CA2375493A1 - Human transport proteins - Google Patents

Abstract

The invention provides human transport proteins (TPPT) and polynucleotides which identify and encode TPPT. The invention also provides expression vectors, host cells, antibodies, agonists, and antagonists. The invention also provides methods for diagnosing, treating, or preventing disorders associated with expression of TPPT.

Description

HUMAN TRANSPORT PROTEINS
TECHNICAL FIELD
This invention relates to nucleic acid and amino acid sequences of human transport proteins and to the use of these sequences in the diagnosis, treatment, and prevention of transport, metabolic, neurological, cardiovascular, reproductive, and immune disorders, and cell proliferative disorders including cancer.
BACKGROUND OF THE INVENTION
Eukaryotic cells are surrounded and subdivided into functionally distinct organelles by hydrophobic lipid bilayer membranes. These membranes act as a barrier to most molecules, and maintain the essential differences between the cytosol, the extracellular environment, and the contents of each intracellular organelle. Transport of essential nutrients, certain metal ions, metabolic waste products, cell signaling molecules, macromolecules, and proteins across lipid membranes and between organelles must be mediated by a variety of transport molecules.
Transport between the cytoplasm and the extracellular environment, and between the cytoplasm and lumenal spaces of cellular organelles requires specific transport proteins. Each transport protein carries a particular class of molecule, such as ions, sugars, or amino acids, and often is specific to a certain molecular species of the class.
Cells and organelles require transport molecules to import and export essential nutrients and metal ions including K+, NH4+, P;, SO.~'--, sugars, and vitamins, as well as various metabolic waste products. Transport proteins also play roles in antibiotic resistance, toxin secretion, ion balance, synaptic neurotransmission, kidney function, intestinal absorption, tumor growth, and other diverse cell functions (Griffith, J. and C. Sansom (1998) The Transporter Facts Book, Academic Press, San Diego CA, pp. 3-29). Transport can occur by a passive concentration-dependent mechanism, or can be linked to an energy source such as ATP hydrolysis or an ion gradient.
Proteins that function in transport include carrier proteins, which bind to a specific solute and undergo a conformational change that transfers the bound solute across the membrane, and channel proteins, which form hydrophilic pores that allow specific solutes to diffuse through the membrane down an electrochemical solute gradient.
Transport proteins are often mufti-pass transmembrane proteins, which either actively transport molecules across the membrane or passively allow them to cross.
Active transport involves directional pumping of a solute across the membrane, usually against an electrochemical gradient.
Active transport is tightly coupled to a source of metabolic energy, such as ATP hydrolysis or an electrochemically favorable ion gradient. Passive transport involves the movement of a solute down its electrochemical gradient. Transport proteins can be further classified as either Garner proteins or channel proteins. Carrier proteins, which can function in active or passive transport, bind to a specific solute to be transported and undergo a conformational change which transfers the bound solute across the membrane. Channel proteins, which only function in passive transport, form hydrophilic pores across the membrane. When the pores open, specific solutes, such as inorganic ions, pass through the membrane and down the electrochemical gradient of the solute. Examples include facilitative transporters, the secondary active symporters and antiporters driven by ion gradients, and active ATP binding cassette transporters involved in multiple-drug resistance and targeting of antigenic peptides to MHC Class I molecules. Transported substrates range from nutrients and ions to a broad variety of drugs, peptides and proteins.
Information on the action of ARL-6 (ADP-ribosylation like factor), an endoplasmic reticulum transmembrane protein, can be found in Greenfield, J.J. and S. High ( 1999; J.
Cell Sci. 112:1477-1486). Information on reduced folate carrier transporter proteins can be found in Dixon, K.H. et al.
( 1994; J. Biol. Chem. 269:17-20) and Moscow, J.A. et al. ( 1995; Cancer Res.
55:5983-5987).
Carrier proteins which transport a single solute from one side of the membrane to the other are called uniporters. In contrast, coupled transporters link the transfer of one solute with simultaneous or sequential transfer of a second solute, either in the same direction (symport) or in the opposite direction (antiport). For example, intestinal and kidney epithelia contain a variety of symporter systems driven by the sodium gradient that exists across the plasma membrane. Sodium moves into the cell down its electrochemical gradient and brings the solute into the cell with it. The sodium gradient that provides the driving force for solute uptake is maintained by the ubiquitous Na+/K+ ATPase. Sodium-coupled transporters include the mammalian glucose transporter (SGLTI ), iodide transporter (NIS), and multivitamin transporter (SMVT). All three transporters have twelve putative transmembrane segments, extracellular glycosylation sites, and cytoplasmically-oriented N-and C-termini.
Mitochondrial carrier proteins are transmembrane-spanning proteins which transport ions and charged metabolites between the cytosol and the mitochondrial matrix. Examples include the ADP, ATP carrier protein; the 2-oxoglutarate/malate carrier; the phosphate carrier protein; the brown fat uncoupling protein which transports protons from the cytosol into the matrix;
the pyruvate carrier; the dicarboxylate carrier which transports malate, succinate, fumarate, and phosphate; the tricarboxylate carrier which transports citrate and malate; and the Grave's disease carrier protein, a protein recognized by IgG in patients with active Grave's disease, an autoimmune disorder resulting in hyperthyroidism (Stryer, L. ( 1995) Biochemistry, W.H. Freeman and Company, New York NY, p.
551; PROSITE PDOC00189 Mitochondrial energy transfer proteins signature;
Online Mendelian Inheritance in Man (OMIM) *275000 Graves Disease).
This class of transporters also includes the mitochondrial uncoupling proteins, which create proton leaks across the inner mitochondria) membrane, thus uncoupling oxidative phosphorylation from ATP synthesis. The result is energy dissipation in the form of heat.
Mitochondria) uncoupling proteins have been implicated as modulators of thermoregulation and metabolic rate, and have been proposed as potential targets for drugs against metabolic diseases such as obesity (Ricquier, D. et al.
( 1999) J. Int. Med. 245:637-642).
A number of metal ions such as iron, zinc, copper, cobalt, manganese, molybdenum, selenium, nickel, and chromium are important as cofactors for a number of enzymes. For example, zinc is required for the function of enzymes such as the extracellular matrix metalloproteinases, and zinc ions stabilize several motifs commonly found in transcription factors, including zinc fingers, zinc clusters, and LIM domains. Zinc and other metal ions must be provided in the diet, and are absorbed by transporters in the gastrointestinal tract. Plasma proteins transport the metal ions to the liver and other target organs, where specific transporters move the ions into cells and cellular organelles as needed. Imbalances in metal ion metabolism have been associated with a number of disease states (banks, D.M. (1986) J. Med. Genet. 23:99-106).
The largest and most diverse family of transport proteins known are the ATP-binding cassette (ABC) transporters. As a family, ABC transporters can transport substances that differ markedly in chemical structure and size, ranging from small molecules such as ions, sugars, amino acids, peptides, and phospholipids, to lipopeptides, large proteins, and complex hydrophobic drugs. ABC proteins consist of four modules: two nucleotide-binding domains (NBD), which hydrolyze ATP to supply the energy required for transport, and two membrane-spanning domains (MSD), each containing six putative transmembrane segments. These four modules may be encoded by a single gene, as is the case for the cystic fibrosis transmembrane regulator (CFTR), or by separate genes. When encoded by separate genes, each gene product contains a single NBD and MSD. These "half molecules" form homo- and heterodimers, such as Tapl and Tap2, the endoplasmic reticulum-based major histocompatibility (MHC) peptide transport system. Several genetic diseases are attributed to defects in ABC transporters, such as the following diseases and their corresponding proteins: cystic fibrosis (CFTR, an ion channel), adrenoleukodystrophy (adrenoleukodystrophy protein, ALDP), Zellweger syndrome (peroxisomal membrane protein-70, PMP70), and hyperinsulinemic hypoglycemia (sulfonylurea receptor, SUR). Overexpression of the multidrug resistance (MDR) protein, another ABC transporter, in human cancer cells makes the cells resistant to a variety of cytotoxic drugs used in chemotherapy (Taglicht, D. and S. Michaelis (1998) Methods Enzymol. 292:131-163).
The nuclear pore complex (NPC) is a large multiprotein complex spanning the nuclear envelope which mediates the transport of proteins and RNA molecules between the nucleus and the cytoplasm, thus contributing to the regulation of gene expression. The NPC
allows passive diffusion of ions, small molecules, and macromolecules under about 60kD, while larger macromolecules are transported by facilitated, energy-dependent pathways. Nuclear localization signals (NLS), consisting of short stretches of amino acids enriched in basic residues, are found on proteins that are targeted to the nucleus, such as the glucocorticoid receptor. The NLS is recognized by the NLS receptor, importin, which then interacts with the monomeric GTP-binding protein Ran.
This NLS
protein/receptor/Ran complex navigates the nuclear pore with the help of the homodimeric protein nuclear transport factor 2 (NTF2) (Nakielny, S. and G. Dreyfuss ( 1997) Curr.
Opin. Cell Biol. 9:420-429; Gorlich, D. ( 1997) Curr. Opin. Cell Biol. 9:412-419). Four O-linked glycoproteins, p62, p58, p54, and p45, exist as a stable "p62 complex" that forms a ring localized on both nucleoplasmic and cytoplasmic surfaces of the NPC. The p62, p58, and p54 proteins all interact directly with the cytosolic transport factors p97 and NTF2, suggesting that the p62 complex is an important ligand binding site near the central gated channel of the NPC (Hu, T. et al. (1996) J. Cell Biol. 134:589-601).
Transport can also occur through intercellular bridges which connect the cytoplasms of sister cells, for example in the male and female germline of species ranging from fruit flies to humans.
These bridges allow passage of cytoplasmic materials between cells during development.
Intercellular bridges have also been found to connect somatic cells. The nurse cells and oocyte of a Drosophila egg chamber, which are derived from a single precursor cell through four rounds of mitosis, are connected to each other through intercellular bridges called ring canals. The cells do not completely separate after mitosis; the mitotic cleavage furrows are transformed into ring canals by the addition of an actin cytoskeleton lining the tunnels between the cells. The Drosophila ketch protein functions in organizing actin in the ring canal. Mutations in ketch cause female sterility in Dros~hila. Ketch contains four protein domains: the NTR domain at the N-terminus, the BTB or POZ domain, the IVR or intervening region; and the ketch repeat domain, which contains six 50-amino acid ketch repeats. The BTB or POZ domain, a 120-amino acid motif that is also found in several zinc-finger containing transcription factors, may be important in dimerization of ketch. Ketch repeats are found in other proteins as well and may be important for actin binding (Robinson, D.N.
and L. Cooley (1997) J. Cell Biol. 138:799-810; Cooley, L. (1998) Cell 93:913-915).
Ion Channels The electrical potential of a cell is generated and maintained by controlling the movement of ions across the plasma membrane. The movement of ions requires ion channels, which form an ion-selective pore within the membrane. Ion channels share common structural and mechanistic themes.
The channel consists of four or five subunits or protein monomers that are arranged like a barrel in the plasma membrane. Each subunit typically consists of six potential transmembrane segments (S 1, S2, S3, S4, S5, and S6). The center of the barrel forms a pore lined by a-helices or ~3-strands. The side chains of the amino acid residues comprising the a-helices or ~3-strands establish the charge (cation or anion) selectivity of the channel. The degree of selectivity, or what specific ions are allowed to pass through the channel, depends on the diameter of the narrowest part of the pore. There are two basic types of ion channels, ion transporters and gated ion channels.
Ion transporters utilize the energy obtained from ATP hydrolysis to actively transport an ion against the ion's concentration gradient. Gated ion channels allow passive flow of an ion down the ion's electrochemical gradient under restricted conditions. Together, these types of ion channels generate, maintain, and utilize an electrochemical gradient that is used in 1) electrical impulse conduction down the axon of a nerve cell, 2) transport of molecules into cells against concentration gradients, 3) initiation of muscle contraction, and 4) endocrine cell secretion.
Transmembrane ATPases are divided into three families. The phosphorylated (P) class ion transporters, including Na+-K+ ATPase, Caz+ ATPase, H'' ATPase, and Cu"' ATPase, are activated by a phosphorylation event. P-class ion transporters are responsible for maintaining resting potential distributions such that cytosolic concentrations of Na+ and Ca2+ are low and cytosolic concentration of K+ is high. The vacuolar (V) class of ion transporters include H+ pumps on intracellular organelles, such as lysosomes and Golgi. V-class ion transporters are responsible for generating the low pH within the lumen of these organelles that is required for function. The coupling factor (F) class consists of H+ pumps in the mitochondria. F-class ion transporters utilize a proton gradient to generate ATP from ADP and inorganic phosphate (P;).
Cu.~ ATPases export copper from cells (PROSITE PDOC00139 E1-E2 ATPases phosphorylation site). Mutations in one Cu-'-' ATPase cause Wilson disease, in which toxic amounts of copper accumulate in a number of organs, particularly the liver and brain (Tanzi, R.E. et al. ( 1993) Nat. Genet. 5:344-350). Mutations in another Cu+' ATPase cause Menkes disease and occipital horn syndrome. Menkes disease mutations block export of copper from the gastrointestinal tract, leading to skeletal abnormalities, severe mental retardation, neurologic degeneration, and mortality in early childhood (Harrison, M.D. and C.T. Dameron (1999) J. Biochem. Mol. Toxicol.
13:93-106).
Occipital horn syndrome mutations cause connective tissue defects (Harrison, suyra; Levinson, B. et al. ( 1996) Hum. Mol. Genet. 5:1737-1742).
The coupling factor (F) class of ion transporters consists of H+ pumps in mitochondria, chloroplasts, and bacteria. For example, the FoF~ ATPase utilizes a proton gradient across the inner mitochondria) membrane to generate ATP from ADP and inorganic phosphate (P;).
The FoF, ATPase is composed of the Fo complex, which is the transmembrane channel through which protons flow, and the F, complex, where ATP synthesis activity resides. Fn has three subunits, A
(also known as protein 6), B, and C (Lodish, H. et al. ( 1995) Molecular Cell BioloQV, Scientific American Books, New York NY, pp. 752-756; PROSITE PDOC00420 ATP synthase a subunit signature).
Voltage-gated Ca'-+ channels are involved in presynaptic neurotransmitter release, and heart and skeletal muscle contraction. The voltage-gated Ca'-+ channels from skeletal muscle (L-type) and brain (N-type) have been purified and, though their functions differ dramatically, they have similar subunit compositions. The channels are composed of three subunits. The a, subunit forms the membrane pore and voltage sensor, while the a,=6 and (3 subunits modulate the voltage-dependence, gating properties, and the current amplitude of the channel. These subunits are encoded by at least six a,, one a"8, and four (3 genes. A fourth subunit, y, has been identified in skeletal muscle (Walker, D. et al. (1998) J. Biol. Chem. 273:2361-2367; and Jay, S.D. et al. (1990) Science 248:490-492). The human ~4 subunit is homologous to the mouse epilepsy gene lethargic, and is a candidate for involvement in neurological disorders including ataxia and absence epilepsy (Escayg, A. et al. ( 1998) Genomics 50:14-22).
Ligand-gated channels open their pores when an extracellular or intracellular mediator binds to the channel. Neurotransmitter-gated channels are channels that open when a neurotransmitter binds to their extracellular domain. These channels exist in the postsynaptic membrane of nerve or muscle cells. There are two types of neurotransmitter-gated channels. Sodium channels open in response to excitatory neurotransmitters, such as acetylcholine, glutamate, and serotonin. This opening causes an influx of Na+ and produces the initial localized depolarization that activates the voltage-gated channels and starts the action potential. Chloride channels open in response to inhibitory neurotransmitters, such as y-aminobutyric acid (GABA) and glycine, leading to hyperpolarization of the membrane and the subsequent generation of an action potential.
Ion channels are expressed in a number of tissues where they are implicated in a variety of processes. CNG channels, while abundantly expressed in photoreceptor and olfactory sensory cells, are also found in kidney, lung, pineal, retinal ganglion cells, testis, aorta, and brain. Calcium-activated K+ channels may be responsible for the vasodilatory effects of bradykinin in the kidney and for shunting excess K+ from brain capillary endothelial cells into the blood. They are also implicated in repolarizing granulocytes after agonist-stimulated depolarization (Ishi, T.M.
et al. ( 1997) Proc. Natl.
Acad. Sci. USA 94:11651-11656). Another transmembrane protein, the leukotrine B4 receptor (BLT) appears to be involved in inflammation responses and host cell defense against infection. BLT also functions as an HIV coreceptor (Izumi, T. et al. ( 1997) Nature 387:620-624;
Martin, V. et al. ( 1999) J. Biol. Chem. 274:8597-8603).
Ion channels have been the target for many drug therapies. Neurotransmitter-gated channels have been targeted in therapies for treatment of insomnia, anxiety, depression, and schizophrenia.
Voltage-gated channels have been targeted in therapies for arrhythmia, ischemic stroke, head trauma, and neurodegenerative disease (Taylor, C.P. and L.S. Narasimhan ( 1997) Adv.
Pharmacol. 39:47-98).
K+ channels are located in all cell types, and may be regulated by voltage, ATP
concentration, or second messengers such as Ca~ and cAMP. In non-excitable tissue, K+ channels are involved in protein synthesis, control of endocrine secretions, and the maintenance of osmotic equilibrium across membranes. In neurons and other excitable cells, in addition to regulating action potentials and repolarizing membranes, K+ channels are responsible for setting resting membrane potential. The cytosol contains non-diffusible anions and, to balance this net negative charge, the cell contains a Na+-K+ pump and ion channels that provide the redistribution of Na+, K+, and Cl-. The pump actively transports Na+ out of the cell and K+ into the cell in a 3:2 ratio. Ion channels in the plasma membrane allow K+ and CI ~ to flow by passive diffusion. Because of the high negative charge within the cytosol, CI - flows out of the cell. The flow of K' is balanced by an electromotive force pulling K+ into the cell, and a K+ concentration gradient pushing K+ out of the cell. Thus, the resting membrane potential is primarily regulated by K+flow (Salkoff, L. and T. Jegla (1995) Neuron 15:489-492). Information on NY-REN-45, a K+ channel integral membrane protein, can be found in Scanlan, M.J. et al. (1998; Int. J. Cancer 76:652-658). The emopamil-binding protein (EBP) shares structural features with both pro- and eukaryotic drug transport proteins (Hanner, M. et al. ( 1995) J.
Biol. Chem. 270:7551-7557). The Na+ channel, transmembrane protein myelin protein zero (MPZ) may be responsible for some sporadic cases of Dejerine-Scottas disease (hereditary motor and sensory neuropathy type III) (Hayasaka, K. et al. ( 1993) Nat. Genet. 5:266-268).
K+ pore-forming subunits generally have six transmembrane-spanning domains with a short region between the fifth and sixth transmembrane regions that senses membrane potential; and the amino and carboxy termini are located intracellularly. In mammalian heart, the duration of ventricular action potential is controlled by a K' current. Thus, the K+
channel is central to the control of heart rate and rhythm. K+ channel dysfunctions are associated with a number of renal diseases including hypertension, hypokalemia, and the associated Banter's syndrome and Getelman's syndrome, as well as neurological disorders including epilepsy. K+ channels have been implicated in Alzheimer's disease by observations that a significant component of senile plaques, beta amyloid or A beta, also blocks voltage-gated potassium channels in hippocampal neurons (Antes, L.M. et al.
(1998) Seminar Nephrol. 18:31-45; Stoffel, M. and L.Y. Jan (1998) Nat. Genet.
18:6-8; Madeja, M.
et al. (1997) Eur. J. Neurosci. 9:390-395; Good, T.A. et al. (1996) Biophys.
J. 70:296-304).
Gated ion channels control ion flow by regulating the opening and closing of pores. These channels are categorized according to the manner of regulating the gating function. Mechanically-gated channels open pores in response to mechanical stress, voltage-gated channels open pores in response to changes in membrane potential, and ligand-gated channels open pores in the presence of a specific ion, nucleotide, or neurotransmitter.
Voltage-gated Na+ channels are responsible for electrical excitability of neurons, skeletal muscle, heart, and neuroendocrine tissues. For example, the sequential opening and closing of voltage-gated Na+ channels results in the propagation of action potentials down neuronal axons. Na+
channels isolated from rat brain tissue are heterotrimeric complexes composed of a 260 kDa pore forming a subunit that associates with two smaller auxiliary subunits, (31 and (32. The ~i2 subunit is an integral membrane glycoprotein that contains an extracellular Ig domain, and its association with a and (31 subunits correlates with increased function of the channel, a change in the channel's gating properties, as well as an increase in whole cell capacitance (Isom, L.L. et al. ( 1995) Cell 83:433-442).

Integral Membrane Proteins The majority of known integral membrane proteins are transmembrane proteins (TM) which are characterized by an extracellular, a transmembrane, and an intracellular domain. TM domains are typically comprised of 15 to 25 hydrophobic amino acids which are predicted to adopt an a-helical conformation. TM proteins are classified as bitopic (Types I and II) and polytopic (Types III and IV) (Singer, S.J. (1990) Annu. Rev. Cell Biol. 6:247-96). Bitopic proteins span the membrane once while polytopic proteins contain multiple membrane-spanning segments. TM proteins that act as cell-surface receptor proteins involved in signal transduction include growth and differentiation factor receptors, and receptor-interacting proteins such as Drosophila pecanex and frizzled proteins, LIV-1 protein, NF2 protein, and GNS1/SUR4 eukaryotic integral membrane proteins. TM
proteins also act as transporters of ions or metabolites, such as gap junction channels (connexins) and ion channels, and as cell anchoring proteins, such as lectins, integrins, and fibronectins.
TM proteins act as vesicle organelle-forming molecules, such as calveolins, or as cell recognition molecules, such as cluster of differentiation (CD) antigens, glycoproteins, and mucins. Information on connexin can be found in Kanter, H.L. et al. (1994; J. Mol. Cell. Cardiol. 26:861-868).
Many membrane proteins (MPs) contain amino acid sequence motifs that target these proteins to specific subcellular sites. Examples of these motifs include PDZ domains, KDEL, RGD, NGR, and GSL sequence motifs, von Willebrand factor A (vWFA) domains, and EGF-like domains. RGD, NGR, and GSL motif-containing peptides have been used as drug delivery agents in cancer treatments which target tumor vasculature (Arap, W. et al. ( 1998) Science, 279:377-380.) Furthermore, MPs may also contain amino acid sequence motifs, such as the carbohydrate recognition domain (CRD), also known as the C-type lectin domain, that mediate interactions with extracellular or intracellular molecules.
G-protein coupled receptors (GPCR) comprise a superfamily of integral membrane proteins which transduce extracellular signals. GPCRs include receptors for biogenic amines, lipid mediators of inflammation, peptide hormones, and sensory signal mediators. The structure of these highly-conserved receptors consists of seven hydrophobic transmembrane regions, an extracellular N-terminus, and a cytoplasmie C-terminus. Three extracellular loops alternate with three intracellular loops to link the seven transmembrane regions. The most conserved parts of these proteins are the transmembrane regions and the first two cytoplasmic loops. Cysteine disulfide bridges connect the second and third extracellular loops. A conserved, acidic-Arg-aromatic residue triplet present in the second cytoplasmic loop may interact with G proteins. A GPCR consensus pattern is characteristic of most proteins belonging to this superfamily (ExPASy PROSITE document PS00237;
and Watson, S. and S. Arkinstall (1994) The G-protein Linked Receytor Facts Book, Academic Press, San Diego CA, pp 2-6). Mutations and changes in transcriptional activation of GPCR-encoding genes have been associated with neurological disorders such as schizophrenia, Parkinson's disease, Alzheimer's disease, drug addiction, and feeding disorders.
Cytochromes are electron-transferring proteins that contain a heme prosthetic group, a porphyrin ring containing a tightly bound iron atom. Cytochromes act as oxidoreductases in such diverse cellular processes as respiration, photosynthesis, fatty acid metabolism, and neurotransmitter biosynthesis. The heme iron atom serves as the actual electron carrier by changing from the ferric to the ferrous oxidation state when accepting an electron. Cytochromes accept electrons from one substrate such as NADH or ascorbate and donate them to other electron carriers such as other cytochromes, ubiquinone, or semidehydroascorbic acid (Lodish, H. et al. ( 1995) Molecular Cell BioloQV, Scientific American Books, New York NY, pp. 759-770, 786-797;
Sperling, P. et al. (1995) Eur. J. Biochem. 232:798-805; and Online Mendelian Inheritance in Man (OMIM) *600019 Cytochrome b561, CYB561).
Cytochrome b5 is an electron donor in membrane-linked redox enzyme systems involved in lipid and drug metabolism. Cytochrome b5 has been found in Golgi, plasma, outer mitochondrial, endoplasmic reticulum (ER), and microbody membranes. Conserved amino acids in cytochrome b5 include eight invariant amino acids at W34, H51, P52, G53, G54, G63, F70, and H74, and fifteen conserved amino acids at L24, I35, S36, V41, Y42, N43, T45, W47, A48, L58, D65, T67, L85, T87, and G88 (numbering based on the sunflower cytochrome b5/delta-6 desaturase fusion protein; GI
1040729, Sperling, supra). The invariant residues HS 1PGG are involved in heme-binding.
Cytochrome b5-like domains have also been found linked to other enzymes. For example, cytochrome b5-like domains are part of delta-9 fatty acid desaturases in yeast and Histoplasma capsulatum, nitrate reductase, sulfite reductase, flavocytochrome b2, Arabido~sis thaliana acyl lipid desaturase, and Borago officinalis (borage) and Helianthus annuus (sunflower) delta-6 desaturases (Sperling, supra; Sayanova, O. et al (1997) Proc. Natl. Acad. Sci. USA 94:4211-4216; and Mitchell, A.G. and C.E. Martin (1997) J. Biol. Chem. 272:28281-28288).
Signal peptides are found on proteins that are targeted to the endoplasmic reticulum (ER).
Signal peptides consist of stretches of amino acids enriched in hydrophobic residues. Signal peptides are usually found at the extreme N-terminus of the protein and are recognized by a cytosolic signal-recognition peptide (SRP). The SRP binds to the signal peptide and to an SRP
receptor, an integral membrane protein in the ER. Once bound to the SRP receptor, the newly formed protein containing the signal peptide is translocated across the ER membrane. Proteins containing signal peptides may end up inserted into the lipid bilayer, or they may end up in the lumen of an organelle or secreted from the cell.
Disease Correlation The etiology of numerous human diseases and disorders can be attributed to defects in the transport of molecules across membranes. Defects in the trafficking of membrane-bound transporters and ion channels are associated with several disorders, e.g. cystic fibrosis, glucose-galactose malabsorption syndrome, hypercholesterolemia, von Gierke disease, and certain forms of diabetes mellitus. Single-gene defect diseases resulting in an inability to transport small molecules across membranes include, e.g., cystinuria, iminoglycinuria, Hartup disease, and Fanconi disease (van't Hoff, W.G. ( 1996) Exp. Nephrol. 4:253-262; Talente, G.M. et al. ( 1994) Ann.
Intern. Med. 120:218-226;
and Chillon, M. et al. (1995) New Engl. J. Med. 332:1475-1480).
Cystinuria is an inherited disease that results from the inability to transport cystine, the disulfide-linked dimer of cysteine, from the urine into the blood.
Accumulation of cystine in the urine leads to the formation of cystine stones in the kidneys.
Transthyretin (TTR), present in human plasma, binds to and transports the thyroid hormone thyroxine. Mutations in TTR result in the conversion of TTR to amyloid, an insoluble fibrillar structure. The resulting amyloid plaques have been shown to be the causative agent in the development of familial amyloid polyneuropathy and senile systemic amyloidosis (Miroy, G.J. et al.
IS (1996) Proc. Natl. Acad. Sci. USA 93:15051-15056).
Stomatin, a 31-kDa erythrocyte integral membrane protein has been linked to the hereditary anemia stomatocytosis. This anemia is characterized by red blood cells that lack stomatin and leak Na+ and K+. Thus, stomatin is presumed to play a role in the regulation of ion transport. Red blood cell ion transport defects are also linked to other disorders such as hypertension (Stewart, G.W.
(1997) Int. J. Biochem. Cell Biol. 29:271-274).
The discovery of new human transport proteins 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 transport, metabolic, neurological, cardiovascular, reproductive, and immune disorders, and cell proliferative disorders including cancer.
SUMMARY OF THE INVENTION
The invention features purified polypeptides, human transport proteins, referred to collectively as "TPPT" and individually as "TPPT-1," "TPPT-2," "TPPT-3," "TPPT-4.," "TPPT-5,"
"TPPT-6," ..~P.h-7," ".LPph-8 " ".LpPT-9,,~ ".hpPT-lp,,> "TPPT-11,~, ".hPPT-12 " "Z.pp.L-13 ,>
"TPPT-14," "TPPT-15," "TPPT-16," "TPPT-17," "TPPT-18," "TPPT-19," "TPPT-20,"
"TPPT-21,"
"TPPT-22," "TPPT-23," "TPPT-24," "TPPT-25," "TPPT-26," "TPPT-27," "TPPT-28,"
"TPPT-29,"
"TPPT-30," "TPPT-31," "TPPT-32," "TPPT-33," "TPPT-34," "TPPT-35," "TPPT-36,"
"TPPT-37,"
"TPPT-38," "TPPT-39," "TPPT-40," "TPPT-41," "TPPT-42," and "TPPT-43." In one aspect, the invention provides an isolated polypeptide comprising an amino acid sequence selected from the group consisting of a) an amino acid sequence selected from the group consisting of SEQ ID NO:1-43, b) a naturally occurring amino acid sequence having at least 90% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NO:I-43, c) a biologically active fragment of an amino acid sequence selected from the group consisting of SEQ
ID NO:1-43, and d) an immunogenic fragment of an amino acid sequence selected from the group consisting of SEQ ID
NO:1-43. In one alternative, the invention provides an isolated polypeptide comprising the amino acid sequence of SEQ ID NO:1-43.
The invention further provides an isolated polynucleotide encoding a polypeptide comprising an amino acid sequence selected from the group consisting of a) an amino acid sequence selected from the group consisting of SEQ ID NO:1-43, b) a naturally occurring amino acid sequence having at least 90% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID
NO:1-43, c) a biologically active fragment of an amino acid sequence selected from the group consisting of SEQ >D NO:1-43, and d) an immunogenic fragment of an amino acid sequence selected from the group consisting of SEQ ID NO:1-43. In one alternative, the polynucleotide encodes a polypeptide selected from the group consisting of SEQ ID NO:1-43. In another alternative, the polynucleotide is selected from the group consisting of SEQ ID N0:44-86.
Additionally, the invention provides a recombinant polynucleotide comprising a promoter sequence operably linked to a polynucleotide encoding a polypeptide comprising an amino acid sequence selected from the group consisting of a) an amino acid sequence selected from the group consisting of SEQ B7 NO:1-43, b) a naturally occurring amino acid sequence having at least 90%
sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NO:1-43, c) a biologically active fragment of an amino acid sequence selected from the group consisting of SEQ )I7 NO:1-43, and d) an immunogenic fragment of an amino acid sequence selected from the group consisting of SEQ )D NO:1-43. 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 comprising an amino acid sequence selected from the group consisting of a) an amino acid sequence selected from the group consisting of SEQ ID NO:1-43, b) a naturally occurring amino acid sequence having at least 90%
sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NO:1-43, c) a biologically active fragment of an amino acid sequence selected from the group consisting of SEQ ID NO:1-43, and d) an immunogenic fragment of an amino acid sequence selected from the group consisting of SEQ ID NO:1-43. 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 comprising an amino acid sequence selected from the group consisting of a) an amino acid sequence selected from the group consisting of SEQ ID NO' 1-43, b) a naturally occurring amino acid sequence having at least 90% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NO:1-43, c) a biologically active fragment of an amino acid sequence selected from the group consisting of SEQ ID NO:1-43, and d) an immunogenic fragment of an amino acid sequence selected from the group consisting of SEQ ID NO:1-43.
The invention further provides an isolated polynucleotide comprising a polynucleotide sequence selected from the group consisting of a) a polynucleotide sequence selected from the group consisting of SEQ ID N0:44-86, b) a naturally occurnng polynucleotide sequence having at least 70%
sequence identity to a polynucleotide sequence selected from the group consisting of SEQ ID N0:44-86, c) a polynucleotide sequence complementary to a), d) a polynucleotide sequence complementary to 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 comprising a polynucleotide sequence selected from the group consisting of a) a polynucleotide sequence selected from the group consisting of SEQ ID N0:44-86, b) a naturally occurring polynucleotide sequence having at least 70%
sequence identity to a polynucleotide sequence selected from the group consisting of SEQ m N0:44-86, c) a polynucleotide sequence complementary to a), d) a polynucleotide sequence complementary to 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 comprising a polynucleotide sequence selected from the group consisting of a) a polynucleotide sequence selected from the group consisting of SEQ ID N0:44-86, b) a naturally occurring polynucleotide sequence having at least 70%
sequence identity to a polynucleotide sequence selected from the group consisting of SEQ ID N0:44-86, c) a polynucleotide sequence complementary to a), d) a polynucleotide sequence complementary to b), and e) an RNA equivalent of a)-d). The method comprises 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.

The invention further provides a pharmaceutical composition comprising an effective amount of a polypeptide comprising an amino acid sequence selected from the group consisting of a) an amino acid sequence selected from the group consisting of SEQ ID NO:I-43, b) a naturally occurring amino acid sequence having at least 90% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NO:1-43, c) a biologically active fragment of an amino acid sequence selected from the group consisting of SEQ )T7 NO:1-43, and d) an immunogenic fragment of an amino acid sequence selected from the group consisting of SEQ ID NO:1-43, and a pharmaceutically acceptable excipient. In one embodiment, the pharmaceutical composition comprises an amino acid sequence selected from the group consisting of SEQ ID NO:1-43. The invention additionally provides a method of treating a disease or condition associated with decreased expression of functional TPPT, comprising administering to a patient in need of such treatment the pharmaceutical composition.
The invention also provides a method for screening a compound for effectiveness as an agonist of a polypeptide comprising an amino acid sequence selected from the group consisting of a) an amino acid sequence selected from the group consisting of SEQ ID NO:1-43, b) a naturally occurring amino acid sequence having at least 90% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NO:1-43, c) a biologically active fragment of an amino acid sequence selected from the group consisting of SEQ m NO:1-43, and d) an immunogenic fragment of an amino acid sequence selected from the group consisting of SEQ
ID NO:1-43. 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 pharmaceutical 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 TPPT, comprising administering to a patient in need of such treatment the pharmaceutical composition.
Additionally, the invention provides a method for screening a compound for effectiveness as an antagonist of a polypeptide comprising an amino acid sequence selected from the group consisting of a) an amino acid sequence selected from the group consisting of SEQ ID NO:1-43, b) a naturally occurring amino acid sequence having at least 90% sequence identity to an amino acid sequence selected from the group consisting of SEQ )D NO:1-43, c) a biologically active fragment of an amino acid sequence selected from the group consisting of SEQ ID NO:1-43, and d) an immunogenic fragment of an amino acid sequence selected from the group consisting of SEQ
ID NO:1-43. 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 pharmaceutical 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 TPPT, comprising administering to a patient in need of such treatment the pharmaceutical composition.
The invention further provides a method of screening for a compound that specifically binds to a polypeptide comprising an amino acid sequence selected from the group consisting of a) an amino acid sequence selected from the group consisting of SEQ ID NO:1-43, b) a naturally occurring amino acid sequence having at least 90% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NO:1-43, c) a biologically active fragment of an amino acid sequence selected from the group consisting of SEQ ID NO:1-43, and d) an immunogenic fragment of an amino acid sequence selected from the group consisting of SEQ ID NO:1-43. 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 comprising an amino acid sequence selected from the group consisting of a) an amino acid sequence selected from the group consisting of SEQ ID NO:1-43, b) a naturally occurnng amino acid sequence having at least 90% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NO:1-43, c) a biologically active fragment of an amino acid sequence selected from the group consisting of SEQ ID NO:1-43, and d) an immunogenic fragment of an amino acid sequence selected from the group consisting of SEQ
ID NO:1-43. 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 sequence selected from the group consisting of SEQ ID N0:44-86, the method comprising a) exposing a sample comprising the target polynucleotide to a compound, and b) detecting altered expression of the target polynucleotide.
