagagagac taaaggtctg gtaaaggtag ggctcaattg attgattttc 1500 aggagttgaa gcctcaaaga ccagggttga tgcaggtctg caggtcttct gcacccccct 1560 caatgaggag tccctcccag aaaggaaact gatctctggg atgtcagctg ctgagaggag 1620 caagcggtag taccacccct tagttgaggg agtcagcaca gtcctttctg cagcttctaa 1680 cccaggacca tgaactcagg tgcctagaga agccaggcag ctaaaggaca,aggaatgctg 1740 ggggctgtgg gaacaggaat gcagataccc tttgaaggag cattcctgct aaaagaagct 1800 gaaaatgtag acctatgtga agtgctctga tttctaaata ttgtgaaggt taagaaaaac 1860 ataaatttag gtctatgggctagatttagc ccacagttgc cagtttctag cgctaccaaa 1920 tgaatgaata aacatgagct tgcgctccta gcctagagat aaatcctgac. tggcatctct 1980 gttcccagcc tgggaaggtc ctgaatacaa attagaagat.attccttgga ggcctttgaa 2040 gaaattcttc ggttaacctc tttgtagtct tgctacactg ataagtagaa gtagctccct 2100 gtctgtgtcc caaatgaata agaattgtgt aaaggacagc acaactcact tggcatctaa 2160 cagtccattt tcattgtttc caaataccat agcaacctct tgccctttgt gttaccccta 2220 gagagat <210> 55 <211> 1652 <212> DNA
<213> Homo sapiens <220>
<221> misc_feature <223> Incyte ID No: 5158619CB1 <400> 55 cggctcgagc aaggcaggag gctaggaggt ctacaaatag cgactgggta gctggtgtga 60 acacaggggg tactgggggg gcttagcccc caaggaagag gaccagtttt tccccagcac 120 caccatcaag gcctcgaggc tcccagctcc ctctacagcc tgtggactga cttagggaat 180 cccgaacgat gacagaaaag gaggtgctgg agtcccctaa gccctccttc ccagcagaga 240 ctcggcaaag tgggctacag cggctaaagc agttactcag gaagggttct acagggacaa 300 aggagatgga acttccccca gagccccagg ccaatgggga ggcagtggga gctgggggtg 360 ggcccatcta ctacatctat gaggaagagg aagaggaaga agaggaggag gaggagccac 420 ccccagaacc tcctaagctg gtcaacgata agccccacaa attcaaagat cacttcttca 480 agaagccaaa gttctgtgat gtctgtgccc ggatgattgt tctcaacaac aagtttgggc 540 ttcgctgtaa gaactgcaaa accaacatcc atgaacactg tcagtcctat gtggaaatgc 600 agagatgctt cggcaagatc ccacctggtt tccatcgggc ctatagttcc ccactctaca 660 gcaaccagca gtacgcttgt gtcaaagatc tctctgctgc caatcgcaat gatcctgtgt 720 ttgaaaccct gcgcactggg gtgatcatgg caaacaagga acggaagaag ggacaggcag 780 ataagaaaaa tcetgtagca gccatgatgg aggaggagec agagtcggcc agaccagagg 840 aaggcaaacc ccaggatgga aaccctgaag gggataagaa ggctgagaag aagacacctg 900 atgacaagca caagcagcct ggcttccagc agtctcatta ctttgtggct ctctatcggt 960 tcaaagccct ggagaaggac gatctggatt tcccgccagg agagaagatc acagtcattg 1020 atgactccaa tgaagaatgg tggcggggga aaatcgggga gaaggtcgga tttttccctc 1080 caaacttcat cattcgggtc cgggctggag aacgtgtgca ccgcgtgacg agatccttcg 1140 tggggaaccg cgagataggg cagatcactc tcaagaagga ccagatcgtg gtgcagaaag 1200 gagacgaagc gggcggctac gtcaaggtct acaccggccg caaggtgggg ctgtttccca 1260 ccgactttct agaggaaatt taggtgtgcg ggcgcctgca agcgggagac acccacaccc 1320 cattctgggc gggcccagtg gagtttgggg aggggggcga aagcaacggg actgctggga 1380 gaggaggggt aggaaggccc gcctgagcgc gacggggctt ccgggaaggg actggttctc 1440 gcccccttcc ccagcctggg gcctcggata cctgctgccc agagcagccc ggacccgaaa 1500 cctttcaggc cccgcttgca agagctggaa aaaaacgcgt atctactagg aggagccagg 1560 gactggggcg gggggcgggg gcgagggagg gcgaactgtc gaatgttgcg aatttattaa 1620 acttttgaca aaacttaaaa aaaaaaaaaa as 1652 <210> 56 <211> 696 <212> DNA
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 2792745CB1 .
<400> 56 tgatgttttt agtttcaatg ttgacatgtt ttcaattctc ttagttatac acttaggaag 60 ttttatatat ttttacctat taaatactcc ttggaaacat ctatttctct ctatctctaa 120 gctaccaaca ttccctcatc tggactaaag caaaagcctc.ctcecttctt ttttccaggc 180 ctcttctcta cctggcccag taatcccttt tttttttttt tttgagatgg agttttgctc 240 ttttgcccag acgggagtgc aacggcgtga tcgtggctca ctgcaacctc cgcctcccaa 300 gttcaagcaa ttctcccacc tcagcctcct gagtagctgg gattattaca ggcacccacc 360 atcacgccca gataattttt ggatttttgt agtgatgggg cttcaccatg ttggcctggc 420 tggtcttcaa ctcctgacct caagtgatcc acccacctcc acctcccaaa gtgctgggat 480 tacaagtgtg aaccaccgct gagtaatcct ttcaggacag gaatctgatc atgttatccc 540 attttgctta aaagttttcc ctgacatccc tgaggccatg aggccttcca tcccacccag 600 gcattctttc tgctcctggg gtgtgccaca ggeccctccc catgggcccc aggcattttc 660 ctctgccttg aaagctttcc tccactctgc ctggtt 696 <210> 57 <211> 600 <212> DNA
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 2827678CB1 <400> 57 tcacccaggc tggaggcagt ggcttgatct tcgctcactg caacctctgc ttcecaggtt 60 caagtgattc tcctgcctca gcctcccaag tagtgcatgc caccatcctc ggctaatttt 120 ttttgtttgt ttgtttttgt ttttgttttg agatggagtt tcgttcttgt tgcccaggct 180 ggagtgcaat ggtgccgtct cggatcaccg caacctctac ctcccaggtt caagcgattt 240 tcctgcctca cccttccaag tagctgggat tacaggtgcc tgccaccacg cccagctaat 300 ttttttgtat ttttagtaga gacggggttt caccatattg cccaggctgg tttccaactc 360 ctgacctcag gtgatccgcc tgccttgtcc tcccaaagtg ctgggattac aggtgtgagc 420 cactgcgcct ggccaacttt tttgtagaga cagggtttca ccatgttggc caggatggtc 480 tcgaactcct gacctcaagt gatctgccct tctgggcctc caccaagtgc tgggattaca 540 ggcctgagcc actgtgtctg gctttgccca ttttttttgg ttttttaatt ggattgtcta 600 <210> 58 <211> 3005 <212> DNA
<213> Homo sapiens <220>
<221> misc_feature <223> Incyte ID No: 790257CB1 <400> 58 aacagggttt gttatgaggc ttgtgggcga taacgtatgt gacagcaatc tataaaccca 60 aaatattgca cacggttgtt tttattatct ttttctagtt gctctgagtt tagattgaga 120 aggtggctgt acttttggag acgtgttcat atctaaggaa gataccaaat ttcggattag 180 gtttttattt cggagacttt tcaaggaaca tgagaagatt atctgagaag attatctgcg 240 cagacaggcc gagcgacgga aagaactcag ttcagctcca tctcaggaaa cacctgatct 300 ccacgagctt ttgaggaagc ccggagccac agagccccac ccacttcttt cccgaccgcg 360 ccatccttcc tcactcccgg cctcctctct agccaatcac cgctcagttt aacgtcatca 420 ccgagcgcca tcagggagcg tgtcagaggc cgaccacgtg tactgtggag actgtcaaag 480 tctcccggag cccaatttcc ggaagcggtg agttctgaaa gaagttcctg caccgtagtt 540 tcccaagtct gcgaatcccc aaccatgagc gcctcgggcg tactgtcctt tacccagcaa 600 ggatgggagc aggtgctggc caaagtgaaa cgggctgtgg tttacctgga cgccgcctgc 660 gccgagagcc tgcactgggg ctgcggatcc acccgtctcc tggaggcggt ggggggtcct 720 gactgtcacc tgcgagagtt cgagcccgac gcaattggtg gtggagccaa gcagcccaag 780 gcagtgtttg tgctgagctg cctgctgaaa ggccggaccg tggagatcct acgggacatc 840 atctgccgca gtcacttcca gtattgtgtg gtggtcacaa ccgtgagcca cgctgtccac 900 ctcacagcta atcatgtccc agcggcggca gcggccgaga tggaggggca gcagccggtg 960 ttcgagcagc tggaggagaa gctgtgtgaa tggatgggca acatgaacta cacggccgag 1020 gtgttccatg tcccgttatt gcttgcccct gttgctcccc actttgcctt gactccagct 1080 tttgcatccc ttttcccact gctaccccag gatgtgcacc tccttaatag cgcccgaccg 1140 gacaagagga agctgggaag cctgggtgat gtggactcca ctacgctaac cccagagctg 1200 ctgctgcaga tcagatgcct agtgtcaggc ctcagttctc tgtgtgaaca tttaggagta 1260 cgggaggagt gttttgctgt aggttcctta agtcaggtca tcgctgcgga tctggccaat 1320 tatgcccctg caaagaacag gaagaagact gctgcaggca gggcatcagt ggtttttgtg 1380 gacagaaccc tggatctcac aggagcagtt ggacatcatg gagacaactt agtagagaag 1440 atcatttcag cacttcccca gctcccaggc cacacaaatg atgtgatggt taacatgata 1500 gcgctcactg cactccatac tgaggaggaa aattataatg tggttgcacc aggctgtctt 1560 tcacaatcca gtgacaccac agccaaagcc ctatgggaag ctttactgaa cactaagcac 1620 aaagaggcag tgatggaagt tcggagacat ctagtggaag cggcaagcag agaaaacctg 1680 ccaatcaaga tgagtatggg gagagtcaca ccgggacagc tcatgtccta tattcagctc 1740 ttcaagaaca acctcaaagc tctaatgaat cattgtggcc tcctccagct tggactggcc 1800 acagctcaaa cgttgaaaca cccacagact gccaagtggg acaactttct ggcttttgaa 1860 aggctccttc ttcagagcat tggggagtca gcaatgtccg ttgtgttaaa tcagctgctg 1920 cccatgatta agcctgtaac ccagagaacc aacgaggact acagccctga ggaactgctg 1980 atccttctca tatatattta ttctgtcact ggagagctca cggtagacaa agacctgtgt 2040 gaagcagaag aaaaagtcaa gaaagcattg gctcaggtct tctgtgagga atctggatcg 2100 tcacctttgc tgcaaaaaat tacggactgg gactcttcaa ttaatctgac atttcacaaa 2160 tccaaaattg ccgtggatga actctttact tcacttcggg atattgctgg agctcggagt 2220 ctcctgaaac agtttaagtc tgtatatgtt cctggaaatc atacccacca ggcatcttat 2280 aagccattgt tgaagcaagt tgtggaggaa atatttcatc ccgagaggcc agattccgtt 2340 gatattgaac acatgtcttc aggcctcact gatctcctta aaactggatt tagcatgttc 2400 atgaaggtga gccggcctca tcctagtgac taccccctcc tgatcctctt tgtggtaggt 2460 ggggtcacag tctctgaagt gaaaatggtc aaagatcttg tggcatcgtt gaagccagga 2520 acccaggtaa tcgtgctgtc cacacgactc ctgaagccac ttaacattcc tgagctgtta 2580 tttgcaactg accgactgca tccagacctt ggcttctgag catccgctaa gaagataaga 2640 cctactcaag ctggaaatgc cgatgcaatt ttctgccacc actccaaata ctcctccaca 2700 accagcgtcc ctgtcactaa ttgcgagaat gatggaattc tgcctgaagg gtcttgatac 2?60 ctactcagtg aggtactttg cttggattgc tgtgattctt aaaaaaaaaa aaaaaaagtt 2820 tttttatttt cctgaaggcg acagtctgtc ttttggcgag tccaattagt gatcgtttcc 2880 atttatcctt aaccctctgt ttgtgatctt cctgattcca cctacaagtt cattagttct 2940 aaataaaagc ctgttggcaa gtgttggtag attaaaaaaa aaaaaaaaaa aaaaaaaaaa 3000 aaaaa 3005 <210> 59 <211> 2530 <212> DNA
<213> Homo Sapiens <220>
<222> misc_feature <223> Incyte ID No: 2617345CB1 <400> 59 ccagcacgca cgcacgtacg tccggcactt ccggccgcgg cggcctcagc gccggcccga 60 aggaccaggc cgccgtcccc agcgagaggc atgcagcgct gaggagcggc gacccagcac 120 ggcggcgcca tgaacctcct gccgtgtaac cctcacggca acgggctgct ctacgccggc 180 ttcaaccagg accacggatg ctttgcgtgt gggatggaaa atggattccg agtctataac 240 actgatccac taaaagaaaa agagaaacaa gaatttctag.aaggaggagt tggccatgtt 300 gaaatgttat ttcgctgcaa ctatttagct ttagttggtg gtggaaaaaa gccgaaatac 360 cctcccaaca aagtaatgat ctgggatgac ctgaagaaga agactgttat tgaaatagaa 420 ttttctacag aagtcaaggc agtcaagctg cggcgagata gaattgtggt ggttttggac 480 tccatgatta aggtgttcac attcacacac aatccccatc agttgcacgt cttcgaaacc 540 tgctataacc ccaaaggcct ctgtgtcctt tgtcccaata gtaacaactc cctcctggcc 600 ~tttccgggca cgcacacggg ccatgtgcag cttgtggacc tggccagcac ggagaagcca 660 cccgtggaca ttcctgcaca cgagggtgtc ctgagctgca ttgcactcaa cctgcaggga 720 acaagaattg caactgcatc cgagaaaggg acgcttataa gaatatttga tacttcatca 780 gggcatttaa tccaggaact gcgaagagga tctcaagcag ccaatattta ctgcatcaac 840 ttcaatcagg atgcgtccct catctgcgta tccagcgacc acggcacagt gcatattttt 900 gcagctgaag atccaaaaag gaataaacag tccagtttgg cctcagccag tttccttcca 960 aaatacttca gttccaagtg gagtttctcc aagtttcagg ttccctcagg ctctccgtgc 1020 atttgtgcct ttggaacaga gccaaacgcc gtcattgcaa tttgtgcaga cggcagctac 1080 tacaaattcc tgttcaaccc caagggggag tgcatccgag atgtctacgc gcagtttcta 1140 gagatgaccg atgacaagct gtgactccag ctgggggcgc cacagcaccc accacctgcc.1200 gccttcagac tctcggggct ggtgccagtg ccccaggggc ctcctgggcc acgggctgga 1260 ggggctgccc agggaccttg gtctcgaagc catacgtggt tgtctgcttt cctaaggact 1320 cccatttcca gtattaaaga gagaatcatc atcaaggcac cgtaggtaac tcagtggctg 1380 tgaccagctc gactggcggc cactggctgt tcccatgagt tcagctgtga cgttagcttc 1440 agtggctccg ccgcatcctc acactgacgg gggctecata cggacctggg gactgggctg 1500 agagggtgga cgagttcagg tttgtttttg cagcagattc cgtcgttctt actgagtctg 1560 cagcggggga gtgaacaagt gtgcagatgt aagttcttac atgataagca gattgaatac 1620 aacaccagca gcttgcctta gaaaaggaga aaggaattcc ttttcccgcc cgaacatgaa 1680 gaaaaacgac ctgaccctgt agagagaaca cagtgtgaat gtttcccctc gtgtgagccc 1740 agcctgtggt cttctccgta cccgcaacgt ggtcatctgt gcccgtgacg tcacctgtgc 1800 ccgtgcgtgg cgtccccgtc tccgttgggg ccattagaat gaggcagaca ccaggccact 1860 ctagaagccg agccgtcaca cctcaggcgt gtgcggggcg gggacggggg gtctcctggt 1920 tacattttgg attaaacctg tttcccggtt atgtgtaggg aacagcagag tgatgcacga 1980 actttgaaca ttcgttatgg ggaaaacatc ctttaacttc ggggtcgtct gccagagcag 2040 ggtctgggag ggtccatgca gttcccgctg gtgtggaggg aaatgccctg gtctggcctc 2100 cgagccccca ggtccaccgt ctcccctccc ctcatttgta agaatagcta cacactaaca 2160 ttttgggaag gagaggcaca taactttttt taacatttgg taactaggtt atgggctcta 2220 cattgtcagc tacttgggat atatatttaa ttttcttaaa ttcccgttaa actctatttt 2280 atggttttga tttcagattg caaacatgta aaacctgcat agcagcgagt cctcggtttt 2340 gccggtttct ttagttcttt actgtcactg tcatgtaatc agctaattct ctgtggatgt 2400 tgctgtaaag tatgcatgtt cctttcatgt gtatttaatc atgatgttta attttgcaca 2460 cttatttgta atgtttcttt taaataaaag tgactaattt tgttgtagtc tggacctgta 2520 aaaaaaaaaa 2530 <210> 60 <211> 1625 <212> DNA