BRIEF DESCRIPTION OF THE TABLES
Table 1 shows polypeptide and nucleotide sequence identification numbers (SEQ
ID NOs), clone identification numbers (clone IDs), cDNA libraries, and cDNA fragments used to assemble full-length sequences encoding TPPT.
Table 2 shows features of each polypeptide sequence, including potential motifs, homologous sequences, and methods, algorithms, and searchable databases used for analysis of TPPT.

Table 3 shows selected fragments of each nucleic acid sequence; the tissue-specific expression patterns of each nucleic acid sequence as determined by northern analysis; diseases, disorders, or conditions associated with these tissues; and the vector into which each cDNA was cloned.
Table 4 describes the tissues used to construct the cDNA libraries from which cDNA clones encoding TPPT were isolated.
Table 5 shows the tools, programs, and algorithms used to analyze the polynucleotides and polypeptides of the invention, along with applicable descriptions, references, and threshold parameters.
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
"TPPT" refers to the amino acid sequences of substantially purified TPPT
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 TPPT. Agonists may include proteins, nucleic acids, carbohydrates, small molecules, or any other compound or composition which modulates the activity of TPPT either by directly interacting with TPPT or by acting on components of the biological pathway in which TPPT
participates.
An "allelic variant" is an alternative form of the gene encoding TPPT. 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 occurnng 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 m a given sequence.
"Altered" nucleic acid sequences encoding TPPT include those sequences with deletions, insertions, or substitutions of different nucleotides, resulting in a polypeptide the same as TPPT or a polypeptide with at least one functional characteristic of TPPT. Included within this definition are polymorphisms which may or may not be readily detectable using a particular oligonucleotide probe of the polynucleotide encoding TPPT, and improper or unexpected hybridization to allelic variants, with a locus other than the normal chromosomal locus for the polynucleotide sequence encoding TPPT. 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 TPPT. 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 TPPT 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 may 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 polymerise chain reaction (PCR) technologies well known in the art.
The term "antagonist" refers to a molecule which inhibits or attenuates the biological activity of TPPT. Antagonists may include proteins such as antibodies, nucleic acids, carbohydrates, small molecules, or any other compound or composition which modulates the activity of TPPT either by directly interacting with TPPT or by acting on components of the biological pathway in which TPPT

participates.
The term "antibody" refers to intact immunoglobulin molecules as well as to fragments thereof, such as Fab, Flab' ),, and Fv fragments, which are capable of binding an epitopic determinant.
Antibodies that bind TPPT 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 animal (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 (KLH). 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 immunize a host animal, numerous regions 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 may compete with the intact antigen (i.e., the immunogen IS used to elicit the immune response) for binding to an antibody.
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" can refer to the antisense 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 TPPT, 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-5'.
A "composition comprising a given polynucleotide sequence" and a "composition comprising a 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 encoding TPPT or fragments of TPPT 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 solution containing salts (e.g., NaCI), 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 (PE Biosystems, Foster City CA) in the 5' and/or 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 Ala Gly, Ser Arg His, Lys Asn Asp, Gln, His Asp Asn, Glu Cys Ala, Ser Gln Asn, Glu, His Glu Asp, Gln, His Gly Ala His Asn, Arg, Gln, Glu Ile Leu, Val Leu Ile, Val Lys Arg, Gln, 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 sequence 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.
A "fragment" is a unique portion of TPPT or the polynucleotide encoding TPPT
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 ID N0:44-86 comprises a region of unique polynucleotide sequence that specifically identifies SEQ ID N0:44-86, for example, as distinct from any other sequence in the genome from which the fragment was obtained. A fragment of SEQ ID N0:44-86 is useful, for example, in hybridization and amplification technologies and in analogous methods that distinguish SEQ ID N0:44-86 from related polynucleotide sequences. The precise length of a fragment of SEQ
ID N0:44-86 and the region of SEQ ID N0:44-86 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 ID NO:I-43 is encoded by a fragment of SEQ ID N0:44-86. A
fragment of SEQ ID NO: I-43 comprises a region of unique amino acid sequence that specifically identifies SEQ ID NO: I-43. For example, a fragment of SEQ ID NO:1-43 is useful as an immunogenic peptide for the development of antibodies that specifically recognize SEQ ID NO:I-43.
The precise length of a fragment of SEQ ID NO:I-43 and the region of SEQ ID NO:I-43 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 NCBI, Bethesda, MD, and on the Internet at http://www.ncbi.nlm.nih.gov/BLAST/. The BLAST software suite includes various sequence analysis programs including "blastn," that is used to align a known polynucleotide sequence with other polynucleotide sequences from 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, for example:
Matrix: BLOSUM62 Reward for match: 1 Penalty for mismatch: -2 Open Gap: S and Extension Gap: 2 penalties Gap x drop-off: SO
Expect: 70 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 ID 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 IS in a nucleic acid sequence can be made using this degeneracy to produce multiple nucleic acid sequences that all encode substantially the same protein.
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, for a pairwise comparison of two polypeptide sequences, one may use the "BLAST 2 Sequences"
tool Version 2Ø12 (Apr-21-2000) with blastp set at default parameters. Such default parameters may be, for example:
Matrix: BLOSUM62 Open Gap: 11 and Extension Gap: 1 penalties Gap x drop-off. SO

Expect: 10 Word 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 hybridization 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 pg/ml 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 (Tm) for the specific sequence at a defined ionic strength and pH. The Tm is the temperature (under defined ionic strength and pH) at which 50% 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 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 SDS 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 pg/ml. 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 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 TPPT
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 TPPT 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 TPPT. For example, modulation may cause an increase or a decrease in protein activity, binding characteristics, or any other biological, functional, or immunological properties of TPPT.
The phrases "nucleic acid" and "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 join two protein coding regions, in the same reading frame.
"Peptide nucleic acid" (PNA) refers to an antisense molecule or anti-gene agent which comprises an 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 or RNA and stop transcript elongation, and may be pegylated to extend their lifespan in the cell.
"Post-translational modification" of an TPPT may involve lipidation, glycosylation, phosphorylation, acetylation, racemization, proteolytic cleavage, and other modifications known in the art. These processes may occur synthetically or biochemically. Biochemical modifications will vary by cell type depending on the enzymatic milieu of TPPT.
"Probe" refers to nucleic acid sequences encoding TPPT, their complements, or fragments 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 DNA strand by a DNA polymerise enzyme. Primer pairs can be used for amplification (and identification) of a nucleic acid sequence, e.g., by the polymerise chain reaction (PCR).
Probes and primers as used in the present invention typically comprise at least 15 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, or at least 150 consecutive nucleotides of the disclosed nucleic acid sequences. Probes and primers 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, may be used.
Methods for preparing and using probes and primers are described in the references, for example Sambrook, J. et al., 1989, Molecular Cloning: A Laborator~Manual, 2"d ed., vol. 1-3, Cold Spring Harbor Press, Plainview NY; Ausubel, F.M. et a1.,1987, Current Protocols in Molecular Bioloey, Greene Publ. Assoc. & Wiley-Intersciences, New York NY; Innis, 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 known sequence, for example, by using computer programs intended for that purpose such as Primer (Version 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 for 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 oligonucleotides 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 selection of primers that hybridize to either 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, supra. 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 may be 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 nucleic acids encoding TPPT, or fragments thereof, or TPPT itself, may comprise a bodily fluid; an extract from 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 can have a variety of surface forms, such as wells, trenches, pins, channels and pores, to which polynucleotides or polypeptides are bound.
A "transcript image" 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 in 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. 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 95% or at least 98% 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 alternative 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 generally will 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 "single 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 95%, or at least 98% or greater sequence identity over a certain defined length of one of the polypeptides.
THE INVENTION
The invention is based on the discovery of new human transport proteins (TPPT), the polynucleotides encoding TPPT, and the use of these compositions for the diagnosis, treatment, or prevention of transport, metabolic, neurological, cardiovascular, reproductive, and immune disorders, and cell proliferative disorders including cancer.
IS Table I lists the Incyte clones used to assemble full length nucleotide sequences encoding TPPT. Columns I and 2 show the sequence identification numbers (SEQ ID NOs) of the polypeptide and nucleotide sequences, respectively. Column 3 shows the clone IDs of the Incyte clones in which nucleic acids encoding each TPPT were identified, and column 4 shows the cDNA
libraries from which these clones were isolated. Column 5 shows Incyte clones and their corresponding cDNA
libraries. Clones for which cDNA libraries are not indicated were derived from pooled cDNA
libraries. In some cases, GenBank sequence identifiers are also shown in column 5. The Incyte clones and GenBank cDNA sequences, where indicated, in column 5 were used to assemble the consensus nucleotide sequence of each TPPT and are useful as fragments in hybridization technologies.
The columns of Table 2 show various properties of each of the polypeptides of the invention:
column 1 references the SEQ ID NO; column 2 shows the number of amino acid residues in each polypeptide; column 3 shows potential phosphorylation sites; column 4 shows potential glycosylation sites; column 5 shows the amino acid residues comprising signature sequences and motifs; column 6 shows homologous sequences as identified by BLAST analysis; and column 7 shows analytical methods and in some cases, searchable databases to which the analytical methods were applied. The methods of column 7 were used to characterize each polypeptide through sequence homology and protein motifs.
The columns of Table 3 show the tissue-specificity and diseases, disorders, or conditions associated with nucleotide sequences encoding TPPT. The first column of Table 3 lists the nucleotide SEQ )D NOs. Column 2 lists fragments of the nucleotide sequences of column 1.
These fragments are useful, for example, in hybridization or amplification technologies to identify SEQ ID N0:44-86 and to distinguish between SEQ ID N0:44-86 and related polynucleotide sequences. The polypeptides encoded by these fragments are useful, for example, as immunogenic peptides. Column 3 lists tissue categories which express TPPT as a fraction of total tissues expressing TPPT. Column 4 lists diseases, disorders, or conditions associated with those tissues expressing TPPT as a fraction of total tissues expressing TPPT. Column 5 lists the vectors used to subclone each cDNA library.
Of particular interest is the expression of SEQ )D NO:50 exclusively in cardiovascular tissue, the expression of SEQ ID N0:56 in nervous and gastrointestinal tissues, the expression of SEQ ID
N0:57 in gastrointestinal tissues, and the expression of SEQ ID N0:66 in nervous system tissues. Of particular note is the tissue-specific expression of SEQ ID N0:75. Over 71 %
of the cDNA libraries expressing SEQ )D N0:75 are derived from lung tissue.
The columns of Table 4 show descriptions of the tissues used to construct the cDNA libraries from which cDNA clones encoding TPPT were isolated. Column 1 references the nucleotide SEQ ID
NOs, column 2 shows the cDNA libraries from which these clones were isolated, and column 3 shows the tissue origins and other descriptive information relevant to the cDNA
libraries in column 2.
SEQ ID N0:44 maps to chromosome 7 within the interval from 38.80 to 42.10 centiMorgans.
SEQ ID N0:48 maps to chromosome X within the interval from 107.90 to 122.80 centiMorgans.
SEQ ID N0:60 maps to chromosome 2 within the interval from 157.0 to 167.0 centiMorgans. SEQ
ID N0:65 maps to chromosome 2 within the interval from 17.4 to 40.7 centiMorgans and to chromosome 5 within the interval from 61.1 to 69.6 centiMorgans. The interval on chromosome 5 from 61.1 to 69.6 centiMorgans also contains genes associated with Cockayne syndrome. SEQ ID
N0:69 maps to chromosome 3 within the interval from 157.40 to 162.00 centiMorgans. SEQ ID
N0:70 maps to chromosome 3 within the interval from 176.40 to 179.80 centiMorgans. SEQ ID
N0:71 maps to chromosome 18 within the interval from the p-terminus to 52.30 centiMorgans. SEQ
ID N0:73 maps to chromosome 17 within the interval from 75.70 to 84.20 centiMorgans, and to chromosome 2 within the interval from 204.70 to 209.30 centiMorgans. SEQ ID
N0:76 maps to chromosome 20 within the interval from 79.00 to 94.40 centiMorgans. SEQ ID
N0:80 maps to chromosome 18 within the interval from 1.60 to 6.20 centiMorgans, and to chromosome 11 within the interval from 117.90 to 126.00 centiMorgans. SEQ ID N0:83 maps to chromosome 17 within the interval from 67.60 to 69.30 centiMorgans, and from 83.8 centiMorgans to the q-terminus, and to chromosome 7 within the interval from 105.20 to 114.50 centiMorgans.
The invention also encompasses TPPT variants. A preferred TPPT variant is one which has at least about 80%, or alternatively at least about 90%, or even at least about 95% amino acid sequence identity to the TPPT amino acid sequence, and which contains at least one functional or structural characteristic of TPPT.
The invention also encompasses polynucleotides which encode TPPT. In a particular embodiment, the invention encompasses a polynucleotide sequence comprising a sequence selected from the group consisting of SEQ ID N0:44-86, which encodes TPPT. The polynucleotide sequences of SEQ ID N0:44-86, 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 TPPT. 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 TPPT. A particular aspect of the invention encompasses a variant of a polynucleotide sequence comprising a sequence selected from the group consisting of SEQ ID N0:44-86 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 )D N0:44-86. Any one of the polynucleotide variants described above can encode an amino acid sequence which contains at least one functional or structural characteristic of TPPT.
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 TPPT, 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 TPPT, and all such variations are to be considered as being specifically disclosed.
Although nucleotide sequences which encode T'PPT and its variants are generally capable of hybridizing to the nucleotide sequence of the naturally occurring TPPT under appropriately selected conditions of stringency, it may be advantageous to produce nucleotide sequences encoding TPPT 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 TPPT 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 TPPT
and TPPT 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 TPPT 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
N0:44-86 and fragments thereof under various conditions of stringency. (See, e.g., Wahl, G.M. and S.L. Berger ( 1987) Methods Enzymol. 152:399-407; Kimmel, 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 (PE
Biosystems, Foster City CA), thermostable T7 polymerase (Amersham Pharmacia Biotech, Piscataway NJ), or combinations of polymerases and proofreading exonucleases such as those found in the ELONGASE amplification system (Life Technologies, Gaithersburg MD).
Preferably, sequence preparation 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 (PE Biosystems). Sequencing is then carried out using either the ABI 373 or 377 DNA sequencing system (PE Biosystems), the MEGABACE 1000 DNA
sequencing system (Molecular Dynamics, Sunnyvale CA), 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 Biolow and Biotechnolo~y, Wiley VCH, New York NY, pp.
856-853.) The nucleic acid sequences encoding TPPT 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 ligations 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 unknown 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, Palo Alto CA) to walk genomic DNA. This procedure avoids the need to screen libraries and is useful in finding intron/exon 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, PE 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 TPPT may be cloned in recombinant DNA molecules that direct expression of TPPT, 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 TPPT.
The nucleotide sequences of the present invention can be engineered using methods generally known in the art in order to alter TPPT-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 Number 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 TPPT, 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 TPPT 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, TPPT 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 may be achieved using the ABI 431A peptide synthesizer (PE Biosystems). Additionally, the amino acid sequence of TPPT, 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 TPPT, the nucleotide sequences encoding TPPT 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 TPPT. Such elements may vary in their strength and specificity.
Specific initiation signals may also be used to achieve more efficient translation of sequences encoding TPPT. Such signals include the ATG initiation codon and adjacent sequences, e.g. the Kozak sequence. In cases where sequences encoding TPPT 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 coding 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 TPPT and appropriate transcriptional and translational control elements. These methods include in vitro recombinant DNA techniques, synthetic techniques, and in vivo genetic recombination. (See, e.g., Sambrook, J. et al. (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 BioloQV, 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 TPPT. 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, sera; Van Heeke, G.
and S.M. Schuster (1989) J. Biol. Chem. 264:5503-5509; Bitter, G.A. et al. (1987) Methods Enzymol. 153:516-544;
Scorer, C.A. et al. (1994) Bio/Technology 12:181-184; 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;
Takamatsu, N.
(1987) EMBO J. 6:307-311; Coruzzi, G. et al. (1984) EMBO J. 3:1671-1680;
Brogue, R. et al. (1984) Science 224:838-843; Winter, J. et al. (1991) Results Probl. Cell Differ.
17:85-105; 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.
Immunol. 31(3):219-226;
and Verma, LM. 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 TPPT. For example, routine cloning, subcloning, and propagation of polynucleotide sequences encoding TPPT can be achieved using a multifunctional E. coli vector such as PBLUESCRIPT (Stratagene, La Jolla CA) or PSPORT1 plasmid (Life Technologies). Ligation of sequences encoding TPPT 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 TPPT are needed, e.g. for the production of antibodies, vectors which direct high level expression of TPPT may be used.
For example, vectors containing the strong, inducible TS or T7 bacteriophage promoter may be used.
Yeast expression systems may be used for production of TPPT. A number of vectors containing constitutive or inducible promoters, such as alpha factor, alcohol oxidase, and PGH
promoters, may be used in the yeast Saccharomvces cerevisiae or Pichia 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, supra; and Scorer, supra.) Plant systems may also be used for expression of TPPT. Transcription of sequences encoding TPPT may be driven 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, supra; Broglie, supra; and Winter, supra.) 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 Technolo~y ( 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 an expression vector, sequences encoding TPPT
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 TPPT 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 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 TPPT in cell lines is preferred. For example, sequences encoding TPPT 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 phosphoribosyltransferase genes, for use in tk and apr cells, respectively.
(See, e.g., Wigler, M. et al.
( 1977) Cell 11:223-232; Lowy, I. et al. ( 1980) Cell 22:817-823.) Also, antimetabolite, antibiotic, or herbicide resistance can be used as the basis for selection. For example, dhfr confers resistance to methotrexate; neo confers resistance to the aminoglycosides neomycin and G-418; and als and pat confer resistance to chlorsulfuron and phosphinotricin acetyltransferase, 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., Hartman, 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), B glucuronidase and its substrate B-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 TPPT is inserted within a marker gene sequence, transformed cells containing sequences encoding TPPT can be identified by the absence of marker gene function. Alternatively, a marker gene can be placed in tandem with a sequence encoding TPPT 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 TPPT
and that express TPPT 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.
Immunological methods for detecting and measuring the expression of TPPT 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 TPPT 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. (1990) Seroloeical Methods, a Laboratory Manual, APS
Press, St. Paul MN, Sect. IV; Coligan, J.E. et al. (1997) Current Protocols in ImmunoloQV, 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 TPPT
include oligolabeling, nick translation, end-labeling, or PCR amplification using a labeled nucleotide.
Alternatively, the sequences encoding TPPT, or any fragments thereof, may be cloned into a vector for the production of an mRNA probe. Such vectors are known in the art, are commercially available, and may be used to synthesize RNA probes in vitro 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 Pharmacia Biotech, 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 TPPT 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 TPPT may be designed to contain signal sequences which direct secretion of TPPT through a prokaryotic or eukaryotic cell membrane.
In 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, HEK293, and WI38) are available from the American Type Culture 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 TPPT 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 TPPT protein containing a heterologous moiety that can be recognized by a commercially available antibody may facilitate the screening of peptide libraries for inhibitors of TPPT 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 immunoaffinity purification 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 proteolytic cleavage site located between the TPPT encoding sequence and the heterologous protein sequence, so that TPPT may be cleaved away from the heterologous moiety following purification.
Methods for fusion protein expression and purification are discussed in Ausubel ( 1995, supra, 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 TPPT 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.
TPPT of the present invention or fragments thereof may be used to screen for compounds that specifically bind to TPPT. At least one and up to a plurality of test compounds may be screened for specific binding to TPPT. Examples of test compounds include antibodies, oligonucleotides, proteins (e.g., receptors), or small molecules.
In one embodiment, the compound thus identified is closely related to the natural ligand of TPPT, e.g., a ligand or fragment thereof, a natural substrate, a structural or functional mimetic, or a natural binding partner. (See, Coligan, J.E. et al. ( 1991 ) Current Protocols in Immunoloay 1 (2):
Chapter S.) Similarly, the compound can be closely related to the natural receptor to which TPPT
binds, or to at least a fragment of the receptor, e.g., the ligand binding site. In either case, the compound can be rationally designed using known techniques. In one embodiment, screening for these compounds involves producing appropriate cells which express TPPT, either as a secreted protein or on the cell membrane. Preferred cells include cells from mammals, yeast, Drosophila, or E.
coli. Cells expressing TPPT or cell membrane fractions which contain TPPT are then contacted with a test compound and binding, stimulation, or inhibition of activity of either TPPT 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 TPPT, either in solution or affixed to a solid support, and detecting the binding of TPPT 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, or natural product mixtures, and the test compounds) may be free in solution or affixed to a solid support.
TPPT of the present invention or fragments thereof may be used to screen for compounds that modulate the activity of TPPT. Such compounds may include agonists, antagonists, or partial or inverse agonists. In one embodiment, an assay is performed under conditions permissive for TPPT
activity, wherein TPPT is combined with at least one test compound, and the activity of TPPT in the presence of a test compound is compared with the activity of TPPT in the absence of the test compound. A change in the activity of TPPT in the presence of the test compound is indicative of a compound that modulates the activity of TPPT. Alternatively, a test compound is combined with an in vitro or cell-free system comprising TPPT under conditions suitable for TPPT activity, and the assay is performed. In either of these assays, a test compound which modulates the activity of TPPT
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 TPPT or their mammalian homologs may be "knocked out" in an 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 phosphotransferase 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) Clin. 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 C57BL/6 mouse strain. The blastocysts are surgically transferred to pseudopregnant 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 TPPT 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 TPPT can also be used to create "knockin" humanized animals (pigs) or transgenic animals (mice or rats) to model human disease. With knockin technology, a region of a polynucleotide encoding TPPT 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 TPPT, e.g., by secreting TPPT 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 TPPT and human transport proteins. In addition, the expression of TPPT is closely associated with neurological, cardiovascular, reproductive, gastrointestinal, and hematopoietic/immune tissues, and inflammation, cell proliferation, and cancer. Therefore, TPPT
appears to play a role in transport, metabolic, neurological, cardiovascular, reproductive, and immune disorders, and cell proliferative disorders including cancer. In the treatment of disorders associated with increased TPPT expression or activity, it is desirable to decrease the expression or activity of TPPT. In the treatment of disorders associated with decreased TPPT expression or activity, it is desirable to increase the expression or activity of TPPT.
Therefore, in one embodiment, TPPT 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 TPPT.
Examples of such disorders include, but are not limited to, a transport disorder such as akinesia, amyotrophic lateral sclerosis, ataxia telangiectasia, cystic fibrosis, Becker's muscular dystrophy, Bell's palsy, Charcot-Marie Tooth disease, diabetes mellitus, diabetes insipidus, diabetic neuropathy, Duchenne muscular dystrophy, hyperkalemic periodic paralysis, normokalemic periodic paralysis, Parkinson's disease, malignant hyperthermia, multidrug resistance, myasthenia gravis, myotonic dystrophy, catatonia, tardive dyskinesia, dystonias, peripheral neuropathy, cerebral neoplasms, prostate cancer; cardiac disorders associated with transport, e.g., angina, bradyarrythmia, tachyarrythmia, hypertension, Long QT syndrome, myocarditis, cardiomyopathy, nemaline myopathy, centronuclear myopathy, lipid myopathy, mitochondria) myopathy, thyrotoxic myopathy, ethanol myopathy, dermatomyositis, inclusion body myositis, infectious myositis, polymyositis;
neurological disorders associated with transport, e.g., Alzheimer's disease, amnesia, bipolar disorder, dementia, depression, epilepsy, Tourette's disorder, paranoid psychoses, and schizophrenia; and other disorders associated with transport, e.g., neurofibromatosis, postherpetic neuralgia, trigeminal neuropathy, sarcoidosis, sickle cell anemia, Wilson's disease, cataracts, infertility, pulmonary artery stenosis, sensorineural autosomal deafness, hyperglycemia, hypoglycemia, Grave's disease, goiter, Cushing's disease, Addison's disease, glucose-galactose malabsorption syndrome, hypercholesterolemia, adrenoleukodystrophy, Zellweger syndrome, Menkes disease, occipital horn syndrome, von Gierke disease, cystinuria, iminoglycinuria, Hartup disease, and Fanconi disease; a metabolic disorder such as Addison's disease, cerebrotendinous xanthomatosis, congenital adrenal hyperplasia, coumarin resistance, cystic fibrosis, diabetes, fatty hepatocirrhosis, fructose-1,6-diphosphatase deficiency, galactosemia, goiter, glucagonoma, glycogen storage diseases, hereditary fructose intolerance, hyperadrenalism, hypoadrenalism, hyperparathyroidism, hypoparathyroidism, hypercholesterolemia, hyperthyroidism, hypoglycemia, hypothyroidism, hyperlipidemia, hyperlipemia, lipid myopathies, lipodystrophies, lysosomal storage diseases, mannosidosis, neuraminidase deficiency, obesity, pentosuria phenylketonuria, and pseudovitamin D-deficiency rickets; 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, 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; a cardiovascular disorder such as arteriovenous fistula, atherosclerosis, hypertension, vasculitis, Raynaud's disease, aneurysms, arterial dissections, varicose veins, thrombophlebitis and phlebothrombosis, vascular tumors, and complications of thrombolysis, balloon angioplasty, vascular replacement, and coronary artery bypass graft surgery, congestive heart failure, ischemic heart disease, angina pectoris, myocardial infarction, hypertensive heart disease, degenerative valvular heart disease, calcific aortic valve stenosis, congenitally bicuspid aortic valve, mitral annular calcification, mitral valve prolapse, rheumatic fever and rheumatic heart disease, infective endocarditis, nonbacterial thrombotic endocarditis, endocarditis of systemic lupus erythematosus, carcinoid heart disease, cardiomyopathy, myocarditis, pericarditis, neoplastic heart disease, congenital heart disease, and complications of cardiac transplantation, congenital lung anomalies, atelectasis, pulmonary congestion and edema, pulmonary embolism, pulmonary hemorrhage, pulmonary infarction, pulmonary hypertension, vascular sclerosis, obstructive pulmonary disease, restrictive pulmonary disease, chronic obstructive pulmonary disease, emphysema, chronic bronchitis, bronchial asthma, bronchiectasis, bacterial pneumonia, viral and mycoplasmal pneumonia, lung abscess, pulmonary tuberculosis, diffuse interstitial diseases, pneumoconioses, sarcoidosis, idiopathic pulmonary fibrosis, desquamative interstitial pneumonitis, hypersensitivity pneumonitis, pulmonary eosinophilia bronchiolitis obliterans-organizing pneumonia, diffuse pulmonary hemorrhage syndromes, Goodpasture's syndromes, idiopathic pulmonary hemosiderosis, pulmonary involvement in collagen-vascular disorders, pulmonary alveolar proteinosis, lung tumors, inflammatory and noninflammatory pleural effusions, pneumothorax, pleural tumors, drug-induced lung disease, radiation-induced lung disease, and complications of lung transplantation; a reproductive disorder such as a disorder of prolactin production, infertility, including tubal disease, ovulatory defects, and endometriosis, a disruption of the estrous cycle, a disruption of the menstrual cycle, polycystic ovary syndrome, ovarian hyperstimulation syndrome, an endometrial or ovarian tumor, a uterine fibroid, autoimmune disorders, an ectopic pregnancy, and teratogenesis; cancer of the breast, fibrocystic breast disease, and galactorrhea; a disruption of spermatogenesis, abnormal sperm physiology, cancer of the testis, cancer of the prostate, benign prostatic hyperplasia, prostatitis, Peyronie's disease, impotence, carcinoma of the male breast, and gynecomastia;
an immune disorder such as inflammation, actinic keratosis, acquired immunodeficiency syndrome (AIDS), Addison's disease, adult respiratory distress syndrome, allergies, ankylosing spondylitis, amyloidosis, anemia, arteriosclerosis, asthma, atherosclerosis, autoimmune hemolytic anemia, autoimmune thyroiditis, autoimmune polyendocrinopathy-candidiasis-ectodermal dystrophy (APECED), bronchitis, bursitis, cholecystitis, cirrhosis, 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, paroxysmal nocturnal hemoglobinuria, hepatitis, hypereosinophilia, irritable bowel syndrome, mixed connective tissue disease (MCTD), multiple sclerosis, myasthenia gravis, myocardial or pericardial inflammation, myelofibrosis, osteoarthritis, osteoporosis, pancreatitis, polycythemia vera, polymyositis, psoriasis, Reiter's syndrome, rheumatoid arthritis, scleroderma, Sjogren's syndrome, systemic anaphylaxis, systemic lupus erythematosus, systemic sclerosis, primary thrombocythemia, thrombocytopenic purpura, ulcerative colitis, uveitis, Werner syndrome, complications of cancer, hemodialysis, and extracorporeal circulation, viral, bacterial, fungal, parasitic, protozoal, and helminthic infections, trauma, and hematopoietic cancer including lymphoma, leukemia, and myeloma; and 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.
In another embodiment, a vector capable of expressing TPPT 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 TPPT including, but not limited to, those described above.
In a further embodiment, a pharmaceutical composition comprising a substantially purified TPPT 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 TPPT
including, but not limited to, those provided above.
In still another embodiment, an agonist which modulates the activity of TPPT
may be administered to a subject to treat or prevent a disorder associated with decreased expression or activity of TPPT including, but not limited to, those listed above.
In a further embodiment, an antagonist of TPPT may be administered to a subject to treat or prevent a disorder associated with increased expression or activity of TPPT.
Examples of such disorders include, but are not limited to, those transport, metabolic, neurological, cardiovascular, reproductive, and immune disorders, and cell proliferative disorders including cancer described above. In one aspect, an antibody which specifically binds TPPT 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 TPPT.
In an additional embodiment, a vector expressing the complement of the polynucleotide encoding TPPT may be administered to a subject to treat or prevent a disorder associated with increased expression or activity of TPPT 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 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 reducing the potential for adverse side effects.
An antagonist of TPPT may be produced using methods which are generally known in the art.
In particular, purified TPPT may be used to produce antibodies or to screen libraries of pharmaceutical agents to identify those which specifically bind TPPT.
Antibodies to TPPT 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.
For the production of antibodies, various hosts including goats, rabbits, rats, mice, humans, and others may be immunized by injection with TPPT 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 Corvnebacterium~arvum are especially preferable.
It is preferred that the oligopeptides, peptides, or fragments used to induce antibodies to TPPT
have an 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 oligopeptides, peptides, or fragments are identical to a portion of the amino acid sequence of the natural protein.
Short stretches of TPPT
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 TPPT 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.
Immunol. 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 TPPT-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 TPPT 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 TPPT and its specific antibody. A two-site, monoclonal-based immunoassay utilizing monoclonal antibodies reactive to two non-interfering TPPT epitopes is generally used, but a competitive binding assay may also be employed (Pound, su ra .
Various methods such as Scatchard analysis in conjunction with radioimmunoassay techniques may be used to assess the affinity of antibodies for TPPT. Affinity is expressed as an association constant, Ka, which is defined as the molar concentration of TPPT-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 TPPT epitopes, represents the average affinity, or avidity, of the antibodies for TPPT. The Ka determined for a preparation of monoclonal antibodies, which are monospecific for a particular TPPT epitope, represents a true measure of affinity. High-affinity antibody preparations with Ka ranging from about 109 to 10'z L/mole are preferred for use in immunoassays in which the TPPT-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 TPPT, 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 York 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 mg specific antibody/ml, preferably 5-10 mg specific antibody/ml, is generally employed in procedures requiring precipitation of TPPT-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 TPPT, 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 TPPT. 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 TPPT. (See, e.g., Agrawal, S., ed. ( 1996) Antisense Therapeutics, Humana Press Inc., Totawa 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 Clin. Immunol. 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 Moms, M.C. et al. ( 1997) Nucleic Acids Res.
25(14):2730-2736.) In another embodiment of the invention, polynucleotides encoding TPPT 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 (SCID)-X 1 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 VIII or Factor IX
deficiencies (Crystal, R.G. (1995) Science 270:404-410; Verma, LM. and Somia, N. (1997) Nature 389:239-242)), (ii) express a conditionally lethal gene product (e.g., in the case of cancers which result from unregulated 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-I 1399), hepatitis B or C virus (HBV, HCV); fungal parasites, such as Candida albicans and Paracoccidioides brasiliensis; and protozoan parasites such as Plasmodium falciparum and Trypanosoma cruzi). In the case where a genetic deficiency in TPPT expression or regulation causes disease, the expression of TPPT from an appropriate population of transduced 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 TPPT are treated by constructing mammalian expression vectors encoding TPPT
and introducing these vectors by mechanical means into TPPT-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 transfection, (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, J-L. and H.
Recipon (1998) Curr.
Opin. Biotechnol. 9:445-450).
Expression vectors that may be effective for the expression of TPPT include, but are not limited to, the PCDNA 3.1, EPITAG, PRCCMV2, PREP, PVAX vectors (Invitrogen, Carlsbad CA), PCMV-SCRIPT, PCMV-TAG, PEGSH/PERV (Stratagene, La Jolla CA), and PTET-OFF, PTET-ON, PTRE2, PTRE2-LUC, PTK-HYG (Clontech, Palo Alto CA). TPPT may be expressed using (i) a constitutively active promoter, (e.g., from cytomegalovirus (CMV), Rous sarcoma virus (RSV), SV40 virus, thymidine kinase (TK), or (3-actin genes), (ii) an inducible promoter (e.g., the tetracycline-regulated promoter (Gossen, M. and H. Bujard (1992) Proc. Natl.
Acad. Sci. USA
89:5547-5551; Gossen, M. et al. ( 1995) Science 268:1766-1769; Rossi, F.M.V.
and H.M. Blau ( 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 FK506/rapamycin inducible promoter; or the RU486/mifepristone inducible promoter (Rossi, F.M.V.
and H.M. Blau, su ra ), or (iii) a tissue-specific promoter or the native promoter of the endogenous gene encoding TPPT from a normal individual.
Commercially available liposome transformation kits (e.g., the PERFECT LIPID
TRANSFECTION KIT, available from 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. In the alternative, transformation is performed using the calcium phosphate method (Graham, F.L. and A.J. Eb ( 1973) Virology 52:456-467), or 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 TPPT expression are treated by constructing a retrovirus vector consisting of (i) the polynucleotide encoding TPPT 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 (Stratagene) and are based on published data (Riviere, I. et al. ( 1995) Proc.
Natl. Acad. 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 cells or a promiscuous envelope protein such as VSVg (Armentano, D. et al.
(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) J. 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 Number 5,910,434 to Rigg ("Method for obtaining retrovirus packaging cell lines producing high transducing efficiency retroviral supernatant") IS 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 TPPT to cells which have one or more genetic abnormalities with respect to the expression of TPPT. 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 Number 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 18:389:239-242, both incorporated by reference herein.
In another alternative, a herpes-based, gene therapy delivery system is used to deliver polynucleotides encoding TPPT to target cells which have one or more genetic abnormalities with respect to the expression of TPPT. The use of herpes simplex virus (HSV)-based vectors may be especially valuable for introducing TPPT 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 Number 5,804,413 to DeLuca ("Herpes simplex virus strains for gene transfer"), which is hereby incorporated by reference. U.S. Patent Number 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 TPPT to target cells. The biology of the prototypic alphavirus, Semliki 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. Biotech. 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, 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 TPPT into the alphavirus genome in place of the capsid-coding region results in the production of a large number of TPPT-coding RNAs and the synthesis of high levels of TPPT 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 (BHK-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 will allow the introduction of TPPT 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 Immunolo i~ c Approaches, Futura Publishing, Mt. Kisco NY, pp.
163-177.) 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 TPPT.
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 IS 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 oligonucleotides such as solid phase phosphoramidite chemical synthesis.
Alternatively, RNA molecules may be generated by in vitro and in vivo transcription of DNA
sequences encoding TPPT. 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 TPPT. 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 TPPT
expression or activity, a compound which specifically inhibits expression of the polynucleotide encoding TPPT may be therapeutically useful, and in the treament of disorders associated with decreased TPPT expression or activity, a compound which specifically promotes expression of the polynucleotide encoding TPPT 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 polynucleotide encoding TPPT is exposed to at least one test compound thus obtained. The sample may comprise, for example, an intact or permeabilized cell, or an in vitro cell-free or reconstituted biochemical system. Alterations in the expression of a polynucleotide encoding TPPT 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 TPPT. 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 Schizosaccharomyces 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 pharmaceutical 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 pharmaceutical compositions may consist of TPPT, antibodies to TPPT, and mimetics, agonists, antagonists, or inhibitors of TPPT.