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 3254666CB1 <220>
<221> unsure <222> 1556 <223> a, t, c, g, or other .<400> 60 cggcgagaag gcggcgccgc tgccctggca gctggactgc actttccccc cgcccggcct 60 cagctgccgc ccgcccagac gccagcaagc ccccctccca cgacagggct gc.tccgggag 120 cttcggagac ccgccccggg cctgagcgca ggctgcctcc gggaccccac ggctgtccgg 180 acgtgccatg ggcgcgcagc tgccgggcaa cgtgttgtgt aaacactaca.cgtgaagagt 240 ggtgaaaggg aacattgatt actgaagtgc cctggagagg gaaagcactg gtcaacatca 300 catggacaaa tttcattgtt ttctaaagat ggcctggaag tagtctttge cactgcttcc 360 tccacaaaca gctcttcata acatgggctg catgaaatca aagcaaactt tcccatttcc 420 taccatatat gaaggtgaga agcagcatga gagtgaagaa ccctttatgc cagaagagag 480 atgtctacct aggatggctt ctccagttaa tgtcaaagag gaagtgaagg aacctccagg 540 gaccaatact gtgatcttgg aatatgcaca ccgcctgtct caggatatct tgtgtgatgc 600 cttgcagcaa tgggcatgca ataacatcaa gtaccatgac attccataca ttgagagtga 660 ggggccttga ggctgtagga tgacaacact ttgactgtgg aggtgctagt ttgaataaat 720 gtgacaaaag caaaaactgg tgtgaaaaag tacaaataac tatctggatt taaaaatgtg 780 tctacgataa tgtcactatt ataagaacaa ctaggatgaa atgcatttta agtacttcta 840 tgttaacagc aatttctgtt tagtcttaga ttttagtcat ctgaagggct gaacagaggt 900 cctgtgacac ccaataatca gctgaatgtc acagcacttc ttcctaagta atggcatcac 960 caaagaaaat gctaaggaat aaaaactgcc ccaaattcca atggttgaag tttatccttt 1020 aaaataacaa tttttgtttg tttataccca aaaaaagtcc agatatgaaa agggcttttc 1080 taaaatttct tggcgaggga atggcactca aatcatagtg attaacagta agtcttgttt 2140 gtttgtcaag gatctctact tcttgacaca aatgaaccct gtctttaata agataagata 1200 tttatttttg tagatgagaa gtgtaactac caccttggac ctcagggccc taactaatta 1260 cagctgttac tggacgactc agactttgtg cctaaagcca tcttagagat aacagtttat 1320 agaagccatg acattagtgt ttattgcatt gaattaagcc cagtgatata actatacaag 1380 aaaacaagta ggggtacctt ttacaaagag caatccaata aatcttaaaa ataacagaaa 1440 cttagtctgc aaggtagaaa gtttcagttt taattctgta ttaagcttta ctatctcaga 1500 ggtacagagg gctggaatat gggcatttat ttccagtttt ttcttgacta gttaangcgg 1560 tcaccattaa aatagaccag ataatgcatg aagatttaca gttgtattgc aaaacgggaa 1620 agata 1625 <210> 61 <211> 1795 <212> DNA
<213> Homo Sapiens <220>
<221> mist feature <223> Incyte ID No: 4159378CB1 <400> 61 gtgcacccaa cgccagtccc ggtagagttg aacccaccac gagagggaga gggggcggga 60 gcgggggccg agcggccgcg ggctcctccc ccggcgcctc cgctatctgg gtcaggacgc 220 gacggccgcg gcgcgggacc ttaggacccg cgggctccag ggctactgtc cgtccgccac 180 tgcgcgccag caggtcctgg tctccgctct ccaacagctg aaaggccggc gcagtgaaca 240 cagaaacgaa aaccaagaaa tgccttattc cacaaacaaa gagttgatac ttggcatcat 300 ggtgggcact gctggaatca gcttgctgct cttgtggtac cacaaggtcc gtaaaccagg 360 gatagcaatg aagttacctg aatttctttc tctgggtaat acatttaatt caataacttt 420 gcaagatgaa atacatgatg accaaggaac aacagtaatc tttcaagaaa ggcaacttca 480 gatactggag aagttaaacg aattactgac aaatatggaa gaactcaaag aggaaatcag 540 atttcttaaa gaagctattc caaagctgga ggaatatata caagatgaac ttggagggaa 600 aataactgtt cataagataa gccctcagca cagagcgaga aaaagaagac tccccacaat 660 tcaaagttca gcaacaagta atagttcaga ggaagcagaa agtgaaggag ggtatattac 720 agctaatact gacacagaag aacagagttt tccagtccct aaggcattta acacacgtgt 780 agaggaatta aatttagatg tccttcttca gaaggtagat catttacgta tgagtgagtc 840 tggcaagtcg gagagttttg aactacttcg tgaccacaaa gaaaagttta gagatgaaat 900 agagtttatg tggcgatttg ctcgtgctta tggagacatg tatgaactat ctacaaacac 960 acaagaaaag aaacattatg ctaatattgg aaaaacttta agtgaaagag ctattaatag 1020 agcacccatg aatggacatt gtcatctgtg gtatgcagtt ttgtgtggct atgtatctga 1080 gtttgagggt ttacaaaaca aaatcaacta tgggcacctc ttcaaggaac atctagatat 1140 agcaatcaaa cttttaccag aggaaccctt tctatattac ctcaaaggga gatactgcta 1200 tactgtctca aaactgagct ggattgagaa aaaaatggct gctactctgt ttggaaaaat 1260 accatcttca actgtacaag aagctttaca caatttcctt aaggctgaag aac.tatgccc 1320 tggttattct aatcccaatt acatgtactt agcaaagtgt tatactgatc ttgaggaaaa 1380 ccagaatgct ttgaagttct gtaatttggc tttattgctt cctactgtta ccaaagagga 1440 taaagaggca cagaaagaga tgcaaaaaat aatgacttcc ttgaagaggt aaataaacga 1500 atttactctt caacaaatca gatgtggtct accaaaattt aaatgaatca aagttgtgct 1560 tttattatcc ttcatttttg atgtaagggt attgtgctta gatttgaagg taaagccatg 1620 tttctgcaga atgcattcca ctagtagcac tacaaaatta attatgttat ttggagaatc 1680 tatatactat aatgtcaata cataatctat aaacatgtat gctttatatt tttcttatca 1740 ataaactgca gccttaagaa tatttcttta aataaaatat ttaaatccaa taaaa 1795 <210> 62 <211> 2080 <212> DNA
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 4317538CB1 <400> 62 cteggaggct ttgactgcag aagcgagagg agggggcaga agggagttca aaggtcacag 60 ggctggtgga aagctggatc agttgcctga aaaaaatgta ctggggttct ctgcagcagt 120 gttttgcagc agttaaatga agtgtgcttt atttctgaga gtgaagattt tacagcgtgt 180 gtgccgtagt cgttatatag tgcacgctac atgtgacagc actgcagcga tgagtggaat 240 tttaaagagg aagtttgaag aagttgacgg ctcctcaccc tgctcctctg tgagggaatc 300 agatgatgaa gtttccagca gtgaaagtgc tgacagtggg gacagtgtca atccatccac 360 ttctagtcat tttacccctt cctccattct caaaagggag aaacgactga ggacaaagaa 420 tgtacatttt agttgtgtca ccgtgtacta cttcaccagg aggcaaggct tcacaagtgt 480 gcccagtcaa gggggaagca ccctggggat gtccagccgc cataacagcg tgcgccagta 540 cactcttggc gagtttgcaa gggagcagga gaggctccac cgggagatgt tgagagaaca 600 ccttagggag gaaaagctga actccttaaa actaaagatg actaagaatg gcacagtaga 660 atcagaagaa gccagcactc ttacactgga tgacatttct gatgatgaca ttgacctgga 720 caacacagag gtagatgagt acttcttcct acaacctttg ccaacaaaaa aacgaagagc 780 tctgctgcgt gcctctggag tgaaaaagat tgacgtggaa gaaaagcacg aactccgagc 840 catccgcctc tcacgagagg actgtggctg tgactgccga gtgttctgtg atccagacac 900 gtgcacctgc agcctggctg gcattaagtg ccaggtggat cgtatgtctt tcccatgcgg 960 ctgcactaaa gaaggatgta gtaacacagc aggtagaatt gaatttaatc ctatccgtgt 1020 tcggactcac tttttgcaca caataatgaa acttgaactg gagaaaaacc gagagcagca 1080 aatccccacg ctgaatggct gccacagtga gataagtgct cacagtagtt ctatgggccc 1140 tgtcgctcac tccgtagaat attcaatcgc agacagtttt gagattgaaa ctgagcccca 1200 ggctgcagtg ctgcacctgc agtcggctga agaattagat tgccaaggag aggaggagga 1260 agaagaggag gatgggagca gcttttgcgg cggagtcaca gattctagca cgcaaagctt 1320 ggcacctagt gagtcagacg aggaggagga ggaagaagaa gaggaagaag aggaggagga 1380 tgacgatgat gacaaaggag atggcttcgt ggaaggtttg ggcacccatg ccgaagttgt 1440 ccctcttcct tcagttcttt gttattctga tggcaccgcc gttcacgaaa gccatgcaaa 1500 gaatgcttct ttttatgcca actcttcaac tctgtattac caaatagata gccacattcc 1560 aggaactcca aatcagatct ctgagaacta ttctgaaaga gacactgtca aaaatggtac 1620 cctttcgctg gtgccttaca ccatgacccc ggagcaattc gtcgactatg cccgacaagc 1680 agaagaggcc tatggtgcct cccactaccc agctgccaac ccctctgtaa tcgtttgctg 1740 ctcctcttcc gaaaatgata gcggtgtgcc ctgcaatagt ttatatcctg aacacaggtc 1800 caatcaccct caagtggaat ctcactcata cttgaaaggc ccctcccaag aagggtttgt 1860 ctctgcattg aatggtgaca gtcacatttc agagcatcct gctgaaaatt ctttgagcct 1920 tgcagaaaag agcatattgc atgaagagtg catcaaatca cccgtggttg agacagtccc 1980 tgtttagtag cttaaattat tctaggacca actcttctct tatttaaggc actgtattta 2040 attggatttc ctgggetcat cattgtttaa actgaagacc 2080 <210> 63 <211> 1599 <212> DNA
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 1881010CB1 <400> 63 gggctgtggc ggcagctgac gcttgtggcg gcggtggctt cggggtgggc gtaagatggc 60 gacagcagcg cagggacccc taagcttgct gtggggctgg ctgtggagcg agcgcttctg 120 gctacccgag aacgtgagct gggctgatct ggaggggccg gccgacggct acggttaccc 180 ccgcggccgg cacatcctct cggtgttccc gctggcggcg ggcatcttct tcgtgaggct 240 gctcttcgag cgatttattg ccaaaccctg tgcactctgt attggcatcg aggacagtgg 300 tccttatcag gcccaaccca atgccatcct tgaaaaggtg ttcatatcta ttaccaagta 360 tcctgataag aaaaggctgg agggcctgtc aaagcagctg gattggaatg tccgaaaaat 420 ccaatgctgg tttcgccatc ggaggaatca ggacaagccc ccaacgctta ctaaattctg 480 tgaaagcatg tggagattca cattttattt atgtatattc tgctatggaa ttagatttct 540 ctggtcgtca ccttggttct gggacatccg acagtgctgg cataactatc catttcagcc 600 tctttcaagt gggctttatc actattatat catggaattg gccttctatt ggtcccttat 660 gttttctcag tttacagaca ttaaaagaaa ggacttcctg atcatgtttg tgcatcactt 720 ggtcaccatt gggcttatct ccttctccta catcaacaat atggttcgag tgggaactct 780 gatcatgtgt ctacatgatg tctcagactt cttgctggag gcagccaaac tggccaatta 840 tgccaagtat cagcggctct gtgacaccct ttttgtgatc ttcagtgctg tttttatggt 900 tacacgacta ggaatctatc cattctggat tctgaacacg accctctttg agagttggga 960 gataatcggg ccttatgctt catggtggct cctcaatggc ctgctgctga ccctacagct 1020 tctgcatgtc atctggtcct acctaattgc acggattgct ttgaaagcct tgatcagggg 1080 aaaggtatcg aaggatgatc gcagtgatgt ggagagcagc tcagaggaag aagatgtgac 1140 cacctgcaca aaaagtccct gtgacagtag ctccagcaat ggtgccaatc gggtgaatgg 1200 tcacatggga ggcagctact gggctgaaga gtaaggtggt tgctataggg acttcagcac 1260 acatggactt gtagggccac tggcaacata ctcctcttgg cccttcccat atctactctt 1320 ctgtgattgg gagactgcaa ggcactgagg agtatcaaag aagcaaatat tttcactttg 1380 aaagaaaact gccattttgt atttaatagc ctccaggttc tttcagtaat gttatttgct 1440 ctgtgtgttt ttgtgtgttt gttgatgtgc gtttgtgcat atgcgtgagt ttcattgccg 1500 gggttggggc acaattgtgg actgggggcc atgaggcctt ccctggtccc catgaaccca 1560 cttagttcca cattggccgc ~atcctgatta tgccgactc 1599 <210> 64 <211> 4137 <212> DNA