The pharmaceutical compositions utilized in this invention may be administered by any number of routes including, but not limited to, oral, intravenous, intramuscular, intra-arterial, intramedullary, intrathecal, intraventricular, pulmonary, transdermal, subcutaneous, intraperitoneal, intranasal, enteral, topical, sublingual, or rectal means.
Pharmaceutical 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 field 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.
Pharmaceutical 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 pharmaceutical compositions may be prepared for direct intracellular delivery of macromolecules comprising TPPT or fragments thereof. For example, liposome preparations containing a cell-impermeable macromolecule may promote cell fusion and intracellular delivery of the macromolecule. Alternatively, TPPT 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 TPPT
or fragments thereof, antibodies of TPPT, and agonists, antagonists or inhibitors of TPPT, 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.
Pharmaceutical 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 pharmaceutical compositions may be administered every 3 to 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 ~g to 100,000 fig, 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 TPPT may be used for the diagnosis of disorders characterized by expression of TPPT, or in assays to monitor patients being treated with TPPT or agonists, antagonists, or inhibitors of TPPT. Antibodies useful for diagnostic purposes may be prepared in the same manner as described above for therapeutics. Diagnostic assays for TPPT include methods which utilize the antibody and a label to detect TPPT
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 TPPT, including ELISAs, RIAs, and FACS, are known in the art and provide a basis for diagnosing altered or abnormal levels of TPPT expression. Normal or standard values for TPPT expression are established by combining body fluids or cell extracts taken from normal mammalian subjects, for example, human subjects, with antibody to TPPT under conditions suitable for complex formation. The amount of standard complex formation may be quantitated by various methods, such as photometric means. Quantities of TPPT
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 TPPT 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 TPPT may be correlated with disease. The diagnostic assay may be used to determine absence, presence, and excess expression of TPPT, and to monitor regulation of TPPT levels during therapeutic intervention.
In one aspect, hybridization with PCR probes which are capable of detecting polynucleotide sequences, including genomic sequences, encoding TPPT or closely related molecules may be used to identify nucleic acid sequences which encode TPPT. 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 TPPT, allelic variants, or related sequences.
Probes may also be used for the detection of related sequences, and may have at least 50%
sequence identity to any of the TPPT encoding sequences. The hybridization probes of the subject invention may be DNA or RNA and may be derived from the sequence of SEQ ID
N0:44-86 or from genomic sequences including promoters, enhaneers, and introns of the TPPT
gene.
Means for producing specific hybridization probes for DNAs encoding TPPT
include the cloning of polynucleotide sequences encoding TPPT or TPPT 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 3ZP or 35S, or by enzymatic labels, such as alkaline phosphatase coupled to the probe via avidin/biotin coupling systems, and the like.
Polynucleotide sequences encoding TPPT may be used for the diagnosis of disorders associated with expression of TPPT. Examples of such disorders include, but are not limited to, a transport disorder such as akinesia, amyotrophic lateral sclerosis, ataxia telangiectasia, cystic fibrosis, Becker's muscular dystrophy, Bell's palsy, Charcot-Marie Tooth disease, diabetes mellitus, diabetes insipidus, diabetic neuropathy, Duchenne muscular dystrophy, hyperkalemic periodic paralysis, S normokalemic periodic paralysis, Parkinson's disease, malignant hyperthermia, multidrug resistance, myasthenia gravis, myotonic dystrophy, catatonia, tardive dyskinesia, dystonias, peripheral neuropathy, cerebral neoplasms, prostate cancer; cardiac disorders associated with transport, e.g., angina, bradyarrythmia, tachyarrythmia, hypertension, Long QT syndrome, myocarditis, cardiomyopathy, nemaline myopathy, centronuclear myopathy, lipid myopathy, mitochondrial myopathy, thyrotoxic myopathy, ethanol myopathy, dermatomyositis, inclusion body myositis, infectious myositis, polymyositis; neurological disorders associated with transport, e.g., Alzheimer's disease, amnesia, bipolar disorder, dementia, depression, epilepsy, Tourette's disorder, paranoid psychoses, and schizophrenia; and other disorders associated with transport, e.g., neurofibromatosis, postherpetic neuralgia, trigeminal neuropathy, sarcoidosis, sickle cell anemia, Wilson's disease, cataracts, infertility, pulmonary artery stenosis, sensorineural autosomal deafness, hyperglycemia, hypoglycemia, Grave's disease, goiter, Cushing's disease, Addispn's disease, glucose-galactose malabsorption syndrome, hypercholesterolemia, adrenoleukodystrophy, Zellweger syndrome, Menkes disease, occipital horn syndrome, von Gierke disease, cystinuria, iminoglycinuria, Hartup disease, and Fanconi disease; a metabolic disorder such as Addison's disease, cerebrotendinous xanthomatosis, congenital adrenal hyperplasia, coumarin resistance, cystic fibrosis, diabetes, fatty hepatocirrhosis, fructose-1,6-diphosphatase deficiency, galactosemia, goiter, glucagonoma, glycogen storage diseases, hereditary fructose intolerance, hyperadrenalism, hypoadrenalism, hyperparathyroidism, hypoparathyroidism, hypercholesterolemia, hyperthyroidism, hypoglycemia, hypothyroidism, hyperlipidemia, hyperlipemia, lipid myopathies, lipodystrophies, lysosomal storage diseases, mannosidosis, neuraminidase deficiency, obesity, pentosuria phenylketonuria, and pseudovitamin D-deficiency rickets; 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, 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; a cardiovascular disorder such as arteriovenous fistula, atherosclerosis, hypertension, vasculitis, Raynaud's disease, aneurysms, arterial dissections, varicose veins, thrombophlebitis and phlebothrombosis, vascular tumors, and complications of thrombolysis, balloon angioplasty, vascular replacement, and coronary artery bypass graft surgery, congestive heart failure, ischemic heart disease, angina pectoris, myocardial infarction, hypertensive heart disease, degenerative valvular heart disease, calcific aortic valve stenosis, congenitally bicuspid aortic valve, mural annular calcification, mitral valve prolapse, rheumatic fever and rheumatic heart disease, infective endocarditis, nonbacterial thrombotic endocarditis, endocarditis of systemic lupus erythematosus, carcinoid heart disease, cardiomyopathy, myocarditis, pericarditis, neoplastic heart disease, congenital heart disease, and complications of cardiac transplantation, congenital lung anomalies, atelectasis, pulmonary congestion and edema, pulmonary embolism, pulmonary hemorrhage, pulmonary infarction, pulmonary hypertension, vascular sclerosis, obstructive pulmonary disease, restrictive pulmonary disease, chronic obstructive pulmonary disease, emphysema, chronic bronchitis, bronchial asthma, bronchiectasis, bacterial pneumonia, viral and mycoplasmal pneumonia, lung abscess, pulmonary tuberculosis, diffuse interstitial diseases, pneumoconioses, sarcoidosis, idiopathic pulmonary fibrosis, desquamative interstitial pneumonitis, hypersensitivity pneumonitis, pulmonary eosinophilia bronchiolitis obliterans-organizing pneumonia, diffuse pulmonary hemorrhage syndromes, Goodpasture's syndromes, idiopathic pulmonary hemosiderosis, pulmonary involvement in collagen-vascular disorders, pulmonary alveolar proteinosis, lung tumors, inflammatory and noninflammatory pleural effusions, pneumothorax, pleural tumors, drug-induced lung disease, radiation-induced lung disease, and complications of lung transplantation; a reproductive disorder such as a disorder of prolactin production, infertility, including tubal disease, ovulatory defects, and endometriosis, a disruption of the estrous cycle, a disruption of the menstrual cycle, polycystic ovary syndrome, ovarian hyperstimulation syndrome, an endometrial or ovarian tumor, a uterine fibroid, autoimmune disorders, an ectopic pregnancy, and teratogenesis; cancer of the breast, fibrocystic breast disease, and galactorrhea; a disruption of spermatogenesis, abnormal sperm physiology, cancer of the testis, cancer of the prostate, benign prostatic hyperplasia, prostatitis, Peyronie's disease, impotence, carcinoma of the male breast, and gynecomastia; an immune disorder such as inflammation, actinic keratosis, acquired immunodeficiency syndrome (AIDS), Addison's disease, adult respiratory distress syndrome, allergies, ankylosing spondylitis, amyloidosis, anemia, arteriosclerosis, asthma, atherosclerosis, autoimmune hemolytic anemia, autoimmune thyroiditis, autoimmune polyendocrinopathy-candidiasis-ectodermal dystrophy (APECED), bronchitis, bursitis, cholecystitis, cirrhosis, 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, paroxysmal nocturnal hemoglobinuria, hepatitis, hypereosinophilia, irritable bowel syndrome, mixed connective tissue disease (MCTD), multiple sclerosis, myasthenia gravis, myocardial or pericardial inflammation, myelofibrosis, osteoarthritis, osteoporosis, pancreatitis, polycythemia vera, polymyositis, psoriasis, Reiter's syndrome, rheumatoid arthritis, scleroderma, Sjogren's syndrome, systemic anaphylaxis, systemic lupus erythematosus, systemic sclerosis, primary thrombocythemia, thrombocytopenic purpura, ulcerative colitis, uveitis, Werner syndrome, complications of cancer, hemodialysis, and extracorporeal circulation, viral, bacterial, fungal, parasitic, protozoal, and helminthic infections, trauma, and hematopoietic cancer including lymphoma, leukemia, and myeloma; and 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. The polynucleotide sequences encoding TPPT 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 TPPT expression. Such qualitative or quantitative methods are well known in the art.
In a particular aspect, the nucleotide sequences encoding TPPT may be useful in assays that detect the presence of associated disorders, particularly those mentioned above. The nucleotide sequences encoding TPPT 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 TPPT 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 TPPT, 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 TPPT, 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 TPPT
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 TPPT, or a fragment of a polynucleotide complementary to the polynucleotide encoding TPPT, 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 TPPT 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 TPPT 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).
Methods which may also be used to quantify the expression of TPPT 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. Immunol. 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 quanritation.
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 in Seilhamer, J.J. et al., "Comparative Gene Transcript Analysis," U.S. Patent No. 5,840,484, incorporated herein by reference. 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 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 profile.
In another embodiment, antibodies specific for TPPT, or TPPT or fragments thereof 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.
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. Sci.
USA 93:10614-10619; Baldeschweiler et al. ( 1995) PCT application W095/251116;
Shalom D. et al.
( 1995) PCT application W095/35505; Heller, R.A. et al. ( 1997) Proc. 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 Microarra~ys: 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 TPPT
may be used to generate hybridization probes useful in mapping the naturally occurring genomic 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 mufti-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 Pl constructions, or single chromosome cDNA libraries. (See, e.g., Harrington, J.J. et al. (1997) Nat.
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, for example, which correlate the inheritance of a disease state with the inheritance of a particular chromosome region or restriction fragment length polymorphism (RFLP).
(See, e.g., Lander, E.S. and D. Botstein (1986) Proc. Natl. Acad. Sci. USA
83:7353-7357.) Fluorescent in situ hybridization (FISH) may be 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 (OMIM) World Wide Web site. Correlation between the location of the gene encoding TPPT on a physical map and a specific disorder, or a predisposition to a specific disorder, may help define the region of DNA 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 for extending genetic maps.
Often the placement of a gene on the chromosome 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 for 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 l 1q22-23, 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 translocation, inversion, etc., among normal, carrier, or affected individuals.
In another embodiment of the invention, TPPT, 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 in 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 TPPT 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., Geysen, et al. ( 1984) PCT
application W084/03564.) In this method, large numbers of different small test compounds are synthesized on a solid substrate. The test compounds are reacted with TPPT, or fragments thereof, and washed. Bound TPPT is then detected by methods well known in the art.
Purified TPPT 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 TPPT specifically compete with a test compound for binding TPPT. In this manner, antibodies can be used to detect the presence of any peptide which shares one or more antigenic determinants with TPPT.
In additional embodiments, the nucleotide sequences which encode TPPT 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 limited to, such 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 preferred specific embodiments are, therefore, to be construed as merely illustrative, and not limitative 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. 60/139,923, U.S. Ser. No. 60/148,177, U.S. Ser. No.
60/149,357, and U.S.
Ser. No. 60/162,287, are hereby expressly incorporated by reference.
EXAMPLES
I. Construction of cDNA Libraries RNA was purchased from Clontech or isolated from tissues described in Table 4.
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
(Life Technologies), a monophasic solution of phenol and guanidine isothiocyanate. The resulting lysates were centrifuged over CsCI 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 oligo 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 UNIZAP
vector system (Stratagene) or SUPERSCRIPT plasmid system (Life Technologies), 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 CL4B column chromatography (Amersham Pharmacia Biotech) 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), PSPORTI plasmid (Life Technologies), pcDNA2.1 plasmid (Invitrogen, Carlsbad CA), or pINCY plasmid (Incyte Genomics, Palo Alto CA).
Recombinant plasmids were transformed into competent E. coli cells including XLI-Blue, XL1-BIueMRF, or SOLR from Stratagene or DHSa, DH10B, or ElectroMAX DHIOB from Life Technologies.
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, 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 lysis 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 (PE 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 Pharmacia Biotech or supplied in ABI sequencing kits such as the ABI
PRISM BIGDYE Terminator cycle sequencing ready reaction kit (PE Biosystems).
Electrophoretic separation of cDNA sequencing reactions and detection of labeled polynucleotides were carried out using the MEGABACE 1000 DNA sequencing system (Molecular Dynamics); the ABI
PRISM 373 or 377 sequencing system (PE Biosystems) in conjunction with standard ABI
protocols and base calling software; or other sequence analysis systems known in the art. Reading frames within the cDNA
sequences were identified using standard methods (reviewed in Ausubel, 1997, supra, unit 7.7). Some of the cDNA sequences were selected for extension using the techniques disclosed in Example VI.
The polynucleotide sequences derived from cDNA sequencing were assembled and analyzed using a combination of software programs which utilize algorithms well known to those skilled in the art. Table 5 summarizes the tools, programs, and algorithms used and provides applicable descriptions, references, and threshold parameters. The first column of Table 5 shows the tools, IS 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, the greater the homology between two sequences). Sequences were analyzed using MACDNASIS PRO
software (Hitachi Software Engineering, South San Francisco CA) and LASERGENE
software (DNASTAR). Polynucleotide and polypeptide sequence alignments were generated using the default parameters specified by the clustal algorithm as incorporated into the MEGALIGN multisequence alignment program (DNASTAR), which also calculates the percent identity between aligned sequences.
The polynucleotide sequences were validated by removing vector, linker, and polyA
sequences and by masking ambiguous bases, using algorithms and programs based on BLAST, dynamic programing, and dinucleotide nearest neighbor analysis. The sequences were then queried against a selection of public databases such as the GenBank primate, rodent, mammalian, vertebrate, and eukaryote databases, and BLOCKS, PRINTS, DOMO, PRODOM, and PFAM to acquire annotation using programs based on BLAST, FASTA, and BLIMPS. The sequences were assembled into full length polynucleotide sequences using programs based on Phred, Phrap, and Consed, and 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 amino acid sequences, and these full length sequences were subsequently analyzed by querying against databases such as the GenBank databases (described above), SwissProt, BLOCKS, PRINTS, DOMO, PRODOM, Prosite, and Hidden Markov Model (HMM)-based protein family databases such as PFAM. HMM is a probabilistic approach which analyzes consensus primary structures of gene families. (See, e.g., Eddy, S.R. (1996) Curr. Opin. Struct. Biol. 6:361-365.) The programs described above for the assembly and analysis of full length polynucleotide and amino acid sequences were also used to identify polynucleotide sequence fragments from SEQ ID
N0:44-86. Fragments from about 20 to about 4000 nucleotides which are useful in hybridization and amplification technologies were described in The Invention section above.
IV. 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, supra, 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 minimum { 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 product 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.
The results of northern analyses are reported as a percentage distribution of libraries in which the transcript encoding TPPT occurred. Analysis involved the categorization of cDNA libraries by organ/tissue and disease. The organ/tissue categories included cardiovascular, dermatologic, developmental, endocrine, gastrointestinal, hematopoietic/immune, musculoskeletal, nervous, reproductive, and urologic. The disease/condition categories included cancer, inflammation, trauma, cell proliferation, neurological, and pooled. For each category, the number of libraries expressing the sequence of interest was counted and divided by the total number of libraries across all categories.
Percentage values of tissue-specific and disease- or condition-specific expression are reported in Table 3.
V. Chromosomal Mapping of TPPT Encoding Polynucleotides The cDNA sequences which were used to assemble SEQ ID N0:44-49 and SEQ ID
N0:54-86 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 )D N0:44-49 and SEQ ID N0:54-86 were assembled into clusters of contiguous and overlapping sequences using assembly algorithms such as Phrap (Table 5). 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.
The genetic map locations of SEQ ID N0:44, SEQ ID N0:48, SEQ ID N0:60, SEQ ID
N0:65, SEQ ID N0:69, SEQ ID N0:70, SEQ ID N0:71, SEQ ID N0:73, SEQ ID N0:76, SEQ ID
N0:80,and SEQ ID N0:83 are described in The Invention as ranges, or intervals, of human chromosomes. More than one map location is reported for SEQ ID N0:65, SEQ ID
N0:73, SEQ ID
N0:80, and SEQ ID N0:83, indicating that previously mapped sequences having similarity, but not complete identity, to SEQ ID N0:65, SEQ ID N0:73, SEQ ID N0:80, and SEQ ID
N0:83 were assembled into their respective clusters. 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. Diseases associated with the public and Incyte sequences located within the indicated intervals are also reported in the Invention section where applicable. 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.govlgenemap/), can be employed to determine if previously identified disease genes map within or in proximity to the intervals indicated above.
VI. Extension of TPPT Encoding Polynucleotides The full length nucleic acid sequences of SEQ ID N0:44-86 were 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, to initiate 3' extension of the known fragment. 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 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 nmol of each primer, reaction buffer containing Mg'-+, (NH4)ZS04, and (3-mercaptoethanol, Taq DNA polymerase (Amersham Pharmacia Biotech), ELONGASE enzyme (Life Technologies), and Pfu DNA polymerase (Stratagene), with the following parameters for primer pair PCI A and PCI B: Step 1: 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 p1 PICOGREEN
quantitation reagent (0.25% (v/v) PICOGREEN; 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 Fluoroskan II
(Labsystems Oy, Helsinki, Finland) to measure the fluorescence of the sample and to quantify the concentration of DNA. A 5 ~1 to 10 /c1 aliquot of the reaction mixture was analyzed by electrophoresis on a 1 % agarose mini-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 sonicated or sheared prior to religation into pUC 18 vector (Amersham Pharmacia Biotech). 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 religated using T4 ligase (New England Biolabs, Beverly MA) into pUC 18 vector (Amersham Pharmacia Biotech), 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 Garb liquid media.

The cells were lysed, and DNA was amplified by PCR using Taq DNA polymerase (Amersham Pharmacia Biotech) 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 times; Step 6: 72°C, 5 min; Step 7: storage at 4°C. DNA was S quantified by PICOGREEN reagent (Molecular Probes) as described above.
Samples with low DNA
recoveries were reamplified 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 Pharmacia Biotech) or the ABI
PRISM
BIGDYE Terminator cycle sequencing ready reaction kit (PE Biosystems).
In like manner, the polynucleotide sequences of SEQ )D N0:44-86 are used to obtain 5' regulatory sequences using the procedure above, along with oligonucleotides designed for such extension, and an appropriate genomic library.
VII. Labeling and Use of Individual Hybridization Probes Hybridization probes derived from SEQ ID N0:44-86 are employed to screen cDNAs, genomic 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 ,uCi of [y-3zP] adenosine triphosphate (Amersham Pharmacia Biotech), and T4 polynucleotide kinase (DuPont NEN, Boston MA). The labeled oligonucleotides are substantially purified using a SEPHADEX G-25 superfine size exclusion dextran bead column (Amersham Pharmacia Biotech). 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 0.5%
sodium dodecyl sulfate.
Hybridization patterns are visualized using autoradiography or an alternative imaging means and compared.
VIII. 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, supra), 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 skill 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 ( 1998) Nat. Biotechnol. 16:27-31.) Full length cDNAs, Expressed Sequence Tags (ESTs), or fragments or oligomers thereof 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 purified using the oligo-(dT) cellulose method. Each poly(A)+
RNA sample is reverse transcribed using MMLV reverse-transcriptase, 0.05 pg/pl oligo-(dT) primer (2lmer), 1X first strand buffer, 0.03 units/pl RNase inhibitor, 500 ~M dATP, 500 LiM dGTP, 500 L~M dTTP, 40 E.iM
dCTP, 40 E.~M dCTP-Cy3 (BDS) or dCTP-Cy5 (Amersham Pharmacia Biotech). 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 O.SM 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.
(CLONTECH), 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 in 14 ~1 SX SSC/0.2% SDS.
Microarrav 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 pg. Amplified array elements are then purified using SEPHACRYL-400 (Amersham Pharmacia Biotech).
Purified array elements are immobilized on polymer-coated glass slides. Glass microscope slides (Corning) are cleaned by ultrasound in 0.1 % 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 US
Patent No. 5,807,522, incorporated herein by reference. 1 p1 of the array element DNA, at an average concentration of 100 ng/pl, 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 p1 of sample mixture consisting of 0.2 pg each of Cy3 and Cy5 labeled cDNA synthesis products in SX 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 cm' 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 SX 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 first 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 Innova 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 positioned between the array and the photomultiplier tubes are used to filter the signals. The emission maxima 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).
IX. Complementary Polynucleotides Sequences complementary to the TPPT-encoding sequences, or any parts thereof, are used to detect, decrease, or inhibit expression of naturally occurring TPPT. 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 designed using OLIGO
4.06 software (National Biosciences) and the coding sequence of TPPT. To inhibit transcription, a complementary oligonucleotide is designed from the most unique 5' sequence and used to prevent promoter binding to the coding sequence. To inhibit translation, a complementary oligonucleotide is designed to prevent ribosomal binding to the TPPT-encoding transcript.
X. Expression of TPPT

Expression and purification of TPPT is achieved using bacterial or virus-based expression systems. For expression of TPPT 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 (tac) hybrid promoter and the TS or T7 bacteriophage promoter in conjunction with the lac operator regulatory element. Recombinant vectors are transformed into suitable bacterial hosts, e.g., BL21 (DE3).
Antibiotic resistant bacteria express TPPT upon induction with isopropyl beta-D-thiogalactopyranoside (IPTG). Expression of TPPT in eukaryotic cells is achieved by infecting insect or mammalian cell lines with recombinant Auto~aphica californica nuclear polyhedrosis virus (AcMNPV), commonly known as baculovirus. The nonessential polyhedrin gene of baculovirus is replaced with cDNA encoding TPPT by either homologous recombination or bacterial-mediated transposition involving transfer plasmid intermediates. Viral infectivity is maintained and the strong polyhedrin promoter drives high levels of cDNA transcription. Recombinant baculovirus is used to infect Spodoptera fru in~erda (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, TPPT 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 Pharmacia Biotech). Following purification, the GST moiety can be proteolytically cleaved from TPPT 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, supra, ch. 10 and 16). Purified TPPT obtained by these methods can be used directly in the assays shown in Examples XI and XV.
XI. Demonstration of TPPT Activity TPPT transport activity is assayed by measuring uptake of labeled substrates into Xenopus laevis oocytes. Oocytes at stages V and VI are injected with TPPT mRNA (10 ng per oocyte) and incubated for 3 days at 18°C in OR2 medium (82.SmM NaCI, 2.5 mM KCI, 1mM CaCI,, 1mM MgCI,, 1mM Na=HP04, 5 mM Hepes, 3.8 mM NaOH , SOpg/ml gentamycin, pH 7.8) to allow expression of TPPT. Oocytes are then transferred to standard uptake medium ( 100mM NaCI, 2 mM KCI, 1 mM
CaCI,, 1mM MgCI=, 10 mM Hepes/T'ris pH 7.5). Uptake of various substrates (e.g., amino acids, sugars, drugs, ions, and neurotransmitters) is initiated by adding labeled substrate (e.g. radiolabeled with jH, fluorescently labeled with rhodamine, etc.) to the oocytes. After incubating for 30 minutes, uptake is terminated by washing the oocytes three times in Na+-free medium, measuring the incorporated label, and comparing with controls. TPPT activity is proportional to the level of internalized labeled substrate.
XII. Functional Assays TPPT function is assessed by expressing the sequences encoding TPPT at physiologically elevated levels in mammalian cell culture systems. cDNA is subcloned into a mammalian expression vector containing a strong promoter that drives high levels of cDNA
expression. Vectors of choice include pCMV SPORT plasmid (Life Technologies) 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 ~g of an additional plasmid 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 fluorescein-conjugated Annexin V protein to the cell surface. Methods in flow cytometry are discussed in Ormerod, M.G. ( 1994) Flow Cvtometr_y, Oxford, New York NY.
The influence of TPPT on gene expression can be assessed using highly purified populations of cells transfected with sequences encoding TPPT 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 TPPT and other genes of interest can be analyzed by northern analysis or microarray techniques.

XIII. Production of TPPT Specific Antibodies TPPT 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 rabbits and to produce antibodies using standard protocols.
Alternatively, the TPPT 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, supra, ch. 11.) Typically, oligopeptides of about 15 residues in length are synthesized using an ABI 431A
peptide synthesizer (PE Biosystems) using FMOC chemistry and coupled to KLH
(Sigma-Aldrich, St.
Louis MO) by reaction with N-maleimidobenzoyl-N-hydroxysuccinimide ester (MBS) to increase immunogenicity. (See, e.g., Ausubel, 1995, supra.) Rabbits are immunized with the oligopeptide-KL.H complex in complete Freund's adjuvant. Resulting antisera are tested for antipeptide and anti-TPPT activity by, for example, binding the peptide or TPPT to a substrate, blocking with 1 % BSA, reacting with rabbit antisera, washing, and reacting with radio-iodinated goat anti-rabbit IgG.
XIV. Purification of Naturally Occurring TPPT Using Specific Antibodies Naturally occurring or recombinant TPPT is substantially purified by immunoaffmity chromatography using antibodies specific for TPPT. An immunoaffinity column is constructed by covalently coupling anti-TPPT antibody to an activated chromatographic resin, such as CNBr-activated SEPHAROSE (Amersham Pharmacia Biotech). After the coupling, the resin is blocked and washed according to the manufacturer's instructions.
Media containing TPPT are passed over the immunoaffinity column, and the column is washed under conditions that allow the preferential absorbance of TPPT (e.g., high ionic strength buffers in the presence of detergent). The column is eluted under conditions that disrupt antibody/TPPT 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 TPPT is collected.
XV. Identification of Molecules Which Interact with TPPT
TPPT, or biologically active fragments thereof, are labeled with '25I 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 mufti-well plate are incubated with the labeled TPPT, washed, and any wells with labeled TPPT complex are assayed. Data obtained using different concentrations of TPPT are used to calculate values for the number, affinity, and association of TPPT with the candidate molecules.
Alternatively, molecules interacting with TPPT 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).
TPPT 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 between the proteins encoded by two large libraries of genes (Nandabalan, K.
et al. (2000) U.S. Patent No. 6,057,101 ).
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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SEQUENCE LISTING
<110> INCYTE GENOMICS, INC.
LAL, Preeti YANG, Junming YUE, Henry HILLMAN, Jennifer L.
TANG, Y. Tom BANDMAN, Olga BURFORD, Neil BAUGHN, Mariah R.
AZIMZAI, Yalda LU, Dyung Aina M.