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 1593038CB1 <400> 64 , gcgcggtact agcgggcggc tccagggcgg gcgcgcgcaa ggatgctcta gggggcggcg 60 gcagtggccg tgtgggttga ggctgcgagg acagcgcggg cccccgggcc tctcccctga 120 gccggccgcc tgggccgcgc ggtcgaggcc tgtcagcctc cgcccgttca gcctcggggc 180 cggggccgcc gccacctctg cccgcggagg ccggggagcc cctgcctggg cccgcagcct 240 cccccgcggg ggtcgggccc gggcgggggt cccgagcagc tgccctcctc gccggcatcc 300 gagcctcatt tcctgctttt tcagtttcct tggggagggg cgggtgggtc tggatgaatt 360 gtctcggggt cccccgatga ggcgacccgg gcggccctgc cctttttaga gggtcccctc 420 ggggcccggt ggggaagggg ttgtctttgc atgggtgggg gacttcccga gtctgccggg 480 accatgacct gaactgtgcg agcgggacgt gtccgaagcc caagaaccag ctcatctgag 540 ccgccccctc cccggccagc atggcatctg aagaagcctc tctcagggca ttggaaagtc 600 tgatgacaga attttttcac gattgtacaa ccaatgaaag aaaacgtgag atagaggagc 660 ttcttaataa ctttgcccag caaataggag cctggagatt ctgcctgtac tttctctcca 720 gcactaggaa tgactatgta atgatgtaca gtttaacagt ttttgagaat ctgatcaata 780 aaatgtggct tggggtccca tctcaggata agatggaaat ccgtagctgt ctgcccaaac 840 tccttttggc tcaccataaa accttacctt actttatccg gaacaagctc tgcaaagtta 900 ttgttgatat tggacgtcag gattggccca tgttctacca cgactttttt actaacattt 960 tacagttgat ccagtcccct gtgacaaccc cccttgggct gatcatgttg aagacaactt 1420 cagaagagct ggcttgtccc cgtgaggacc tcagtgtggc tcggaaggag gagttgcgga 1080 agctgctact ggaccaggtg cagacagtgc ttgggctact gacaggtatc ttggagactg 1140 tctgggacaa acacagtgtt actgctgcca ctccaccacc atccccgacc tcaggagaaa 1200 gtggtgactt actgagtaac ctgttgcaga gtcccagttc agccaaactg ttgaatcagc 1260 caattcccat ccttgatgtg gagagtgagt atatctgttc cctggctttg gagtgcctgg 1320 cccatctctt cagttggatt cctctgtctg ccagcatcac cccatccctc cttaccacca 1380 tcttccactt tgcacgattt ggctgtgaca tccgggccag aaagatggcg tcagttaacg 1440 gcagcagcca gaactgtgtc tcgggtcagg agcgcggccg gctgggggtc ctggccatgt 1500 cctgcatcaa tgaactcatg tccaagaact gtgtgcctat ggaatttgag gagtatttac 1560 tgcgtatgtt ccagcagact ttctacctcc tgcagaaaat caccaaggat aacaatgccc 1620 acacagtgaa gagcaggcta gaagagctcg atgagagcta tatcgagaag tttactgact 1680 ttcttcggct ctttgtgagt gttcacctaa gaagaatcga gtcttactcc cagttccctg 1740 tggtggagtt tttgacactt ttgttcaagt acacatttca tcagcctact catgaaggtt 1800 acttctcttg tttggatatc tggacgctgt ttttggacta tctgacaagt aaaattaaaa 1860 gtcgtcttgg agacaaggaa gcagttctca acaggtacga agatgccctg gtgctcctgc 1920 tcacagaggt gttgaatcga atccagttca gatacaacca agcccagctg gaggagttgg 1980 atgatgagac tctggatgac gatcagcaga cggagtggca gcggtactta cggcagagct 2040 tggaggtggt ggccaaagtg atggagctcc tgcccacgca cgccttctcc acactgttcc 2100 ctgttcttca ggacaattta gaagtttatt tgggattaca acagtttata gtcacttcag 2160 ggtcaggaca caggttgaac atcacggcgg agaacgactg ccggcggctg cactgctccc 2220 tgagagactt gagctccctg ctgcaggccg tgggccgcct ggccgagtac tttatcgggg 2280 atgtgtttgc tgcacggttc aatgatgccc tcacagtcgt ggaaaggttg gtcaaagtca 2340 ctctgtacgg atctcagata aaattgtaca acattgaaac tgctgtgcca tcagtattga 2400 aacctgacct cattgatgtg catgctcagt ccctggctgc gctgcaggct tactctcact 2460 ggttagcaca gtattgcagt gaagttcacc ggcagaacac gcagcagttc gtgacactca 2520 tctctactac catggatgca atcacacctc taatcagcac caaggtccaa gacaagctgc 2580 tgctatctgc gtgccactta ctggtctcac tggccaccac cgtgcggccc gtctttctga 2640 tcagcatccc tgcagtgcag aaagtattca acagaatcac tgatgcctct gccctgcgac 2700 ttgtcgataa ggcccaggtg ttggtgtgcc gagccctctc taacatcttg ctgcttccgt 2760 ggccaaacct tccagagaat gagcagcagt ggcccgtgcg ctccatcaac cacgccagcc 2820 tcatctctgc actctcccgg gactatcgca acctgaagcc cagtgctgtt gccccacaga 2880 gaaagatgcc actggatgac accaaactga ttatccacca gacactcagc gtcttagaag 2940 atattgtgga gaatatctcg ggggagtcca ccaagtctcg acagatttgc taccagtcgc 3000 tgcaggaatc tgttcaggtc tccctggccc tctttccagc ttttatccat cagtcagatg 3060 tgactgatga gatgctgagc ttcttcctca ctctgtttcg aggccttaga gtacagatgg 3120 gtgtgccttt cactgagcaa atcatacaga ctttcctcaa catgtttacc agagagcagt 3180 tagccgagag catcctccac gagggcagca caggctgccg ggtggtggag aagtttctga 3240 agatcctgca ggtggtggtc caggagccag gccaggtgtt caagcccttc ctccccagca 3300 tcatcgccct gtgcatggag caagtgtatc ccatcattgc cgagcgtccc tcccctgatg 3360 tgaaggccga gctgtttgag ctccttttcc ggacgctcca tcacaactgg aggtacttct 3420 tcaagtccac cgtgctggcc agtgtccaga gggggatcgc tgaggagcag atggagaatg 3480 agccccagtt cagtgccatc atgcaggctt tcggacagtc ctttctccag cccgacatcc 3540 acctttttaa acaaaatctc ttctacttgg agactctcaa caccaagcag aagctgtacc 3600 acaagaagat cttccggact gccatgctgt tccagtttgt gaacgtgctg ctccaggtcc 3660 tggtccacaa gtcccatgat cttctgcagg aggagattgg catcgccatc tacaacatgg 372 0 cctcagtcga ctttgatggc ttctttgccg ccttcctccc agagttcctg accagctgtg 3780 atggtgtgga tgccaaccag aaaagtgtgc tggggcggaa tttcaagatg gatcgggacc 3840 tgccctcatt cacccagaat gtgcacaggc tggtcaacga cctgcgctac tacagactct 3900 gcaacgacag cctgccccct ggcactgtga agctctaggc ctgctactgc ctgggggaca 3960 cggacttctg ctgctgccac ctgcgccagc cctaccttcc accacagatg tctccccaga 4020 tgggccttgg tcacactcct tggcttctcc ccaccgcaag caacgctgcc tgctcttgcg 4080 tcctccacat cttggcgctg ccagcagagc tggcttctgg gtccacctga gcacctg 4137 <210> 65 <211> 4019 <212> DNA:
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 7494930CB1 <400> 65 tggtaccagc cactgcaaaa acataccaaa ttataaagat catcaacact atgaagaaac 60 tgcatcaact aatgggcaag aataaccagc taacatcata atgacaggat ccgatttaca 120 catcaccgta ttgaccttac atgtaagtgg gactaatgac ccaattaaaa ggcacagact 180 ggacaattgg ataaaaagtc aagacccatc ggtgtgctgt attcaggaga cccatctcac 240 gtggcaagac acgcacgggg ctcaagtaaa ggggtggaaa aaggcatttc atgcaaatgg 300 acaccaaaaa caagcaggag tagctattct tatatcagac aaaacaaact gtaaagcaac 360 agcagttaaa aaagacaaag agggacatta cataagggta aaaggtcttg tgcaacagga 420 aaatatcaca gtcctaaaca tatatgcact taacactgga gcttccaaat ttataaaaca 480 attactaata gacctaagaa atgagacaga cagcaacaca atagtagtgg gggacttcaa 540 tgctccactg acaacagtag acaggtcatc aagacagaaa gtcaacaaag aaacaatggg 600 tttaaactac accttggaac aaatggactt aacagatgta tacagaacat tccatccagc 660 aaccagagaa tacacattct attcaacagt gcatggaact ttctccaaga tagaccatat 720 gataggccac aaaacaaatc tcaatacatt taagaaaact gaaattatat caaggactcc 780 ttcagaccac agtgtaatca aattagaact cagaattaag aaactcactc aaaaccgcac 840 aactacgtgg aaattgaaca acttgctgct gaatgactct tgggtaaata atgaaattaa 900 ggcagaaatt aagaagttgt ttgaaaccaa tgagagcaaa gagacaatgt accagaatct 960 ctgggacaca gttacagcag tgtttagagg aaaatttata gcactaaatg cccatgggag 1020 aaagcaggaa agatctaaaa tcaataccct aacatcacaa ttaaaagaac tagagaagca 1080 aaagcaaata aattcaaaag ctagcagaag acaagaaata actaagatca gggcagaact 1140 gaaggagaca gagacacaaa aaacccttca aaaaataaat gaaacaaaaa gctggttctt 1200 tgaaaaaaat aaaattgata gaccattagc aagattcatc cagacgaaaa gagataagaa 1260 ccaaacagac acaataaaaa atgataaagt ggctatcacc actgatccca cagaaataca 1320 aactaccatc agagaatact ataaacacct ctatgcaaaa aaactagata atctagaaga 1380 aatggataaa ttcctggact catacaccct tccaagacta aatcaggaag aaacagaaaa 1440 cctgaacaga acaataatga gtagtaaaat tgaatcagtt ataaaaagtc tcccaacaaa 1500 gaaaagtcca agactggatg gctttactgc tgaattttac caaacttata aagaactaat 1560 actaattatt ctcaaacaat ttcaaaagat agagaaagac ggaaccctcc ctaattcatt 1620 ctatgaagcc agcatcacca taataccaaa accaggaaag gacataacca aaaaagaaaa 1680' ctatagactc atatccctga tgaacataga tgctaaaatc cttaacaaaa tgctagctaa 1740 ccaaatccaa caacatatca aaaagataat ccaccatgat caagtgggtt tcataccagg 1800 gatgcaggaa tggtttaata cacacatgtc aataaatgtg atgtaccacg taaacagaat 1860 ..
taaaacaaaa aatcacttga tcatctcaat agatgcagaa aaagcatttg ataaaatcca 1920 gcattgcttt atgattaaaa ctctcagtaa aatcggtgga caagggacat acttcaacat 1980 aataaaagcc atctatgaca aaagcacagc caacataata ctgaatgggg aaaagttgaa 2040 agcattccct ctgagaaatg gaaaaagaca aggatgccca ctctcaccac tcctcttcaa 2100 catagtactg gaagtcctag ccagagcaat cagacaagag aaagaaataa.agggcatcca 2160 aattggtaaa gaagaagtca aactgtcact gtttgcagaa gacatgattg tatatttaga 222 0 aaaccccatc atctcagccc aaaatctcct taagctgata agcaacttca. gcaaaatctc 2280 aggatacaaa atcaatgtgc aaaaatcagt agctctccta tgcgccaaca~gcgaccaagc 2340 agagaatcaa atcaagaact caaccccttt tacaatagct gccaaaaaaa acaaacactt 2400 acgaatatac cttaccaaag aggagaaaga cttctacaag gaaaactaca aaacactgct 2460 gaaagaaatc atacatgaca caaacaaatg gaaacatatc ccatgctcat ggataggtag 2520 aatcaatatt gtgaaaatgg ccatactgcc caaagtaatt tatagaatca atactatccc 2580 catcaagcta ccattgactt tcttcacaga attagaaaaa acaactttaa atttcatatg 2640 gaaccaaaaa aagagcctgt atagccaaga caatcctaaa caaaagaaca aagctggagg 2700 catcatgcta cctaacttca aactacacta caaggctaca gtaaccaaaa cagcatggta 2760 ctggtaccaa aacagatata tagaccaatg gaacagcaca gaggcctcag aaataacacc 2820 agacatctac aaccatctga tctttgacaa acctgacaaa aacaagaaat ggggaaagga 2880 ttccctattt aataaatggt gttgggaaaa ctggctagcc atatgcagaa aactgaaact 2940 ggatcccttc cttacaactt atacaaaaat taactcaaga tggattaaag acttaaacct 3000 aagacctaaa accataaaaa ccctagaaga aaacccaggc aataccattc aggacatagg 3060 catgggtaaa gacttcatga ctaaaacacc aaaagcaatt gcaacaaaag ccaaaattga 3120 caaatgggat cttattaaac taaagagctt ctgcacagca aaagaaacta tcatcagagt 3180 gaacaggcaa cctacagaat gggagaaaat ttttgcaatc tgtccatctg acaaagggct 3240 aatatccaga atctacaagg agcttaaaca aatttacaag aaaaaaacaa acaatctatt 3300 aaaaaaatgg gcaaaagaca tgaacagaca cttctcaaaa gaagacatac aagcagccag 3360 caaacatatg gaaaaatgct caatatcact aatcatcaga gaaatgcaaa tcaaaaccac 3420 aatgatacct aaccttactc ctgtaagaat ggccataagc aaagaagcaa aaaacagtgg 3480 atgtttgcat ggatgtggtg atcagggaac acttttacac tgctggcggc aatgtaaact 3540 agtacagcca ctatggaaaa cagtgtggag atttcttaaa gagctaaaag tagacccacc 3600 atttgatcca gcaatcccac tactgggtat ctacccagag gaaaagaagt cattatacaa -3660 aaaagacact'tgcacacaca tgtctatagc agcacgattc gcaattgcaa aattgtggaa 3720 ccagccaaat tgcccatcaa ccaaggagtg gataaagaaa atgtggcaga tgtatacctt 3780 ggaatactat gcagccataa aaaaggatga gatcatgtcc tttgcaggga catggatgat 3840 gctggaaacc atcattctca gcaaactaac acaggaacag caaaccaaac actgcatgtt 3900 ctcactcata agtgggagtt gaacaatgag aacacatgga cacagggaag ggaatatcac 3960 acactggggt ctgttttggg tggggggcta gggagcgata gcattaggag aaataccta 4019 <210> 66 <211> 1965 <212> DNA
<213> Homo sapiens <220>
<221> misc_feature <223> Incyte ID No: 7497349CB1 <400> 66 tccaacccta tgccagtgag ctccacgaag cagtgacgcc tgcagagact ctggccacag 60 gaagtgtcag tatcctttgg tgcagctgtg aatggtgccc aggagctgag cgtcttgtag 120 tgcctgaagg gcgcattggt ggccaccccc cgtgagcgag aggacatcca gcccccacct 180 caggggactc agtggttgga gaaagttcct gattcagctc cacgtggctc tccatggaga 240 agcagaaaga gaatcgtgtt ttcttgcaga ggcattgagg ctttgggaac gatcactgca 300 gtgcctgtca cgggtcctca ggtcagctcc ttgcagaggt tggccgggca aggagcggca 360 gtgctacctc aggttaggcc aaagactctg attccagaca gcctccccgt tgccccgggc 420 cgggaccggc cacccaagca gcccccaaca ttccagaagg ccaccgtggt cagcgtcaag 480 aaccccagcc cagccctccc caccgccaac aacactgtca gccatgtgcc agcgcccggc 540 agccagcccc aggccctcgc cgagc~cgcc gccctcgcct ctccgctgag cagtgcgggg 600 gtggcctacg ccatcatctc cacctccccc agcaatgccg ccgccatggc ccccagcacc 660 gccgtgtctg tggtcagtga cagcatcaaa gtccagcccc tcctcatcag tgctgacaac 720 aaggtcatca tcattcagcc tcaagtgcag~acgcagcccg.agagcacggc agagtcgcgg 780 ccgcccacag aggagccatc tcagggagct caggccacca.aaaagaagaa ggaagaccgg 840 cccccgaccc aggagaaccc cgagaaaatc gccttcatgg tagcgctagg cctggttacc 900 acggaacatt tggaagaaat ccagagcaag cgacaggagc ggaagagaag aagcacagcc 960 aaccctgcct acagcggcct cctggagacc gagaggaaac ggctggcctc caactatctc 1020 aacaaccccc tgttcctcac agcgagagcc aatgaggacc cctgctggaa gaacgagatc 1080 acccacgatg agcactgtgc cgcctgcaag cgaggggcca acctgcagcc ctgcggcacc 114 0 tgcccggggg cctaccacct cagctgcctg gagccgcccc tcaagacggc gcccaagggc 1200 gtgtgggtgt gccccaggtg ccagcagaag gccttaaaga aagacgaggg tgtgccctgg 1260 actgggatgc tggccatcgt gcactcttat gtcacccaca agacagtcaa agaagaggag 1320 aagcagaagc tgctgcaacg aggcagtgag ctgcagaacg agcaccagca gctggaggag 1380 cgggaccggc ggctggcgtc agcagtgcag aaatgcctgg agttgaagac aagcctgctg 1440 gcccgccaga ggggcaccca gtcatccctg gaccgcctgc gggccctcct gagactgata 1500 cagggcgagc agctgctcca ggtcaccatg acgaccacta gccctgcccc actgctggcc 1560 gggccctgga ccaagccctc agtggcagcc acacacccca ccgtccagca cccccagggc 1620 cacaactgac cccgagggac cagtcttcat acccacgccg ggtgaggagg acgcaatgct 1680 cttcccctct ggaggagctg ttggaagcag agagcagccc tcaccgcaca gccaacctat 1740 ggtgccttcg tcttggactt cccagcctcc agaactcatt ggtacctgca acccacaggc 1800 gtgttttcta cttgcgagtg actgtggacc ggcccagccc agccaactgg gcaaacctca 1860 gcacccagag ggcttgcagc tgctccttct ccaaagaggt ctgaatctca gctttgggga 1920 tgggcttccc cttcgaaatt gttcctcccc aggatgctct catgt 1965 <210> 67 <211> 499 <212> DNA