AU-YOUNG, Janice PATTERSON, Chandra <120> HUMAN TRANSPORT PROTEINS
<130> PF-0709 PCT
<140> To Be Assigned <141> Herewith <150> 60/139,923; 60/148,177; 60/149,357; 60/162,287 <151> 1999-06-17; 1999-08-10; 1999-08-18; 1999-10-28 <160> 86 <170> PERL Program <210> 1 <211> 623 <212> PRT
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 264114CD1 <400> 1 Met Ser Thr Gln Asp Glu Arg Gln Ile Asn Thr Glu Tyr Ala Val Ser Leu Leu Glu Gln Leu Lys Leu Phe Tyr Glu Gln Gln Leu Phe Thr Asp Ile Val Leu Ile Val Glu Gly Thr Glu Phe Pro Cys His Lys Met Val Leu Ala Thr Cys Ser Ser Tyr Phe Arg Ala Met Phe Met Ser Gly Leu Ser Glu Ser Lys Gln Thr His Val His Leu Arg Asn Val Asp Ala Ala Thr Leu Gln Ile Ile Ile Thr Tyr Ala Tyr Thr Gly Asn Leu Ala Met Asn Asp Ser Thr Val Glu Gln Leu Tyr Glu Thr Ala Cys Phe Leu Gln Val Glu Asp Val Leu Gln Arg Cys Arg Glu Tyr Leu Ile Lys Lys Ile Asn Ala Glu Asn Cys Val Arg Leu Leu Ser Phe Ala Asp Leu Phe Ser Cys Glu Glu Leu Lys Gln Ser Ala Lys Arg Met Val Glu His Lys Phe Thr Ala Val Tyr His Gln Asp Ala Phe Met Gln Leu Ser His Asp Leu Leu Ile Asp Ile Leu Ser Ser Asp Asn Leu Asn Val Glu Lys Glu Glu Thr Val Arg Glu Ala Ala Met Leu Trp Leu Glu Tyr Asn Thr Glu Ser Arg Ser Gln Tyr Leu Ser Ser Val Leu Ser Gln Ile Arg Ile Asp Ala Leu Ser Glu Val Thr Gln Arg Ala Trp Phe Gln Gly Leu Pro Pro Asn Asp Lys Ser Val Val Val Gln Gly Leu Tyr Lys Ser Met Pro Lys Phe Phe Lys Pro Arg Leu_Gly Met Thr Lys Glu Glu Met Met Ile Phe Ile Glu Ala Ser Ser Glu Asn Pro Cys Ser Leu Tyr Ser Ser Val Cys Tyr Ser Pro Gln Ala Glu Lys Val Tyr Lys Leu Cys Ser Pro Pro Ala Asp Leu His Lys Val Gly Thr Val Val Thr Pro Asp Asn Asp Ile Tyr Ile Ala Gly Gly Gln Val Pro Leu Lys Asn Thr Lys Thr Asn His Ser Lys Thr Ser Lys Leu Gln Thr Ala Phe Arg Thr Val Asn Cys Phe Tyr Trp Phe Asp Ala Gln Gln Asn Thr Trp Phe Pro Lys Thr Pro Met Leu Phe Val Arg Ile Lys Pro Ser Leu Val Cys Cys Glu Gly Tyr Ile Tyr Ala Ile Gly Gly Asp Ser Val Gly Gly Glu Leu Asn Arg Arg Thr Val Glu Arg Tyr Asp Thr Glu Lys Asp Glu Trp Thr Met Val Ser Pro Leu Pro Cys Ala Trp Gln Trp Ser Ala Ala Val Val Val His Asp Cys Ile Tyr Val Met Thr Leu Asn Leu Met Tyr Cys Tyr Phe Pro Arg Ser Asp Ser Trp Val Glu Met Ala Met Arg Gln Thr Ser Arg Ser Phe Ala Ser Ala Ala Ala Phe Gly Asp Lys Ile Phe Tyr Ile Gly Gly Leu His Ile Ala Thr Asn Ser Gly Ile Arg Leu Pro Ser Gly Thr Val Asp Gly Ser Ser Val Thr Val Glu Ile Tyr Asp Val Asn Lys Asn Glu Trp Lys Met Ala Ala Asn Ile Pro Ala Lys Arg Tyr Ser Asp Pro Cys Val Arg Ala Val Val Ile Ser Asn Ser Leu Cys Val Phe Met Arg Glu Thr His Leu Asn Glu Arg Ala Lys Tyr Val Thr Tyr Gln Tyr Asp Leu Glu Leu Asp Arg Trp Ser Leu Arg Gln His Ile Ser Glu Arg Val Leu Trp Asp Leu Gly Arg Asp Phe Arg Cys Thr Val Gly Lys Leu Tyr Pro Ser Cys Leu Glu Glu Ser Pro Trp Lys Pro Pro Thr Tyr Leu Phe Ser Thr Asp Gly Thr Glu Glu Phe Glu Leu Asp Gly Glu Met Val Ala Leu Pro Pro Val <210> 2 <211> 99 <212> PRT
<213> Homo sapiens <220>
<221> misc_feature <223> Incyte ID No: 1455669CD1 <400> 2 Met Ala Ala Pro Ala Glu Pro Cys Ala Gly Gln Gly Val Trp Asn Gln Thr Glu Pro Glu Pro Ala Ala Thr Ser Leu Leu Ser Leu Cys Phe Leu Arg Thr Ala Gly Val Trp Val Pro Pro Met Tyr Leu Trp Val Leu Gly Pro Ile Tyr Leu Leu Phe Ile His His His Gly Arg Gly Tyr Leu Arg Met Ser Pro Leu Phe Lys Ala Lys Met Val Ala Ala Ile Pro Gly Ser Leu Glu Pro Gly Asn Val Arg Gly Arg Gln Gly Thr Gly Trp Asn Leu Val Lys Ser <210> 3 <211> 374 <212> PRT
<213> Homo sapiens <220>
<221> misc_feature <223> Incyte ID No: 2084989CD1 <400> 3 Met Glu Ser Lys Met Gly Glu Leu Pro Leu Asp Ile Asn Ile Gln Glu Pro Arg Trp Asp Gln Ser Thr Phe Leu Gly Arg Ala Arg His Phe Phe Thr Val Thr Asp Pro Arg Asn Leu Leu Leu Ser Gly Ala Gln Leu Glu Ala Ser Arg Asn Ile Val Gln Asn Tyr Arg Ala Gly Val Val Thr Pro Gly Ile Thr Glu Asp Gln Leu Trp Arg Ala Lys Tyr Val Tyr Asp Ser Ala Phe His Pro Asp Thr Gly Glu Lys Val so a5 90 Val Leu Ile Gly Arg Met Ser Ala Gln Val Pro Met Asn Met Thr Ile Thr Gly Cys Met Leu Thr Phe Tyr Arg Lys Thr Pro Thr Val Val Phe Trp Gln Trp Val Asn Gln Ser Phe Asn Ala Ile Val Asn Tyr Ser Asn Arg Ser Gly Asp Thr Pro Ile Thr Val Arg Gln Leu Gly Thr Ala Tyr Val Ser Ala Thr Thr Gly Ala Val Ala Thr Ala Leu Gly Leu Lys Ser Leu Thr Lys His Leu Pro Pro Leu Val Gly Arg Phe Val Pro Phe Ala Ala Val Ala Ala Ala Asn Cys Ile Asn Ile Pro Leu Met Arg Gln Arg Glu Leu Gln Val Gly Ile Pro Val Ala Asp Glu Ala Gly Gln Arg Leu Gly Tyr Ser Val Thr Ala Ala Lys Gln Gly Ile Phe Gln Val Val Ile Ser Arg Ile Cys Met Ala Ile Pro Ala Met Ala Ile Pro Pro Leu Ile Met Asp Thr Leu Glu Lys Lys Asp Phe Leu Lys Val Gly Asp Cys Thr Ser Leu Val Leu Glu Trp Ala Met Ala Gly Arg Ser Asp Gln Ala Pro Thr Leu Ser Pro Ala Ser Pro Asp Ser Leu Arg Leu Ala Ser Pro Ser Pro Asp Pro Cys Thr Ala Ser Ser Thr Phe Val His Ser Ala Arg Met Asn Trp Ala Gly Val Lys Glu Leu Cys Arg Gly Arg Arg Arg Gly Gln Arg Lys Glu Thr Asn Phe Ile Ser Val Thr Pro Val Ala Ser Asp Thr Gln Lys Gly Thr Val Ile Val Met Leu Asp Leu Met Leu Ile Leu Leu Pro Pro Ser Ala Ser Ile Leu Arg Gly Thr His Gly <210> 4 <211> 271 <212> PRT
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 2501034CD1 <400> 4 Met Gly Asn Gly Gly Arg Ser Gly Leu Gln Gln Gly Lys Gly Asn Val Asp Gly Val Ala Ala Thr Pro Thr Ala Ala Ser Ala Ser Cys Gln Tyr Arg Cys Ile Glu Cys Asn Gln Glu Ala Lys Glu Leu Tyr Arg Asp Tyr Asn His Gly Val Leu Lys Ile Thr Ile Cys Lys Ser Cys Gln Lys Pro Val Asp Lys Tyr Ile Glu Tyr Asp Pro Val Ile Ile Leu Ile Asn Ala Ile Leu Cys Lys Ala Gln Ala Tyr Arg His Ile Leu Phe Asn Thr Gln Ile Asn Ile His Gly Lys Leu Cys Ile Phe Cys Leu Leu Cys Glu Ala Tyr Leu Arg Trp Trp Gln Leu Gln Asp Ser Asn Gln Asn Thr Ala Pro Asp Asp Leu Ile Arg Tyr Ala Lys Glu Trp Asp Phe Tyr Arg Met Phe Ala Ile Ala Ala Leu Glu Gln Thr Ala Tyr Phe Ile Gly Ile Phe Thr Phe Leu Trp Val Glu Arg Pro Met Thr Ala Lys Lys Lys Pro Asn Phe Ile Leu Leu Leu Lys Ala Leu Leu Leu Ser Ser Tyr Gly Lys Leu Leu Leu Ile Pro Ala Val Ile Trp Glu His Asp Tyr Thr Ser Val Cys Leu Lys Leu Ile Lys Val Phe Val Leu Thr Ser Asn Phe Gln Ala Ile Arg Val Thr Leu Asn Ile Asn Arg Lys Leu Ser Phe Leu Ala Val Leu Ser Gly Leu Leu Leu Glu Ser Ile Met Val Tyr Phe Phe Gln Ser Met Glu Trp Asp Val Gly Ser Asp Tyr Ala Ile Phe Lys Ser Gln Asp Phe <210> 5 <211> 323 <212> PRT
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 2745212CD1 <400> 5 Met Ala Pro Lys Gln Asp Pro Lys Pro Lys Phe Gln Glu Gly Glu Arg Val Leu Cys Phe His Gly Pro Leu Leu Tyr Glu Ala Lys Cys Val Lys Val Ala Ile Lys Asp Lys Gln Val Lys Tyr Phe Ile His Tyr Ser Gly Trp Asn Lys Asn Trp Asp Glu Trp Val Pro Glu Ser Arg Val Leu Lys Tyr Val Asp Thr Asn Leu Gln Lys Gln Arg Glu Leu Gln Lys Ala Asn Gln Glu Gln Tyr Ala Glu Gly Lys Met Arg Gly Ala Ala Pro Gly Lys Lys Thr Ser Gly Leu Gln Gln Lys Asn Val Glu Val Lys Thr Lys Lys Asn Lys Gln Lys Thr Pro Gly Asn Gly Asp Gly Gly Ser Thr Ser Glu Thr Pro Gln Pro Pro Arg Lys Lys Arg Ala Arg Val Asp Pro Thr Val Glu Asn Glu Glu Thr Phe Met Asn Arg Val Glu Val Lys Val Lys Ile Pro Glu Glu Leu Lys Pro Trp Leu Val Asp Asp Trp Asp Leu Ile Thr Arg Gln Lys Gln Leu Phe Tyr Leu Pro Ala Lys Lys Asn Val Asp Ser Ile Leu Glu Asp Tyr Ala Asn Tyr Lys Lys Ser Arg Gly Asn Thr Asp Asn Lys Glu Tyr Ala Val Asn Glu Val Val Ala Gly Ile Lys Glu Tyr Phe Asn Val Met Leu Gly Thr Gln Leu Leu Tyr Lys Phe Glu Arg Pro Gln Tyr Ala Glu Ile Leu Ala Asp His Pro Asp Ala Pro Met Ser Gln Val Tyr Gly Ala Pro His Leu Leu Arg Leu Phe Val Arg Ile Gly Ala Met Leu Ala Tyr Thr Pro Leu Asp Glu Lys Ser Leu Ala Leu Leu Leu Asn Tyr Leu His Asp Phe Leu Lys Tyr Leu Ala Lys Asn Ser Ala Thr Leu Phe Ser Ala Ser Asp Tyr Glu Val Ala Pro Pro Glu Tyr His Arg Lys Ala Val <210> 6 <211> 274 <212> PRT
<213> Homo sapiens <220>
<221> misc_feature <223> Incyte ID No: 4833111CD1 <400> 6 Met Asp Arg Pro Gly Phe Val Ala Ala Leu Val Ala Gly Gly Val Ala Gly Val Ser Val Asp Leu Ile Leu Phe Pro Leu Asp Thr Ile Lys Thr Arg Leu Gln Ser Pro Gln Gly Phe Ser Lys Ala Gly Gly Phe His Gly Ile Tyr Ala Gly Val Pro Ser Ala Ala Ile Gly Ser Phe Pro Asn Ala Ala Ala Phe Phe Ile Thr Tyr Glu Tyr Val Lys Trp Phe Leu His Ala Asp Ser Ser Ser Tyr Leu Thr Pro Met Lys His Met Leu Ala Ala Ser Ala Gly Glu Val Val Ala Cys Leu Ile Arg Val Pro Ser Glu Val Val Lys Gln Arg Ala Gln Val Ser Ala Ser Thr Arg Thr Phe Gln Ile Phe Ser Asn Ile Leu Tyr Glu Glu Gly Ile Gln Gly Leu Tyr Arg Gly Tyr Lys Ser Thr Val Leu Arg Glu Ile Pro Phe Ser Leu Val Gln Phe Pro Leu Trp Glu Ser Leu Lys Ala Leu Trp Ser Trp Arg Gln Asp His Val Val Asp Ser Trp Gln Ser Ala Val Cys Gly Ala Phe Ala Gly Gly Phe Ala Ala Ala Val Thr Thr Pro Leu Asp Val Ala Lys Thr Arg Ile Thr Leu Ala Lys Ala Gly Ser Ser Thr Ala Asp Gly Asn Val Leu Ser Val Leu His Gly Val Trp Arg Ser Gln Gly Leu Ala Gly Leu Phe Ala Gly Val Phe Pro Arg Met Ala Ala Ile Ser Leu Gly Gly Phe Ile Phe Leu Gly Ala Tyr Asp Arg Thr His Ser Leu Leu Leu Glu Val Gly Arg Lys Ser Pro <210> 7 <211> 291 <212> PRT
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 876677CD1 <400> 7 Met Asp Ser Arg Val Ser Ser Pro Glu Lys Gln Asp Lys Glu Asn Phe Val Gly Val Asn Asn Lys Arg Leu Gly Val Cys Gly Trp Ile Leu Phe Ser Leu Ser Phe Leu Leu Val Ile Ile Thr Phe Pro Ile Ser Ile Trp Met Cys Leu Lys Ile Ile Lys Glu Tyr Glu Arg Ala Val Val Phe Arg Leu Gly Arg Ile Gln Ala Asp Lys Ala Lys Gly Pro Gly Leu Ile Leu Val Leu Pro Cys Ile Asp Val Phe Val Lys Val Asp Leu Arg Thr Val Thr Cys Asn Ile Pro Pro Gln Glu Ile Leu Thr Arg Asp Ser Val Thr Thr Gln Val Asp Gly Val Val Tyr Tyr Arg Ile Tyr Ser Ala Val Ser Ala Val Ala Asn Val Asn Asp Val His Gln Ala Thr Phe Leu Leu Ala Gln Thr Thr Leu Arg Asn Val Leu Gly Thr Gln Thr Leu Ser Gln Ile Leu Ala Gly Arg Glu Glu Ile Ala His Ser Ile Gln Thr Leu Leu Asp Asp Ala Thr Glu Leu Trp Gly Ile Arg Val Ala Arg Val Glu Ile Lys Asp Val Arg Ile Pro Val Gln Leu Gln Arg Ser Met Ala Ala Glu Ala Glu Ala Thr Arg Glu Ala Arg Ala Lys Val Leu Ala Ala Glu Gly Glu Met Asn Ala Ser Lys Ser Leu Lys Ser Ala Ser Met Val Leu Ala Glu Ser Pro Ile Ala Leu Gln Leu Arg Tyr Leu Gln Thr Leu Ser Thr Val Ala Thr Glu Lys Asn Ser Thr Ile Val Phe Pro Leu Pro Met Asn Ile Leu Glu Gly Ile Gly Gly Val Ser Tyr Asp Asn His Lys Lys Leu Pro Asn Lys Ala <210> 8 <211> 381 <212> PRT
<213> Homo sapiens <220>
<221> misc_feature <223> Incyte ID No: 2326143CD1 <400> 8 Met Ser Arg His Glu Gly Val Ser Cys Asp Ala Cys Leu Lys Gly Asn Phe Arg Gly Arg Arg Tyr Lys Cys Leu Ile Cys Tyr Asp Tyr Asp Leu Cys Ala Ser Cys Tyr Glu Ser Gly Ala Thr Thr Thr Arg His Thr Thr Asp His Pro Met Gln Cys Ile Leu Thr Arg Val Asp Phe Asp Leu Tyr Tyr Gly Gly Glu Ala Phe Ser Val Glu Gln Pro Gln Ser Phe Thr Cys Pro Tyr Cys Gly Lys Met Gly Tyr Thr Glu Thr Ser Leu Gln Glu His Val Thr Ser Glu His Ala Glu Thr Ser Thr Glu Val Ile Cys Pro Ile Cys Ala Ala Leu Pro Gly Gly Asp Pro Asn His Val Thr Asp Asp Phe Ala Ala His Leu Thr Leu Glu His Arg Ala Pro Arg Asp Leu Asp Glu Ser Ser Gly Val Arg His Val Arg Arg Met Phe His Pro Gly Arg Gly Leu Gly Gly Pro Arg Ala Arg Arg Ser Asn Met His Phe Thr Ser Ser Ser Thr Gly Gly Leu Ser Ser Ser Gln Ser Ser Tyr Ser Pro Ser Asn Arg Glu Ala Met Asp Pro Ile Ala Glu Leu Leu Ser Gln Leu Ser Gly Val Arg Arg Ser Ala Gly Gly Gln Leu Asn Ser Ser Gly Pro Ser Ala Ser Gln Leu Gln Gln Leu Gln Met Gln Leu Gln Leu Glu Arg Gln His Ala Gln Ala Ala Arg Gln Gln Leu Glu Thr Ala Arg Asn Ala Thr Arg Arg Thr Asn Thr Ser Ser Val Thr Thr Thr Ile Thr Gln Ser Thr Ala Thr Thr Asn Ile Ala Asn Thr Glu Ser Ser Gln Gln Thr Leu Gln Asn Ser Gln Phe Leu Leu Thr Arg Leu Asn Asp Pro Lys Met Ser Glu Thr Glu Arg Gln Ser Met Glu Ser Glu Arg Ala Asp Arg Ser Leu Phe Val Gln Glu Leu Leu Leu Ser Thr Leu Val Arg Glu Glu Ser Ser Ser Ser Asp Glu Asp Asp Arg Gly Glu Met Ala Asp Phe Gly Ala Met Gly Cys Val Asp Ile Met Pro Leu Asp Val Ala Leu Glu Asn Leu Asn Leu Lys Glu Ser Asn Lys Gly Asn Glu Pro Pro Pro Pro Pro Leu <210> 9 <211> 190 <212> PRT
<213> Homo sapiens <220>
<221> misc_feature <223> Incyte ID No: 2786302CD1 <400> 9 Met Lys Tyr Gly Asn Glu Ile Met Asn Lys Asp Pro Val Phe Arg Ile Ser Pro Arg Ser Arg Glu Thr His Pro Asn Pro Glu Glu Pro Glu Glu Glu Asp Glu Asp Val Gln Ala Glu Arg Val Gln Ala Ala Asn Ala Leu Thr Ala Pro Asn Leu Glu Glu Glu Pro Val Ile Thr Ala Ser Cys Leu His Lys Glu Tyr Tyr Glu Thr Lys Lys Ser Cys Phe Ser Thr Arg Lys Lys Lys Ile Ala Ile Arg Asn Val Ser Phe Cys Val Lys Lys Gly Glu Val Leu Gly Leu Leu Gly His Asn Gly Ala Gly Lys Ser Thr Ser Ile Lys Met Ile Thr Gly Cys Thr Lys Pro Thr Ala Gly Val Val Val Leu Gln Gly Ser Arg Ala Ser Val Arg Gln Gln His Asp Asn Ser Leu Lys Phe Leu Gly Tyr Cys Pro Gln Glu Asn Ser Leu Trp Pro Lys Leu Thr Met Lys Glu His Leu Glu Leu Tyr Ala Ala Val Glu Arg Leu Gly Gln Lys Arg Cys Cys Ser Gln Tyr Phe Thr Ile Gly Gly Arg Ser <210> 10 <211> 297 <212> PRT
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 3735780CD1 <400> 10 Met Met Asp Ser Glu Ala His Glu Lys Arg Pro Pro Ile Leu Thr Ser Ser Lys Gln Asp Ile Ser Pro His Ile Thr Asn Val Gly Glu Met Lys His Tyr Leu Cys Gly Cys Cys Ala Ala Phe Asn Asn Val Ala Ile Thr Phe Pro Ile Gln Lys Val Leu Phe Arg Gln Gln Leu Tyr Gly Ile Lys Thr Arg Asp Ala Ile Leu Gln Leu Arg Arg Asp Gly Phe Arg Asn Leu Tyr Arg Gly Ile Leu Pro Pro Leu Met Gln Lys Thr Thr Thr Leu Ala Leu Met Phe Gly Leu Tyr Glu Asp Leu Ser Cys Leu Leu His Lys His Val Ser Ala Pro Glu Phe Ala Thr Ser Gly Val Ala Ala Val Leu Ala Gly Thr Thr Glu Ala Ile Phe Thr Pro Leu Glu Arg Val Gln Thr Leu Leu Gln Asp His Lys His His Asp Lys Phe Thr Asn Thr Tyr Gln Ala Phe Lys Ala Leu Lys Cys His Gly Ile Gly Glu Tyr Tyr Arg Gly Leu Val Pro Ile Leu Phe Arg Asn Gly Leu Ser Asn Val Leu Phe Phe Gly Leu Arg Gly Pro Ile Lys Glu His Leu Pro Thr Ala Thr Thr His Ser Ala His Leu Val Asn Asp Phe Ile Cys Gly Gly Leu Leu Gly Ala Met Leu Gly Phe Leu Phe Phe Pro Ile Asn Val Val Lys Thr Arg Ile Gln Ser Gln Ile Gly Gly Glu Phe Gln Ser Phe Pro Lys Val Phe Gln Lys Ile Trp Leu Glu Arg Asp Arg Lys Leu Ile Asn Leu Phe Arg Gly Ala His Leu Asn Tyr His Arg Ser Leu Ile Ser Trp Gly Ile Ile Asn Ala Thr Tyr Glu Phe Leu Leu Lys Val Ile <210> 11 <211> 89 <212> PRT
<213> Homo sapiens <220>
<221> misc_feature <223> Incyte ID No: 039026CD1 <400> 11 Met Ala Ala Gln Ile Pro Glu Ser Asp Gln Ile Lys Gln Phe Lys Glu Phe Leu Gly Thr Tyr Asn Lys Leu Thr Glu Thr Cys Phe Leu Asp Cys Val Lys Asp Phe Thr Thr Arg Glu Val Lys Pro Glu Glu Thr Thr Cys Ser Glu His Cys Leu Gln Lys Tyr Leu Lys Met Thr Gln Arg Ile Ser Met Arg Phe Gln Glu Tyr His Ile Gln Gln Asn Glu Ala Leu Ala Ala Lys Ala Gly Leu Leu Gly Gln Pro Arg <210> 12 <211> 115 <212> PRT
<213> Homo sapiens <220>
<221> misc_feature <223> Incyte ID No: 260607CD1 <400> 12 Met Ala Leu Ile Pro Ser Arg Val Trp Leu Pro Phe Ala Val Trp Val Val Asp Ser Ala Pro Val Arg Gly Leu Val Arg Arg Glu Pro Phe Leu Arg Thr Gly Ser Phe Ile Ala Leu Phe Tyr Phe Pro Pro Leu Leu Pro Val Leu Ile Asn Leu Phe Ser Phe Phe Leu Thr Pro Ser Phe Trp Arg Gln Leu Gly Ala Ile Leu Val Tyr Ala Ser Leu Leu Ala Glu Lys Thr Pro Phe Lys Thr Gln Arg Thr Leu Glu Gly Asp Ala Leu Val Gly Ser Val Ser Ile Phe Leu Cys Ala Lys Asp Arg Gln Thr Glu Ala Glu Arg Gly Cys Ser <210> 13 <211> 675 <212> PRT
<213> Homo sapiens <220>
<221> misc feature <223> Incyte ID No: 1429651CD1 <400> 13 Met Glu Ser Gly Thr Ser Ser Pro Gln Pro Pro Gln Leu Asp Pro Leu Asp Ala Phe Pro Gln Lys Gly Leu Glu Pro Gly Asp Ile Ala Val Leu Val Leu Tyr Phe Leu Phe Val Leu Ala Val Gly Leu Trp Ser Thr Val Lys Thr Lys Arg Asp Thr Val Lys Gly Tyr Phe Leu Ala Gly Gly Asp Met Val Trp Trp Pro Val Gly Ala Ser Leu Phe Ala Ser Asn Val Gly Ser Gly His Phe Ile Gly Leu Ala Gly Ser Gly Ala Ala Thr Gly Ile Ser Val Ser Ala Tyr Glu Leu Asn Gly Leu Phe Ser Val Leu Met Leu Ala Trp Ile Phe Leu Pro Ile Tyr Ile Ala Gly Gln Val Thr Thr Met Pro Glu Tyr Leu Arg Lys Arg Phe Gly Gly Ile Arg Ile Pro Ile Ile Leu Ala Val Leu Tyr Leu Phe Ile Tyr Ile Phe Thr Lys Ile Ser Val Asp Met Tyr Ala Gly Ala Ile Phe Ile Gln Gln Ser Leu His Leu Asp Leu Tyr Leu Ala Ile Val Gly Leu Leu Ala Ile Thr Ala Val Tyr Thr Val Ala Gly Gly Leu Ala Ala Val Ile Tyr Thr Asp Ala Leu Gln Thr Leu Ile Met Leu Ile Gly Ala Leu Thr Leu Met Gly Tyr Ser Phe Ala Ala Val Gly Gly Met Glu Gly Leu Lys Glu Lys Tyr Phe Leu Ala Leu Ala Ser Asn Arg Ser Glu Asn Ser Ser Cys Gly Leu Pro Arg Glu Asp Ala Phe His Ile Phe Arg Asp Pro Leu Thr Ser Asp Leu Pro Trp Pro Gly Val Leu Phe Gly Met Ser Ile Pro Ser Leu Trp Tyr Trp Cys Thr Asp Gln Val Ile Val Gln Arg Thr Leu Ala Ala Lys Asn Leu Ser His A1a Lys Gly Gly Ala Leu Met Ala Ala Tyr Leu Lys Val Leu Pro Leu Phe Ile Met Val Phe Pro Gly Met Val Ser Arg Ile Leu Phe Pro Asp Gln Val Ala Cys Ala Asp Pro Glu Ile Cys Gln Lys Ile Cys Ser Asn Pro Ser Gly Cys Ser Asp Ile Ala Tyr Pro Lys Leu Val Leu Glu Leu Leu Pro Thr Gly Leu Arg Gly Leu Met Met Ala Val Met Val Ala Ala Leu Met Ser Ser Leu Thr Ser Ile Phe Asn Ser Ala Ser Thr Ile Phe Thr Met Asp Leu Trp Asn His Leu Arg Pro Arg Ala Ser Glu Lys Glu Leu Met Ile Val Gly Arg Val Phe Val Leu Leu Leu Val Leu Val Ser Ile Leu Trp Ile Pro Val Val Gln Ala Ser Gln Gly Gly Gln Leu Phe Ile Tyr Ile Gln Ser Ile Ser Ser Tyr Leu Gln Pro Pro Val Ala Val Val Phe Ile Met Gly Cys Phe Trp Lys Arg Thr Asn Glu Lys Gly Ala Phe Trp Gly Leu Ile Ser Gly Leu Leu Leu Gly Leu Val Arg Leu Val Leu Asp Phe Ile Tyr Val Gln Pro Arg Cys Asp Gln Pro Asp Glu Arg Pro Val Leu Val Lys Ser Ile His Tyr Leu Tyr Phe Ser Met Ile Leu Ser Thr Val Thr Leu Ile Thr Val Ser Thr Val Ser Trp Phe Thr Glu Pro Pro Ser Lys Glu Met Val Ser His Leu Thr Trp Phe Thr Arg His Asp Pro Val Val Gln Lys Glu Gln Ala Pro Pro Ala Ala Pro Leu Ser Leu Thr Leu Ser Gln Asn Gly Met Pro Glu Ala Ser Ser Ser Ser Ser Val Gln Phe Glu Met Val Gln Glu Asn Thr Ser Lys Thr His Ser Cys Asp Met Thr Pro Lys Gln Ser Lys Val Val Lys Ala Ile Leu Trp Leu Cys Gly Ile Gln Glu Lys Gly Lys Glu Glu Leu Pro Ala Arg Ala Glu Ala Ile Ile Val Ser Leu Glu Glu Asn Pro Leu Val Lys Thr Leu Leu Asp Val Asn Leu Ile Phe Cys Val Ser Cys Ala Ile Phe Ile Trp Gly Tyr Phe Ala <210> 14 <211> 320 <212> PRT
<213> Homo sapiens <220>
<221> misc_feature <223> Incyte ID No: 2069971CD1 <400> 14 Met Tyr His Cys His Ser Gly Ser Lys Pro Thr Glu Lys Gly Ala Asn Glu Tyr Ala Tyr Ala Lys Trp Lys Leu Cys Ser Ala Ser Ala Ile Cys Phe Ile Phe Met Ile Ala Glu Val Val Gly Gly His Ile Ala Gly Ser Leu Ala Val Val Thr Asp Ala Ala His Leu Leu Ile Asp Leu Thr Ser Phe Leu Leu Ser Leu Phe Ser Leu Trp Leu Ser Ser Lys Pro Pro Ser Lys Arg Leu Thr Phe Gly Trp His Arg Ala Glu Ile Leu Gly Ala Leu Leu Ser Ile Leu Cys Ile Trp Val Val Thr Gly Val Leu Val Tyr Leu Ala Cys Glu Arg Leu Leu Tyr Pro Asp Tyr Gln Ile Gln Ala Thr Val Met Ile Ile Val Ser Ser Cys Ala Val Ala Ala Asn Ile Val Leu Thr Val Val Leu His Gln Arg Cys Leu Gly His Asn His Lys Glu Val Gln Ala Asn Ala Ser Val Arg Ala Ala Phe Val His Ala Leu Gly Asp Leu Phe Gln Ser Ile Ser Val Leu Ile Ser Ala Leu Ile Ile Tyr Phe Lys Pro Glu Tyr Lys Ile Ala Asp Pro Ile Cys Thr Phe Ile Phe Ser Ile Leu Val Leu Ala Ser Thr Ile Thr Ile Leu Lys Asp Phe Ser Ile Leu Leu Met Glu Gly Val Pro Lys Ser Leu Asn Tyr Ser Gly Val Lys Glu Leu Ile Leu Ala Val Asp Gly Val Leu Ser Val His Ser Leu His Ile Trp Ser Leu Thr Met Asn Gln Val Ile Leu Ser Ala His Val Ala Thr Ala Ala Ser Arg Asp Ser Gln Val Val Arg Arg Glu Ile Ala Lys Ala Leu Ser Lys Ser Phe Thr Met His Ser Leu Thr Ile Gln Met Glu Ser Pro Val Asp Gln Asp Pro Asp Cys Leu Phe Cys Glu Asp Pro Cys Asp <210> 15 <211> 462 <212> PRT
<213> Homo sapiens <220>
<221> misc_feature <223> Incyte ID No: 2329339CD1 <400> 15 Met Ala Glu Glu Gln Glu Phe Thr Gln Leu Cys Lys Leu Pro Ala Gln Pro Ser His Pro His Cys Val Asn Asn Thr Tyr Arg Ser Ala Gln His Ser Gln Ala Leu Leu Arg Gly Leu Leu Ala Leu Arg Asp Ser Gly Ile Leu Phe Asp Val Val Leu Val Val Glu Gly Arg His Ile Glu Ala His Arg Ile Leu Leu Ala Ala Ser Cys Asp Tyr Phe Arg Gly Met Phe Ala Gly Gly Leu Lys Glu Met Glu Gln Glu Glu Val Leu Ile His Gly Val Ser Tyr Asn Ala Met Cys Gln Ile Leu His Phe Ile Tyr Thr Ser Glu Leu Glu Leu Ser Leu Ser Asn Val Gln Glu Thr Leu Val Ala Ala Cys Gln Leu Gln Ile Pro Glu Ile Ile His Phe Cys Cys Asp Phe Leu Met Ser Trp Val Asp Glu Glu Asn Ile Leu Asp Val Tyr Arg Leu Ala Glu Leu Phe Asp Leu Ser Arg Leu Thr Glu Gln Leu Asp Thr Tyr Ile Leu Lys Asn Phe Val Ala Phe Ser Arg Thr Asp Lys Tyr Arg Gln Leu Pro Leu Glu Lys Val Tyr Ser Leu Leu Ser Ser Asn Arg Leu Glu Val Ser Cys Glu Thr Glu Val Tyr Glu Gly Ala Leu Leu Tyr His Tyr Ser Leu Glu Gln Val Gln Ala Asp Gln Ile Ser Leu His Glu Pro Pro Lys Leu Leu Glu Thr Val Arg Phe Pro Leu Met Glu Ala Glu Val Leu Gln Arg Leu His Asp Lys Leu Asp Pro Ser Pro Leu Arg Asp Thr Val Ala Ser Gly Leu Met Tyr His Arg Asn Glu Ser Leu Gln Pro Ser Leu Gln Ser Pro Gln Thr Glu Leu Arg Ser Asp Phe Gln Cys Val Val Gly Phe Gly Gly Ile His Ser Thr Pro Ser Thr Val Leu Ser Asp Gln Ala Lys Tyr Leu Asn Pro Leu Leu Gly Glu Trp Lys His Phe Thr Ala Ser Leu Ala Pro Arg Met Ser Asn Gln Gly Ile Ala Val Leu Asn Asn Phe Val Tyr Leu Ile Gly Gly Asp Asn Asn Val Gln Gly Phe Arg Ala Glu Ser Arg Cys Trp Arg Tyr Asp Pro Arg His Asn Arg Trp Phe Gln Ile Gln Ser Leu Gln Gln Glu His Ala Asp Leu Ser Val Cys Val Val Gly Arg Tyr Ile Tyr Ala Val Ala Gly Arg Asp Tyr His Asn Asp Leu Asn Ala Val Glu Arg Tyr Asp Pro Ala Thr Asn Ser Trp Ala Tyr Val Ala Pro Leu Lys Arg Glu Val Tyr Ala His Ala Gly Ala Thr Leu Glu Gly Lys Met Tyr Ile Thr Cys Gly Arg Lys Leu Ile Pro Phe Ser Glu Gly <210> 16 <211> 98 <212> PRT
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 2540219CD1 <400> 16 Met Arg Ala Cys Ala Val Trp Leu Ala Gly Gly Met Ala Gly Ala Ile Ser Trp Gly Thr Ala Thr Pro Met Asp Val Val Lys Ser Arg Leu Gln Ala Asp Gly Val Tyr Leu Asn Lys Tyr Lys Gly Val Leu Asp Cys Ile Ser Gln Ser Tyr Gln Lys Glu Gly Leu Lys Val Phe Phe Arg Gly Ile Thr Val Asn Ala Val Arg Gly Phe Pro Met Ser Ala Ala Met Phe Leu Gly Tyr Glu Leu Ser Leu Gln Ala Ile Arg Gly Asp His Ala Val Thr Ser Pro <210> 17 <211> 748 <212> PRT
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 2722462CD1 <400> 17 Met Asn Tyr Gln Glu Ala Ala Ile Tyr Leu Gln Glu Gly Glu Asn Asn Asp Lys Phe Phe Thr His Pro Lys Asp Ala Lys Ala Leu Ala Ala Tyr Leu Phe Ala His Asn His Leu Phe Tyr Leu Met Glu Leu Ala Thr Ala Leu Leu Leu Leu Leu Leu Ser Leu Cys Glu Ala Pro Ala Val Pro Ala Leu Arg Leu Gly Ile Tyr Val His Ala Thr Leu Glu Leu Phe Ala Leu Met Val Val Val Phe Glu Leu Cys Met Lys Leu Arg Trp Leu Gly Leu His Thr Phe Ile Arg His Lys Arg Thr Met Val Lys Thr Ser Val Leu Val Val Gln Phe Val Glu Ala Ile Val Val Leu Val Arg Gln Met Ser His Val Arg Val Thr Arg Ala Leu Arg Cys Ile Phe Leu Val Asp Cys Arg Tyr Cys Gly Gly Val Arg Arg Asn Leu Arg Gln Ile Phe Gln Ser Leu Pro Pro Phe Met Asp Ile Leu Leu Leu Leu Leu Phe Phe Met Ile Ile Phe Ala Ile Leu Gly Phe Tyr Leu Phe Ser Pro Asn Pro Ser Asp Pro Tyr Phe Ser Thr Leu Glu Asn Ser Ile Val Ser Leu Phe Val Leu Leu Thr Thr Ala Asn Phe Pro Asp Val Met Met Pro Ser Tyr Ser Arg Asn Pro Trp Ser Cys Val Phe Phe Ile Val Tyr Leu Ser Ile Glu Leu Tyr Phe Ile Met Asn Leu Leu Leu Ala Val Val Phe Asp Thr Phe Asn Asp Ile Glu Lys Arg Lys Phe Lys Ser Leu Leu Leu His Lys Arg Thr Ala Ile Gln His Ala Tyr Arg Leu Leu Ile Ser Gln Arg Arg Pro Ala Gly Ile Ser Tyr Arg Gln Phe Glu Gly Leu Met Arg Phe Tyr Lys Pro Arg Met Ser Ala Arg Glu Arg Tyr Leu Thr Phe Lys Ala Leu Asn Gln Asn Asn Thr Pro Leu Leu Ser Leu Lys Asp Phe Tyr Asp Ile Tyr Glu Val Ala Ala Leu Lys Trp Lys Ala Lys Lys Asn Arg Glu His Trp Phe Asp Glu Leu Pro Arg Thr Ala Leu Leu Ile Phe Lys Gly Ile Asn Ile Leu Val Lys Ser Lys Ala Phe Gln Tyr Phe Met Tyr Leu Val Val Ala Val Asn Gly Val Trp Ile Leu Val Glu Thr Phe Met Leu Lys Gly Gly Asn Phe Phe Ser Lys His Val Pro Trp Ser Tyr Leu Val Phe Leu Thr Ile Tyr Gly Val Glu Leu Phe Leu Lys Val Ala Gly Leu Gly Pro Val Glu Tyr Leu Ser Ser Gly Trp Asn Leu Phe Asp Phe Ser Val Thr Val Phe Ala Phe Leu Gly Leu Leu Ala Leu Ala Leu Asn Met Glu Pro Phe Tyr Phe Ile Val Val Leu Arg Pro Leu Gln Leu Leu Arg Leu Phe Lys Leu Lys Glu Arg Tyr Arg Asn Val Leu Asp Thr Met Phe Glu Leu Leu Pro Arg Met Ala Ser Leu Gly Leu Thr Leu Leu Ile Phe Tyr Tyr Ser Phe Ala Ile Val Gly Met Glu Phe Phe Cys Gly Ile Val Phe Pro Asn Cys Cys Asn Thr Ser Thr Val Ala Asp Ala Tyr Arg Trp Arg Asn His Thr Val Gly Asn Arg Thr Val Val Glu Glu Gly Tyr Tyr Tyr Leu Asn Asn Phe Asp Asn Ile Leu Asn Ser Phe Val Thr Leu Phe Glu Leu Thr Val Val Asn Asn Trp Tyr Ile Ile Met Glu Gly Val Thr Ser Gln Thr Ser His Trp Ser Arg Leu Tyr Phe Met Thr Phe Tyr Ile Val Thr Met Val Val Met Thr Ile Ile Val Ala Phe Ile Leu Glu Ala Phe Val Phe Arg Met Asn Tyr Ser Arg Lys Asn Gln Asp Ser Glu Val Asp Gly Gly Ile Thr Leu Glu Lys Glu Ile Ser Lys Glu Glu Leu Val Ala Val Leu Glu Leu Tyr Arg Glu Ala Arg Gly Ala Ser Ser Asp Val Thr Arg Leu Leu Glu Thr Leu Ser Gln Met Glu Arg Tyr Gln Gln His Ser Met Val Phe Leu Gly Arg Arg Ser Arg Thr Lys Ser Asp Leu Ser Leu Lys Met Tyr Gln Glu Glu Ile Gln Glu Trp Tyr Glu Glu His Ala Arg Glu Gln Glu Gln Gln Arg Gln Leu Ser Ser Ser Ala Ala Pro Ala Ala Gln Gln Pro Pro Gly Ser Arg Gln Arg Ser Gln Thr Val Thr <210> 18 <211> 507 <212> PRT