<213> Homo.sapiens <220>
<221> misc_feature <223> Incyte ID No: 5510805CB1 <400> 67 ggagcgagga acatgccaag gaggactgga gagtgcagag gaggagcaag gaacatagca 60 ggaagggggc caagagaagg gcctcctgtc ttcccttccc cagtgcaggt ttccagaccc 120 acagccgctc aagctcacag agaggaaaag tttctctctg aatgaatcct gaagctaata 180 atgttatatt agactaggcc gggcatttta attttgtaaa tgatactatt tgtgatgcga 240 aagagccaga aaaaaagcta tgttatcttc aaaaatcagt gtgattctgt ttagaactag 300 acagagtttt gttctaaagg gacagctgga gaaaagaata aggctgtttt atgattgttc 360 ctggagtcta acttacccat ataacacaag tgaattcatg aaagggtgga gatcatgtct 420 aatgcttatt gaagctgcta aggtactttg cacattacac atcattttcg ttagagctta 480 tttgattgca agaaaatag 499 <210> 68 <211> 3760 <212> DNA
<213> Homo sapiens <220>
<221> misc_feature <223> Incyte ID No: 1577482CB1 <400> 68 cggcaccgtc gtaggtgcgg gccgcatgaa tggagcgccg ggcgtaaggc aaagcctggc 60 accgtctgcg cggccgctat ctgctcccgg agcgtgagtg cggggtgtgg ggcgtgcgcg 120 tgcgcgctca gagggggctc aaggcgagcg cgccgggcag ttgcgggcgc gtggctgctg 180 aggttggcgg cggtgccgcg cgcccgacgg gccggtggtt gcggggcctc ccgcctcgac 240 ccgggctggg ggcagccgtg gcgggcgccg gggaccgcaa ggggcggagg aaaggagggg 300 gccggtcccg gcacgcagag gagcagccga ccatgccccg agacaacatg gcctccttga 360-tccaacggat cgcccgccag gcttgcctca ccttccgggg cagcgggggc ggccgcggcg 420 cttccgatcg cgacgcggct tctggcccgg aggcgccgat gcagccgggc ttccccgaga 480 acctgagcaa gctgaagagc ctcctgaccc agctccgcgc-cgaggacttg aacatcgccc 540 cgcgcaaggc cacactgcag ccgctgccgc ccaacctgcc gccagtcacc tacatgcaca 600 tctacgagac ggacggcttc agcctgggcg tgttcctgct,caagagcggc acgtccatcc 660 cgctgcacga ccacccgggc atgcacggca tgctcaaggt gctgtacggc accgtgcgca~720 tcagctgcat ggacaagcta gacgcgggcg gcgggcaacg gccgcgggcc ttgccgcccg 780 agcagcagtt cgagccgccg ctgcagcccc gggagcgaga agccgtgcgg ccgggcgtgc 840 tgcgttcgcg ggccgagtac accgaggcca gcggcccctg catcctcaca ccgcaccggg 900 acaacctgca ccagatcgac gccgtggaag ggcctgccgc cttcctggac atcctggccc 960 cgccctacga cccggacgat ggccgggact gccactatta ccgggtgctg gagccggtca 1020 ggcccaagga ggcctccagc tcggcctgtg acctgcctcg agaggtgtgg ctcctggaga 1080 ccccacaggc cgatgacttc tggtgcgagg gagaacccta tccaggtccc aaggtcttcc 1140 cttgaagcca ctggcgccca ggagcggtgg gccgaagacg tgccctaccc taccacaagg 1200 gctgtgtctc taccccctag cctgggcgtt ggatctactg gaatgagcag cagccgcttc 1260 ctcggcagcc ttgggaagca cgggcgactg gacagcagcc gccgggcacg gttatggggg 1320 cggggtgggc ggggaggcta gattgtttcc tggtactgtc actgccactg gggctttgat 1380 ttggaggaat ggggcagggg actatctgaa gcgcttccat cctaaagcca taatgaaaat 1440 atcttcctct cttccccatt ctatacaaaa tactaagtgg ttttcttgct cccactccct 1500 accccttagt taaatagggt ttattttcca ctcatgccct tatgcctttt tttcttatag 1560 ttttttaact tattgactgt gcatgaccca gtggtttgaa ttgtttttag ttcaagtcat 1620 tggtaaaaac taggtttaag gagatgagct actgtttaaa gtgagctggc ctgcctaatt 1680 aattccttgt gaaaactaaa tgattttttc agtttgggga tcattctcac aacataacta 1740 tgcatgtaga ggacaagatt tattttcttt tctccctttg cccagtagcc acatctggtt 1800 tactcaggca gcatctacta agaaattcag cacctgcata tctctgtgac atggtcactt 1860 agagcttatc ttccctatga atctccagat ctgtgagtcg agcagatttc atgttgcaga 1920 ttcaccttta atgcaaagac tgtattatcc tcacatgact ttttttcttg tcttactgta 1980 ccttaaaagg tgatagagta attctgtatt ttctaacggg aagattcaaa ggagctgaat 2040 gtgttatgct tccaaacaac tgaatgtaaa acactcctag ccagttgttg cattccctat 2100 atttatttac ttccaatatt ttactgtaaa agtagggaga aatattatgt tgatagttgt 2160 ttcatattct ctcaggaact ttaatgttcc cgactcgggt gattccagct gtgttgctgg 2220 cagtgttgtc tcaaccctct ccctaaaatg actgagccct gggttcatct aatgtggttt 2280 tccttaggaa gagatagaag gcacagaaga tcacagctag agaattgaga attaactata 2340 ctactagcca ttttagggca ccaaaacttg ggattaaaca cttcctactt cccactccca 2400 actcctgaaa tgaagtcttg ctatctgtga ctagttttat ttttgtgctt ttaatagtcc 2460 gagcagtctt accttgttta cacatgtatt gacaccattt gcttcaggcc atggagcact 2520 gtttctccct ttttactatt tataggattc cgttttttca caagactttt aataaaaaga 2580 aattgtagaa ataaacacat taaaatttgc ccagctgtcg tctaagccct cttgattgac 2640 ttgcccaagt ggcaatagag ttctaatatc tataataaag ggagatttgc tattattagt 2700 ggaatgttga ccccgtatgt agagaaacag tacgtgcctt tgcctcttta tgcacacaga 2760 gacccagggt gagggagtat ttgttcccag tttttaagat agtataaaaa agcaaatact 2820 tggtgagtga tttaaaaata aaaccaaaac aaaacacaaa aagatattcc acaggacatg 2880 ccactttatt ataaaacctg acacaggcat agtaccaagt atttcctgca ttgttgctaa 2940 aattgtttta ttgtagctcc acattctggt gtagtttaaa atgcctttgg gggcagtttg 3000 aagcagttct tcatgccact tagtttgaaa ataaaattct agatatgcaa atgattttct 3060 tagaaaactt cacaaaataa aagatcttgt ttttttttcc atagcacagt aatgaatgtg 3120 gttatcaatc acatactttt ttggattata ttgtagcaaa aagttgatta gcttaccaag 3180 attattaata gcaatgtatg tgttataata caacttagta cattaaagct acgaaaactc 3240 atcctggctg taggatagta ataaaggaag aattatgact tcattatgaa aaaaagaagt 3300 tttaaagttt tcaattacga gcaatttgga agaaaaaacc taaggtgctt ttcaaaagag 3360 taactgaaat tgttgcaggc caaaacagca atatgatatc tcagatttag ttcaataaga 3420 acagtgaaac ttttggttca ctaataaatt ctgagtaaat tagtggtgaa gacaaaatat 3480 aacttgtttt agtgagccac tgaggaaaga atatgcttat tacaaagaca aaatgtggtg 3540 cagaaactat cttgcacctg tgtgcataaa ctgttagtcg tgactgactt ggtgtgttgc 3600 tattgtgttt ctatatactc cgtccaatat agataatgtt ttaataacaa ctgtgggata 3660 aaagttatct tccccttgga aagactaatg agcacaatga tattaatcac ttttatggtg 3720 aataataaat gcaataattg cctcatgggt gaaaaaaaaa 3760 <210> 69 <211> 2800 <212> DNA
<213> Homo sapiens <220>
<221> misc_feature <223> Incyte ID No: 1505054CB1 <400> 69 ttgatactag gtggccacct ggcgataagg actgcagggc ctaagaaagt acaaaagatg 60 acctttagta ggagctccca acccagaacc acaggaaatc aaaaaaagcc cgcacgaact 120 aggaaagaaa atacacaacc ggccatccgt gcgatcaccc atctccctca ccaggaaagt 180 agctccaaac cgccaatcag cggcgacgct ggacgtagac gtcctacgcc gtgatattaa 240 agcaagatgg ccgcgccctg cagattgtct cttgttgcgt aagttttttt gaccgtcact 300 cgtgtcagct tcaaagtcag atagattttt ctcccagcat gttctacttc cgaggctgtg 360 gccgttgggt cgcggtttcc ttcaccaagc agcaatttcc gttggcacgg ttgagcagtg 420 acagcgcggc gccccggact ccgcacttcg acgtgatagt cattggtgga ggacatgccg 480 ggactgaggc agccaccgcc gccgctcggt gcggctctcg gactctgctc ctcactcacc 540 gcgtggacac gatcggtcag atgtcatgta atccttcctt tggtggcatc ggaaagggac 600 atttaatgag ggaagtagat gccttggatg gcctgtgttc tcgcatctgt gaccagtctg 660 gtgtacatta taaagtatta aaccggcgta agggaccagc tgtgtggggt ctgagagctc 720 agattgatag gaaactctat aaacagaaca tgcagaaaga aatcttgaat acaccactgc 780 ttactgttca ggagggagct gtagaagatc ttattcttac agaaccagag cctgaacaca 840 ctgggaaatg ccgtgtcagt ggggttgttt tggtggatgg aagcacagta tatgcagaga 900 gtgtgattct gactactggg acatttctga gaggcatgat tgtaattgga ttggagacgc 960 atccagcagg acgtttaggg gatcagcctt ctataggatt ggctcagaca ctggagaagt 1020 tagggtttgt ggtgggaagg ttgaagactg ggactccacc ccgaattgcc aaagagtcca 1080 ttaatttcag tattctaaac aagcatatac cggacaatcc atccatacca ttcagcttta 1140 ccaatgagac agtatggatt aagccagaag atcagctgcc atgttacttg actcacacca 1200 accctagagt ggatgagatt gtccttaaga accttcacct taatagtcat gttaaagaaa 1260 cgacaagagg acctcgatac tgtccctcca ttgaatcaaa agttttgcgt tttccaaacc 1320 gtctacatca ggtttggttg gaacctgaag gaatggattc tgaccttatc tacccacagg 1380 ggttatctat gacgctacca gctgagttac aagagaaaat gatcacatgc atcagaggct 1440 tggagaaagc taaagtgatt cagccaggct acggtgttca gtatgattac ttagatcccc 1500 gtcagatcac cccttccttg gagactcatt tggttcaacg actcttcttt gctggacaga 1560 tcaatggcac cactggttat gaggaagctg cagctcaagg tgtgatagcc ggaatcaacg 1620 ccagtcttcg ggtcagtcgc aagcctccct ttgtggttag ccgaacagaa ggttacatag 1680 gagtcttgat tgatgacctc actactctgg gcaccagtga accataccgc atgtttacca 1740 gccgagtaga gttccgtttg tcactgcgcc ctgataatgc tgacagccgg ctcacactgc 1800 gagggtataa agacgctggc tgtgtgtccc aacaacgata tgaaagagct tgttggatga 1860 agtcttcttt agaagaaggc atttctgtgt tgaaatctat tgagtttttg agctctaaat 1920 ggaaaaaatt aatcccagag gcttctataa gtactagtag aagtctgcct gtcagagctc 1980 tcgatgttct gaagtatgag gaagttgaca tggattcatt agccaaggct gttccagagc 2040 ccttgaagaa gtatactaaa tgtagagagc tggctgaaag actgaaaata gaagccactt 2100 atgaatcagt gttgttccat caactacaag aaataaaggg agttcagcaa gatgaagctc 2160 tccaactgcc aaaagaccta gattatttga ctatcaggga tgtgtctttg tcccatgaag 2220 ttcgagagaa actacatttt agtcgtccac agacgatcgg ggctgctagt cgcatacccg 2280 gagtaacacc tgccgccatc atcaatctgc tgagatttgt gaagaccact caacgaagac 2340 agtcggctat gaatgaatca tccaagactg atcaatactt atgtgatgca gacagacttc 2400 aagagagaga gttatagctt tcaattcata aaagattttt aaagagcata taaataattt 2460 gatcaataca acagtataga taaaagaatt atttagcaca tgttaaaata gctttattag 2520 gttactatgg gtttgccatt aatttctgag tgggacagaa attataattg tgctttttcg 2580 tgtatatgaa aaaactagtc gtaaacaatt tgtactcttt ctttaaggag ctgtaataca 2640 aataactttg tgcagtgttc atcaaagaga gagacagtga acctaaaact gaacctggaa 2700 taaaactcaa catgcagatt tgcctactca ta.gggacttt gcctattaag tctaccaaat 2'l60 taaaagtctt atcattcagc gtgaaaaaaa aaaaaaaaaa 2800 .
<210> 70 <211> 1314 <212> DNA
<213> Homo Sapiens <220>
<221> misc feature <223> Incyte ID No: 7492708CB1 <400> 70 caagatccgc tgttctacag ccactgctgc tgatacccag gcaaacaggg tctggagtgg 60 acctctagca aactccaaca gacctgcagc tgagggtcct gtctgttaga aggaaaacta 120 acaaacagaa aggacatcca caccaaaaac cctctgtaca tcaccatcat caaagaccaa 180 aagtagataa aaccacaaag atggggaaaa aacagagcag aaaaactgga aactctaaaa 240 agcagagcgc ctctcctcct ccaaaggaac gcagctcctc actagcaacg gaacaaagcc 300 ggacggaaaa tgagtttgac gagctgagag aagaaggctt cagacgatca aactactccg 360 agctacagga ggaaattcaa accaaaggca aagaagttga aaactttgaa aaaaatttag 420 atgaatgtat aactagaata accaatacag agaagtgctt aaaggagctg atggagctga 480 aagccaaggc tcgagaactg cgtgaagaat acagaagcct caggagccga tgcgatcaac 540 tggaagaaag gctatcaatg atggaagatg aaatgaatga aatgaagcga gaagggaagt 600 ttagagaaaa aagaataaaa agaaacgaac aaagccccca agaaatatgg gactatgtga 660 aaagaccaaa tctatgtctg attgttgtac ctgaaagtga cggggagaat ggaaccaagt 720 tggaaaacac tctgcgaaat attttccagg agaacttccc caatctagca aggcaggcca 780 acattcagat tcaggaaata cagagaacgc cacaaagata ctcctcgaga agagcaactc 840 caagactcat aattgtcaga ttcaccaaag ttgaaatgaa ggaaaaaatg ttaagggcag 900 ccagagagaa aggtcaggtt acccacaaag ggaagaccat cagactaaca gcagatctct 960 cggcagaaac tctacaagcc agaagagagt gggggccaat attcaacatt cttaaagaaa 1020 agaattttca acccagaatt tcatatccag ccaaactaag cttcataagt gaaggagaaa 1080 taaaatactt tacagacaag caaatgctga gagattttgt caccaccagg cctgccctaa 1140 aagagctcct gaaggaagca ctaaatatgg aaaggaacaa ccagtaccag ccactgcaaa 1200 atcatgccaa attgtaaaga ccatcgagac taggaagaaa ctgcatcaac taatgagcaa 1260 aataaccagc taacatcata atgacgggat caaattcaca cataaaaata ttaa 1314 <210> 71 <211> 3594 <212> DNA
<213> Homo Sapiens <220>
<221> misc-feature <223> Incyte ID No: 7490847CB1 <400> 71 aataggtggg atttctctac acccagccaa cttaatgttt tataatgtct cttattttct 60 tatttaacct cgttattggg atgttgattt acagtgaaga atccattgca ttagtaaggg 120 tgatatctta gtattttgct tcctgggtga gctagctttg cttatttctg tttgttaact 180 aggccttttg atatttttat agacaatctg cttcatcctg attctgcggc attccaaact 240 gtggtatcct tgggggttct cttaacattg ctatggtgca gaagatttaa aatcagctga 300 tgttcacaag gaatagagtc acgtgatgta tgaagggaaa catatacact tctctgaggt 360 tgacaataag cccttgtgct catatagccc caaactgtgc aagcagaggc gactcaacgg 420 ctacgccttc tgtatcagac acgttctgga ggacaagact gcccccttca agcaatgtga 480 atatgtggcc aagtataaca gccaacgctg caccaacccc atccccaaat cagaggatcg 540 taggtactgc aacagccact tgcaggtact tggctttatc ccgaaaaaag agaggaagaa 600 aaagaatgat cctatagatg aggtgaaggt caggcaccag atggatacca tggcctttag 660 cctgacagtg cccacgttgg ccttgaagat gcccaacgga ctggatggaa tgtccctctc 720 tccacctggg gcaagggtcc ctctccacta cctggaaact gaattggaag acccatttgc 780 tttcaatgag gaagatgatg acctaaagaa aggggcaact gtgagaaaga agttgcagag 840 caagttggcc cagaatcggc agcgccagag agagacagag attttaaaag ttcgacaaga 900 gcactttagt ccccctcctg caccttcaca gcagcagcct ccgcagcagc actcccacct 960 gtcaccttta tctacttctt taaaacctcc agcgccaccg cagggttcag tctgcaagtc 1020 acctcaacct cagaacacca gcctaccaat gcagggagtg gcacccacca cacacactat 1080 agcacaagca cggcagttgt ctcacaagag gcctctgccc ctcctgccat ccagtagggc 1140 tcccactgtg gacccaccca ggactgaccg gatcctcatg aaagccacag ccttctctcc 1200 acacttctca tgtataagcc gactgcagag actggtgaaa ctgtgcaccc agaaacatca 1260 gttggacact gatctgtttc cccatttagg tttggactgg tctgaagaga gcggagagga 1320 accagaggac tcagagcagg cctcgcccta ccaggttgca tggtccatcc gggaaaccct 1380 cagatatcaa agacatgcgt cagatgatga tgatgcggag agtaggagct ccggggtgac 1440 tcaactttgc acttactttc agcagaaata taagcacctc tgccgcctgg agcgggcaga 1500 atctcgtcaa aagaaatgcc ggcatacgtt taggaaagct ttgctgcagg cggccagtaa 1560 agaaccagaa tgcactggtc agttaataca agaactgcgg agagctgcat gcagtcgaac 1620 cagcataagc cggaccaagc tgagggaggt ggaaccagca gcatgcagtg gaaccgtgaa 1680 gggtgaacag tgcgctaaca~aagcccttcc attcaccaga cattgtttcc aacatatcct 1740 cttgaaccac tctcagcagc tcttctcaag ttgcacagcc aagtttgcag atggacagca 1800 gtgctctgtg ccagtttttg acattacaca tcagacacct ctgtgtgaag aacatgccaa 1860 aaaaatggat aatttcttga gaggagataa ctcccgtaaa gttcagcacc agcagcagag 1920 gaaacccagg aaaaaaacca agcctcctgc actaaccaaa aaacacaaga agaagagaag 1980 gcgtggacct cgtcgacccc aaaaacccat tccacctgca gtcccccaag ggaacctcag 2040 catgcctgcc agcgtctcac tgccagtgga ggcctctcac atccggagcc catccacgcc 2100 agagctgagt gctgatgagt tgccggatga cattgccaat gagatcactg acattccaca 2160 tgacttggaa ttgaaccagg aggacttttc agatgtcctg ccacggctac ctgatgactt 2220 acaagatttt gatttttttg aagggaagaa tggagacctc ctcccaacta ccgaagaggc 2280 tgaggagctt gaacgggcct tgcaggctgt aacttctctc gagtgcctga gtaccattgg 2340 ggtccttgcc cagtcagatg gtgtgccagt ccaggagttg tcagatagag gaataggggt 2400 gttctccaca ggtactggag cttcaggaat acaatccttg agccgagagg tgaacacaga 2460 cctaggggag ctattgaatg ggcgtatagt acatgataat ttttctagtc tagagctgga 2520 tgagaacctg ctccgttctg ctaccttgtc aaacccacct acacccctgg cagggcagat 2580 ccaggggcag ttctctgccc cagccaacgt tggccttact~tctgccactc tgatcagcca 2640 gagtgcactt ggggagagag ccttcccagg acagtttcat ggacttcatg acggcagcca 2700 tgcctcccag aggccacatc ctgcccagct gctgagcaag gcagatgacc taatcacctc 2760 acgacagcaa tacagcagtg atcactcaca ctcctcaccc catggaagcc attatgatag 2820 tgagcatgtg ccgtctccct acagtgacca tatcacctct ccccacacaa catcgtactc 2880 tggtgataat atggcagcta ccttttcagc agagatgccc atcatggcgc agcacttgct 2940 cccaacccaa cttgaggtgc cacttggagg cgtggtaaac cccagaactc actggggcaa 3000 tctccctgtc aaccttggag acccctctcc atttagcaac cttctcggcg cagatggaca 3060 tcttctttcc acttccctat ccacgccacc caccacttcg aactcagaga ccacacagcc 3120 tgccttcgcc accgtgaccc ccagcagctc cagtgtgctt ccggggttac cacagaccag 3180 cttcagtggc atggggcctt ctgctgaact aatggcctcc acctctccca agcagcaact 3240 ccctcagttc agcgcagcct ttggccacca gctgagttct cacagtggca ttcctaagga 3300 cctgcagccc agccacagct ctatagcccc tcctacaggc ttcacagtaa caggtgccac 3360 agctacaagt accaataatg catcttctcc ctttccctcc cctaactgag ggtgtctgtg 3420 tgtttgcagg caggtgg,gga acccagtgtt ttctatcttt gtttcctctt tagcacccct 3480 ctcccctttt cctaaagaag atataacaga tggggatcag aggggcaccc'cccttcccag 3540 ttcaatgagt ggccctgcag tggacctcgc tacagtgttc acatgtgctc ctta 3594 <210> 72 <211> 4123 <212> DNA
<213> Homo Sapiens <22.0>
<221> misc feature <223> Incyte ID No: 7493059CB1 <400> 72 ataaaatcct ttacagacaa gcaaatgctg agagattttg tcaccaccag gcctgcccta 60 caacagctcc tgatggaagc actaaacatg gaaaggaaca actggtacca gccactgcaa 120 aaacatgcca aattataaag accatcgagg ctaggaagaa actgcaacta atgagcaaaa 180 taaccagcta acatcataat gacaggatca aattcacaca taacaatatt aaccttaaat 240 gtaaatgggc taaatgctcc aattaaaata cactggctgg caaattggat aaagagtcaa 300 gacccatcag tgtgctatat tcaggaaacc catttcacgt gcagagacac acataggctc 360 aaaataaagg gatggaggaa gatctaccaa gcaaatggaa aacaaaaaaa ggcaggggtt 420 gcaatcctag tctcggataa aacagacttt aaaccaacaa agatcaaaag agacaaagaa 480 ggccattgca taatggtaaa gggatcaatt caacaagaag agctaactat cctaaatata 540 tatgcaccca acacaggagg acccagattc ataaagcaag tccttagaga gaggaatcag 600 tcattagact cccacacaat aataatggga gactttaaca ccccactgtc aacattagac 660 agatcaacaa gacagaaagt taacaaggat atccaggaat tgaactcagc tctgcaccaa 720 gcagacctaa tagacatcta cagaactctc cacccccaat caacagaata tacattcttc 780 tcagcaccac accgcactta ttccaaaatt gatcacatag ttggaagtaa agcactcctc 840 agcaaatgta aaagaacaga aattataaca aactgtctct cagaccacag tgcaattaaa 900 caagaactca cgattaagaa actcactcaa aaccactcaa ctacatggaa actgaacaac 960 ctagtcctga atgactactg ggtacataac aaaatgaagg cagaaataaa gatgttcttt 1020 gaaaccaacg aggacaaaga cacaacacac cagaatctct gggacacatt caaagcagtg 1080 tgtagaggga aatttatagc actaaatgct cacaagagaa agcaggaaag atctaatatt 1140 gacaccctga catcacaatt aaaagaacta gagaagcaag agcaaacata ttcaaaagct 1200 agcagaaggc aagaaataac taagatcagg gcagaactga aggaaataga gacccaaaaa 1260 acccttcaaa aaaaaataaa tgaatccagg agctggtttt ttgaaaagat caacaaaatt 1320 gatagaccac tagcaagact aataaagaag aaaagagaga aaaatcaaat agacacaata 1380 aaaaatcata aaggggatat caccaccggt cccacagaaa tacaaactac catcagagaa 1440 tactataaac acctctacgc gaataaacta gaaaatctag aagaaatgga taaattcctc 1500 aacacataca ccctcccaag actaaatcag gaagaagttg aatctctgaa tagaccaata 1560 acaggttctg aaattgaggc aataattagt agcttaccaa cccaaaaaag tccaggacca 1620 gatggattca cagccgaatt ctaccagagg tacaaagaga agctggtacc attccttctg 1680 aaactattcc aatcaacaga aaaagaggga atcctcccta actcatttta tgaggccagc 1740 atcatcctga taccaaagcc tggcagagac acaacaaaaa aagagaattt tagaccaata 1800 gccctgatga acagcaatgc aaaaatcctc aatagaacac tggcaaaccg aatccagcag 1860 cacatcaaga agcttatcca ccatgatcaa gtgggcttca tccctgagat gcaaggctgg 1920 ttcaacatac acaaatcaat aaacgtaatc catcacataa acagaaccaa cgacaaaaac 1980 cacatgatta tctcaataga tgcagaaaag gcctttgaca aaattcagcc cttcatgcta 2040 aaaactctca gtaaactagg tatcgatgga acatatctca aaataataag agctatttat 2100 gacaaaccca cagccaatat catactgaat gggcaaaaac tggaagcatt ccatttgaca 2160 actggcgcaa gacaaggatg ccctctctta ctactcctat tcaacgtagt attggaagtt 2220 ctggccaggg caatcaggca agagaaagaa ataaagggtg ttcagatagg aagggaggaa 2280 gtcaaattgt ctctgtttac agatggcatg attgtatatt taggaaaccc catcgtctca 2340 gcccaaaatc tccttaagct gagaagcaac ttcagcaaag tctcgggata caaaatcaat 2400 gtgcaaaaat cacaagcatt cctatacacc aataacagac aaacagagag ccaaatcatg 2460 agtgaactcc cattcacaat tgcttcaaag agaataaaat acctaggaat ccaacttaca 2520 agggatgtga aggacctctt caaggagaac tacaaaccac tgctcaatga aataaaagag 2580 gacacaaaca aatggaagaa cattccatgc tcatgggtag gaagaatcaa tatcatgaaa 2640 atggccatac tgcccaaggt aatttataga ttcaatggca tccccatcaa gctaccaatg 2700 actttcttta cagaattgga aaaaactact ttaaagttca tatggaacca aaaaagagcc 2760 cacattgcca agacaatcct aagccaaaag aacaaagctg gaggcatcat gctacctgac 2820 ttcaaactat actacaaggc tacagtaacc aaaacagcat ggtactggta ccaaaacaga 2880 gatatagacc aacggaacag aacagagccc tcacaaataa tgccgcataa ctacaaccat 2940 ctcatctttg acaaacctaa caaaaacaag aaatggggaa agcattccct atttaataaa 3000 tggtgctggg aaaactggct agccatatgt agaaagctga aactggatc.c cttccttaca 3060 ccttacacaa aaattaattc aagatggatt aaagacttac atgttagacc taaaaccata 3120 aaaaccctag aagaaaacct aggcaatatc attcaggaca taggcatggg caaggacttc 3180 atgtcgaaaa caccaaaagc aatggcaaca aaagccaaaa ttgacaaatg ggatctactt 3240 aaactaaaga gcttctttac agcaaaagaa actaccatca gagtgaacag gcaacctaca 3300 gaatgggaga aaatttttgc aacctactca tctgacagag ggctaatatc cagaatctac 3360 aataaactca aacaaattta taagaaaaaa acaaacaacc ccatcaaaaa gtgggtgaag 3420 gatatgaaca gacacttctc aaaagaagac atttatgcag ccaaaagaca catgaaaaaa 3480 tgctcatcat caatggccat cagagaaatg caaatcaaaa cctcaatgag ataccatctc 3540 acaccagtta gaatggcaat cattaaaaag tcaggaaaca acaggtgctg gagagaatgt 3600 ggagaaactg gaatactttt acactgttgg tgggactgta aactagttca accattgtgg 3660 aagtcagtgt ggcgattcct cagggatcta gaacaagaaa taccatttga cccagccatc 3720 ccactactgg gtatataccc aaaggattat aaatcatgct gcaataaaga cacgtgcaca 3780 cctatgttta ttgcagcact attcacaata gcaaagactt ggaaccaacc caaatgtcca 3840 acaatggtag agtggatgaa gaaaatgtgg cacatataca ccatgcaata ctatgcagcc 3900 ataaaaaatg atgcgttcat gtcctttgta gggacatgga tgaaattgga aatcatcatt 3960 ctcagtaaac tatcgcaaga acaaaaaacc aaacaccgta tactctcact cataggtggg 4020 aattgaacaa tgagaacaca tggacacagg aaggggaaca tcacactctg gggactgttg 4080 tggggtgggg ggagggggga aagatagcat tgggagatat acc 4123 <210> 73 <211> 2569 <212> DNA
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 2321130CB1 <220>