<213> Homo sapiens <220>
<221> misc_feature <223> Incyte ID No: 2739264CD1 <400> 18 Met Ala Phe Asn Phe Gly Ala Pro Ser Gly Thr Ser Gly Thr Ala Ala Ala Thr Ala Ala Pro Ala Gly Gly Phe Gly Gly Phe Gly Thr Thr Ser Thr Thr Ala Gly Ser Ala Phe Ser Phe Ser Ala Pro Thr Asn Thr Gly Thr Thr Gly Leu Phe Gly Gly Thr Gln Asn Lys Gly Phe Gly Phe Gly Thr Gly Phe Gly Thr Thr Thr Gly Thr Ser Thr Gly Leu Gly Thr Gly Leu Gly Thr Gly Leu Gly Phe Gly Gly Phe Asn Thr Gln Gln Gln Gln Gln Thr Thr Leu Gly Gly Leu Phe Ser Gln Pro Thr Gln Ala Pro Thr Gln Ser Asn Gln Leu Ile Asn Thr Ala Ser Ala Leu Ser Ala Pro Thr Leu Leu Gly Asp Glu Arg Asp Ala Ile Leu Ala Lys Trp Asn Gln Leu Gln Ala Phe Trp Gly Thr Gly Lys Gly Tyr Phe Asn Asn Asn Ile Pro Pro Val Glu Phe Thr Gln Glu Asn Pro Phe Cys Arg Phe Lys Ala Val Gly Tyr Ser Cys Met Pro Ser Asn Lys Asp Glu Asp Gly Leu Val Val Leu Val Phe Asn Lys Lys Glu Thr Glu Ile Arg Ser Gln Gln Gln Gln Leu Val Glu Ser Leu His Lys Val Leu Gly Gly Asn Gln Thr Leu Thr Val Asn Val Glu Gly Thr Lys Thr Leu Pro Asp Asp Gln Thr Glu Val Val Ile Tyr Val Val Glu Arg Ser Pro Asn Gly Thr Ser Arg Arg Val Pro Ala Thr Thr Leu Tyr Ala His Phe Glu Gln Ala Asn Ile Lys Thr Gln Leu Gln Gln Leu Gly Val Thr Leu Ser Met Thr Arg Thr Glu Leu Ser Pro Ala Gln Ile Lys G1n Leu Leu Gln Asn Pro Pro Ala Gly Val Asp Pro Ile Ile Trp Glu Gln Ala Lys Val Asp Asn Pro Asp Ser Glu Lys Leu Ile Pro Val Pro Met Val Gly Phe Met Thr Phe Tyr Ile Val Thr Met Val Val Met Thr Ile Ile V

Lys Glu Leu Leu Arg Arg Leu Lys Val Gln Asp Gln Met Thr Lys Gln His Gln Thr Arg Leu Asp Ile Ile Ser Glu Asp Ile Ser Glu Leu Gln Lys Asn Gln Thr Thr Ser Val Ala Lys Ile Ala Gln Tyr Lys Arg Lys Leu Met Asp Leu Ser His Arg Thr Leu Gln Val Leu Ile Lys Gln Glu Ile Gln Arg Lys Ser Gly Tyr Ala Ile Gln Ala Asp Glu Glu Gln Leu Arg Val Gln Leu Asp Thr Ile Gln Gly Glu Leu Asn Ala Pro Thr Gln Phe Lys Gly Arg Leu Asn Glu Leu Met Ser Gln Ile Arg Met Gln Asn His Phe Gly Ala Val Arg Ser Glu Glu Arg Tyr Tyr Ile Asp Ala Asp Leu Leu Arg Glu Ile Lys Gln His Leu Lys Gln Gln Gln Glu Gly Leu Ser His Leu Ile Ser Ile Ile Lys Asp Asp Leu Glu Asp Ile Lys Leu Val Glu His Gly Leu Asn Glu Thr Ile His Ile Arg Gly Gly Val Phe Ser <210> 19 <211> 592 <212> PRT
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 2758310CD1 <400> 19 Met Trp Phe Cys Gly Gln Ser Thr Pro Phe Gly Cys Glu Leu His Asp Thr Cys Val Gln Leu Cys His Phe His Ser Ala Leu Leu His Arg Arg Gln Lys Pro Trp Pro Ser Pro Ala Val Phe Phe Arg Arg Asn Val Arg Gly Leu Pro Pro Arg Phe Ser Ser Pro Thr Pro Leu Trp Arg Lys Val Leu Ser Thr Ala Val Val Gly Ala Pro Leu Leu Leu Gly Ala Arg Tyr Val Met Ala Glu Ala Arg Glu Lys Arg Arg Met Arg Leu Val Val Asp Gly Met Gly Arg Phe Gly Arg Ser Leu Lys Val Gly Leu Gln Ile Ser Leu Asp Tyr Trp Trp Cys Thr Asn Val Val Leu Arg Gly Trp Lys Ser Pro Gly Tyr Leu Glu Val Met Ser Ala Cys His Gln Arg Ala Ala Asp Ala Leu Val Ala Gly Ala Ile Ser Asn Gly Gly Leu Tyr Val Lys Leu Gly Gln Gly Leu Cys Ser Phe Asn His Leu Leu Pro Pro Glu Tyr Thr Arg Thr Leu Arg Val Leu Glu Asp Arg Ala Leu Lys Arg Gly Phe Gln Glu Val Asp Glu Leu Phe Leu Glu Asp Phe Gln Ala Leu Pro His Glu Leu Phe Gln Glu Phe Asp Tyr Gln Pro Ile Ala Ala Ala Ser Leu Ala Gln Val His Arg Ala Lys Leu His Asp Gly Thr Ser Val Ala Val Lys Val Gln Tyr Ile Asp Leu Arg Asp Arg Phe Asp Gly Asp Ile His Thr Leu Glu Leu Leu Leu Arg Leu Val Glu Val Met His Pro Ser Phe Gly Phe Ser Trp Val Leu Gln Asp Leu Lys Gly Thr Leu Ala Gln Glu Leu Asp Phe Glu Asn Glu Gly Arg Asn Ala Glu Arg Cys Ala Arg Glu Leu Ala His Phe Pro Tyr Val Val Val Pro Arg Val His Trp Asp Lys Ser Ser Lys Arg Val Leu Thr Ala Asp Phe Cys Ala Gly Cys Lys Val Asn Asp Val Glu Ala Ile Arg Ser Gln Gly Leu Ala Val His Asp Ile Ala Glu Lys Leu Ile Lys Ala Phe Ala Glu Gln Ile Phe Tyr Thr Gly Phe Ile His Ser Asp Pro His Pro Gly Asn Val Leu Val Arg Lys Gly Pro Asp Gly Lys Ala Glu Leu Val Leu Leu Asp His Gly Leu Tyr Gln Phe Leu Glu Glu Lys Asp Arg Ala Ala Leu Cys Gln Leu Trp Arg Ala Ile Ile Leu Arg Asp Asp Ala Ala Met Arg Ala His Ala Ala Ala Leu Gly Val Gln Asp Tyr Leu Leu Phe Ala Glu Met Leu Met Gln Arg Pro Val Arg Leu Gly Gln Leu Trp Gly Ser His Leu Leu Ser Arg Glu Glu Ala Ala Tyr Met Val Asp Met Ala Arg Glu Arg Phe Glu Ala Val Met Ala Val Leu Arg Glu Leu Pro Arg Pro Met Leu Leu Val Leu Arg Asn Ile Asn Thr Val Arg Ala Ile Asn Val Ala Leu Gly Ala Pro Val Asp Arg Tyr Phe Leu Met Ala Lys Arg Ala Val Arg Gly Trp Ser Arg Leu Ala Gly Ala Thr Tyr Arg Gly Val Tyr Gly Thr Ser Leu Leu Arg His Ala Lys Val Val Trp Glu Met Leu Lys Phe Glu Val Ala Leu Arg Leu Glu Thr Leu Ala Met Arg Leu Thr Ala Leu Leu Ala Arg Ala Leu Val His Leu Ser Leu Val Pro Pro Ala Glu Glu Leu Tyr Gln Tyr Leu Glu Thr <210> 20 <211> 841 <212> PRT
<213> Homo sapiens <220>
<221> misc_feature <223> Incyte ID No: 2762348CD1 <400> 20 Met Ala Ser Val Phe Arg Ser Glu Glu Met Cys Leu Ser Gln Leu Phe Leu Gln Val Glu Ala Ala Tyr Cys Cys Val Ala Glu Leu Gly Glu Leu Gly Leu Val Gln Phe Lys Asp Leu Asn Met Asn Val Asn Ser Phe Gln Arg Lys Phe Val Asn Glu Val Arg Arg Cys Glu Ser Leu Glu Arg Ile Leu Arg Phe Leu Glu Asp Glu Met Gln Asn Glu Ile Val Val Gln Leu Leu Glu Lys Ser Pro Leu Thr Pro Leu Pro Arg Glu Met Ile Thr Leu Glu Thr Val Leu Glu Lys Leu Glu Gly Glu Leu Gln Glu Ala Asn Gln Asn Gln Gln Ala Leu Lys Gln Ser Phe Leu Glu Leu Thr Glu Leu Lys Tyr Leu Leu Lys Lys Thr Gln Asp Phe Phe Glu Thr Glu Thr Asn Leu Ala Asp Asp Phe Phe Thr Glu Asp Thr Ser Gly Leu Leu Glu Leu Lys Ala Val Pro Ala Tyr Met Thr Gly Lys Leu Gly Phe Ile Ala Gly Cys Asp Pro Thr Gly Lys Arg Met Ala Ser Phe Glu Arg Leu Leu Trp Arg Val Cys Arg Gly Asn Val Tyr Leu Lys Phe Ser Glu Met Asp Ala Pro Leu Glu Asp Pro Val Thr Lys Glu Glu Ile Gln Lys His Ile Phe Ile Ile Phe Tyr Gln Gly Glu Gln Leu Arg Gln Lys Ile Lys Lys Ile Cys Asp Gly Phe Arg Ala Thr Val Tyr Pro Cys Pro Glu Pro Ala Val Glu Arg Arg Glu Met Leu Glu Ser Val Asn Val Arg Leu Glu Asp Leu Ile Thr Val Ile Thr Gln Thr Glu Ser His Arg Gln Arg Leu Leu Gln Glu Ala Ala Ala Asn Trp His Ser Trp Leu Ile Lys Val Gln Lys Met Lys Ala Val Tyr His Ile Leu Asn Met Cys Asn Ile Asp Val Thr Gln Gln Cys Val Ile Ala Glu Ile Trp Phe Pro Val Ala Asp Ala Thr Arg Ile Lys Arg Ala Leu Glu Gln Gly Met Glu Leu Ser Gly Ser Ser Met Ala Pro Ile Met Thr Thr Val Gln Ser Lys Thr Ala Pro Pro Thr Phe Asn Arg Thr Asn Lys Phe Thr Ala Gly Phe Gln Asn Ile Val Asp Ala Tyr Gly Val Gly Ser Tyr Arg Glu Ile Asn Pro Ala Pro Tyr Thr Ile Ile Thr Phe Pro Phe Leu Phe Ala Val Met Phe Gly Asp Cys Gly His Gly Thr Val Met Leu Leu Ala Ala Leu Trp Met Ile Leu Asn Glu Arg Arg Leu Leu Ser Gln Lys Thr Asp Asn Glu Ile Trp Asn Thr Phe Phe His Gly Arg Tyr Leu Ile Leu Leu Met Gly Ile Phe Ser Ile Tyr Thr Gly Leu Ile Tyr Asn Asp Cys Phe Ser Lys Ser Leu Asn Ile Phe Gly Ser Ser Trp Ser Val Gln Pro Met Phe Arg Asn Gly Thr Trp Asn Thr His Val Met Glu Glu Ser Leu Tyr Leu Gln Leu Asp Pro Ala Ile Pro Gly Val Tyr Phe Gly Asn Pro Tyr Pro Phe Gly Ile Asp Pro Ile Trp Asn Leu Ala Ser Asn Lys Leu Thr Phe Leu Asn Ser Tyr Lys Met Lys Met Ser Val Ile Leu Gly Ile Val Gln Met Val Phe Gly Val Ile Leu Ser Leu Phe Asn His Ile Tyr Phe Arg Arg Thr Leu Asn Ile Ile Leu Gln Phe Ile Pro Glu Met Ile Phe Ile Leu Cys Leu Phe Gly Tyr Leu Val Phe Met Ile Ile Phe Lys Trp Cys Cys Phe Asp Val His Val Ser Gln His Ala Pro Ser Ile Leu Ile His Phe Ile Asn Met Phe Leu Phe Asn Tyr Ser Asp Ser Ser Asn Ala Pro Leu Tyr Lys His Gln Gln Glu Val Gln Ser Phe Phe Val Val Met Ala Leu Ile Ser Val Pro Trp Met Leu Leu Ile Lys Pro Phe Ile Leu Arg Ala Ser His Arg Lys Ser Gln Leu Gln Ala Ser Arg Ile Gln Glu Asp Ala Thr Glu Asn Ile Glu Gly Asp Ser Ser Ser Pro Ser Ser Arg Ser Gly Gln Arg Thr Ser Ala Asp Thr His Gly Ala Leu Asp Asp His Gly Glu Glu Phe Asn Phe Gly Asp Val Phe Val His Gln Ala Ile His Thr Ile Glu Tyr Cys Leu Gly Cys Ile Ser Asn Thr Ala Ser Tyr Leu Arg Leu Trp Ala Leu Ser Leu Ala His Ala Gln Leu Ser Glu Val Leu Trp Thr Met Val Met Asn Ser Gly Leu Gln Thr Arg Gly Trp Gly Gly Ile Val Gly Val Phe Ile Ile Phe Ala Val Phe Ala Val Leu Thr Val Ala Ile Leu Leu Ile Met Glu Gly Leu Ser Ala Phe Leu His Ala Leu Arg Leu His Trp Val Glu Phe Gln Asn Lys Phe Tyr Val Gly Asp Gly Tyr Lys Phe Ser Pro Phe Ser Phe Lys His Ile Leu Asp Gly Thr Ala Glu Glu <210> 21 <211> 253 <212> PRT
<213> Homo sapiens <220>
<221> misc_feature <223> Incyte ID No: 3715961CD1 <400> 21 Met Ser Glu Cys Pro Leu Ile Leu Tyr Ile His Lys His Ile Asp Thr Tyr Ser Gln Ser Tyr Leu Phe Asn Asp Leu Phe Tyr Pro Val Tyr Ser Gly Gly Arg Met Val Thr Tyr Glu His Leu Arg Glu Val Val Phe Gly Lys Ser Glu Asp Glu His Tyr Pro Leu Trp Lys Ser Val Ile Gly Gly Met Met Ala Gly Val Ile Gly Gln Phe Leu Ala Asn Pro Thr Asp Leu Val Lys Val Gln Met Gln Met Glu Gly Lys Arg Lys Leu Glu Gly Lys Pro Leu Arg Phe Arg Gly Val His His Ala Phe Ala Lys Ile Leu Ala Glu Gly Gly Ile Arg Gly Leu Trp Ala Gly Trp Val Pro Asn Ile Gln Arg Ala Ala Leu Val Asn Met Gly Asp Leu Thr Thr Tyr Asp Thr Val Lys His Tyr Leu Val Leu Asn Thr Pro Leu Glu Asp Asn Ile Met Thr His Gly Leu Ser Ser Leu Cys Ser Gly Leu Val Ala Ser Ile Leu Gly Thr Pro Ala Asp WO 00/78953 PCT/~JS00/16668 Val Ile Lys Ser Arg Ile Met Asn Gln Pro Arg Asp Lys Gln Gly Arg Gly Leu Leu Tyr Lys Ser Ser Thr Asp Cys Leu Ile Gln Ala Val Gln Gly Glu Gly Phe Met Ser Leu Tyr Lys Gly Phe Leu Pro Ser Trp Leu Arg Met Thr Pro Trp Ser Met Val Phe Trp Leu Thr Tyr Glu Lys Ile Arg Glu Met Ser Gly Val Ser Pro Phe <210> 22 <211> 229 <212> PRT
<213> Homo sapiens <220>
<221> misc_feature <223> Incyte ID No: 5108194CD1 <400> 22 Met Gly Asn Gly Val Lys Glu Gly Pro Val Arg Leu His Glu Asp Ala Glu Ala Val Leu Ser Ser Ser Val Ser Ser Lys Arg Asp His Arg Gln Val Leu Ser Ser Leu Leu Ser Gly Ala Leu Ala Gly Ala Leu Ala Lys Thr Ala Val Ala Pro Leu Asp Arg Thr Lys Ile Ile Phe Gln Val Ser Ser Lys Arg Phe Ser Ala Lys Glu Ala Phe Arg Val Leu Tyr Tyr Thr Tyr Leu Asn Glu Gly Phe Leu Ser Leu Trp Arg Gly Asn Ser Ala Thr Met Val Arg Val Val Pro Tyr Ala Ala Ile Gln Phe Ser Ala His Glu Glu Tyr Lys Arg Ile Leu Gly Ser Tyr Tyr Gly Phe Arg Gly Glu Ala Leu Pro Pro Trp Pro Arg Leu Phe Ala Gly Ala Leu Ala Gly Thr Thr Ala Ala Ser Leu Thr Tyr Pro Leu Asp Leu Val Arg Ala Arg Met Ala Val Thr Pro Lys Glu Met Tyr Ser Asn Ile Phe His Val Phe Ile Arg Ile Ser Arg Glu Glu Gly Leu Lys Thr Leu Tyr His Gly Phe Met Pro Thr Val Leu Gly Val Ile Pro Tyr Ala Gly Leu Ser Phe Phe Thr Tyr Glu Thr Leu Lys Ser Leu His Arg Glu Tyr Ser Gly Arg Lys Leu Ile Pro Phe Ser Glu Gly <210> 23 <211> 170 <212> PRT
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 5503122CD1 <400> 23 Met Tyr Asp Asn Leu Tyr Leu His Gly Ile Glu Asp Ser Glu Ala Gly Ser Ala Asp Ser Tyr Thr Ser Arg Pro Ser Asp Ser Asp Val Ser Leu Glu Glu Asp Arg G1u Ala Ile Arg Gln Glu Arg Glu Gln Gln Ala Ala Ile Gln Leu Glu Arg Ala Lys Ser Lys Pro Val Ala Phe Ala Val Lys Thr Asn Val Ser Tyr Cys Gly Ala Leu Asp Glu Asp Val Pro Val Pro Ser Thr Ala Ile Ser Phe Asp Ala Lys Asp Phe Leu His Ile Lys Glu Lys Tyr Asn Asn Asp Trp Trp Ile Gly Arg Leu Val Lys Glu Gly Cys Glu Ile Gly Phe Ile Pro Ser Pro Leu Arg Leu Glu Asn Ile Arg Ile Gln Gln Glu Gln Lys Arg Gly Arg Phe His Gly Gly Lys Ser Ser Gly Asn Ser Ser Ser Ser Leu Gly Glu Met Val Ser Gly Thr Phe Arg Ala Thr Pro Thr Ser Thr Gly Glu Gly Cys Ser <210> 24 <211> 655 <212> PRT
<213> Homo sapiens <220>
<221> misc_feature <223> Incyte ID No: 5517972CD1 <400> 24 Met Ser Ser Ser Asn Val Glu Val Phe Ile Pro Val Ser Gln Gly Asn Thr Asn Gly Phe Pro Ala Thr Ala Ser Asn Asp Leu Lys Ala Phe Thr Glu Gly Ala Val Leu Ser Phe His Asn Ile Cys Tyr Arg Val Lys Leu Lys Ser Gly Phe Leu Pro Cys Arg Lys Pro Val Glu Lys Glu Ile Leu Ser Asn Ile Asn Gly Ile Met Lys Pro Gly Leu Asn Ala Ile Leu Gly Pro Thr Gly Gly Gly Lys Ser Ser Leu Leu Asp Val Leu Ala Ala Arg Lys Asp Pro Ser Gly Leu Ser Gly Asp Val Leu Ile Asn Gly Ala Pro Arg Pro Ala Asn Phe Lys Cys Asn Ser Gly Tyr Val Val Gln Asp Asp Val Val Met Gly Thr Leu Thr 125 i30 135 Val Arg Glu Asn Leu Gln Phe Ser Ala Ala Leu Arg Leu Ala Thr Thr Met Thr Asn His Glu Lys Asn Glu Arg Ile Asn Arg Val Ile Gln Glu Leu Gly Leu Asp Lys Val Ala Asp Ser Lys Val Gly Thr Gln Phe Ile Arg Gly Val Ser Gly Gly Glu Arg Lys Arg Thr Ser Ile Gly Met Glu Leu Ile Thr Asp Pro Ser Ile Leu Phe Leu Asp Glu Pro Thr Thr Gly Leu Asp Ser Ser Thr Ala Asn Ala Val Leu Leu Leu Leu Lys Arg Met Ser Lys Gln Gly Arg Thr Ile Ile Phe Ser Ile His Gln Pro Arg Tyr Ser Ile Phe Lys Leu Phe Asp Ser Leu Thr Leu Leu Ala Ser Gly Arg Leu Met Phe His Gly Pro Ala Gln Glu Ala Leu Gly Tyr Phe Glu Ser Ala Gly Tyr His Cys Glu Ala Tyr Asn Asn Pro Ala Asp Phe Phe Leu Asp Ile Ile Asn Gly Val Gln Gly Glu Gly Phe Met Ser Leu Tyr Lys Gly Phe Leu Pro Asp Ser Thr Ala Val Ala Leu Asn Arg Glu Glu Asp Phe Lys Ala Thr Glu Ile Ile Glu Pro Ser Lys Gln Asp Lys Pro Leu Ile Glu Lys Leu Ala Glu Ile Tyr Val Asn Ser Ser Phe Tyr Lys Glu Thr Lys Ala Glu Leu His Gln Leu Ser Gly Gly Glu Lys Lys Lys Lys Ile Thr Val Phe Lys Glu Ile Ser Tyr Thr Thr Ser Phe Cys His Gln Leu Arg Trp Val Ser Lys Arg Ser Phe Lys Asn Leu Leu Gly Asn Pro Gln Ala Ser Ile Ala Gln Ile Ile Val Thr Val Val Leu Gly Leu Val Ile Gly Ala Ile Tyr Phe Gly Leu Lys Asn Asp Ser Thr Gly Ile Gln Asn Arg Ala Gly Val Leu Phe Phe Leu Thr Thr Asn Gln Cys Phe Ser Ser Val Ser Ala Val Glu Leu Phe Val Val Glu Lys Lys Leu Phe Ile His Glu Tyr Ile Ser Gly Tyr Tyr Arg Val Ser Ser Tyr Phe Leu Gly Lys Leu Leu Ser Asp Leu Leu Pro Met Arg Met Leu Pro Ser Ile Ile Phe Thr Cys Ile Val Tyr Phe Met Leu Gly Leu Lys Pro Lys Ala Asp Ala Phe Phe Val Met Met Phe Thr Leu Met Met Val Ala Tyr Ser Ala Ser Ser Met Ala Leu Ala Ile Ala Ala Gly Gln Ser Val Val Ser Val Ala Thr Leu Leu Met Thr Ile Cys Phe Val Phe Met Met Ile Phe Ser Gly Leu Leu Val Asn Leu Thr Thr Ile Ala Ser Trp Leu Ser Trp Leu Gln Tyr Phe Ser Ile Pro Arg Tyr Gly Phe Thr Ala Leu Gln His Asn Glu Phe Leu Gly Gln Asn Phe Cys Pro Gly Leu Asn Ala Thr Gly Asn Asn Pro Cys Asn Tyr Ala Thr Cys Thr Gly Glu Glu Tyr Leu Val Lys Gln Gly Ile Asp Leu Ser Pro Trp Gly Leu Trp Lys Asn His Val Ala Leu Ala Cys Met Ile Val Ile Phe Leu Thr Ile Ala Tyr Leu Lys Leu Leu Phe Leu Lys Lys Tyr Ser <210> 25 <211> 184 <212> PRT
<213> Homo sapiens <220>
<221> misc_feature <223> Incyte ID No: 5593114CD1 <400> 25 Met Trp Val Phe Gly Tyr Gly Ser Leu Ile Trp Lys Val Asp Phe Pro Tyr Gln Asp Lys Leu Val Gly Tyr Ile Thr Asn Tyr Ser Arg Arg Phe Trp Gln Gly Ser Thr Asp His Arg Gly Val Pro Gly Lys Pro Gly Arg Val Val Thr Leu Val Glu Asp Pro Ala Gly Cys Val Trp Gly Val Ala Tyr Arg Leu Pro Val Gly Lys Glu Glu Glu Val Lys Ala Tyr Leu Asp Phe Arg Glu Lys Gly Gly Tyr Arg Thr Thr Thr Val Ile Phe Tyr Pro Lys Asp Pro Thr Thr Lys Pro Phe Ser Val Leu Leu Tyr Ile Gly Thr Cys Asp Asn Pro Asp Tyr Leu Gly Pro Ala Pro Leu Glu Asp Ile Ala Glu Gln Ile Phe Asn Ala Ala Gly Pro Ser Gly Arg Asn Thr Glu Tyr Leu Phe Glu Leu Ala Asn Ser Ile Arg Asn Leu Val Pro Glu Glu Ala Asp Glu His Leu Phe Ala Leu Glu Lys Leu Val Lys Glu Arg Leu Glu Gly Lys Gln Asn Leu Asn Cys Ile <210> 26 <211> 154 <212> PRT
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 044775CD1 <400> 26 Met Gly Ala Phe Glu Cys Val Arg Lys Val Tyr Gln Thr Asp Gly Leu Lys Gly Phe Tyr Arg Gly Met Ser Ala Ser Tyr Ala Gly Ile Ser Glu Thr Val Ile His Phe Val Ile Tyr Glu Ser Ile Lys Gln Lys Leu Leu Glu Tyr Lys Thr Ala Ser Thr Met Glu Asn Asp Glu Glu Ser Val Lys Glu Ala Ser Asp Phe Val Gly Met Met Leu Ala Ala Ala Thr Ser Lys Thr Cys Ala Thr Thr Ile Ala Tyr Pro His Glu Val Val Arg Thr Arg Leu Arg Glu Glu Gly Thr Lys Tyr Arg Ser Phe Phe Gln Thr Leu Ser Leu Leu Val Gln Glu Glu Gly Tyr Gly Ser Leu Tyr Arg Gly Leu Thr Thr His Leu Val Arg Gln Ile Pro Asn Thr Ala Ile Met Met Ala Thr Tyr Glu Leu Val Val Tyr Leu Leu Asn Gly <210> 27 <211> 438 <212> PRT
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 116588CD1 <400> 27 Met Leu Leu Val Thr Pro Arg Pro Glu Arg Gly Gly Arg Gly Thr Glu Leu Gly Glu Phe Cys Gly Thr Pro Leu Leu Phe Ser Ser Tyr Phe Cys Tyr Asp Asn Pro Ala Ala Leu Gln Thr Gln Val Lys Arg Asp Met Gln Val Asn Thr Thr Lys Phe Met Leu Leu Tyr Ala Trp Tyr Ser Trp Pro Asn Val Val Leu Cys Phe Phe Gly Gly Phe Leu Ile Asp Arg Val Phe Gly Ile Arg Trp Gly Thr Ile Ile Phe Ser Cys Phe Val Cys Ile Gly Gln Val Val Phe Ala Leu Gly Gly Ile Phe Asn Ala Phe Trp Leu Met Glu Phe Gly Arg Phe Val Phe Gly Ile Gly Gly Glu Ser Leu Ala Val Ala Gln Asn Thr Tyr Ala Val Ser Trp Phe Lys Gly Lys Glu Leu Asn Leu Val Phe Gly Leu Gln Leu Ser Met Ala Arg Ile Gly Ser Thr Val Asn Met Asn Leu Met Gly Trp Leu Tyr Ser Lys Ile Glu Ala Leu Leu Gly Ser Ala Gly His Thr Thr Leu Gly Ile Thr Leu Met Ile Gly Gly Val Thr Cys Ile Leu Ser Leu Ile Cys Ala Leu Ala Leu Ala Tyr Leu Asp Gln Arg Ala Glu Arg Ile Leu His Lys Glu Gln Gly Lys Thr Gly Glu Val Ile Lys Leu Thr Asp Val Lys Asp Phe Ser Leu Pro Leu Trp Leu Ile Phe Ile Ile Cys Val Cys Tyr Tyr Val Ala Val Phe Pro Phe Ile Gly Leu Gly Lys Val Phe Phe Thr Glu Lys Phe Gly Phe Ser Ser Gln Ala Ala Ser Ala Ile Asn Ser Val Val Tyr Val Ile Ser Ala Pro Met Ser Pro Val Phe Gly Leu Leu Val Asp Lys Thr Gly Lys Asn Ile Ile Trp Val Leu Cys Ala Val Ala Ala Thr Leu Val Ser His Met Met Leu Ala Phe Thr Met Trp Asn Pro Trp Ile Ala Met Cys Leu Leu Gly Leu Ser Tyr Ser Leu Leu Ala Cys Ala Leu Trp Pro Met Val Ala Phe Val Val Pro Glu His Gln Leu Gly Thr Ala Tyr Gly Phe Met Gln Ser Ile Gln Asn Leu Gly Leu Ala Ile Ile Ser Ile Ile Ala Gly Met Ile Leu Asp Ser Arg Gly Tyr Leu Phe Leu Glu Val Phe Phe Ile Ala Cys Val Ser Leu Ser Leu Leu Ser Val Val Leu Leu Tyr Leu Val Asn Arg Ala Gln Gly Gly Asn Leu Asn Tyr Ser Ala Arg Gln Arg Glu Glu Ile Lys Phe Ser His Thr Glu <210> 28 <211> 237 <212> PRT
<213> Homo sapiens <220>
<221> misc_feature <223> Incyte ID No: 875369CD1 <400> 28 Met Ala His Val Gly Ser Arg Lys Arg Ser Arg Ser Arg Ser Arg Ser Arg Gly Arg Gly Ser Glu Lys Arg Lys Lys Lys Ser Arg Lys Asp Thr Ser Arg Asn Cys Ser Ala Ser Thr Ser Gln Gly Arg Lys Ala Ser Thr Ala Pro Gly Ala Glu Ala Ser Pro Ser Pro Cys Ile Thr Glu Arg Ser Lys Gln Lys Ala Arg Arg Arg Thr Arg Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Asp Gly Arg Lys Lys Arg Gly Lys Tyr Lys Asp Lys Arg Arg Lys Lys Lys Lys Lys Arg Lys Lys Leu Lys Lys Lys Gly Lys Glu Lys Ala Glu Ala Gln Gln Val Glu Ala Leu Pro Gly Pro Ser Leu Asp Gln Trp His Arg Ser Ala Gly Glu Glu Glu Asp Gly Pro Val Leu Thr Asp Glu Gln Lys Ser Arg Ile Gln Ala Met Lys Pro Met Thr Lys Glu Glu Trp Asp Ala Arg Gln Ser Ile Ile Arg Lys Val Val Asp Pro Glu Thr Gly Arg Thr Arg Leu Ile Lys Gly Asp Gly Glu Val Leu Glu Glu Ile Val Thr Lys Glu Arg His Arg Glu Ile Asn Lys Gln Ala Thr Arg Gly Asp Cys Leu Ala Phe Gln Met Arg Ala Gly Leu Leu Pro <210> 29 <211> 219 <212> PRT
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 1325518CD1 <400> 29 Met Lys Leu Leu Leu Trp Ala Cys Ile Val Cys Val Ala Phe Ala Arg Lys Arg Arg Phe Pro Phe Ile Gly Glu Asp Asp Asn Asp Asp Gly His Pro Leu His Pro Ser Leu Asn Ile Pro Tyr Gly Ile Arg Asn Leu Pro Pro Pro Leu Tyr Tyr Arg Pro Val Asn Thr Val Pro Ser Tyr Pro Gly Asn Thr Tyr Thr Asp Thr Gly Leu Pro Ser Tyr Pro Trp Ile Leu Thr Ser Pro Gly Phe Pro Tyr Val Tyr His Ile Arg Gly Phe Pro Leu Ala Thr Gln Leu Asn Val Pro Pro Leu Pro Pro Arg Gly Phe Pro Phe Val Pro Pro Ser Arg Phe Phe Ser Ala Ala Ala Ala Pro Ala Ala Pro Pro Ile Ala Ala Glu Pro Ala Ala Ala Ala Pro Leu Thr Ala Thr Pro Val Ala Ala Glu Pro Ala Ala Gly Ala Pro Val Ala Ala Glu Pro Ala Ala Glu Ala Pro Val Gly Ala Glu Pro Ala Ala Glu Ala Pro Val Ala Ala Glu Pro Ala Ala Glu Ala Pro Val Gly Val Glu Pro Ala Ala Glu Glu Pro Ser Pro Ala Glu Pro Ala Thr Ala Lys Pro Ala Ala Pro Glu Pro His Pro Ser Pro Ser Leu Glu Gln Ala Asn Gln <210> 30 <211> 707 <212> PRT
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 2060987CD1 <400> 30 Met Ala Ala Ala Ala Thr Ala Ala Glu Gly Val Pro Ser Arg Gly Pro Pro Gly Glu Val Ile His Leu Asn Val Gly Gly Lys Arg Phe Ser Thr Ser Arg Gln Thr Leu Thr Trp Ile Pro Asp Ser Phe Phe Ser Ser Leu Leu Ser Gly Arg Ile Ser Thr Leu Lys Asp Glu Thr Gly Ala Ile Phe Ile Asp Arg Asp Pro Thr Val Phe Ala Pro Ile Leu Asn Phe Leu Arg Thr Lys Glu Leu Asp Pro Arg Gly Val His Gly Ser Ser Leu Leu His Glu Ala Gln Phe Tyr Gly Leu Thr Pro Leu Val Arg Arg Leu Gln Leu Arg Glu Glu Leu Asp Arg Ser Ser Cys Gly Asn Val Leu Phe Asn Gly Tyr Leu Pro Pro Pro Val Phe Pro Val Lys Arg Arg Asn Arg His Ser Leu Val Gly Pro Gln Gln Leu Gly Gly Arg Pro Ala Pro Val Arg Arg Ser Asn Thr Met Pro Pro Asn Leu Gly Asn Ala Gly Leu Leu Gly Arg Met Leu Asp Glu Lys Thr Pro Pro Ser Pro Ser Gly Gln Pro Glu Glu Pro Gly Met Val Arg Leu Val Cys Gly His His Asn Trp Ile Ala Val Ala Tyr Thr Gln Phe Leu Val Cys Tyr Arg Leu Lys Glu Ala Ser Gly Trp Gln Leu Val Phe Ser Ser Pro Arg Leu Asp Trp Pro Ile Glu Arg Leu Ala Leu Thr Ala Arg Val His Gly Gly Ala Leu Gly Glu His Asp Lys Met Val Ala Ala Ala Thr Gly Ser Glu Ile Leu Leu Trp Ala Leu Gln Ala Glu Gly Gly Gly Ser Glu Ile Gly Val Phe His Leu Gly Val Pro Val Glu Ala Leu Phe Phe Val Gly Asn Gln Leu Ile Ala Thr Ser His Thr Gly Arg Ile Gly Val Trp Asn Ala Val Thr Lys His Trp Gln Val Gln Glu Val Gln Pro Ile Thr Ser Tyr Asp Ala Ala Gly Ser Phe Leu Leu Leu Gly Cys Asn Asn Gly Ser Ile Tyr Tyr Val Asp Val Gln Lys Phe Pro Leu Arg Met Lys Asp Asn Asp Leu Leu Val Ser Glu Leu Tyr Arg Asp Pro Ala Glu Asp Gly Val Thr Ala Leu Ser Val Tyr Leu Thr Pro Lys Thr Ser Asp Ser Gly Asn Trp Ile Glu Ile Ala Tyr Gly Thr Ser Ser Gly Gly Val Arg Val Ile Val Gln His Pro Glu Thr Val Gly Ser Gly Pro Gln Leu Phe Gln Thr Phe Thr Val His Arg Ser Pro Val Thr Lys Ile Met Leu Ser Glu Lys His Leu Ile Ser Val Cys Ala Asp Asn Asn His Val Arg Thr Trp Ser Val Thr Arg Phe Arg Gly Met Ile Ser Thr Gln Pro Gly Ser Thr Pro Leu Ala Ser Phe Lys Ile Leu Ala Leu Glu Ser Ala Asp Gly His Gly Gly Cys Ser Ala Gly Asn Asp Ile Gly Pro Tyr Gly Glu Arg Asp Asp Gln Gln Val Phe Ile Gln Lys Val Val Pro Ser Ala Ser Gln Leu Phe Val Arg Leu Ser Ser Thr Gly Gln Arg Val Cys Ser Val Arg Ser Val Asp Gly Ser Pro Thr Thr Ala Phe Thr Val Leu Glu Cys Glu Gly Ser Arg Arg Leu Gly Ser Arg Pro Arg Arg Tyr Leu Leu Thr Gly Gln Ala Asn Gly Ser Leu Ala Met Trp Asp Leu Thr Thr Ala Met Asp Gly Leu Gly Gln Ala Pro Ala Gly Gly Leu Thr Glu Gln Glu Leu Met Glu Gln Leu Glu His Cys Glu Leu Ala Pro Pro Ala Pro Ser Ala Pro Ser Trp Gly Cys Leu Pro Ser Pro Ser Pro Arg Ile Ser Leu Thr Ser Leu His Ser Ala Ser Ser Asn Thr Ser Leu Ser Gly His Arg Gly Ser Pro Ser Pro Pro Gln Ala Glu Ala Arg Arg Arg Gly Gly Gly Ser Phe Val Glu Arg Cys Gln Glu Leu Val Arg Ser Gly Pro Asp Leu Arg Arg Pro Pro Thr Pro Ala Pro Trp Pro Ser Ser Gly Leu Gly Thr Pro Leu Thr Pro Pro Lys Met Lys Leu Asn Glu Thr Ser Phe <210> 31 <211> 279 <212> PRT