<221> unsure <222> 2536 <223> a, t, c, g, or other <400> 73 tggactatga gagtaggacg tttattgccg cgtacgtaag ctcggacgca tgctcgagct 60 gggaaaggaa gtccgggacc tccctgctct cggtcctcct ccgcttcctg cctcatgcct 120 caccttgtcc ccagcgcctg gactccccct taactgcttg ggaaatgtga cctttgctct 180 ggggggcctg gccctgcagg ccccaacctt ccctcatctc tggcggccct cttgggcctc 240 tgacccagcc cctccccggg ccaggctcac agaagctggc ttctgggact gtcctgggcc 300 caagtgggca cctgcgccag ccccacctgt gcctgggctg tggccccttc ctacagggcg 360 ctcaccatgg ccccgccgct cctgctgctg ctgctggcca gtggagcggc cgcctgcccg 420 ctgccctgcg tctgccagaa cctgtccgag tcgctcagca ccctctgtgc ccaccgaggc 480 ctgctgtttg tgccgcccaa cgtggaccgg cgcacagtgg agctgcggct ggctgacaac 540 ttcatccagg ccctggggcc ccctgacttc cgcagcatga cgggactggt ggacctgaca 600 ctgtctcgca atgccatcac ccgcattggg gcccgcgcct ttggggacct cgagagcctg 660 cgttccctec accttgacgg caacaggctg gtggagctgg gcaccgggag cctccggggc 720 cccgtcaatc tgcagcacct catcctcagc ggcaaccagc tgggccgcat cgcgccggga 780 gccttcgacg acttcctaga gagcctggag gacctggacc tgtcctacaa caacctccgg 840 caggtgccct gggccggcat cggcgccatg cctgccctgc acaccctcaa cctggaccat 900 aaccttattg acgcactgcc cccaggcgcc ttcgcccagc tcggtcagct ctcccgcctg 960 gacctcacct ccaaccgcct ggccacgctg gctccggacc cgcttttctc tcgtgggcgt 1020 gatgcagagg cctctcccgc ccccctggtg ctgagcttta gcgggaaccc cctgcactgc 1080 aactgtgagc tgctgtggct gcggcggctg.gcgcggccgg acgacctgga aacgtgcgcc 1140 tccccgcccg gcctggccgg ccgctacttc~tgggcagtgc ccgagggcga gttctcctgt 1200 gagccgcccc tcattgcccg ccacacgcag cgcctctggg tgctggaagg.ccagcgggcc 1260 acgctgcggt gccgggccct gggtgacccc gcgcctacca tgcactgggt cggtcctgac 1320 gaccggttgg ttggcaactc ctcccgagcc cgggctttcc ccaacgggac cttagagatt 1380 ggggtgaccg gcgctgggga cgctgggggc tacacctgca tcgccaccaa ccctgctggt 1440 gaggccacag cccgagtaga actgcgggtg ctggccttgc cccatggtgg gaacagcagt 1500 gccgaggggg gccgccccgg gccctcggac atcgccgcct ccgctcgcac tgctgccgag 1560 ggtgagggga cgctggagbc tgagccagcc gtgcaggtga cggaggtgac cgccacctca 1620 gggctggtga gctggggtcc cgggcggcca gccgacccag tgtggatgtt ccaaatccag 1680 tacaacagca gcgaagatga gaccctcatc taccggattg tcccagcctc cagccaccac 1740 ttcctgctga agcacctcgt ccccggcgct gactatgacc tctgcctgct ggccttgtca 1800 ccggccgctg ggccctctga cctcacggcc accaggctgc tgggctgtgc ccatttctcc 1860 acgctgccgg cctcgcccct gtgccacgcc etgcaggccc acgtgctggg cgggaccctg 1920 accgtggccg tggggggtgt gctggtggct gccttactgg tcttcactgt ggccttgctg 1980 gttcggggcc ggggggccgg aaatggccgc ctccccctca agctcagcca cgtccagtcc 2040 cagaccaatg gaggccccag ccccacaccc aaggcccacc cgccgcggag ccccccgccc 2100 cggccgcagc gcagctgctc tctggacctg ggagatgccg ggtgctacgg ttatgccagg 2160 cgcctgggag gagcttgggc ccgacggagc cactctgtgc atggggggct gctcggggca 2220 gggtgccggg gggtaggagg cagcgccgag cggctggaag agagtgtggt gtgatggacg 2280 ggcagcttcc tgtgtgctcc aagggatgag cctcgtgggg cagagggccc ggggccgccg 2340 cctggcctgg gggtccctcc ctggttttta ttctcagtac ctcaggctcc cctgtgtact 2400 tggaggggca gggagccctt tcctcggttc tggcctccag accagggtaa gggcaggccc 2460 ctccaacagg tgctcacagc caccgaggca gggccgcagc cacccactgg gagtctgttt 2520 ttatttataa taaaantatg cgggcagcac atgagcttca ccgcgattg 2569 <210> 74 <211> 1066 <212> DNA
<213> Homo Sapiens <220>
<221> misc feature <223> Incyte ID No: 2008365CB1 <400> 74 gtgaggcata actcatcgct tcactcttct caacggttcc ctcttcggaa ggaagcgcag 60 tccgcagcgt cgtccgaggg cgtgatcgcc tgcgtcgcac ccgcgtctcc agtggccacc 120 ccttgtccta caggaacgcg catctgctta gcaggcgctg agcccttgag gaggcctaaa 180 atgaatgaac gcggctgtgc ctcctgagca ggctcacagt tgtgggtggg ggacagaagg 240 gtgtccatgc ctaaggtcga cggcgataag acagacgttc ttcccaggag gagatcagtt 300 ccagaacagg tggagaggta tgaagaacga ggaacactgc ccggggagtt tcttcctctg 360 caaaataagg gagtgtgtac tgaattaccg tttccagctc caacatccag gatttcagca 420 ctacctgcag agtagtggga ggagagatag aggaagaagt gaagataaga aaccactcga 480 agccggtgtc tggtgctggg accgtggtgg ctgggacggc agcagtcgtg ctgtccatct 540 gctcttccga ggggtggcac atcctagcct ctatcttttc cccagagaag atccccctcg 600 tctgctcttc ccccgtctca gccttttagt gtgtgaacag ttctggtgtt actcagcaac 660 gcttttgtta gcgcctttac cagcaagtac ttgttaaaca cttaaatctt acaagtggaa 720 agaaaattcc gtggtaaatt taagtacaaa agtcaaccaa aattctgagt ctcggacaag 780 actggttaaa aatcgttcta caatgaaact aataaattaa atcatataaa acttttattg 840 ctttgcaagg aagctttagt attaaatgta agcattttca tctggtgatt cttcccagtc 900 tcctgtggta ataatacctg gacagggaag aaatcttttg agataactaa actatactgg 960 gaaatgggcc ccttgctaag tttcctgtgg cagtgacacc ccctctgttg caaatatttt 1020 ctccgatgcg tggtttcagt tccaagccaa acccgttcac ctgccc 1066 <210> 75 <211> 1817 <212> DNA
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 3580778CB1 <400> 75 atgacttcga aagaggaaag caggaggcag~cagcccacag ctggtcctgc agggcaggga 60 aagttaccct cgccctccga gccacaactc cccacgccgc caactcggtc tttacatcat 120 tttcgacgcc ccctaagtcc ctcccgagag gcgcaggcgc acatcgcccc ttctagcgaa 180 ctacatctcc cacaatccca atcggccgga ccacctccgc tcggggcggg gacggaggtg 240 gagctggtgg tccccggtcg ggacgaaggc tcccgaggtg ccctgcctgg gtcctccggg 300 gtaaagttcg tttggcggaa gattgtccgt tttcctggtc attgctagga agaactctgg 360 gtacaataat gaatacaatg tatgtgatga tggctcagat cttaagatct cacctgataa 420 aggctacagt gattcctaat cgagtgaaaa tgcttccata ttttggtatc attagaaata 480 gaatgatgtc aacccataaa tccaaaaaga agatcagaga atattataga ctgctgaacg 540 tggaggaagg atgctctgca gatgaagtca gggaatcttt tcataagctt gccaagcaat 600 atcatcctga cagtggctct aatactgctg attctgcaac atttataagg attgaaaaag 660 cttatagaaa ggtgctctcc catgtgatag aacaaacaaa tgccagtcag agtaaaggtg 720 aagaagaaga agatgtagaa aaattcaaat ataaaacacc ccaacaccga cattatttaa 780 gttttgaagg tattggtttt gggactccaa ctcaacgaga gaagcattat aggcaattta 840 gggcagaccg tgctgctgaa caagtgatgg aatatcaaaa gcagaaacta caaagccagt 900 attttcctga tagtgtaatt gttaaaaata taagacagag caaacagcaa aagataacgc 960 aagctataga acgtttagtg gaggacctca ttcaagaatc catggcaaaa ggagactttg 1020 acaatctcag tgggaaagga aaacctctga aaaagttttc tgactgttct tacattgatc 1080 ccatgactca caacctgaac cgaatactga tcgataatgg ataccaacca gaatggatcc 1140 ttaagcaaaa ggaaataagc gatactattg agcaactcag agaggcaatt ttagtgtcta 1200 ggaaaaaact tgggaatcca atgacaccaa ctgaaaagaa acagtggaac catgtttgtg 1260 agcagtttca agaaaacatc agaaaattaa acaagcgaat taatgatttt aatttaattg 1320 ttcccatcct gaccaggcaa aaagtccatt ttgatgctca gaaagaaatt gtcagagccc 1380 agaaaatata cgagaccctt ataaaaacaa aagaagtcac agatagaaac ccaaataacc 1440 ttgatcaagg agaaggagag aaaacacctg aaatcaagaa aggtttttta aactggatga 1500 atctgtggaa atttattaaa atacgatcat tttgatgttt actatcataa atcattctta 1560 gttccactga cactttacat ggaaaatgag atttattgct ataatacaag aatttaagaa 1620 ttgtgccatt gtacttatca caaaactaat cacatagcca atgatgtgtg agtgagaaac 1680 ctatcaggtt tgtcctgagg atatagcaag aaaagaaaat aactgaaact cctttttttt 1740 tgagacggcg tctcgctctg tcacccaggc tggagtgcaa tggcacaatc taggctcact 1800 gcaacctctg cttccca 1817 <210> 76 <211> 2391 <212> DNA
<213> Homo Sapiens <220>
<221> mist feature <223> Incyte ID No: 7948785CB1 <400> 76 tggtacgagc tcgcctcact agtacggctg catgtgctgg caaagtcgtg acccgggcag 60 caggcactat atttgtatgt gtcttgtaga acccacgctt ggaaatgctg acagcaggct 120 tcaggacagc tgagccccac taaacaccaa gaaaacccat ggctgtggct ccatctttca 180 acatgaccaa tccacagcct gccatagaag gaggaatttc tgaagttgag atcatctccc 240 aacaagtaga cgaagaaacc aagagcattg ctcctgtgca gctggtgaac tttgcctatc 300 gggacttgcc cctggctgct gtcgatctct ccacggcggg ctcgcagctc ctgtc~aatc 360 tggacgaaga ttaccaaaga gaagggtcta actggctgaa gccgtgctgt gggaagagag 420 cagccgtgtg gcagtttcca gcgcattcca gtttgctggc gtgattcact ggatcagcct 480 ggtcattctg tccgtgttct tctcagagac tgttctacgg atagtggtag cttgggatct 540 gggattactt cgaaaacaaa atagaggtgt ttgacggggc tgtgatca c ctatctttgg 600 ctccgatggt ggcatccact gtggccaatg gacccaggag cccctgggac gccatcagcc 660 tcatcatcat gctccggatc tggagggtga.agagggtcat tgatgcctac gtcctgccag 720 tgaagctgga gatggagatg gttatccagc agtacgagaa ggccaaggtc atccaagacg 780 agcagctgga gaggctgacg cagatctgtc aggagcaagg gtttgagatc cggcagctgc 840 gcgcgcacct ggcgcagcag gacctggacc tggctgccga gcgcgaagcg gcgctccagg 900 ccccgcacgt gctcagccag ccgcgcagcc gcttcaaagt gttggaggcc ggcacgtggg 960 acgaggagac ggcggccgag agcgtcgtgg aggagctgca gccctcgcaa gaagccacga 1020 tgaaggacga catgaacagc tacatcagtc agtattacaa tgggcccagc agtgacagcg 1080 gtgtcccaga gccagctgtg tgtatggtca ccacggccgc aatagacatt caccagccca 1140 acatctcctc ggacctcttc tctctggaca tgcccctcaa actcggcggt aatggcacca 1200 gcgccacctc ggagagtgcc tcccgcagct cagtcacccg ggcccagagt gacagcagcc 1260 agacgctggg ctcctccatg gactgcagca ctgcccgcga ggagccgtcc tctgagcccg 1320 gcccttctcc cccgccgctg ccatcccagc agcaggtgga ggaggccaca gtccaggacc 1380 tgctgtcctc cctgtcggag gacccctgcc cttcccagaa ggccttggac ccagcccccc 1440 tcgcccggcc cagcccagcg ggctcggccc aaaccagccc cgagctggaa cacagggtaa 1500 gtctgttcaa ccagaagaac caggagggct tcactgtctt tcagatcagg cctgtcatcc 1560 acttccagcc cactgtgccc atgctggagg acaagttcag atctttggaa tccaaagagc 1620 aaaagctgca cagggtccct gaggcctaga gcctgccatg ggctgggtga gatgagggga 1680 gacagccatc tcaaagctct cctgggaccc tggaggctgc caagggccac acgcggggcc 1740 caggagccca cctggcctcc ctcagggtgc tgcctgcctc cagggaggcg acgccaggcc 1800 aggaggccac aagcttcaga cctcaaagcc cagagctggc gcctcttctc gccctgctca 1860 ggggagggtg gtgctcgtgg ctgggttttc tttttaacca tttttacaaa aaccagcctg 1920 tggcccagct tcagcagggt agagtgtggg ggggccagct cagcctcttc cgttgccttc 1980 gttcctgacg cccaccctgg actctaggga acagcactgt gagcaggggc tgtccagccc 2040 cacccctaag ccgtctttcc caggaatcct gggtggagtc caacacaatc acacggagac 2100 caccatctga gcctatgtca tttgtcctca ttctcattcc agcatgagcg tttctgagtc 2160 tcttcaagac gaatctagtt ttcaccttca caggatataa ggggatcacc tagaggtggg 2220 tgggaggggc ccaaaaaggc agggaaaaca cccattttcc aaccctgggt tttattatta 2280 aaaaccagtt ggctgagaaa aaaaaaaaaa agggggcccg gataggaagc tcttacccgg 2340 aaataatccg gaccgaccgg agggttttcc acccaggcaa aaaataaatg g 2391 <210> 77 <211> 1314 <212> DNA
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 7494415CB1 <400> 77 ctggtgatac ccaggcaaac agggtctgga atggacctcc agcaaagtcc agcagacctg 60 cagcagaggg gactgagtgt tagaaggaaa actaacaaac agaaaggatt agcatcaaca 120 tcaacaaaaa ggacgtccac acagaaaccc catccaaagg gcaccatcat caaaaaccaa 180 aggtagataa atccatgaag atgaggaaaa accagcccca aaagactgaa aattccaaaa 240 aacagaacgg ctcttctcct tcaaaggatc atgacttctc tccagcaagg gaacaaacct 300 ggatggagaa tgagtttgac aaagggacag aagaaggctt cagaaataac aaaccctctg 360 agctacagga gcatgttcta acccaatgca aggaagctaa gaaccttgaa aaagggttag 420 aggaattgct aactagaata accagtttag agaagaacac aaatgacctg atagagctga 480 aaaacacagc acaggaactt catgaagcat acacgagtat caatagccca attgatcaag 540 cagaagaaag gatatcagag attgaagatc aacttaataa aataaagtgt gaagggaagt 600 ttagagaaaa aagaataaaa agaaataaac.aaagcctcca agaaatatgg gactatgtga 660 aaagaccaaa tctgcatctg attggtgtac ccgaaagtga cggggagaat ggaaccaagt 720 tggaaaacac tctgcaggat attatccagg agaacttccc caatctggca aggcaggcca 780 .
acattcagat tcaggaaata cagagaacgc tacaaagata ctcctcgaga agagcaactc 840 caagacacat aattgtcaga ttcaccaaag ttgaaatgaa ggaa~aaaatg ttaagggcag 900 ccagagagaa aggtcgggtt accctcaaat ggaagcccat cagactaaca gtggatctct 960 cagcaggaac tctcctagcc agaagagagt ggggaccaat attc.aacatt cttaaagaaa 1020 agaattttca atccagaatt tcatatccag ccaaactaag, cttcataagg gaaggagaaa 1080 taaaatactt tacagacaag caaatgctga gagattttgt caccaccagg cctgccctaa 1140 aagagctcct gaaggaagcg ctaaacatgg aaaggaacaa ccggtaccag ctgctgcaaa 1200 atcatgccaa aatgtaaaca ccattgagac taagaagaaa ttgcatcaac taacgagcaa 1260 aataaccagc taacatcata atgacaggat caaattcaca cataacaata ttaa 1314 <210> 78 <211> 2076 <212> DNA
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 2234223CB1 <400> 78 gcctgacgcc ctgcttcgtc gcctcctttc tctcccaggt gctggaccag ggactgagcg 60 tcccccggag agggtccggt gtgaccccga caagaagcag aaatggggaa gaaactggat 120 ctttccaagc tcactgatga agaggcccag catgtcttgg aagttgttca acgagatttt 180 gacctccgaa ggaaagaaga ggaacggcta gaggcgttga agggcaagat taagaaggaa 240 agctccaaga gggagctgct ttccgacact gcccatctga acgagaccca ctgcgcccgc 300 tgcctgcagc cctaccagct gcttgtgaat agcaaaaggc agtgcctgga atgtggcctc 360 ttcacctgca aaagctgtgg ccgcgtccac ccggaggagc agggctggat ctgtgacccc 420 tgccatctgg ccagagtcgt gaagatcggc tcactggagt ggtactatga gcatgtgaaa 480 gcccgcttca agaggttcgg aagtgccaag gtcatccggt ccctccacgg gcggctgcag 540 ggtggagctg ggcctgaact gatatctgaa gagagaagtg gagacagcga ccagacagat 600 gaggatggag aacctggctc agaggcccag gcccaggccc agccctttgg cagcaaaaaa 660 aagcgcctcc tctccgtcca cgacttcgac ttcgagggag actcagatga ctccactcag 720 cctcaaggtc actccctgca cctgtcctca gtccctgagg ccagggacag cccacagtcc 780 ctcacagatg agtcctgctc agagaaggca gcccctcaca aggctgaggg cctggaggag 840 gctgatactg gggcctctgg gtgccactcc catccggaag agcagccgac cagcatctca 900 ccttccagac acggcgccct ggctgagctc tgcccgcctg gaggctccca caggatggcc 960 ctggggactg ctgctgcact cgggtcgaat gtcatcagga atgagcagct gcccctgcag 1020 tacttggccg atgtggacac ctctgatgag gaaagcatcc gggctcacgt gatggcctcc 1080 caccattcca agcggagagg ccgggcgtct tctgagagtc agggtctagg tgctggagcg 1140 cgcacggagg ccgatgtaga ggaggaggcc ctgaggagga agctggagga gctgaccagc 1200 aacgtcagtg accaggagac ctcgtccgag gaggaggagt ccaaggacga aaaggcagag 1260 cccaacaggg acaaatcagt tgggcctctc ccccaggcgg acccggaggt ttcagacatt 1320 gaatccagga ttgcagccct gagggccgca gggctcacgg tgaagccctc gggaaagccc 1380 cggaggaagt caaacctccc gatatttctc cctcgagtgg ctgggaaact tggcaagaga 1440 ccagaggacc caaatgcaga cccttcaagt gaggccaagg caatggctgt gccctatctt 1500 ctgagaagaa agttcagtaa ttccctgaaa agtcaaggta aagatgatga ttcttttgat 1560 cggaaatcag tgtaccgagg ctcgctgaca cagagaaacc ccaacgcgag gaaaggaatg 1620 gccagccaca ccttcgcgaa acctgtggtg gcccaccagt cctaacggga caggacagag 1680 agacagagca gccctgcact gttttccctc caccacagcc atcctgtccc tcattggctc 1740 tgtgctttcc actgtacaca gtcaccgtcc caatgagaaa caagaaggag caccctccac 1800 atggactccc acctgcaagt ggacagcgac attcagtcct gcactgctca cctgggttta 1860 ctgatgactc ctggctgccc caccatcctc tctgatctgt gagaaacagc taagctgctg 1920 tgacttccct ttaggacaat gttgtgtaaa tctttgaagg acacaccgaa gacctttata 1980 ctgtgatctt ttaccccttt cactcttggc tttcttatgt tgctttcatg aatggaatgg 2040 aaaaaagatg actcagttaa ggcacccaaa aaaaaa 2076