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 2172064CD1 <400> 31 Met Cys Gly Arg Phe Leu Arg Arg Leu Leu Ala Glu Glu Ser Arg Arg Ser Thr Pro Val Gly Arg Leu Leu Leu Pro Val Leu Leu Gly Phe Arg Leu Val Leu Leu Ala Ala Ser Gly Pro Gly Val Tyr Gly Asp Glu Gln Ser Glu Phe Val Cys His Thr Gln Gln Pro Gly Cys Lys Ala Ala Cys Phe Asp Ala Phe His Pro Leu Ser Pro Leu Arg Ser Trp Val Phe Gln Val Ile Leu Val Ala Val Pro Ser Ala Leu Tyr Met Gly Phe Thr Leu Tyr His Val Ile Trp His Trp Glu Leu Ser Gly Lys Gly Lys Glu Glu Glu Thr Leu Ile Gln Gly Arg Glu Gly Asn Thr Asp Val Pro Gly Ala Gly Ser Leu Arg Leu Leu Trp Ala Tyr Val Ala Gln Leu Gly Ala Arg Leu Val Leu Glu Gly Ala Ala Leu Gly Leu Gln Tyr His Leu Tyr Gly Phe Gln Met Pro Ser Ser Phe Ala Cys Arg Arg Glu Pro Cys Leu Gly Ser Ile Thr Cys Asn Leu Ser Arg Pro Ser Glu Lys Thr Ile Phe Leu Lys Thr Met Phe Gly Val Ser Gly Phe Cys Leu Leu Phe Thr Phe Leu Glu Leu Val Leu Leu Gly Leu Gly Arg Trp Trp Arg Thr Trp Lys His Lys Ser Ser Ser Ser Lys Tyr Phe Leu Thr Ser Glu Ser Thr Arg Arg His Lys Lys Ala Thr Asp Ser Leu Pro Val Val Glu Thr Lys Glu Gln Phe Gln Glu Ala Val Pro Gly Arg Ser Leu Ala Gln Glu Lys Gln Arg Pro Val Gly Pro Arg Asp Ala <210> 32 <211> 154 <212> PRT
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 2219267CD1 <400> 32 Met Val Thr Gly Leu Ala Ser Leu Leu Leu Leu Ala Gly Ala Gln Tyr Leu Pro Gly Trp Thr Val Leu Phe Leu Ser Val Leu Gly Leu Leu Ala Ser Arg Ala Val Ser Ala Leu Ser Ser Leu Phe Ala Ala Glu Val Phe Pro Thr Val Ile Arg Gly Ala Gly Leu Gly Leu Val Leu Gly Ala Gly Phe Leu Gly Gln Ala Ala Gly Pro Leu Asp Thr Leu His Gly Arg Gln Gly Phe Phe Leu Gln Gln Val Val Phe Ala Ser Leu Ala Val Leu Ala Leu Leu Cys Val Leu Leu Leu Pro Glu Ser Arg Ser Arg Gly Leu Pro Gln Ser Leu Gln Asp Ala Asp Arg Leu Arg Arg Ser Pro Leu Leu Arg Gly Arg Pro Arg Gln Asp His Leu Pro Leu Leu Pro Pro Ser Asn Ser Tyr Trp Ala Gly His Thr Pro Glu Gln His <210> 33 <211> 289 <212> PRT
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 2308629CD1 <400> 33 Met Val Ala Gly Ala Val Ala Gly Ile Leu Glu His Cys Val Met Tyr Pro Ile Asp Cys Val Lys Thr Arg Met Gln Ser Leu Gln Pro Asp Pro Ala Ala Arg Tyr Arg Asn Val Leu Glu Ala Leu Trp Arg Ile Ile Arg Thr Glu Gly Leu Trp Arg Pro Met Arg Gly Leu Asn Val Thr Ala Thr Gly Ala Gly Pro Ala His Ala Leu Tyr Phe Ala Cys Tyr Glu Lys Leu Lys Lys Thr Leu Ser Asp Val Ile His Pro Gly Gly Asn Ser His Ile Ala Asn Gly Ala Ala Gly Cys Val Ala Thr Leu Leu His Asp Ala Ala Met Asn Pro Ala Glu Val Val Lys Gln Arg Met Gln Met Tyr Asn Ser Pro Tyr His Arg Val Thr Asp Cys Val Arg Ala Val Trp Gln Asn Glu Gly Ala Gly Ala Phe Tyr Arg Ser Tyr Thr Thr Gln Leu Thr Met Asn Val Pro Phe Gln Ala Ile His Phe Met Thr Tyr Glu Phe Leu Gln Glu His Phe Asn Pro Gln Arg Arg Tyr Asn Pro Ser Ser His Val Leu Ser Gly Ala Cys Ala Gly Ala Val Ala Ala Ala Ala Thr Thr Pro Leu Asp Val Cys Lys Thr Leu Leu Asn Thr Gln Glu Ser Leu Ala Leu Asn Ser His Ile Thr Gly His Ile Thr Gly Met Ala Ser Ala Phe Arg Thr Val Tyr Gln Val Gly Gly Val Thr Ala Tyr Phe Arg Gly Val Gln Ala Arg Val Ile Tyr Gln Ile Pro Ser Thr Ala Ile Ala Trp Ser Val Tyr Glu Phe Phe Lys Tyr Leu Ile Thr Lys Arg Gln Glu Glu Trp Arg Ala Gly Lys <210> 34 <211> 300 <212> PRT
<213> Homo sapiens <220>
<221> misc_feature <223> Incyte ID No: 2660038CD1 <400> 34 Met Asp Phe Leu Met Ser Gly Leu Ala Ala Cys Gly Ala Cys Val Phe Thr Asn Pro Leu Glu Val Val Lys Thr Arg Met Gln Leu Gln Gly Glu Leu Gln Ala Pro Gly Thr Tyr Gln Arg His Tyr Arg Asn Val Phe His Ala Phe Ile Thr Ile Gly Lys Val Asp Gly Leu Ala Ala Leu Gln Lys Gly Leu Ala Pro Ala Leu Leu Tyr Gln Phe Leu Met Asn Gly Ile Arg Leu Gly Thr Tyr Gly Leu Ala Glu Ala Gly Gly Tyr Leu His Thr Ala Glu Ala Thr His Ser Pro Ala Arg Ser Ala Ala Ala Gly Ala Met Ala Gly Val Met Gly Ala Tyr Leu Gly Ser Pro Ile Tyr Met Val Lys Thr His Leu Gln Ala Gln Ala Ala Ser Glu Ile Ala Val Gly His Gln Tyr Lys His Gln Gly Met Phe Gln Ala Leu Thr Glu Ile Gly Gln Lys His Gly Leu Val Gly Leu Trp Arg Gly Ala Leu Gly Gly Leu Pro Arg Val Ile Val Gly Ser Ser Thr Gln Leu Cys Thr Phe Ser Ser Thr Lys Asp Leu Leu Ser Gln Trp Glu Ile Phe Pro Pro Gln Ser Trp Lys Leu Ala Leu Val Ala Ala Met Met Ser Gly Ile Ala Val Val Leu Ala Met Ala Pro Phe Asp Val Ala Cys Thr Arg Leu Tyr Asn Gln Pro Thr Asp Ala Gln Gly Lys Gly Leu Met Tyr Arg Gly Ile Leu Asp Ala Leu Leu Gln Thr Ala Arg Thr Glu Gly Ile Phe Gly Met Tyr Lys Gly Ile Gly Ala Ser Tyr Phe Arg Leu Gly Pro His Thr Ile Leu Ser Leu Phe Phe Trp Asp Gln Leu Arg Ser Leu Tyr Tyr Thr Asp Thr Lys <210> 35 <211> 382 <212> PRT
<213> Homo sapiens <220>
<221> misc_feature <223> Incyte ID No: 2670745CD1 <400> 35 Met Leu Arg Trp Thr Val His Leu Glu Gly Gly Pro Arg Arg Val Asn His Ala Ala Val Ala Val Gly His Arg Val Tyr Ser Phe Gly Gly Tyr Cys Ser Gly Glu Asp Tyr Glu Thr Leu Arg Gln Ile Asp Val His Ile Phe Asn Ala Val Ser Leu Arg Trp Thr Lys Leu Pro Pro Val Lys Ser Ala Ile Arg Gly Gln Ala Pro Val Val Pro Tyr Met Arg Tyr Gly His Ser Thr Val Leu Ile Asp Asp Thr Val Leu Leu Trp Gly Gly Arg Asn Asp Thr Glu Gly Ala Cys Asn Val Leu Tyr Ala Phe Asp Val Asn Thr His Lys Trp Phe Thr Pro Arg Val Ser Gly Thr Val Pro Gly Ala Arg Asp Gly His Ser Ala Cys Val Leu Gly Lys Ile Met Tyr Ile Phe Gly Gly Tyr Glu Gln Gln Ala Asp Cys Phe Ser Asn Asp Ile His Lys Leu Asp Thr Ser Thr Met Thr Trp Thr Leu Ile Cys Thr Lys Gly Ser Pro Ala Arg Trp Arg Asp Phe His Ser Ala Thr Met Leu Gly Ser His Met Tyr Val Phe Gly Gly Arg Ala Asp Arg Phe Gly Pro Phe His Ser Asn Asn Glu Ile Tyr Cys Asn Arg Ile Arg Val Phe Asp Thr Arg Thr Glu Ala Trp Leu Asp Cys Pro Pro Thr Pro Val Leu Pro Glu Gly Arg Arg Ser His Ser Ala Phe Gly Tyr Asn Gly Glu Leu Tyr Ile Phe Gly Gly Tyr Asn Ala Arg Leu Asn Arg His Phe His Asp Leu Trp Lys Phe Asn Pro Val Ser Phe Thr Trp Lys Lys Ile Glu Pro Lys Gly Lys Gly Pro Cys Pro Arg Arg Arg Gln Cys Cys Cys Ile Val Gly Asp Lys Ile Val Leu Phe Gly Gly Thr Ser Pro Ser Pro Glu Glu Gly Leu Gly Asp Glu Phe Asp Leu Ile Asp His Ser Asp Leu His Ile Leu Asp Phe Ser Pro Ser Leu Lys Thr Leu Cys Lys Leu Ala Val Ile Gln Tyr Asn Leu Asp Gln Ser Cys Leu Pro His Asp Ile Arg Trp Glu Leu Asn Ala Met Thr Thr Asn Ser Asn Ile Ser Arg Pro Ile Val Ser Ser His Gly <210> 36 <211> 287 <212> PRT
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 2676443CD1 <400> 36 Met Ala Ala Glu Ala Arg Val Ser Arg Trp Tyr Phe Gly Gly Leu Ala Ser Cys Gly Ala Ala Cys Cys Thr His Pro Leu Asp Leu Leu Lys Val His Leu Gln Thr Gln Gln Glu Val Lys Leu Arg Met Thr Gly Met Ala Leu Arg Val Val Arg Thr Asp Gly Ile Leu Ala Leu Tyr Ser Gly Leu Ser Ala Ser Leu Cys Arg Gln Met Thr Tyr Ser Leu Thr Arg Phe Ala Ile Tyr Glu Thr Val Arg Asp Arg Val Ala Lys Gly Ser Gln Gly Pro Leu Pro Phe His Glu Lys Val Leu Leu Gly Ser Val Ser Gly Leu Ala Gly Gly Phe Val Gly Thr Pro Ala Asp Leu Val Asn Val Arg Met Gln Asn Asp Val Lys Leu Pro Gln Gly Gln Arg Arg Asn Tyr Ala His Ala Leu Asp Gly Leu Tyr Arg Val Ala Arg Glu Glu Gly Leu Arg Arg Leu Phe Ser Gly Ala Thr Met Ala Ser Ser Arg Gly Ala Leu Val Thr Val Gly Gln Leu Ser Cys Tyr Asp Gln Ala Lys Gln Leu Val Leu Ser Thr Gly Tyr Leu Ser Asp Asn Ile Phe Thr His Phe Val Ala Ser Phe Ile Ala Gly Gly Cys Ala Thr Phe Leu Cys Gln Pro Leu Asp Val Leu Lys Thr Arg Leu Met Asn Ser Lys Gly Glu Tyr Gln Gly Val Phe His Cys Ala Val Glu Thr Ala Lys Leu Gly Pro Leu Ala Phe Tyr Lys Gly Leu Val Pro Ala Gly Ile Arg Leu Ile Pro His Thr Val Leu Thr Phe Val Phe Leu Glu Gln Leu Arg Lys Asn Phe Gly Ile Lys Val Pro Ser <210> 37 <211> 497 <212> PRT
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 3295764CD1 <400> 37 Met Asp Val Pro Gly Pro Val Ser Arg Arg Ala Ala Ala Ala Ala Ala Thr Val Leu Leu Arg Thr Ala Arg Val Arg Arg Glu Cys Trp Phe Leu Pro Thr Ala Leu Leu Cys Ala Tyr Gly Phe Phe Ala Ser Leu Arg Pro Ser Glu Pro Phe Leu Thr Pro Tyr Leu Leu Gly Pro Asp Lys Asn Leu Thr Glu Arg Glu Val Phe Asn Glu Ile Tyr Pro Val Trp Thr Tyr Ser Tyr Leu Val Leu Leu Phe Pro Val Phe Leu Ala Thr Asp Tyr Leu Arg Tyr Lys Pro Val Val Leu Leu Gln Gly Leu Ser Leu Ile Val Thr Trp Phe Met Leu Leu Tyr Ala Gln Gly Leu Leu Ala Ile Gln Phe Leu Glu Phe Phe Tyr Gly Ile Ala Thr Ala Thr Glu Ile Ala Tyr Tyr Ser Tyr Ile Tyr Ser Val Val Asp Leu Gly Met Tyr Gln Lys Val Thr Ser Tyr Cys Arg Ser Ala Thr Leu Val Gly Phe Thr Val Gly Ser Val Leu Gly Gln Ile Leu Val Ser Val Ala Gly Trp Ser Leu Phe Ser Leu Asn Val Ile Ser Leu Thr Cys Val Ser Val Ala Phe Ala Val Ala Trp Phe Leu Pro Met Pro Gln Lys Ser Leu Phe Phe His His Ile Pro Ser Thr Cys Gln Arg Val Asn Gly Ile Lys Val Gln Asn Gly Gly Ile Val Thr Asp Thr Pro Ala Ser Asn His Leu Pro Gly Trp Glu Asp Ile Glu Ser Lys Ile Pro Leu Asn Met Glu Glu Pro Pro Val Glu Glu Pro Glu Pro Lys Pro Asp Arg Leu Leu Val Leu Lys Val Leu Trp Asn Asp Phe Leu Met Cys Tyr Ser Ser Arg Pro Leu Leu Cys Trp Ser Val Trp Trp Ala Leu Ser Thr Cys Gly Tyr Phe Gln Val Val Asn Tyr Thr Gln Gly Leu Trp Glu Lys Val Met Pro Ser Arg Tyr Ala Ala Ile Tyr Asn Gly Gly Val Glu Ala Val Ser Thr Leu Leu Gly Ala Val Ala Val Phe Ala Val Gly Tyr Ile Lys Ile Ser Trp Ser Thr Trp Gly Glu Met Thr Leu Ser Leu Phe Ser Leu Leu Ile Ala Ala Ala Val Tyr Ile Met Asp Thr Val Gly Asn Ile Trp Val Cys Tyr Ala Ser Tyr Val Val Phe Arg Ile Ile Tyr Met Leu Leu Ile Thr Ile Ala Thr Phe Gln Ile Ala Ala Asn Leu Ser Met Glu Arg Tyr Ala Leu Val Phe Gly Val Asn Thr Phe Ile Ala Leu Ala Leu Gln Thr Leu Leu Thr Leu Ile Val Val Asp Ala Ser Gly Leu Gly Leu Glu Ile Thr Thr Gln Phe Leu Ile Tyr Ala Ser Tyr Phe Ala Leu Ile Ala Val Val Phe Leu Ala Ser Gly Ala Val Ser Val Met Lys Lys Cys Arg Lys Leu Glu Asp Pro Gln Ser Ser Ser Gln Val Thr Thr Ser <210> 38 <211> 228 <212> PRT
<213> Homo sapiens <220>
<221> misc_feature <223> Incyte ID No: 3438320CD1 <400> 38 Met Pro Arg Arg Gly Leu Val Ala Gly Pro Asp Leu Glu Tyr Phe Gln Arg Arg Tyr Phe Thr Pro Ala Glu Val Ala Gln His Asn Arg Pro Glu Asp Leu Trp Val Ser Tyr Leu Gly Arg Val Tyr Asp Leu Thr Ser Leu Ala Gln Glu Tyr Lys Gly Asn Leu Leu Leu Lys Pro Ile Val Glu Val Ala Gly Gln Asp Ile Ser His Trp Phe Asp Pro Lys Thr Arg Asp Ile Arg Lys His Ile Asp Pro Leu Thr Gly Cys Leu Arg Tyr Cys Thr Pro Arg Gly Arg Phe Val His Val Pro Pro Gln Leu Pro Cys Ser Asp Trp Ala Asn Asp Phe Gly Lys Pro Trp Trp Gln Gly Ser Tyr Tyr Glu Val Gly Arg Leu Ser Ala Lys Thr Arg Ser Ile Arg Ile Ile Asn Thr Leu Thr Ser Gln Glu His Thr Leu Glu Val Gly Val Leu Glu Ser Ile Trp Glu Ile Leu His Arg Tyr Leu Pro Tyr Asn Ser His Ala Ala Ser Tyr Thr Trp Lys Tyr Glu Gly Lys Asn Leu Asn Met Asp Phe Thr Leu Glu Glu Asn Gly Ile Arg Asp Glu Glu Glu Glu Phe Asp Tyr Leu Ser Met Asp Gly Thr Leu His Thr Pro Ala Ile Leu Leu Tyr Phe Asn Asp Asp Leu Thr Glu Leu <210> 39 <211> 273 <212> PRT
<213> Homo sapiens <220>
<221> misc_feature <223> Incyte ID No: 3986488CD1 <400> 39 Met Ala Ala Thr Ile Met Ile Leu Tyr Val Ser Lys Leu Asn Lys Ile Ile His Phe Pro Asp Phe Asp Lys Lys Ile Pro Val Lys Leu Phe Pro Leu Pro Leu Leu Tyr Val Gly Asn His Ile Ser Gly Leu Ser Ser Thr Ser Lys Leu Ser Leu Pro Met Phe Thr Val Leu Arg Lys Phe Thr Ile Pro Leu Thr Leu Leu Leu Glu Thr Ile Ile Leu Gly Lys Gln Tyr Ser Leu Asn Ile Ile Leu Ser Val Phe Ala Ile Ile Leu Gly Ala Phe Ile Ala Ala Gly Ser Asp Leu Ala Phe Asn Leu Glu Gly Tyr Ile Phe Val Phe Leu Asn Asp Ile Phe Thr Ala Ala Asn Gly Val Tyr Thr Lys Gln Lys Met Asp Pro Lys Glu Leu Gly Lys Tyr Gly Val Leu Phe Tyr Asn Ala Cys Phe Met Ile Ile Pro Thr Leu Ile Ile Ser Val Ser Thr Gly Asp Leu Gln Gln Ala Thr Glu Phe Asn Gln Trp Lys Asn Val Val Phe Ile Leu Gln Phe Leu Leu Ser Cys Phe Leu Gly Phe Leu Leu Met Tyr Ser Thr Val Leu Cys Ser Tyr Tyr Asn Ser Ala Leu Thr Thr Ala Val Val Gly Ala Ile Lys Asn Val Ser Val Ala Tyr Ile Gly Ile Leu Ile Gly Gly Asp Tyr Ile Phe Ser Leu Leu Asn Phe Val Gly Leu Asn Ile Cys Met Ala Gly Gly Leu Arg Tyr Ser Phe Leu Thr Leu Ser Ser Gln Leu Lys Pro Lys Pro Val Gly Glu Glu Asn Ile Cys Leu Asp Leu Lys Ser <210> 40 <211> 206 <212> PRT
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 4378816CD1 <400> 40 Met Gly Ala Glu Trp Glu Leu Gly Ala Glu Ala Gly Gly Ser Leu Leu Leu Cys Ala Ala Leu Leu Ala Ala Gly Cys Ala Leu Gly Leu Arg Leu Gly Arg Gly Gln Gly Ala Ala Asp Arg Gly Ala Leu Ile Trp Leu Cys Tyr Asp Ala Leu Val His Phe Ala Leu Glu Gly Pro Phe Val Tyr Leu Ser Leu Val Gly Asn Val Ala Asn Ser Asp Gly Leu Ile Ala Ser Leu Trp Lys Glu Tyr Gly Lys Ala Asp Ala Arg Trp Val Tyr Phe Asp Pro Thr Ile Val Ser Val Glu Ile Leu Thr Val Ala Leu Asp Gly Ser Leu Ala Leu Phe Leu Ile Tyr Ala Ile Val Lys Glu Lys Tyr Tyr Arg His Phe Leu Gln Ile Thr Leu Cys Val Cys Glu Leu Tyr Gly Cys Trp Met Thr Phe Leu Pro Glu Trp Leu Thr Arg Ser Pro Asn Leu Asn Thr Ser Asn Trp Leu Tyr Cys Trp Leu Tyr Leu Phe Phe Phe Asn Gly Val Trp Val Leu Ile Pro Gly Leu Leu Leu Trp Gln Ser Trp Leu Glu Leu Lys Lys Met His Gln Lys Glu Thr Ser Ser Val Lys Lys Phe Gln <210> 41 <211> 235 <212> PRT
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 4797137CD1 <400> 41 Met Gln Gln Arg Gly Ala Ala Gly Ser Arg Gly Cys Ala Leu Phe Pro Leu Leu Gly Val Leu Phe Phe Gln Gly Val Tyr Ile Val Phe Ser Leu Glu Ile Arg Ala Asp Ala His Val Arg Gly Tyr Val Gly Glu Lys Ile Lys Leu Lys Cys Thr Phe Lys Ser Thr Ser Asp Val Thr Asp Lys Leu Thr Ile Asp Trp Thr Tyr Arg Pro Pro Ser Ser Ser His Thr Val Ser Ile Phe His Tyr Gln Ser Phe Gln Tyr Pro Thr Thr Ala Gly Thr Phe Arg Asp Arg Ile Ser Trp Val Gly Asn Val Tyr Lys Gly Asp Ala Ser Ile Ser Ile Ser Asn Pro Thr Ile Lys Asp Asn Gly Thr Phe Ser Cys Ala Val Lys Asn Pro Pro Asp Val His His Asn Ile Pro Met Thr Glu Leu Thr Val Thr Glu Arg Gly Phe Gly Thr Met Leu Ser Ser Val Ala Leu Leu Ser Ile Leu Val Phe Val Pro Ser Ala Val Val Val Ala Leu Leu Leu Val Arg Met Gly Arg Lys Ala Ala Gly Leu Lys Lys Arg Ser Arg Ser Gly Tyr Lys Lys Ser Ser Ile Glu Val Ser Asp Asp Thr Asp Gln Glu Glu Glu Glu Ala Cys Met Ala Arg Leu Cys Val Arg Cys Ala Glu Cys Leu Asp Ser Asp Tyr Glu Glu Thr Tyr <210> 42 <211> 147 <212> PRT
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 5470806CD1 <400> 42 Met Ala Ser Leu Arg Leu Phe Leu Leu Cys Leu Ala Val Leu Ile Phe Ala Ser Glu Ala Gly Pro Gly Gly Ala Gly Glu Ser Lys Cys Pro Leu Met Val Lys Val Leu Asp Ala Val Arg Gly Ser Pro Ala Val Asp Val Ala Val Lys Val Phe Lys Lys Thr Ala Asp Gly Ser Trp Glu Pro Phe Ala Ser Gly Lys Thr Ala Glu Ser Gly Glu Leu His Gly Leu Thr Thr Asp Glu Lys Phe Thr Glu Gly Val Tyr Arg Val Glu Leu Asp Thr Lys Ser Tyr Trp Lys Ala Leu Gly Ile Ser Pro Phe His Glu Tyr Ala Glu Val Val Phe Thr Ala Asn Asp Ser Gly His Arg His Tyr Thr Ile Ala Ala Leu Leu Ser Pro Tyr Ser Tyr Ser Thr Thr Ala Val Val Ser Asn Pro Gln Asn <210> 43 <211> 147 <212> PRT
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 5473242CD1 <400> 43 Met Val His Leu Thr Asp Ala Glu Lys Ala Thr Val Asn Gly Leu Trp Gly Lys Val Asn Pro Val Glu Ile Gly Ala Glu Ser Leu Ala Ser Leu Leu Ile Val Tyr Pro Trp Thr Gln Arg Tyr Phe Ser Lys Phe Gly Asp Leu Ser Ser Val Ser Ala Ile Met Gly Asn Pro Gln Val Lys Ala His Gly Glu Lys Val Ile Asn Ala Phe Asp Asp Gly Leu Lys His Leu Asp Asn Leu Lys Gly Thr Phe Ala Ser Leu Ser Glu Leu His Cys Asp Lys Leu His Val Asp Pro Glu Asn Phe Arg Leu Leu Gly Asn Met Ile Val Ile Met Met Gly His His Leu Gly Lys Glu Phe Thr Pro Ser Ala Gln Ala Ala Phe Gln Lys Val Val Ala Gly Val Ala Ser Ala Leu Ala His Lys Tyr His <210> 44 <211> 2701 <212> DNA
<213> Homo sapiens <220>
<221> misc_feature <223> Incyte ID No: 264114CB1 <400> 44 gcggcggcgc cagcttcctc ggccggaggg gaggcgagac cccaggcgag gccgcggcgg 60 gagggccacg cccccgacgc cgcgccggag gggcccagtg tggacggggc caccggctgg 120 agcggatccc acacctccgg accgagggac gcggttactc cacaggatcc gctgaacata 180 ggatgttgcc acaaaatcta cctcgtgtat ttttctcttt cactcatgag ctgcacaatt 240 gcagatttga gcacaatgtc tgcagactgt gttgaaaaac tctgaagaac ctaattaaca 300 caggatgacc taggagtgat tctaagtctg tgtaacaaga tattactcat tagtgaatgt 360 gtcagtcttg gtactgaatg ctgcagataa cagcaagtag gttctccttt atttctgaag 420 tattcacttg accttccatc agtaagacgg acttttctaa tctgttcctg gagatattaa 480 tggaatacag tcatgtccac tcaagacgag aggcagatca atactgaata tgctgtgtca 540 ttgttggaac agttgaaact gttttatgaa cagcagttgt ttactgacat agtgttaatt 600 gttgagggca ctgaattccc ttgtcataag atggttcttg caacatgtag ctcttatttc 660 agggccatgt ttatgagtgg actaagtgaa agcaaacaaa cccatgtaca cctgaggaat 720 gtcgatgctg ccaccttaca gataataata acttatgcat acacgggtaa cttggcaatg 780 aatgacagca ctgtagaaca gctttatgaa acagcttgct tcctacaggt agaagatgtg 840 ttacaacgtt gtcgagaata tttaattaaa aaaataaatg cagagaattg tgtacgattg 900 ttgagttttg ctgatctctt cagttgtgag gaattaaaac agagtgctaa aagaatggtg 960 gagcacaagt tcactgctgt gtatcatcag gacgcgttca tgcagctgtc acatgaccta 1020 ctgatagata ttctcagtag tgacaattta aatgtagaaa aggaagaaac cgttcgagaa 1080 gctgctatgc tgtggctaga gtataacaca gaatcacgat cccagtattt gtcttctgtt 1140 cttagccaaa tcagaattga tgcactttca gaagtaacac agagagcttg gtttcaaggt 1200 ctgccaccca atgataagtc agtggtggtt caaggtctgt ataagtccat gcccaagttt 1260 ttcaaaccaa gacttgggat gactaaagag gaaatgatga ttttcattga agcatcttca 1320 gaaaatcctt gtagtcttta ctcttctgtc tgttacagcc cccaagcaga aaaagtttac 1380 aagttatgta gcccaccagc tgatttgcat aaggttggga ccgttgtaac tcctgataat 1440 gatatctaca tagcaggggg tcaagttcct ctgaaaaaca caaaaacaaa tcacagtaaa 1500 acaagcaaac ttcagactgc cttcagaact gtgaattgct tttattggtt tgatgcacag 1560 caaaatacct ggtttccaaa gaccccaatg ctttttgtcc gcataaagcc atctttggtt 1620 tgctgtgaag gctatatcta tgcaattgga ggagatagcg taggtggaga acttaatcgg 1680 aggaccgtag aaagatacga cactgagaaa gatgagtgga cgatggtaag ccctttacct 1740 tgtgcttggc aatggagtgc agcagttgtg gttcatgact gcatttatgt gatgacactg 1800 aacctcatgt actgttattt tccaaggtct gactcatggg tagaaatggc catgagacag 1860 actagtaggt cctttgcttc agctgcagct tttggtgata aaattttcta tattggaggg 1920 ttgcatattg ctaccaattc cggcataaga ctcccctctg gcactgtaga tgggtcttca 1980 gtaactgtgg aaatttatga tgtgaataaa aatgagtgga aaatggcagc caacatccct 2040 gctaagaggt actctgaccc ctgtgttaga gctgttgtga tctcaaattc tctatgtgtg 2100 tttatgcgag aaacccactt aaatgagcga gctaaatacg tcacctacca atatgacctg 2160 gaacttgacc ggtggtctct gcggcagcat atatctgaac gtgtactgtg ggacttgggg 2220 agagattttc gatgcactgt ggggaaactc tatccatcct gccttgaaga gtctccatgg 2280 aaaccaccaa cttatctttt ttcaacggat gggacagaag agtttgaact ggatggagaa 2340 atggttgcac taccacctgt atagtgggga agttcaggga gtgcacgcct gagttatgtg 2400 ctttgtcatt ttctttgcta aacaaaagag gctatgaaag aactaaatat gagtacataa 2460 aattctatct ttgataaatt ttatttttat gccctactta atatttgcat cagtataata 2520 tatatcagtg agtcttacag aaagatatgc ttccataata tgaaatagat tattcaataa 2580 ttgagaaact ttatgtgtaa tcatgagagt ataagaatct ggattatcta acattgttag 2640 ccctgtgtat gtacagttca aaaagttcat ttataaaagt agtttcctgt tcctagttga 2700 t 2701 <210> 45 <211> 736 <212> DNA
<213> Homo sapiens <220>
<221> misc_feature <223> Incyte ID No: 1455669CB1 <400> 45 gagacttagc gacagacaga cgctgggacc cacgacgaca gaaggcgccg atggccgcgc 60 ctgctgagcc ctgcgcgggg cagggggtct ggaaccagac agagcctgaa cctgccgcca 120 ccagcctgct gagcctgtgc ttcctgagaa cagcaggggt ctgggtaccc cccatgtacc 180 tctgggtcct tggtcccatc tacctcctct tcatccacca ccatggccgg ggctacctcc 240 ggatgtcccc actcttcaaa gccaagatgg tagctgccat ccctgggagc ctggaaccag 300 gcaatgttcg ggggaggcag gggacaggct ggaacctggt gaagtcttaa agtagactcc 360 tcctatcggg gtgtagaagg gaatctgtta atcaaacaga gcaatattag aaaggctaca 420 gaggtcaact cagtggaaca cggttctccc aaacagattt tgtaattccg aaaatccacg 480 catgcgcaaa catacgcata cactcccatg ttcctggaca gtttatagct accataacct 540 ggcattttcc aaaacatacc atgtagactc ttggatacac aaggtaattt tagagccaca 600 ttaggatgaa ccttttaaaa agttatgcat ttatttttat gttcccccac tggctgtatt 660 ataggacaat ttttatatgt gatatgtatt taccttagtg tgttaaataa acactggcat 720 tccaagtgtg aaaaaa 736 <210> 46 <211> 1826 <212> DNA
<213> Homo sapiens <220>
<221> misc_feature <223> Incyte ID No: 2084989CB1 <400> 46 ccttaggcgc cagggacagc cgagcgttac ctggtcccgg gcagcggagt tctttaccca 60 ccccagttct ggttctgacg ccctagctca ttccgcaaat ttagggcttg ggtctggctt 120 gttcccctcc ggctcgaacc acctcttctc tgagccgagc cagctaccgg ggctcctgga 180 attgccaccc ctccctgggc acccttgagg cctccgtgga gggacgtcac ggggcagagc 240 gggacgtgag cctgagtttg ctgcaggcgt gctctgtgtg gtggctgggt tctgccaatc 300 cccgtgccca ccgggtgggc gcggccggga agctcctgcc cctccctgct ggtcggcgtc 360 acgcgtgacg tcccgcgtga tggctgggag ggcccggcgg cgacagcgga ggcagagagg 420 aaggcggttc tgagagcttc agagagcgat ggaaagcaaa atgggtgaat tgcctttaga 480 catcaacatc caggaacctc gctgggacca aagtactttc ctgggcagag cccggcactt 540 tttcactgtt actgatcctc gaaatctgct gctgtccggg gcacagctgg aagcttctcg 600 gaacatcgtg cagaactaca gggccggcgt ggtgacccca gggatcaccg aggaccagct 660 gtggagggcc aagtatgtgt atgactccgc cttccatccg gacacagggg agaaggtggt 720 cctgattggc cgcatgtcag cccaggtgcc catgaacatg accatcactg gctgcatgct 780 cacattctac aggaagaccc caaccgtggt gttctggcag tgggtgaatc agtccttcaa 840 tgccattgtt aactactcca accgcagtgg tgacactccc atcactgtga ggcagctggg 900 gacagcctat gtgagtgcca ccactggagc tgtggccacg gccctgggac tcaaatccct 960 caccaagcac ctgcccccct tggtcggcag atttgtgccc tttgcagcag tggcagctgc 1020 caactgcatc aacatccccc tgatgaggca gagagagctg caggtgggca tcccggtggc 1080 tgatgaggca ggtcagaggc ttggctactc ggtgactgca gccaagcagg gaatcttcca 1140 ggtggtgatt tcaagaatct gcatggcgat tcctgccatg gccatcccac cactgatcat 1200 ggacactctg gagaagaaag acttcctgaa ggtaggcgac tgtacctctc ttgtcctgga 1260 atgggcgatg gctgggagaa gtgaccaggc cccaactctc tctccagcct cgcctgattc 1320 tctaagactt gccagccctt ctcctgaccc ctgcaccgcc tcctccacct tcgttcattc 1380 agcaagaatg aactgggctg gggtgaagga actctgcagg ggcaggagga gaggacaaag 1440 gaaggaaacc aacttcatca gtgttactcc agtggcttct gacacacaga aggggactgt 1500 catagtcatg cttgatctca tgctcattct tttaccccct agtgcctcca tactgagagg 1560 tacacacggg tgaacacgca cacacagaca tgaacaggac acgaaagcaa agcacaggaa 1620 caagctctgg ctcattcaca gaatcattta ttcacaaatg tattgagtgc catgcaccag 1680 gcatgtttta gggctgagga gatggcactg aacacaatgg ttatggcccc tgtcctcatg 1740 aagtttatag tctgatgcag. aaaccaataa acaaggaggc acccacataa atacattctt 1800 agaaagtgta aaaataaaaa aaaaaa 1826 <210> 47 <211> 1325 <212> DNA
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 2501034CB1 <400> 47 ccacgggtcc ggttctggac tgcagttgag tggaaatggg caacggcggg cggagcggcc 60 tgcagcaggg gaaggggaac gtggatgggg tggcagcgac tcctactgct gcctcggcct 120 cctgccagta caggtgcatc gaatgcaacc aggaggccaa agagttgtac cgagactata 180 accacggtgt gctgaagata accatctgta aatcctgcca gaaacctgta gacaaatata 240 tcgagtatga tcctgttatc atcttgatta atgctatatt gtgcaaagct caggcctaca 300 gacatattct tttcaatact caaataaata tccatggaaa actctgcata ttttgtttgc 360 tttgtgaagc atacctgagg tggtggcagc ttcaagattc caaccagaat actgcccctg 420 atgacttgat cagatatgct aaggaatggg atttctatag aatgtttgcg attgctgctt 480 tagaacaaac tgcctatttt attggcattt ttaccttcct gtgggtagaa cggcccatga 540 cggcaaaaaa aaagcccaac ttcattttgc tgctgaaagc attattatta tctagctacg 600 gaaaactctt gctgattcca gctgtcattt gggaacatga ctacacatct gtgtgcctca 660 aactcattaa agtatttgtt cttacatcaa attttcaggc aattagagtg accctaaaca 720 tcaaccgtaa gctctccttc ttggccgtgt tgagtggctt actgctggaa agcatcatgg 780 tctacttctt ccagagtatg gaatgggatg ttggaagtga ttatgccatc tttaaatctc 840 aggacttctg aagagtttta ttcttcttca ctatctgtgg catgaccagc tgtatctgaa 900 agagaaaaga catgaaatat aaaccaacct cctcatttct gttgagtaaa atgaagcaaa 960 gattggaaac actttctgaa aaagaaagca atgataatag cggtggatac ccacccccac 1020 aaatgcaccc aagagacaag ccatttacat acagatattc acagtcacac atagaaacac 1080 ccacatggac acaaggaatg ttgctgcaga gactgaatga catgcaacag gtgaaggttt 1140 atacgttata cacaaggcca ggtaagcgct cataattcac acataataaa acatctaggt 1200 ttcattcctt tgacatgttt atatcttttt aatttaaatg ttgttactgg cttaaaatat 1260 tttgtgttct tacaatagaa acgcttttaa taaagtcttt cagaataaac caaaaaaaaa 1320 aaaaa 1325 <210> 48 <211> 1832 <212> DNA
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 2745212CB1 <400> 48 tgggctgtcg ttggctggag cagcggctgc gcgggtcgcg gtgctgtgag gtctgcgggc 60 gctggcaaat ccggcccagg atgtagagct ggcagtgcct gacggcgcgt ctgacgcgga 120 gttgggtggg gtagagagta gggggcggta gtcgggggtg gtgggagaag gaggaggcgg 180 cgaatcactt ataaatggcg ccgaagcagg acccgaagcc taaattccag gagggtgagc 240 gagtgctgtg ctttcatggg cctcttcttt atgaagcaaa gtgtgtaaag gttgccataa 300 aggacaaaca agtgaaatac ttcatacatt acagtggttg gaataaaaat tgggatgaat 360 gggttccgga gagcagagta ctcaaatacg tggacaccaa tttgcagaaa cagcgagaac 420 ttcaaaaagc caatcaggag cagtatgcag aggggaagat gagaggggct gccccaggaa 480 agaagacatc tggtctgcaa cagaaaaatg ttgaagtgaa aacgaaaaag aacaaacaga 540 aaacacctgg aaatggagat ggtggcagta ccagtgagac ccctcagcct cctcggaaga 600 aaagggcccg ggtagatcct actgttgaaa atgaggaaac attcatgaac agagttgaag 660 ttaaagtaaa gattcctgaa gagctaaaac cgtggcttgt tgatgactgg gacttaatta 720 ccaggcaaaa acagctcttt tatcttcctg ccaagaagaa tgtggattcc attcttgagg 780 attatgcaaa ttacaagaaa tctcgtggaa acacagataa taaggagtat gcggttaatg 840 aagttgtggc agggataaaa gaatacttca acgtaatgtt gggtacccag ctactctata 900 aatttgagag accacagtat gctgaaattc ttgcagatca tcccgatgca cccatgtccc 960 aggtgtatgg agcgccacat ctcctgagat tatttgtacg aattggagca atgttggctt 1020 atacacctct ggatgagaag agccttgctt tattactcaa ttatcttcac gatttcctaa 1080 agtacctggc aaagaattct gcaactttgt tcagtgccag cgattatgaa gtggctcctc 1140 ctgagtacca tcggaaagct gtgtgagagg cactctcact cacttatgtt tggatctccg 1200 taaacacatt tttgttctta gtctatctct tgtacaaacg atgtgctttg aagatgttag 1260 tgtataacaa ttgatgtttg ttttctgttt gattttaaac agagaaaaaa taaaaggggg 1320 taatagctcc ttttttcttc tttctttttt tttttcattt caaaattgct gccagtgttt 1380 tcaatgatgg acaacagagg gatatgctgt agagtgtttt attgcctagt tgacaaagct 1440 gcttttgaat gctggtggtt ctattccttt gacactacgc acttttataa tacatgttaa 1500 tgctatatga caaaatgctc tgattcctag tgccaaaggt tcaattcagt gtatataact 1560 gaacacactc atccatttgt gcttttgttt ttttttatgg tgcttaaagt aaagagccca 1620 tcctttgcaa gtcatccatg ttgttactta ggcattttat cttggctcaa attgttgaag 1680 aatggtggct tgtttcatgg tttttgtatt tgtgtctaat gcacgtttta acatgataga 1740 cgcaatgcat tgtgtagcta gttttctgga aaagtcaatc ttttaggaat tgtttttcag 1800 atcttcaata aattttttct ttaaatttca as 1832 <210> 49 <211> 1211 <212> DNA
<213> Homo sapiens <220>
<221> misc_feature <223> Incyte ID No: 4833111CB1 <400> 49 gacagacccg cctcaaacat ggcggcgccc agcgcgcgag gacgtgatcc gcttctgctc 60 cggcttggat tgtagccttg acgaggtctg agcgaccatg gaccggccgg ggttcgtggc 120 agcgctggtg gctggtgggg tagcaggtgt ttctgttgac ttgatattat ttcctctgga 180 taccattaaa accaggctgc agagtcccca aggatttagt aaggctggtg gttttcatgg 240 aatatatgct ggcgttcctt ctgctgctat tggatccttt cctaatgctg ctgcattttt 300 tatcacctat gaatatgtga agtggttttt gcatgctgat tcatcttcgt atttgacacc 360 tatgaaacat atgttggctg cctctgctgg agaagtggtt gcctgcctga ttcgagttcc 420 atctgaagtg gttaagcaga gggcacaggt atctgcttct acaagaacat ttcagatttt 480 ctctaacatc tr.atatgaag agggtatcca agggttgtat cgaggctata aaagcacagt 540 tttaagagag attccttttt ctttggtcca gtttccctta tgggagtcct taaaagccct 600 ctggtcctgg aggcaggatc atgtggtgga ttcttggcag tcagcagtct gtggagcttt 660 tgcaggtgga tttgccgctg cagtcaccac ccctctagac gtggcaaaga caagaattac 720 gctggcaaag gctggctcca gcactgctga tgggaatgtg ctctctgtcc tgcatggggt 780 ctggcggtca caggggctgg caggattatt tgcaggtgtc ttccctcgaa tggcagccat 840 cagtctggga ggtttcatct ttctgggggc ttatgaccga acgcacagct tgctgttgga 900 agttggcaga aagagtcctt gaagcagaga caagcctcac ctccacttct gtcaagagag 960 gggcctgcag tgcaaaccct cttccgctga gcagctgtct gaactatagg ccccagtgct 1020 gaagaccagt tgtgctaaga taccggcatg gagattgtgc catccgtggt ataggctggc 1080 tggtatgaag tcattggcct gtatgccaga gagctaagag aagaaaacgg ggtctgtggc 1140 ggtactctga acaatttcct cagaacctct taataaataa gtttggtaat gctgagaaaa 1200 aaaaaaaaaa a 1211 <210> 50 <211> 1046 <212> DNA
<213> Homo sapiens <220>
<221> misc_feature <223> Incyte ID No: 876677CB1 <400> 50 cccacgcgtc cgggaatgtc tttcactctc tcatactttc tcctctcccc tctcccaagc 60 acatctgagt tgctgcctgt tcttcacact tagctccaaa cccatgaaaa attgccaagt 120 ataaaagctt ctcaagaatg agatggattc tagggtgtct tcacctgaga agcaagataa 180 agagaatttc gtgggtgtca acaataaacg gcttggtgta tgtggctgga tcctgttttc 240 cctctctttc ctgttggtga tcattacctt ccccatctcc atatggatgt gcttgaagat 300 cattaaggag tatgaacgtg ctgttgtatt ccgtctggga cgcatccaag ctgacaaagc 360 caaggggcca ggtttgatcc tggtcctgcc atgcatagat gtgtttgtca aagttgacct 420 ccgaacagtt acttgcaaca ttcctccaca agagatcctc accagagact ccgtaactac 480 tcaggtagat ggagttgtct attacagaat ctatagtgct gtctcagcag tggctaatgt 540 caacgatgtc catcaagcaa catttctgct ggctcaaacc actctgagaa atgtcttagg 600 gacacagacc ttgtcccaga tcttagctgg acgagaagag atcgcccata gcatccagac 660 tttacttgat gatgccaccg aactgtgggg gatccgggtg gcccgagtgg aaatcaaaga 720 tgttcggatt cccgtgcagt tgcagagatc catggcagcc gaggctgagg ccacccggga 780 agcgagagcc aaggtccttg cagctgaagg agaaatgaat gcttccaaat ccctgaagtc 840 agcctccatg gtgctggctg agtctcccat agctctccag ctgcgctacc tgcagacctt 900 gagcacggta gccaccgaga agaattctac gattgtgttt cctctgccca tgaatatact 960 agagggcatt ggtggcgtca gctatgataa ccacaagaag cttccaaata aagcctgagg 1020 tcctcttgcg gtagtcagct attgca 1046 <210> 51 <211> 1660 <212> DNA
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 2326143C81 <400> 51 gctcccctgc ccaccccgcc cccgtggccg agcccgggag tcgagtggga gtcggccggc 60 cggcgcgggc agccgtgacc cccgcggggg acactgcagc cggagcccgg gaggggccgc 120 gccgccaccg tctgaactag gatgtcccga catgaaggtg tcagctgtga tgcatgttta 180 aaaggaaatt ttcgaggtcg cagatataag tgtttaattt gctacgatta cgatctttgt 240 gcatcttgtt atgaaagtgg tgcaacaaca acaaggcata caactgacca cccaatgcag 300 tgcatattaa caagggtaga ttttgattta tactatggtg gggaagcttt ctctgtagag 360 cagccacagt cttttacttg tccctattgt ggaaaaatgg gctatacgga gacatctctt 420 caagaacatg ttacttctga acatgcagaa acatcaacag aagtgatttg tccaatatgt 480 gcagcgttac ctggaggcga tcctaatcat gtcacggatg actttgcagc tcatcttaca 540 cttgaacaca gagcccctag agatttagat gaatcgagtg gtgttcgaca tgtacgtaga 600 atgtttcacc ctggccgggg attaggaggt cctcgtgctc gtagatcaaa catgcacttt 660 actagcagtt ctactggtgg actttcttct tctcagagtt catattctcc aagcaatagg 720 gaagccatgg atcctatagc tgagctttta tctcagttat caggagtgag acgttctgca 780 ggaggacagc ttaattcctc tggcccttcc gcttctcagt tacaacaact gcagatgcag 840 ctgcagctag aacggcagca tgcccaggca gcacggcaac aactggagac cgcacgcaac 900 gcaacccggc gtactaacac aagcagtgtc accactacaa tcacacaatc cacagcaaca 960 accaacatag ctaatacaga aagcagtcag cagactctac agaattccca gtttctttta 1020 acaaggttga atgatcctaa aatgtctgaa acggagcgcc agtccatgga aagcgagcgt 1080 gcagaccgca gcctgtttgt ccaagagctc cttctgtcca ctttagtgcg tgaagagagc 1140 tcatcctcag atgaggatga tcggggggag atggcagatt ttggtgctat gggctgtgta 1200 gatattatgc ctttagatgt tgctttagaa aacctaaatt taaaagagag taataaagga 1260 aatgagcctc caccacctcc tctttgatga catcccaatt cgcagacaat gtcctctgtg 1320 ctgtatttgc caatgaaagt ggacaacaac tatcttgggt ttgtttggtg attgtaattt 1380 caggtctgtc actcttgtta cattgtgtac attcaaaagg aagagagaaa atatatatga 1440 taatcatttc cacttaacta atttttactt ctagcaggta aatgtaggta gcagtgcagg 1500 ggtgatctct gcttcctgta ccttgacatg caaaaggctc tcctaatact ccacattcaa 1560 actgaagagg aaaattgaaa tctctaatga agctgctgtg tgtatttatg aatattaatg 1620 aataaaaact gcttggatgg tttaccttaa aaaaaaaaaa 1660 <210> 52 <211> 1110 <212> DNA
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 2786302CB1 <400> 52 cctttattag agaaagagtg acctatctac cccttacttg gggaagactc tctgcttcaa 60 cagtttcttc tgtcttctgc tcctggacat ggcatgaaaa cggaagtttg agtatttcaa 120 ggataaagaa taactctagg acaagagttg ctttggggct aaacctatct cctgtattca 180 tttagatttg gctttaaaag cccatactat tttattagct cttctatgct ttttaaaagt 240 ttttaaaaaa attaatgtgg gctttttaat tctttctcag tgggagggtg agtttgaata 300 aacctttctt ccacatgaga agtattttac aagttgcttg tcaaatttaa aagaaaatga 360 tcaaatcttc aagaaaatga tcaaatcttc tgtgacaaaa aaatggacaa atattcacca 420 ttgagtgtga atgccatggt gatgctaatt ctgattattt ttcttttctt ttagccatac 480 cttcagagtg ttattttcct ttttgtcata aggtgtctgg aaatgaagta tggaaatgaa 540 ataatgaata aagacccagt tttcagaatc tctccacgga gtagagaaac tcatcccaat 600 ccggaagagc ccgaagaaga agatgaagat gttcaagctg aaagagtcca agcagcaaat 660 gcactcactg ctccaaactt ggaggaggaa ccagtcataa ctgcaagctg tttacacaag 720 gaatattatg agacaaagaa aagttgcttt tcaacaagaa agaagaaaat agccatcaga 780 aatgtttcct tttgtgttaa aaaaggtgaa gttttgggat tactaggaca caatggagct 840 ggtaaaagta cttccattaa aatgataact gggtgcacaa agccaactgc aggagtggtg 900 gtgttacaag gcagcagagc atcagtaagg caacagcatg acaacagcct caagttcttg 960 gggtactgcc ctcaggagaa ctcactgtgg cccaagctta ccatgaaaga gcacttggag 1020 ttgtatgcag ctgtggaaag actggggcaa aaaagatgct gctctcagta tttcacgatt 1080 ggtgggaggt cttaagctcc aggaacaact 1110 <210> 53 <211> 1120 <212> DNA
<213> Homo sapiens <220>
<221> misc_feature <223> Incyte ID No: 3735780CB1 <400> 53 gaaatccagt tatcaaaatt gactcaagaa gagagaacct aacagaacaa taacaatgga 60 agaaattggg aacattatca caaagctatc atcctgccaa actccaggct cagatgtcac 120 aggttaaaaa aagtccttca tgaaaaagaa agatcttaag cagcatgatg gattcagaag 180 ctcatgaaaa gaggccacca atactaacat cttcaaaaca agatatatca cctcatatta 240 caaatgttgg tgagatgaag cattacttgt gtggctgctg tgcagccttc aacaatgtcg 300 caatcacatt tcccattcag aaggtcctct ttcgacaaca gctgtatggc atcaaaaccc 360 gggatgcaat acttcagttg agaagggatg gatttcgaaa tttgtatcgt ggaatccttc 420 ccccattgat gcagaagaca actacgcttg cacttatgtt tggtctgtat gaggatttat 480 cctgccttct ccacaagcat gtcagtgctc cagagtttgc aaccagtggc gtggcggcag 540 tgcttgcagg gacaacagaa gcaattttca ctccactgga aagagttcag acattgcttc 600 aagaccacaa acatcatgac aaatttacca acacttacca ggctttcaag gcactgaaat 660 gtcatggaat tggagagtat tatcgaggct tggtgcccat tcttttccgg aatggactca 720 gcaatgtctt gtttttcggc cttcgaggtc ccattaagga gcatctgcct accgcaacga 780 ctcacagtgc tcatctggtc aatgatttta tctgtggagg tctattgggt gccatgttgg 840 gattcttgtt ttttccaatt aatgttgtaa aaactcgcat acagtctcag attggtgggg 900 aatttcagtc tttccccaag gttttccaaa aaatctggct ggaacgggac agaaaactga 960 taaatctttt cagaggtgcc catctgaatt accatcggtc cctcatctct tggggcataa 1020 tcaatgcaac ttatgagttc ttgttaaagg ttatatgaaa aaaccatcag ttaagtgcca 1080 tttatcaact gaatagacct tctaagaaga aaaaaaaaaa 1120 <210> 54 <211> 886 <212> DNA
<213> Homo sapiens <220>
<221> misc_feature <223> Incyte ID No: 039026CB1 <400> 54 ggccgcggct cctgtccaga ccctgaccct ccctcccaag gctcaaccgt cccccaacaa 60 ccgccagcct tgtactgatg tcggatgcga gagcctgtgc ttaagtaaga atcaggcctt 120 attggagaca ttcaagcaaa ggttggacaa ctacttttcc agaacagaaa ggaaactcat 180 gcatcagaaa aggtgactaa taaaggtacc agaagaatat ggctgcacaa ataccagaat 240 ctgatcagat aaaacagttt aaggaatttc tggggaccta caataaactt acagagacct 300 gctttttgga ctgtgttaaa gacttcacaa caagagaagt aaaacctgaa gagaccacct 360 gttcagaaca ttgcttacag aaatatttaa aaatgacaca aagaatatcc atgagatttc 420 aggaatatca tattcagcag aatgaagccc tggcagccaa agcaggactc cttggccaac 480 cacgatagag aagtcctgat ggatgaactt ttgatgaaag attgccaaca gctgctttat 540 tggaaatgag gactcatctg atagaatccc ctgaaagcag tagccaccat gttcaaccat 600 ctgtcatgac tgtttggcaa atggaaaccg ctggagaaac aaaattgcta tttaccagga 660 ataatcacaa tagaaggtct tattgttcag tgaaataata agatgcaaca tttgttgagg 720 ccttatgatt cagcagcttg gtcacttgat tagaaaaata aaccattgtt tcttcaattg 780 tgactgttaa ttttaaagca acttatgtgt tcgatcatgt atgagataga aaaattttta 840 ttactcaaag taaaataaat ggaaatatca ctgaaaaaaa aaaaaa 886 <210> 55 <211> 2336 <212> DNA
<213> Homo sapiens <220>
<221> misc_feature <223> Incyte ID No: 260607CB1 <400> 55 taatacgctc actataggga atttggccct cgagcagtaa ttcggcacga ggaccatctc 60 tttaggatat atttttaaat tctttgaaac acataaccaa aatggtttga ttcactgact 120 gactttgaag ctgcatctgc cagttacacc ccaaatggct ttaatcccct ctcgggtctg 180 gttgcctttt gcagtttggg ttgtggactc agctcctgtg aggggtctgg ttaggagaga 240 gccattttta aggacaggga gttttatagc ccttttctac tttcctcccc tcctcccagt 300 ccttatcaat cttttttcct ttttcctgac cccctccttc tggaggcagt tgggagctat 360 ccttgtttat gcctcactat tggcagaaaa gaccccattt aaaacccaga gaacactgga 420 gggggatgct ctagttggtt ctgtgtccat tttcctctgt gccaaagaca gacagacaga 480 ggctgagaga ggctgttcct gaatcaaagc aatagccagc tttcgacaca tacctggctg 540 tctgaggagg aaggcctcct ggaaactggg agctaagggc gaggcccttc ccttcagagg 600 ctcctggggg attagggtgt ggtgtttgcc aagccaaggg gtagggagcc gagaaattgg 660 tctgtcggct cctggttgca ctttggggaa ggagaggaag tttggggctc caggtagctc 720 cctgttgtgg gactgctctg tcccctgccc ctactgcaga gatagcactg ccgagttccc 780 ttcaggcctg gcagacgggc agtgaggagg ggcctcagtt agctctcaag ggtgccttcc 840 cctcctccca acccagacat accctctgcc aaactgggaa ccagcagtgc tagtaactac 900 ctcacagagc cccagagggc ctgcttgagc cttcttgctc cacaggagaa gctggtgcct 960 ctaggcaacc ccttcctccc acctctcatc aggggtgggg gttctccttt ctttcccctg 1020 aagtgtttat ggggagatcc tagtggcttt gccattcaaa ccactcgact gtttgcctgt 1080 ttcttgaaaa ccagtagaag ggaaacagca cagcctgtca cagtaattgc aggaagattg 1140 aagaaaaatc ctcatcaatg ccaggggaca taaaagccat ttcccttcca aatactcgac 1200 aatttagatg cagaacattt ctctgtattc agacttagag taacaccagc tgaaaactgc 1260 agtttctttc ctttggatac ataaggcttc tctatcgggg tacgggacag ggaggaggcc 1320 tcatgtctga agggggattt aggggcgaga gccgcagccc tgaccctcgg tcctgtgcac 1380 cgctttgggg cacagtctga tggcgccttt gctggcgcct tagtatggtt gactccggat 1440 ggacaaaaga aaaaaaattt tttttcttga atgaaatagc aggaagctcc tcgggagcat 1500 gtgttttgat taaccgtagt gatggatgct acgagtataa atggattaac tacctcaatc 1560 cttacagtaa gattggaact aagggcaggg actcatgcat aagggtatga atcccagcca 1620 ggacaagtga gttgaggctt gtgccacaaa aggtttgtcc ttggggaaca ggcaggcctg 1680 ccaggatccc ccccatatcg attgggctgg gagggctggc cgtgaggtcc ccactttctg 1740 ctttccttgc ccatgtgtca cccctttggc ctccagcttg tccctctctc actttctata 1800 gctttgttgg accagatggt gaggaaagga atggcctctt cccttctaga gggggctggc 1860 tggagtgaga cctggggctt ggcctggaac ccaccacaca gccccaaagt caggaagcct 1920 ggggaaacca gagctgagac ctcttcaaca gggtttcttt gagatcctac acctccattg 1980 ggcccttttt cagtcttcaa tgggggccca gttggctcta gaaggagaag aggtgaagca 2040 ggatcctttg ccctggggga gtctgagggc gcggtccttg gactcattca ggccgtcttt 2100 gtagttgggg gagttccact gggcgatccc agcccctccc cacccaccct ctaatggacc 2160 tcctcataga agccccattt cacttttgtt ttatctacct cttagcaaaa caatagataa 2220 attaggtagt ggcagctcca cttgcttagg ttaggggggg aaaaagattt ctttttccaa 2280 aggaaaaaaa tattaccttg agaatacttt ccaaaaaata aaatttaaaa aaaaaa 2336 <210> 56 <211> 2200 <212> DNA
<213> Homo sapiens <220>
<221> misc_feature <223> Incyte ID No: 1429651CB1 <400> 56 gagaacttta cgcctggatc tcatctaact gacacagaaa ccctgtaagg atccagaggt 60 ctcgttcagg accatggaga gcggcaccag cagccctcag cctccacagt tagatcccct 120 ggatgcgttt ccccagaagg gcttggagcc tggggacatc gcggtgctag ttctgtactt 180 cctctttgtc ctggctgttg gactatggtc cacagtgaag accaaaagag acacagtgaa 240 aggctacttc ctggctggag gggacatggt gtggtggcca gtgggtgcat ccttgtttgc 300 cagcaatgtt ggaagtggac atttcattgg cctggcaggg tcaggtgctg ctacgggcat 360 ttctgtatca gcttatgaac ttaatggctt gttttctgtg ctgatgttgg cctggatctt 420 cctacccatc tacattgctg gtcaggtcac cacgatgcca gaatacctac ggaagcgctt 480 cggtggcatc agaatcccca tcatcctggc tgtactctac ctatttatct acatcttcac 540 caagatctcg gtagacatgt atgcaggtgc catcttcatc cagcagtctt tgcacctgga 600 tctgtacctg gccatagttg ggctactggc catcactgct gtatacacgg ttgctggtgg 660 cctggctgct gtgatctaca cggatgccct gcagacgctg atcatgctta taggagcgct 720 caccttgatg ggctacagtt tcgccgcggt tggtgggatg gaaggactga aggagaagta 780 cttcttggcc ctggctagca accggagtga gaacagcagc tgcgggctgc cccgggaaga 840 tgccttccat attttccgag atccgctgac atctgatctc ccgtggccgg gggtcctatt 900 tggaatgtcc atcccatccc tctggtactg gtgcacggat caggtgattg tccagcggac 960 tctggctgcc aagaacctgt cccatgccaa aggaggtgct ctgatggctg catacctgaa 1020 ggtgctgccc ctcttcataa tggtgttccc tgggatggtc agccgcatcc tcttcccaga 1080 tcaagtggcc tgtgcagatc cagagatctg ccagaagatc tgcagcaacc cctcaggctg 1140 ttcggacatc gcgtatccca aactcgtgct ggaactcctg cccacagggc tccgtgggct 1200 gatgatggct gtgatggtgg cggctctcat gtcctccctc acctccatct ttaacagtgc 1260 cagcaccatc ttcaccatgg acctctggaa tcacctccgg cctcgggcat ctgagaagga 1320 gctcatgatt gtgggcaggg tgtttgtgct gctgctggtc ctggtctcca tcctctggat 1380 ccctgtggtc caggccagcc agggcggcca gctcttcatc tatatccagt ccatcagctc 1440 ctacctgcag ccgcctgtgg cggtggtctt catcatggga tgtttctgga agaggaccaa 1500 tgaaaagggt gccttctggg gcctgatctc gggcctgctc ctgggcttgg ttaggctggt 1560 cctggacttt atttacgtgc agcctcgatg cgaccagcca gatgagcgcc cggtcctggt 1620 gaagagcatt cactacctct acttctccat gatcctgtcc acggtcaccc tcatcactgt 1680 ctccaccgtg agctggttca cagagccacc ctccaaggag atggtcagcc acctgacctg 1740 gtttactcgt cacgaccccg tggtccagaa ggaacaagca ccaccagcag ctcccttgtc 1800 tcttaccctc tctcagaacg ggatgccaga ggccagcagc agcagcagcg tccagttcga 1860 gatggttcaa gaaaacacgt ctaaaaccca cagctgtgac atgaccccaa agcagtccaa 1920 agtggtgaag gccatcctgt ggctctgtgg aatacaggag aagggcaagg aagagctccc 1980 ggccagagca gaagccatca tagtttccct ggaagaaaac cccttggtga agaccctcct 2040 ggacgtcaac ctcattttct gcgtgagctg cgccatcttt atctggggct attttgctta 2100 gtgtggggtg aacccagggg tccaaactct gtttctcttc agtgctccat ttttttaatg 2160 aaagaaaaaa taataaagct tttgtttacc aaaaaaaaaa 2200 <210> 57 <211> 2823 <212> DNA
<213> Homo sapiens <220>
<221> misc_feature <223> Incyte ID No: 2069971CB1 <400> 57 gaaagacata cacacttcat gtaatgctac ctgcaagtct ccctagaaaa gcagtttttg 60 taggtgaaaa caatgaagcc aggtaatatt gcaaggaggc tgtaatttta gcagacctac 120 caacaacact gatgtaggaa gctcattatt ttaatttctg gagcctttta attttttctt 180 tagaaagtgt ataaataatt gcagtgctgc tttgcttcca aaactgggca gtgagttcaa 240 caacaacgac aacaacagcc gcagctcatc ctggccgtca tggagtttct tgaaagaacg 300 tatcttgtga atgataaagc tgccaagatg tatgctttca cactagaaag aaggagctgc 360 aaatgaacac ttcatagcaa tgtggaactc caacagaaac cggtgaataa agatcagtgt 420 cccagagaga gaccagagga gctggagtca ggaggcatgt accactgcca cagtggctcc 480 aagcccacag aaaagggggc gaatgagtac gcctatgcca agtggaaact ctgttctgct 540 tcagcaatat gcttcatttt catgattgca gaggtcgtgg gtgggcacat tgctgggagt 600 cttgctgttg tcacagatgc tgcccacctc ttaattgacc tgaccagttt cctgctcagt 660 ctcttctccc tgtggttgtc atcgaagcct ccctctaagc ggctgacatt tggatggcac 720 cgagcagaga tccttggtgc cctgctctcc atcctgtgca tctgggtggt gactggcgtg 780 ctagtgtacc tggcatgtga gcgcctgctg tatcctgatt accagatcca ggcgactgtg 840 atgatcatcg tttccagctg cgcagtggcg gccaacattg tactaactgt ggttttgcac 900 cagagatgcc ttggccacaa tcacaaggaa gtacaagcca atgccagcgt cagagctgct 960 tttgtgcatg cccttggaga tctatttcag agtatcagtg tgctaattag tgcacttatt 1020 atctacttta agccagagta taaaatagcc gacccaatct gcacattcat cttttccatc 1080 ctggtcttgg ccagcaccat cactatctta aaggacttct ccatcttact catggaaggt 1140 gtgccaaaga gcctgaatta cagtggtgtg aaagagctta ttttagcagt cgacggggtg 1200 ctgtctgtgc acagcctgca catctggtct ctaacaatga atcaagtaat tctctcagct 1260 catgttgcta cagcagccag ccgggacagc caagtggttc ggagagaaat tgctaaagcc 1320 cttagcaaaa gctttacgat gcactcactc accattcaga tggaatctcc agttgaccag 1380 gaccccgact gccttttctg tgaagacccc tgtgactagc tcagtcacac cgtcagtttc 1440 ccaaatttga caggccacct tcaaacatgc tgctatgcag tttctgcatc atagaaaata 1500 aggaaccaaa ggaagaaatt catgtcatgg tgcaatgcac attttatcta tttatttagt 1560 tccattcacc atgaaggaag aggcactgag atccatcaat caattggatt atatactgat 1620 cagtagctgt gttcaattgc aggaatgtgt atatagatta ttcctgagtg gagccgaagt 1680 aacagctgtt tgtaactatc ggcaatacca aattcatctc ccttccaata atgcatcttg 1740 agaacacata ggtaaatttg aactcaggaa agtcttacta gaaatcagtg gaagggacaa 1800 atagtcacaa aattttacca aaacattaga aacaaaaaat aaggagagcc aagtcaggaa 1860 taaaagtgac tctgtatgct aacgccacat tagaacttgg ttctctcacc aagctgtaat 1920 gtgatttttt tttctactct gaattggaaa tatgtatgaa tatacagaga agtgcttaca 1980 actaattttt atttacttgt cacattttgg caataaatcc ctcttatttc taaattctaa 2040 cttgtttatt tcaaaacttt atataatcac tgttcaaaag gaaatatttt cacctaccag 2100 agtgcttaaa cactggcacc agccaaagaa tgtggttgta gagacccaga agtcttcaag 2160 aacagccgac aaaaacattc gagttgaccc caccaagttg ttgccacaga taatttagat 2220 atttacctgc aggaaggaat aaagcagatg caaccaattc attcagtcca cgagcatgat 2280 gtgagcactg ctttgtgcta gacattgggc ttagcattga aactataaag aggaatcaga 2340 cgcagcaagt gcttctgtgt tctggtagca actcaacact atctgtggag agtaaactga 2400 agatgtgcag gccaacattc tggaaatcct atgtcactgg gtttggtttg gaaacctgga 2460 cttctgcatt tttaaaagtt acccagagat gcttctaaag atgagccata gtctagaaga 2520 ttgtcaacca caggagttca ttgagtggga cagctagata catacattgg cagctacaat 2580 agtatcatga attgcaatga tgtagtgggg tataaaagga aagcgatgga tattgccgga 2640 tgggcatggc cagtgatgtt tcacgtcatt gaggtgacag ctctgctgga ctttgaatta 2700 catatggagg ctctccagga agacgaagaa gagaaggaca ttctaggcaa aaagaagact 2760 aggcacaagg cacacttatg tttgtctgtt agcttttagt tgaaaaagca agatacaggg 2820 tcg 2823 <210> 58 <211> 1491 <212> DNA
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 2329339CB1 <400> 58 cgcctccctc cagctgcgag tgcggcctcg gctggcggcg gcaccaggcc acagttgtaa 60 gggatcttgt ggctgtcagg atggcagagg agcaggagtt cacccagctc tgcaagttgc 120 ctgcacagcc ctcacaccca cactgcgtga acaacaccta ccgcagcgca cagcactccc 180 aggctctgct ccgagggctg ctggctctcc gggacagcgg aatcctcttc gatgttgtgc 240 tggtggtgga gggcagacac atcgaggccc atcgcatcct gctggctgcg tcctgcgatt 300 acttcagagg aatgtttgct gggggattga aggagatgga acaggaagag gtcctgatcc 360 acggtgtgtc ctacaatgct atgtgccaaa tcctacattt catatacacc tccgagctgg 420 agctcagcct gagcaatgta caagagacac tggtggctgc ctgccagctt cagatcccag 480 aaattatcca tttctgctgt gatttcctca tgtcctgggt ggacgaagag aacattctcg 540 atgtctaccg gctggcagag ctgtttgact tgagccgcct gactgagcaa ctggacacct 600 atatcctcaa aaactttgtg gccttctctc ggactgacaa gtaccgccag cttccattgg 660 agaaggtcta ctccctcctc agcagcaatc gcctggaggt ctcctgcgag accgaggtat 720 atgagggggc ccttctctac cattatagcc tggagcaggt gcaggctgac cagatctcgc 780 tgcacgagcc cccaaagctc cttgagacag tgcggtttcc gctgatggaa gctgaggtcc 840 tgcagcggct gcatgacaag ctggacccca gccctttgag ggacacagtg gccagcggcc 900 tcatgtacca ccggaacgag agcctacagc ccagcctgca gagcccgcaa acggagctgc 960 ggtcggactt ccagtgcgtt gtgggcttcg ggggcattca ctccacgccg tccactgtcc 1020 tcagcgacca ggccaagtat ctaaacccct tactgggaga gtggaagcac ttcactgcct 1080 ccctggcccc ccgcatgtcc aaccagggca tcgcggtgct caacaacttc gtatacttga 1140 ttggagggga caacaatgtc caaggatttc gagcagagtc ccgatgctgg aggtatgacc 1200 cacggcacaa ccgctggttc cagatccagt ccctgcagca ggagcacgcc gacctgtccg 1260 tgtgtgttgt aggcaggtac atctacgctg tggcgggccg tgactaccac aatgacctga 1320 atgctgtgga gcgctacgac cctgccacca actcctgggc atacgtggcc ccactcaaga 1380 gggaggtgta tgcccacgca ggcgcgacgc tggaggggaa gatgtatatc acctgcggac 1440 gcaagcttat tccctttagt gagggttaat tttagcttgc actggccgtc g 1491 <210> 59 <211> 986 <212> DNA
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 2540219CB1 <400> 59 ggacgccaac ctcggtttga agtccagggc agtggctcct gcggacagcc agcataccag 60 gggccagtgc actgcattac aaccattgtg aggaatgagg gcctgtgccg tgtggctggc 120 gggcggcatg gcaggagcaa tttcttgggg gacagcgact cctatggatg tcgtgaaaag 180 tcgactccaa gctgatgggg tttatttaaa caaatataaa ggtgtcctgg actgtatctc 240 ccagagttac cagaaggaag gtcttaaagt gtttttcaga ggcatcactg tgaacgcggt 300 gcggggcttc cccatgagtg cggccatgtt ccttgggtac gagctgtcgc tgcaggctat 360 ccgcggggac cacgcagtga cgagcccata agcgccagga ggtgaacaca ggatgactac 420 agtgttcccc tgggcctcat ctctgcatgt gaagccctga gagctgcaga tgtttgccct 480 ttggacctcc aagtggacat caattagcaa gcgtgggcta ggatggtgca gacactgacg 540 tggcccttct gatgcctggg atgcctcatg agtcactgat tcaagccctc caaggttctg 600 atccccaatg cccactctgc taggctggca tcaaagagct ttccaagaaa tgtttggtcc 660 agctgagaag tcctgaccat gagcaccagg gagccagaaa ccacccagag aaacgttgct 720 tcactcctct gtctgaggat ggggaggggc cagtgagctc tgggctcagc cactccctcc 780 agtctcaagt aacacgtccc cgtgcctcca gtctcctctc agcaccgacc aggtttttcc 840 ccgctcctgc acccgtggat cctgaggaca gcggtagcgc cttcctcacc gcacgctgag 900 tccagtgcgt gctcctcact gtgcacttat tagtgtctgt tgagtgatta aatcacatcc 960 tcaggtctgc agcaaataaa tgaaag 986 <210> 60 <211> 4023 <212> DNA