Claims (133)

What is claimed is:

1. An isolated polypeptide selected from the group consisting of:

a) a polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO:1-39, b) a polypeptide comprising a naturally occurring amino acid sequence at least 90%
identical to an amino acid sequence selected from the group consisting of SEQ
ID
NO:2-5, SEQ ID NO:7, SEQ ID NO:10-13, SEQ ID NO:15-20, SEQ ID NO:22-32, SEQ ID NO:34-36 and SEQ ID NO:38, c) a polypeptide comprising a naturally occurring amino acid sequence at least 92%
identical to an amino acid sequence selected from the group consisting of SEQ
ID
NO:14 and SEQ ID NO:33, d) a polypeptide comprising a naturally occurring amino acid sequence at least 96%
identical to the amino acid sequence of SEQ ID NO:37, e) a polypeptide consisting essentially of a naturally occurring amino acid sequence at least 90% identical to the amino acid sequence of SEQ ID NO:39, f) a biologically active fragment of a polypeptide having an amino acid sequence selected from the group consisting of SEQ ID NO:1-39, and g) an immunogenic fragment of a polypeptide having an amino acid sequence selected from the group consisting of SEQ ID NO:1-39.

2. An isolated polypeptide of claim 1 comprising an amino acid sequence selected from the group consisting of SEQ ID NO:1-39.

3. An isolated polynucleotide encoding a polypeptide of claim 1.

4. An isolated polynucleotide encoding a polypeptide of claim 2.

5. An isolated polynucleotide of claim 4 comprising a polynucleotide sequence selected from the group consisting of SEQ ID NO:40-78.

6. A recombinant polynucleotide comprising a promoter sequence operably linked to a polynucleotide of claim 3.

7. A cell transformed with a recombinant polynucleotide of claim 6.

8. A transgenic organism comprising a recombinant polynucleotide of claim 6.

9. A method of producing a polypeptide of claim 1, the method comprising:

a) culturing a cell under conditions suitable for expression of the polypeptide, wherein said cell is transformed with a recombinant polynucleotide, and said recombinant polynucleotide comprises a promoter sequence operably linked to a polynucleotide encoding the polypeptide of claim 1, and b) recovering the polypeptide so expressed.

10. A method of claim 9, wherein the polypeptide comprises an amino acid sequence selected from the group consisting of SEQ ID NO:1-39.

11. An isolated antibody which specifically binds to a polypeptide of claim 1.

12. An isolated polynucleotide selected from the group consisting of:

a) a polynucleotide comprising a polynucleotide sequence selected from the group consisting of SEQ ID NO:40-78, b) a polynucleotide comprising a naturally occurring polynucleotide sequence at least 90% identical to a polynucleotide sequence selected from the group consisting of SEQ
ID NO:42-46, SEQ ID NO:48-59, SEQ ID NO:61-71, SEQ ID NO:73-75 and SEQ
ID NO:77, c) a polynucleotide comprising a naturally occurring polynucleotide sequence at least 92% identical to the polynucleotide sequence of SEQ ID NO:72, d) a polynucleotide comprising a naturally occurring polynucleotide sequence at least 96% identical to the polynucleotide sequence of SEQ ID NO:76, e) a polynucleotide consisting essentially of a naturally occurring polynucleotide sequence at least 90% identical to the polynucleotide sequence of SEQ ID
NO:78, f) a polynucleotide complementary to a polynucleotide of a), g) a polynucleotide complementary to a polynucleotide of b), h) a polynucleotide complementary to a polynucleotide of c), i) a polynucleotide complementary to a polynucleotide of d), j) a polynucleotide complementary to a polynucleotide of e), and k) an RNA equivalent of a)-j).

13. An isolated polynucleotide comprising at least 60 contiguous nucleotides of a polynucleotide of claim 12.

14. A method of detecting a target polynucleotide in a sample, said target polynucleotide having a sequence of a polynucleotide of claim 12, the method comprising:

a) hybridizing the sample with a probe comprising at least 20 contiguous nucleotides comprising a sequence complementary to said target polynucleotide in the sample, and which probe specifically hybridizes to said target polynucleotide, under conditions whereby a hybridization complex is formed between said probe and said target polynucleotide or fragments thereof, and b) detecting the presence or absence of said hybridization complex, and, optionally, if present, the amount thereof.

15. A method of claim 14, wherein the probe comprises at least 60 contiguous nucleotides.

16. A method of detecting a target polynucleotide in a sample, said target polynucleotide having a sequence of a polynucleotide of claim 12, the method comprising:

a) amplifying said target polynucleotide or fragment thereof using polymerase chain reaction amplification, and b) detecting the presence or absence of said amplified target polynucleotide or fragment thereof, and, optionally, if present, the amount thereof.

17. A composition comprising a polypeptide of claim 1 and a pharmaceutically acceptable excipient.

18. A composition of claim 17, wherein the polypeptide comprises an amino acid sequence selected from the group consisting of SEQ ID NO:1-39.

19. A method for treating a disease or condition associated with decreased expression of functional MDDT, comprising administering to a patient in need of such treatment the composition of claim 17.

20. A method of screening a compound for effectiveness as an agonist of a polypeptide of claim 1, the method comprising:

a) exposing a sample comprising a polypeptide of claim 1 to a compound, and b) detecting agonist activity in the sample.

21. A composition comprising an agonist compound identified by a method of claim 20 and a pharmaceutically acceptable excipient.

22. A method for treating a disease or condition associated with decreased expression of functional MDDT, comprising administering to a patient in need of such treatment a composition of claim 21.

23. A method of screening a compound for effectiveness as an antagonist of a polypeptide of claim 1, the method comprising:

a) exposing a sample comprising a polypeptide of claim 1 to a compound, and b) detecting antagonist activity in the sample.

24. A composition comprising an antagonist compound identified by a method of claim 23 and a pharmaceutically acceptable excipient.

25. A method for treating a disease or condition associated with overexpression of functional MDDT, comprising administering to a patient in need of such treatment a composition of claim 24.

26. A method of screening for a compound that specifically binds to the polypeptide of claim 1, the method comprising:

a) combining the polypeptide of claim 1 with at least one test compound under suitable conditions, and b) detecting binding of the polypeptide of claim 1 to the test compound, thereby identifying a compound that specifically binds to the polypeptide of claim 1.

27. A method of screening for a compound that modulates the activity of the polypeptide of claim 1, the method comprising:

a) combining the polypeptide of claim 1 with at least one test compound under conditions permissive for the activity of the polypeptide of claim 1, b) assessing the activity of the polypeptide of claim 1 in the presence of the test compound, and c) comparing the activity of the polypeptide of claim 1 in the presence of the test compound with the activity of the polypeptide of claim 1 in the absence of the test compound, wherein a change in the activity of the polypeptide of claim 1 in the presence of the test compound is indicative of a compound that modulates the activity of the polypeptide of claim 1.

28. A method of screening a compound for effectiveness in altering expression of a target polynucleotide, wherein said target polynucleotide comprises a sequence of claim 5, the method comprising:
a) exposing a sample comprising the target polynucleotide to a compound, under conditions suitable for the expression of the target polynucleotide, b) detecting altered expression of the target polynucleotide, and c) comparing the expression of the target polynucleotide in the presence of varying amounts of the compound and in the absence of the compound.

29. A method of assessing toxicity of a test compound, the method comprising:
a) treating a biological sample containing nucleic acids with the test compound, b) hybridizing the nucleic acids of the treated biological sample with a probe comprising at least 20 contiguous nucleotides of a polynucleotide of claim 12 under conditions whereby a specific hybridization complex is formed between said probe and a target polynucleotide in the biological sample, said target polynucleotide comprising a polynucleotide sequence of a polynucleotide of claim 12 or fragment thereof, c) quantifying the amount of hybridization complex, and d) comparing the amount of hybridization complex in the treated biological sample with the amount of hybridization complex in an untreated biological sample, wherein a difference in the amount of hybridization complex in the treated biological sample is indicative of toxicity of the test compound.

30. A diagnostic test for a condition or disease associated with the expression of MDDT in a biological sample, the method comprising:
a) combining the biological sample with an antibody of claim 11, under conditions suitable for the antibody to bind the polypeptide and form an antibody:polypeptide complex, and b) detecting the complex, wherein the presence of the complex correlates with the presence of the polypeptide in the biological sample.

31. The antibody of claim 11, wherein the antibody is:
a) a chimeric antibody, b) a single chain antibody, c) a Fab fragment, d) a F(ab')2 fragment, or e) a humanized antibody.

32. A composition comprising an antibody of claim 11 and au acceptable excipient.

33. A method of diagnosing a condition or disease associated with the expression of MDDT
in a subject, comprising administering to said subject an effective amount of the composition of claim 32.

34. A composition of claim 32, wherein the antibody is labeled.

35. A method of diagnosing a condition or disease associated with the expression of MDDT
in a subject, comprising administering to said subject an effective amount of the composition of claim 34.

36. A method of preparing a polyclonal antibody with the specificity of the antibody of claim 11, the method comprising:
a) immunizing au animal with a polypeptide consisting of an amino acid sequence selected from the group consisting of SEQ ID NO:1-39, or an immunogenic fragment thereof, under conditions to elicit an antibody response, b) isolating antibodies from said animal, and c) screening the isolated antibodies with the polypeptide, thereby identifying a polyclonal antibody which specifically binds to a polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO:1-39.

37. A polyclonal antibody produced by a method of claim 36.

38. A composition comprising the polyclonal antibody of claim 37 and a suitable carrier.

39. A method of making a monoclonal antibody with the specificity of the antibody of claim 11, the method comprising:
a) immunizing an animal with a polypeptide consisting of an amino acid sequence selected from the group consisting of SEQ ID NO:1-39, or an immunogenic fragment thereof, under conditions to elicit an antibody response, b) isolating antibody producing cells from the animal, c) fusing the antibody producing cells with immortalized cells to form monoclonal antibody-producing hybridoma cells, d) culturing the hybridoma cells, and e) isolating from the culture monoclonal antibody which specifically binds to a polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO:1-39.

40. A monoclonal antibody produced by a method of claim 39.

41. A composition comprising the monoclonal antibody of claim 40 and a suitable carrier.

42. The antibody of claim 11, wherein the antibody is produced by screening a Fab expression library.

43. The antibody of claim 11, wherein the antibody is produced by screening a recombinant immunoglobulin library.

44. A method of detecting a polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO:1-39 in a sample, the method comprising:
a) incubating the antibody of claim 11 with a sample under conditions to allow specific binding of the antibody and the polypeptide, and b) detecting specific binding, wherein specific binding indicates the presence of a polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO:1-39 in the sample.

45. A method of purifying a polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO:1-39 from a sample, the method comprising:
a) incubating the antibody of claim 11 with a sample under conditions to allow specific binding of the antibody and the polypeptide, and b) separating the antibody from the sample and obtaining the purified polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID
NO:1-39.

46. A microarray wherein at least one element of the microarray is a polynucleotide of claim 13.

47. A method of generating an expression profile of a sample which contains polynucleotides, the method comprising:
a) labeling the polynucleotides of the sample, b) contacting the elements of the microarray of claim 46 with the labeled polynucleotides of the sample under conditions suitable for the formation of a hybridization complex, and c) quantifying the expression of the polynucleotides in the sample.

48. An array comprising different nucleotide molecules affixed in distinct physical locations on a solid substrate, wherein at least one of said nucleotide molecules comprises a first oligonucleotide or polynucleotide sequence specifically hybridizable with at least 30 contiguous nucleotides of a target polynucleotide, and wherein said target polynucleotide is a polynucleotide of claim 12.

49. An array of claim 48, wherein said first oligonucleotide or polynucleotide sequence is completely complementary to at least 30 contiguous nucleotides of said target polynucleotide.

50. An array of claim 48, wherein said first oligonucleotide or polynucleotide sequence is completely complementary to at least 60 contiguous nucleotides of said target polynucleotide.

51. An array of claim 48, wherein said first oligonucleotide or polynucleotide sequence is completely complementary to said target polynucleotide.

52. An array of claim 48, which is a microarray.

53. An array of claim 48, further comprising said target polynucleotide hybridized to a nucleotide molecule comprising said first oligonucleotide or polynucleotide sequence.

54. An array of claim 48, wherein a linker joins at least one of said nucleotide molecules to said solid substrate.

55. An array of claim 48, wherein each distinct physical location on the substrate contains multiple nucleotide molecules, and the multiple nucleotide molecules at any single distinct physical location have the same sequence, and each distinct physical location on the substrate contains nucleotide molecules having a sequence which differs from the sequence of nucleotide molecules at another distinct physical location on the substrate.

56. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID
NO:1.

57. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID
NO:2.

58. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID
NO:3.

59. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID
NO:4.

60. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID
NO:5.

61. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID
NO:6.

62. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID
NO:7.

63. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID
NO:8.

64. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID
NO:9.

65. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID
NO:10.

66. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID
NO:11.

67. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID
NO:12.

68. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID
NO:13.

69. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID
NO:14.

70. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID
NO:15.

71. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID
NO:16.

72. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID
NO:17.

73. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID
NO:18.

74. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID
NO:19.

75. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID
NO:20.

76. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID
NO:21.

77. A polypeptide of claim 1, comprising,the amino acid sequence of SEQ ID
NO:22.

78. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID
NO:23.

79. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID
NO:24.

80. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID
NO:25.

81. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID
NO:26.

82. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID
NO:27.

83. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID
NO:28.

84. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID
NO:29.

85. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID
NO:30.

86. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID
NO:31.

87. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID
NO:32.

88. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID
NO:33.

89. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID
NO:34.

90. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID
NO:35.

91. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID
NO:36.

92. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID
NO:37.

93. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID
NO:38.

94. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID
NO:39.

95. A polynucleotide of claim 12, comprising the polynucleotide sequence of SEQ ID NO:40.

96. A polynucleotide of claim 12, comprising the polynucleotide sequence of SEQ ID NO:41.

97. A polynucleotide of claim 12, comprising the polynucleotide sequence of SEQ ID NO:42.

98. A polynucleotide of claim 12, comprising the polynucleotide sequence of SEQ ID NO:43.

99. A polynucleotide of claim 12, comprising the polynucleotide sequence of SEQ ID NO:44.

100. A polynucleotide of claim 12, comprising the polynucleotide sequence of SEQ ID
NO:45.

101. A polynucleotide of claim 12, comprising the polynucleotide sequence of SEQ ID
NO:46.

102. A polynucleotide of claim 12, comprising the polynucleotide sequence of SEQ ID
NO:47.

103. A polynucleotide of claim 12, comprising the polynucleotide sequence of SEQ ID
NO:48.

104. A polynucleotide of claim 12, comprising the polynucleotide sequence of SEQ ID
NO:49.

105. A polynucleotide of claim 12, comprising the polynucleotide sequence of SEQ ID
NO:50.

106. A polynucleotide of claim 12, comprising the polynucleotide sequence of SEQ ID
NO:51.

107. A polynucleotide of claim 12, comprising the polynucleotide sequence of SEQ ID
NO:52.

108. A polynucleotide of claim 12, comprising the polynucleotide sequence of SEQ ID
NO:53.

109. A polynucleotide of claim 12, comprising the polynucleotide sequence of SEQ ID
NO:54.

110. A polynucleotide of claim 12, comprising the polynucleotide sequence of SEQ ID

NO:55.

111. A polynucleotide of claim 12, comprising the polynucleotide sequence of SEQ ID
NO:56.

112. A polynucleotide of claim 12, comprising the polynucleotide sequence of SEQ ID
NO:57.

113. A polynucleotide of claim 12, comprising the polynucleotide sequence of SEQ ID
NO:58.

114. A polynucleotide of claim 12, comprising the polynucleotide sequence of SEQ ID
NO:59.

115. A polynucleotide of claim 12, comprising the polynucleotide sequence of SEQ ID
NO:60.

116. A polynucleotide of claim 12, comprising the polynucleotide sequence of SEQ ID
NO:61.

117. A polynucleotide of claim 12, comprising the polynucleotide sequence of SEQ ID
NO:62.

118. A polynucleotide of claim 12, comprising the polynucleotide sequence of SEQ ID
NO:63.

119. A polynucleotide of claim 12, comprising the polynucleotide sequence of SEQ ID
NO:64.

120. A polynucleotide of claim 12, comprising the polynucleotide sequence of SEQ ID
NO:65.

121. A polynucleotide of claim 12, comprising the polynucleotide sequence of SEQ ID

NO:66.

122. A polynucleotide of claim 12, comprising the polynucleotide sequence of SEQ ID
NO:67.

123. A polynucleotide of claim 12, comprising the polynucleotide sequence of SEQ ID
NO:68.

124. A polynucleotide of claim 12, comprising the polynucleotide sequence of SEQ ID
NO:69.

125. A polynucleotide of claim 12, comprising the polynucleotide sequence of SEQ ID
NO:70.

126. A polynucleotide of claim 12, comprising the polynucleotide sequence of SEQ ID
NO:71.

127. A polynucleotide of claim 12, comprising the polynucleotide sequence of SEQ ID
NO:72.

128. A polynucleotide of claim 12, comprising the polynucleotide sequence of SEQ ID
NO:73.

129. A polynucleotide of claim 12, comprising the polynucleotide sequence of SEQ ID
NO:74.

130. A polynucleotide of claim 12, comprising the polynucleotide sequence of SEQ ID
NO:75.

131. A polynucleotide of claim 12, comprising the polynucleotide sequence of SEQ ID
NO:76.

132. A polynucleotide of claim 12, comprising the polynucleotide sequence of SEQ ID

NO:77.

133. A polynucleotide of claim 12, comprising the polynucleotide sequence of SEQ ID
NO:78.