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 2722462CB1 <400> 60 gtccggggcg gcgcgtatgg cggcactatg ccatccacga ctcccaggcc cccagtctca 60 gctctggggg tgagagttcc ccctccagcc ccgcacacaa ctgggagatg aattaccaag 120 aggcagcaat ctacctccag gaaggcgaga acaacgacaa gttcttcacc caccccaagg 180 atgccaaggc gctggcggcc tacctctttg cacacaatca cctcttctac ctgatggagc 240 tggccacggc cctgctgctg ctgctgctct ccctgtgcga ggcccccgcc gtccccgcac 300 tccggcttgg catctatgtc cacgccaccc tggagctgtt tgccctgatg gtggtagtgt 360 ttgaactctg catgaagtta cgctggctgg gcctccacac cttcatccgg cacaagcgga 420 ccatggtcaa gacctcggtg ctggtggtgc agtttgtcga ggccatcgtg gtgttggtac 480 ggcagatgtc ccatgtgcgg gtgacccgag cactgcgctg cattttcctg gtggactgtc 540 ggtattgcgg tggcgtccgg cgcaacctgc ggcagatctt ccagtccctg ccgcccttca 600 tggacatcct cctgctgctg ctgttcttca tgatcatctt tgccatcctc ggtttctact 660 tgttctcccc taacccttca gacccctact tcagcaccct ggagaacagc atcgtcagtc 720 tgtttgtcct tctgaccaca gccaatttcc cagatgtgat gatgccctcc tactcccgga 780 acccctggtc ctgcgtcttc ttcatcgtgt acctctccat cgagctgtat ttcatcatga 840 acctgcttct ggctgtggtg ttcgacacct tcaatgacat tgagaaacgc aagttcaagt 900 ctttgctact gcacaagcga accgctatcc agcatgccta ccgcctgctc atcagccaga 960 ggaggcctgc cggcatctcc tacaggcagt ttgaaggcct catgcgcttc tacaagcccc 1020 ggatgagtgc cagggagcgc tatcttacct tcaaggccct gaatcagaac aacacacccc 1080 tgctcagcct aaaggacttt tacgatatct acgaagttgc tgctttgaag tggaaggcca 1140 agaaaaacag agagcactgg tttgatgagc ttcccaggac ggcgctcctc atcttcaaag 1200 gtattaatat ccttgtgaag tccaaggcct tccagtattt catgtacttg gtggtggcag 1260 tcaacggggt ctggatcctc gtggagacat ttatgctgaa aggtgggaac ttcttctcca 1320 agcacgtgcc ctggagttac ctcgtctttc taactatcta tggggtggag ctgttcctga 1380 aggttgccgg cctgggccct gtggagtact tgtcttccgg atggaacttg tttgacttct 1440 ccgtgacagt gttcgccttc ctgggactgc tggcgctggc cctcaacatg gagcccttct 1500 atttcatcgt ggtcctgcgc cccctccagc tgctgaggtt gtttaagttg aaggagcgct 1560 accgcaacgt gctggacacc atgttcgagc tgctgccccg gatggccagc ctgggcctca 1620 ccctgctcat cttttactac tccttcgcca tcgtgggcat ggagttcttc tgcgggatcg 1680 tcttccccaa ctgctgcaac acgagtacag tggcagatgc ctaccgctgg cgcaaccaca 1740 ccgtgggcaa caggaccgtg gtggaggaag gctactatta tctcaataat tttgacaaca 1800 tcctcaacag ctttgtgacc ctgtttgagc tcacagttgt caacaactgg tacatcatca 1860 tggaaggcgt cacctctcag acctcccact ggagccgcct ctacttcatg accttttaca 1920 ttgtgaccat ggtggtgatg acgatcattg tcgcctttat cctcgaggcc ttcgtcttcc 1980 gaatgaacta cagccgcaag aaccaggact cggaagttga tggtggcatc acccttgaga 2040 aggaaatctc caaagaagag ctggttgccg tcctggagct ctaccgggag gcacgggggg 2100 cctcctcgga tgtcaccagg ctgctggaga ccctctccca gatggagaga taccagcaac 2160 attccatggt gtttctggga cggcgatcaa ggaccaagag cgacctgagc ctgaagatgt 2220 accaggagga gatccaggag tggtatgagg agcatgccag ggagcaagag cagcagcgac 2280 aactcagcag cagtgcagcc cccgccgccc agcagccccc aggcagccgc cagcgctccc 2340 agaccgttac ctagcccagc gcccgaaagc cgtctcttct atgcaataac acaatagtat 2400 tactctactg cgatgtacgg aactgcggtg tgtgtacaca tactcacgta tatgcacata 2460 tttatataca ggaagaaaaa agacagacaa gatggggctt ggtttataac caccttgccc 2520 tgtcttcctt aactccagaa gccagtttgg tgaggggtgg gggtgcggcc accaggtctg 2580 agctcttcct actgtggaag gctccagaag gcccttcaca aggagacccc tcacctggat 2640 ccagtcgact gcggggcttg cccctcatgt gggctggcct ccatcggcca cgtccaaagc 2700 tgtcactgct actgcttcag gctcacatcc ccccgacctg atggcgtgcc cgccccctct 2760 ccctgcggcc catgccacag gtttctgtgt tttgctttag ggacagaacc acttaggaaa 2820 gaaagaactc ccggtctcca gggtggtatt tcagtgtctg tgataatgtc acgcaacacc 2880 tcttcgggga ccagtgccca ggatctaatg gaagcggaat tggggcaact gggcccgtgt 2940 ggccagagct cagttagcca gtgccgggcg gccacagatt acactgacca atctcctccc 3000 ttggctctgc aagcctccca cccagccttc tctggcttaa cccttgttgg cgaaaactct 3060 tccacagtgg cctccttggg gacccagaac ccggaggaag gggcatgagg caggaagtgg 3120 ggccgatgtc tgcaacccag accacttcgt ggaatgggct cttgaccaaa tccctttttt 3180 ttgcgattta cccgttcaag caaaacaacg ttttggttaa ctaaggattg tgctaaagcc 3240 gataccaggt ccttcacacg tgtgcactag gaacaggagc gaacagcaca gagagacgct 3300 ccctgtggga cgcagcagcc ccgtggcccc ggcccagttc ccagccaccc tccctggctc 3360 tgctcacacc agagatttcc atagcaggag cggttggtgc agaagtaggt tcagatgaac 3420 ctcagttaac gtcgccaccc ctcctcccac catggtaccc tgtaggagcc ctgtatgaca 3480 tctgagcgtg gtggaggtag gagggttgcc agctgcagtg accctgccac agaggcaggg 3540 tcagtgcaga ggtcgctttg gttccgcttc cctgggccac agaacggaac acagcatagg 3600 ttctgcagca ggagccgcag tggcaggatg gagggtgcga agggcaagga gtgcactgct 3660 gggcattcct ggccagcccc ggccctctgg tgcctgcttc ctgtgacttc agaaggcagg 3720 tggacagagc ctccctctgg ccttgtcctc ttcccagcca cagaacgggc agggtggcac 3780 ccgaccccag gggagcagta cctggtcccc caccccctcc tcccaaccac ctccaaggcc 3840 aagctgggtc ccatagccag cacggcatgg ttctcccctt ccccccttcc caggtcaggg 3900 gagttggaca agtagcaggt gtttgttttt aaagcacagc cctttgggaa agcaacacat 3960 tattgagact cactgtgatt cccccgggag tcagactggc tttgtctctt tctctctgga 4020 ggg 4023 <210> 61 <211> 2345 <212> DNA
<213> Homo sapiens <220>
<221> misc_feature <223> Incyte ID No: 2739264081 <400> 61 gaaaggaggc ggaactcggt gatctgactg gcggtttccc ggccggactg agaaggggag 60 cgcgctgcgc gtcgcaggag taacctactt ggtctcctgc tttcgcgaca tggccttcaa 120 ttttggggct ccctcgggca cctccggtac cgctgcagcc accgcggccc ccgcgggtgg 180 gtttggagga tttgggacaa catctacaac tgcaggttct gcattcagct tttctgcccc 240 aactaacaca ggcactactg gactctttgg tggtactcag aacaaaggtt ttggatttgg 300 tactggtttt ggcacaacaa cgggaactag tactggttta ggtactggtt tgggaactgg 360 actgggattt ggaggattta atacacagca gcagcagcaa actacattag gtggtctctt 420 cagtcagcct acacaagctc ctacccagtc caaccagctg ataaatactg cgagtgctct 480 ttctgctcca acgctgttgg gagatgagag agatgctatt ttggcaaaat ggaatcaact 540 gcaggccttt tggggaacag gaaaagggta tttcaacaat aatattccgc cagtggaatt 600 cacacaagaa aatccctttt gccgatttaa ggcagtaggt tatagttgca tgcccagtaa 660 taaagatgaa gatgggctag tggttttagt tttcaacaaa aaagaaacag agattcgaag 720 ccaacaacaa cagttggtag aatcattgca taaagttttg ggaggaaacc agacccttac 780 tgtaaatgta gagggcacta aaacattgcc agatgatcag acagaagttg ttatttatgt 840 tgttgagcgt tcgccaaatg gtacttcaag aagagttcca gctacaacgc tatatgccca 900 ttttgaacaa gccaatataa aaacacaatt gcagcaactt ggtgtaaccc tttctatgac 960 tagaacagaa ctttctcctg cacagatcaa acagctttta cagaatcctc ctgctggtgt 1020 tgatcctatt atctgggaac aggccaaggt agataaccct gattctgaaa agttaattcc 1080 tgtaccaatg gtgggtttta aggaacttct ccgaagactg aaggttcaag atcagatgac 1140 taagcagcat caaaccagat tagatatcat atctgaagat attagtgagc tacaaaagaa 1200 tcaaactaca tctgtagcca aaattgcaca atacaagagg aaactcatgg atctttccca 1260 tagaacttta caggtcctaa tcaaacagga aattcaaagg aagagtggtt atgccattca 1320 ggctgatgaa gagcagttgc gagttcagct ggatacgatt cagggtgaac taaatgcacc 1380 tactcagttc aagggccgac taaatgaatt gatgtctcaa atcaggatgc agaatcattt 1440 tggagcagtc agatctgaag aaaggtatta catagatgca gatctgttac gagaaatcaa 1500 gcagcatttg aaacaacaac aggaaggcct tagccatttg attagcatca ttaaagacga 1560 tctagaagat ataaagctgg tcgaacatgg attgaatgaa accatccaca tcagaggtgg 1620 tgtctttagt tgacagttca caaacttgtg taaaggtttg tgaaatgcat cttcttactg 1680 catcagacct tccttaagaa tgaaaccgac cacatggagg gaaaaagaaa acaattcttt 1740 cttggattgg ttttttgaga agtttactga caaattactg ttcatcaaat ctgaaatagt 1800 cacctcacag ctcttcaaag aaaacctttg aaagatttat atctaaaagc tgtatttact 1860 ttaaaagaag tgcataatta ccaaaattgt atgtactatt gtacattttt acaacagcat 1920 tttcttaaac ataatctgtg tttaatgatt attgtccatt gagcctgtac tctgctttcc 1980 ataccaagta aatatgaaat aatctacttt gcacataaca gaacaaacta taattacttg 2040 gctgttggag atttgtactt gagtataaat gtacaccagt ttttgtattt gtgaactcat 2100 ctgtgggagg agtaaagaaa atccaaaagc atttaatgtt ttgtttttgt tctataaaga 2160 tatgaaaatg tatttttata ttattttact tatttggaat ttacagagca cacctaagca 2220 attaggatat aacaaaacta cttaaccatt tttgcaacca ttttgttttt taagcctttt 2280 tatttctaaa aagatgaaaa cttataaata aattcttaat ttgtaattac ttttaaaaaa 2340 aaaaa 2345 <210> 62 <211> 2085 <212> DNA
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 2758310CB1 <400> 62 cggagatgtg gcgaccgttt ctggcatcat tctgagactc ggcagttgct tctcactgct 60 gcggccgggc ctgtctgtgg gagctgcatc ctcctcatct gcaggcgctg gaaaaccaga 120 cacgatcgga catgcatgtg gttctgcggc caaagcacgc cctttggttg tgaacttcat 180 gatacctgtg tgcagctctg tcatttccac tctgctctgc tgcacagaag gcagaagccc 240 tggccgtccc ctgctgtgtt cttcaggaga aacgtcaggg gccttcctcc aaggttctcc 300 agccccacac ccctgtggag gaaggtgctc tccaccgcgg tagtgggggc gcccctgctc 360 ctcggagccc gctatgtcat ggcagaggca cgggagaaga ggaggatgcg gctcgtggtg 420 gatggcatgg ggcgctttgg caggtctctg aaggtcggcc tgcagatctc cctggactac 480 tggtggtgca ccaatgttgt ccttcgaggg tggaagagcc caggctactt ggaggtgatg 540 tctgcgtgtc accagcgggc ggctgatgcc ctggtggcag gggccatcag caacgggggc 600 ctctacgtga agctgggcca ggggctgtgc tccttcaacc acctgcttcc ccccgagtat 660 acccggaccc tgcgcgtgct agaggacagg gccctcaagc ggggcttcca ggaggtggat 720 gagttgttcc ttgaggactt ccaggccctc ccccacgagc tcttccagga gtttgactac 780 cagccaattg ctgccgccag cctggcacag gtgcacagag ccaagctgca cgatggcacc 840 agcgtggctg tgaaggtgca gtacatcgac ctgcgggacc gctttgatgg ggacatccac 900 accctggagc tcctgctgcg gctcgttgag gtcatgcacc ccagctttgg cttcagctgg 960 gtcctccagg acctgaaggg gaccctggcc caggagctgg acttcgagaa tgagggccgc 1020 aacgcagagc gctgtgcgcg ggagctggcg cacttcccct acgtcgtggt gccccgcgtg 1080 cactgggaca agtccagcaa gcgcgtgctc actgccgact tctgcgccgg ctgcaaggtc 1140 aacgatgtgg aggccatcag gagccagggg ctggcagtgc atgacatagc agaaaagctc 1200 atcaaggcct ttgctgagca gatattttac accggcttca tccactcgga cccacatcct 1260 ggcaacgttc tggtgcggaa aggcccggac gggaaagcgg agctggtgct gctggaccac 1320 gggctctacc agttcctgga ggagaaggac cgcgcagccc tctgccagct gtggcgggcc 1380 atcatcctgc gggacgacgc cgccatgagg gcgcacgcag ccgcactggg ggtgcaagac 1440 tacctcctgt tcgccgagat gctcatgcag cgccccgtgc gcctggggca gctgtggggc 1500 tcgcacctac tgagccgcga agaggcggcc tacatggtgg acatggcccg cgagcgcttc 1560 gaggccgtca tggcggtgct cagggagctg ccgcggccca tgctgctggt gctgcgcaac 1620 atcaacaccg tgcgcgctat caacgtggcc ctcggcgccc ccgtggaccg ctacttcctt 1680 atggctaaaa gggctgtccg gggctggagc cgcctggcgg gcgccacgta tcggggtgtc 1740 tacggcacca gcctcctgcg ccacgccaag gtcgtctggg agatgctcaa gtttgaagtg 1800 gcgctcaggc tggagacctt ggccatgcgg ctgaccgccc tcctggctcg tgctctggtc 1860 cacctgagcc tcgtgccccc agcggaggag ctctaccagt acctggagac ctagggtgca 1920 gccgcccagg gccggcgggg cccttttcac cttgggctga cggaggtggc ggggctagag 1980 gtgtagacac cccgagcccc gtgggcactc gcactggggg gctgtgacag cagctgggcc 2040 aggaggccgt gtaatgacca cacactcctc tcaagcaaaa aaaaa 2085 <210> 63 <211> 3014 <212> DNA
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 2762348CB1 <400> 63 ggaggcaaag gccaggctga ggatcagggt ggcccgggtg gcagcgggga ggcgctgcat 60 gctggaggct gtgctgagtg cccggtgcag gtgagccggt cctgcggagt tgtgccgagt 120 gcctgctgca gaccgaggct gggccaagat ggcgtctgtg tttcgaagcg aggagatgtg 180 tttgtcacaa ctgtttctcc aggtggaagc tgcatattgc tgtgtggctg agctcggaga 240 gctcggattg gttcagttca aagatttaaa tatgaatgtg aacagctttc aaaggaaatt 300 tgtgaatgaa gtcagaaggt gtgaatcact ggagagaatc ctccgttttc tggaagacga 360 gatgcaaaat gagattgtag ttcagttgct cgagaaaagc ccactgaccc cgctcccacg 420 ggaaatgatt accctggaga ctgttctaga aaaactggaa ggagagttac aggaagccaa 480 ccagaaccag caggccttga aacaaagctt cctagaactg acagaactga aatacctcct 540 gaagaaaacc caagacttct ttgagacgga aaccaattta gctgatgatt tctttactga 600 ggacacttct ggcctcctgg agttgaaagc agtgcctgca tatatgaccg gaaagttggg 660 gttcatagcc gggtgtgatc caacagggaa gaggatggct tcctttgagc ggttactgtg 720 gcgagtctgc cgaggaaacg tgtacttgaa gttcagtgag atggacgccc ctctggagga 780 tcctgtgacg aaagaagaaa ttcagaagca catattcatc atattttacc aaggagagca 840 gctcaggcag aaaatcaaga agatctgtga tgggtttcga gccactgtct acccttgccc 900 agagcctgcg gtggagcgca gagagatgtt ggagagcgtc aatgtgaggc tggaagattt 960 aatcaccgtc ataacacaaa cagagtctca ccgccagcgc ctgctgcagg aagccgctgc 1020 caactggcac tcctggctca tcaaggtgca gaagatgaaa gctgtctacc acatcctgaa 1080 catgtgcaac atcgacgtca cccagcagtg tgtcatcgcc gagatctggt tcccggtggc 1140 agatgccaca cgtatcaaga gggcactgga gcaaggcatg gaactaagtg gctcctccat 1200 ggcccccatc atgaccacag tgcaatctaa aacagcccct cccacattta acaggaccaa 1260 taaattcaca gctggcttcc agaatattgt tgatgcctat ggtgtcggca gctaccggga 1320 gataaaccca gccccctaca ccatcatcac tttccccttc ctgttcgctg tgatgtttgg 1380 agactgtggt catggaaccg tgatgctcct ggctgcactt tggatgattc tgaatgagag 1440 acgcttgctc tcccagaaga cagacaatga gatttggaac accttcttcc acgggcgcta 1500 tctgatccta cttatgggca tcttctccat ctacacgggt ttgatctaca atgactgctt 1560 ctccaagtcc ttgaacatct ttggctcttc ttggagtgtc caacccatgt tcagaaacgg 1620 cacatggaat actcatgtaa tggaggaaag tctatatctg cagctggacc cagccatacc 1680 aggagtgtat tttggaaatc catacccgtt tgggattgat ccgatttgga acttggcttc 1740 aaacaaactc acatttctga actcgtataa aatgaagatg tcggtgatcc tgggaattgt 1800 ccagatggtt tttggtgtca tcctcagcct tttcaatcac atatacttca gaagaactct 1860 caacatcatt ctgcaattta tccctgagat gatttttatc ctgtgtctgt ttggatacct 1920 ggttttcatg atcattttca aatggtgctg ctttgacgtc cacgtatctc agcacgcccc 1980 cagcatcctc atccacttca tcaacatgtt tctgtttaac tacagtgact cttccaacgc 2040 acccctctac aaacatcagc aagaagtcca aagtttcttt gtggttatgg ctttgatttc 2100 tgtgccgtgg atgcttctga ttaagccgtt tattcttaga gccagtcatc ggaaatccca 2160 gctgcaggca tccaggatcc aagaagatgc cactgagaac attgaaggtg atagctccag 2220 cccttctagc cgttctggcc agaggacttc tgcagatacc cacggggctc tggacgacca 2280 tggagaagag ttcaactttg gagacgtctt tgtccaccaa gccatccaca ccatcgagta 2340 ctgcctgggc tgcatttcaa acacagcctc ctacctgcgg ctctgggccc tcagcctggc 2400 tcatgcacaa ctgtctgaag tgctctggac tatggtgatg aacagcggcc ttcagacgcg 2460 aggctgggga ggaatcgtcg gggtttttat tatttttgcc gtatttgctg tcctgacagt 2520 agccatcctt ctgatcatgg agggcctctc tgctttcctg cacgccctgc gactgcactg 2580 ggttgagttc cagaacaagt tctatgtcgg ggatggttac aagttttctc cattctcctt 2640 taaacacatc ctggatggca cagccgagga gtaggctgag ggctgcacct cccacggtgg 2700 tcaccatgcc aatgaaggaa gttcagtctt gtctttgata tcagcccctg caaggcgctc 2760 aatgggaagg ttgttcttgg ctcacctgaa gcatgaaact gtgtattatt tggacgtcag 2820 cctgtggatt tgatacgact taaccacgtc agaggaagga ctttggcaag tgatattgtc 2880 ttcatgtggg gtattaattc tcaaataata aagtaattga caaatgaggg gagaatgcta 2940 aacagatgtc ttcttgcaat attttaaata ttgtatttga gaaaataaac atctgagtca 3000 ttcaaaaaaa aaaa 3014 <210> 64 <211> 1726 <212> DNA
<213> Homo sapiens <220>
<221> misc_feature <223> Incyte ID No: 3715961CB1 <400> 64 tgaaaatgcc tgctgcgtac caaggtatgt actagggcat ctggggtaag taaaaacaaa 60 cacatagagc ctgcctggag aagctcatgg tctgatggaa agataagcaa gaagagttaa 120 tttctaatca atatgataaa aaggtcagag agcagtttct gaaaaacatg tttttgagtt 180 gagtcctgaa agacaaggag atgttagtaa agcagagaag ggagaattca ttctagaaag 240 atcagacagt gtgtgggaag ggcagagtct gaaaagagca tgccccattt ggagaagcat 300 caagaagccc acgtgttaga agcaccggcc ccatgagaca aagacacagc tagagagatt 360 gactaggcca tgtcggaatg tcctcttatt ttatacatac ataagcatat agatacatat 420 agccaaagtt acctttttaa tgatcttttt tacccagtgt attctggagg tcgaatggtc 480 acatatgaac atctccgaga ggttgtgttt ggcaaaagtg aagatgagca ttatcccctt 540 tggaaatcag tcattggagg gatgatggct ggtgttattg gccagttttt agccaatcca 600 actgacctag tgaaggttca gatgcaaatg gaaggaaaaa ggaaactgga aggaaaacca 660 ttgcgatttc gtggtgtaca tcatgcattt gcaaaaatct tagctgaagg aggaatacga 720 gggctttggg caggctgggt acccaatata caaagagcag cactggtgaa tatgggagat 780 ttaaccactt atgatacagt gaaacactac ttggtattga atacaccact tgaggacaat 840 atcatgactc acggtttatc aagtttatgt tctggactgg tagcttctat tctgggaaca 900 ccagccgatg tcatcaaaag cagaataatg aatcaaccac gagataaaca aggaagggga 960 cttttgtata aatcatcgac tgactgcttg attcaggctg ttcaaggtga aggattcatg 1020 agtctatata aaggcttttt accatcttgg ctgagaatga ccccttggtc aatggtgttc 1080 tggcttactt atgaaaaaat cagagagatg agtggagtca gtccatttta aacccctaaa 1140 gatgcaaccc ttaaagatac agtgttcagt attattgaaa tatgggcatc tgcaacacat 1200 accccctatt atttctacct ctttaggaag acacctattc cacagagact gatttatagg 1260 gggcagcact ttattttttt ctggaaaccc aagttctctt tgactcctct ttttgtccaa 1320 aagtgatctg gtcggatctc acaaggccat ccaatgagac cccgcacagc attttctaaa 1380 gaagaatcga agcctgacca ctttcacctt gggcaagaag gtttggcctt tgagttgcta 1440 ttctatgctg aagagcctgc ttagaggagg agtaccagga gggagccagc atttcagatc 1500 tgaagtagac gataggaatg tggaagaaca catacatagt gcttaagaaa tacatttaac 1560 ctgttatgtc agtatttatc aatgaagttt gataattcac ttttctgtca ttgttaaagc 1620 gtacatactg taaattaaag ggaggtgaat ggaaattaat gaataaacat tttgagtttc 1680 cctagtgttg aaggaaggtg tactttttct tgtcagaaag ataaaa 1726 <210> 65 <211> 899 <212> DNA
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 5108194CB1 <400> 65 gcggcggcga cgcgtccggg ccggtgaggg ggcgcggggg gcgccggggg ggcccaagcg 60 tcagcgcccg cgcctgtcgg gctgaactga ggaccgagtc tcctgccatt ccgagcaggc 120 ctggtatggg taatggtgtg aaggaaggcc cggtgcgatt gcatgaggat gctgaggctg 180 tcctgtcctc gtccgtctca tcaaagcgtg accacaggca agtgctcagc tccctgctgt 240 ctggggccct ggctggtgcc cttgccaaaa cagcggtagc tcccctggac cgaaccaaaa 300 tcatcttcca agtgtcttca aaaagatttt ctgccaagga ggccttccgg gtcctctact 360 acacctacct caacgaggga tttctcagct tgtggcgcgg gaactcggcc accatggtgc 420 gcgtggtgcc ctacgccgcc atccagttca gcgcacacga ggagtacaag cgcatcctgg 480 gcagctacta tggcttccgt ggagaagccc tgcccccttg gcctcgcctc ttcgccggcg 540 cactggctgg aacgacagcc gcttcactga cctaccccct ggacctggtc agagcgcgga 600 tggccgtaac cccgaaggaa atgtacagca acatctttca tgtcttcatc cgcatctcga 660 gagaagaggg gctgaagact ctctaccatg gatttatgcc caccgtgctg ggggtcattc 720 cctacgctgg cctgagcttc ttcacctatg agacgctcaa gagcttgcac agagagtaca 780 gcggccgcaa gcttattccc tttagtgagg gttaatttta gcttggcact ggccgtcgtt 840 ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcaca 899 <210> 66 <211> 643 <212> DNA
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 5503122CB1 <400> 66 ctttaagctg tagctgtggt ttctgcagca attttgtttt tgccttgaaa gaggtgctct 60 ggattatcac acctccatgt atgacaattt gtacctgcat ggaattgaag actcggaggc 120 tggttcagcg gattcctaca caagcaggcc gtctgactcc gatgtctctt tggaagagga 180 ccgggaagca attcgacagg agagagaaca gcaagcagct atccagcttg agagagcaaa 240 gtccaaacct gtagcatttg ccgtgaagac aaatgtgagc tactgcggcg ccctggacga 300 ggatgtgcct gttccaagca cagctatctc ctttgatgct aaagactttc tacatattaa 360 agagaaatat aacaatgatt ggtggatagg aaggctggtg aaagagggct gtgaaattgg 420 cttcattcca agtccactca gattggagaa catacggatc cagcaagaac aaaaaagagg 480 acgttttcac ggagggaaat caagtggaaa ttcttcttca agtcttggag aaatggtatc 540 tgggacattc cgagcaactc ccacatcaac aggtgagggt tgtagttaaa ctctttttca 600 tacactgtat tccttttaaa aatatttgaa cacacatgca agc 643 <210> 67 <211> 2574 <212> DNA
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 5517972CB1 <400> 67 gcgcctccca cgccggccgc cgccgacgtg atcgctcggg cgcgccgggc gtggttgggg 60 gaaggggttg tgccgcgcga cggtctgcgt gctgtgccca ctcaaaaggt tccgggcgcg 120 caggacggaa gacgcagtgc tcgccactcc cactgagatt gagagacgcg gcaaggaggc 180 agcctgtgga ggaactgggt aggatttagg aacgcaccgt gcacatgctt ggtggtcttg 240 ttaagtggaa actgctgctt tagagtttgt ttggaaggtc cgggtgactc atcccaacat 300 ttacatcctt aattgttaaa gcgctgcctc cgagcgcacg catcctgaga tcctgagcct 360 ttggttaaga ccgagctcta ttaagctgaa aagataaaaa ctctccagat gtcttccagt 420 aatgtcgaag tttttatccc agtgtcacaa ggaaacacca atggcttccc cgcgacagct 480 tccaatgacc tgaaggcatt tactgaagga gctgtgttaa gttttcataa catctgctat 540 cgagtaaaac tgaagagtgg ctttctacct tgtcgaaaac cagttgagaa agaaatatta 600 tcgaatatca atgggatcat gaaacctggt ctcaacgcca tcctgggacc cacaggtgga 660 ggcaaatctt cgttattaga tgtcttagct gcaaggaaag atccaagtgg attatctgga 720 gatgttctga taaatggagc accgcgacct gccaatttca aatgtaattc aggttacgtg 780 gtacaagatg atgttgtgat gggcactctg acggtgagag aaaacttaca gttctcagca 840 gctcttcggc ttgcaacaac tatgacgaat catgaaaaaa acgaacggat taacagggtc 900 attcaagagt taggtctgga taaagtggca gactccaagg ttggaactca gtttatccgt 960 ggtgtgtctg gaggagaaag aaaaaggact agtataggaa tggagcttat cactgatcct 1020 tccatcttgt tcttggatga gcctacaact ggcttagact caagcacagc aaatgctgtc 1080 cttttgctcc tgaaaaggat gtctaagcag ggacgaacaa tcatcttctc cattcatcag 1140 cctcgatatt ccatcttcaa gttgtttgat agcctcacct tattggcctc aggaagactt 1200 atgttccacg ggcctgctca ggaggccttg ggatactttg aatcagctgg ttatcactgt 1260 gaggcctata ataaccctgc agacttcttc ttggacatca ttaatggaga ttccactgct 1320 gtggcattaa acagagaaga agactttaaa gccacagaga tcatagagcc ttccaagcag 1380 gataagccac tcatagaaaa attagcggag atttatgtca actcctcctt ctacaaagag 1440 acaaaagctg aattacatca actttccggg ggtgagaaga agaagaagat cacagtcttc 1500 aaggagatca gctacaccac ctccttctgt catcaactca gatgggtttc caagcgttca 1560 ttcaaaaact tgctgggtaa tccccaggcc tctatagctc agatcattgt cacagtcgta 1620 ctgggactgg ttataggtgc catttacttt gggctaaaaa atgattctac tggaatccag 1680 aacagagctg gggttctctt cttcctgacg accaaccagt gtttcagcag tgtttcagcc 1740 gtggaactct ttgtggtaga gaagaagctc ttcatacatg aatacatcag cggatactac 1800 agagtgtcat cttatttcct tggaaaactg ttatctgatt tattacccat gaggatgtta 1860 ccaagtatta tatttacctg tatagtgtac ttcatgttag gattgaagcc aaaggcagat 1920 gccttcttcg ttatgatgtt tacccttatg atggtggctt attcagccag ttccatggca 1980 ctggccatag cagcaggtca gagtgtggtt tctgtagcaa cacttctcat gaccatctgt 2040 tttgtgttta tgatgatttt ttcaggtctg ttggtcaatc tcacaaccat tgcatcttgg 2100 ctgtcatggc ttcagtactt cagcattcca cgatatggat ttacggcttt gcagcataat 2160 gaatttttgg gacaaaactt ctgcccagga ctcaatgcaa caggaaacaa tccttgtaac 2220 tatgcaacat gtactggcga agaatatttg gtaaagcagg gcatcgatct ctcaccctgg 2280 ggcttgtgga agaatcacgt ggccttggct tgtatgattg ttattttcct cacaattgcc 2340 tacctgaaat tgttatttct taaaaaatat tcttaaattt ccccttaatt cagtatgatt 2400 tatcctcaca taaaaaagaa gcactttgat tgaagtattc aatcaagttt ttttgttgtt 2460 ttctgttccc ttgccatcac actgttgcac agcagcaatt gttttaaaga gatacatttt 2520 tagaaatcac aacaaactga attaaacatg aaagaaccca agaaaaaaaa aaaa 2574 <210> 68 <211> 1571 <212> DNA
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 5593114CB1 <400> 68 ggcgcggcga cagctagggt tcacggccac tggggcagag gagccgcgag aagatgtggg 60 tttttggtta cgggtccctg atctggaagg tggatttccc ctatcaggac aagctggtcg 120 gatacatcac caactacagc aggcgcttct ggcagggcag cacggaccac cgcggggtcc 180 ccggcaagcc tggaagagtt gtgactcttg ttgaagatcc tgcgggatgt gtatggggtg 240 ttgcttacag attgccagta ggaaaggaag aagaagtaaa agcatacctt gacttcagag 300 aaaaaggagg ctacagaacc acaacagtca ttttttatcc aaaagatccc acaacaaaac 360 cattcagtgt attgctatat attggaacat gtgataatcc tgattatctt ggtcctgcac 420 ctctggaaga cattgctgaa caaattttta atgcagctgg tccaagtgga agaaatacag 480 aatatctttt tgaacttgca aattctatta ggaaccttgt gccagaagaa gcagatgagc 540 atcttttcgc tttggaaaaa ttagtaaagg aacgtttaga agggaaacag aacctcaatt 600 gcatataatt tagtcttcag agaattaact tcagtgcaca atgacaatat gatttggaaa 660 tacgtttact taaagatctt atttttaatg tagtgaggat attatttaaa cttttatttt 720 aactggaaat gtcctgaaac acatatttaa aatattggga tacagtgaaa gaaaaattca 780 aattttaata acataaagat ttcctaactt tatgttattg aacacttact cactagaagt 840 gagttcttta gaaaaataca gtgaaggact cagttcagtc ttgtttttat cagagtgata 900 atcatcctgt ttcacatccc aatactattt tgaaattcta aacaattaaa ccaaaattcc 960 aataaatata aggttatgcc ttcaatatat tcctatacaa ttctgtaacc atggtttaaa 1020 atacacaagc ttaaaataac atgcttagaa atacacaata atatgaacag tatttcagcc 1080 ttaattgtga atttccttgt tattcaagta ttaaatgaaa tcttttgagt ttttagccaa 1140 aaattggcat ttttaaaata cgaaaatttc cttggaatta taatgtactg tacctcttct 1200 tttttaaata aaggcatttt actatatgga aaataactca ctaaagcata aattacatta 1260 tacaaatcat gatcactaat gatgtagtct gtcattcact ttgtattaat cttataccaa 1320 aactgaaaaa gatgggctga tactacaaat taatggcaca tataatgaaa atttagtttt 1380 taaaacagct tttggaattc tttgtctgtc actatctcaa tttgtgtgtg tgtgtgtgta 1440 tatacataaa tatacatata aaattttttt tttctttgca gcctgcgtct ggccatccca 1500 caggctggaa agtgtaacct ctggcagaag ccaagaacag gcacctcctg gaattataat 1560 tttgttttgt t 1571 <210> 69 <211> 1549 <212> DNA
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 044775C81 <400> 69 cgcgcttagg caggcggtgg cgcggctgga gtgccgcggg gagggctgtg ccggttgctt 60 tctgcagccg catctcggcc agctctcctc gccgtccccg gggcgctgtg cgtctccagt 120 ccgggaccga agccgcctgc cgtagcgggc ggccagatcc gcgtcccgcc tcagcggccg 180 gaggacatgc gggagagaga atgagccaga gggacacgct ggtgcatctg tttgccggag 240 gatgtggtgg tacagtggga gctattctga catgtccact ggaagttgta aaaacacgac 300 tgcagtcatc ttctgtgacg ctttatattt ctgaagttca gctgaacacc atggctggag 360 ccagtgtcaa ccgagtagtg tctcccggac ctcttcattg cctaaaggtg atcttggaaa 420 aagaagggcc tcgttccttg tttagaggac taggccccaa tttagtgggg gtagcccctt 480 ccagagcaat atactttgct gcttattcaa actgcaagga aaagttgaat gatgtatttg 540 atcctgattc tacccaagta catatgattt cagctgcaat ggcaggcatt tactgtacat 600 ttctcccgag aaaagagtga gatcgtgtca tctcatgctc cccatccgca ggtcacttcc 660 tgtagaaata tggactaact taaacctcgt tttactgcaa tcacagcaac caaccccatt 720 tggcttataa agactcggtt acagcttgat gcaaggaacc gcggggaaag gcgaatgggt 780 gcttttgaat gtgttcgtaa agtgtatcag acagatggac taaaaggatt ttataggggc 840 atgtctgctt catatgctgg tatatcagag actgttatcc attttgttat ttatgaaagt 900 ataaaacaaa aactactgga atataagact gcttctacaa tggaaaatga tgaagagtct 960 gtgaaagaag catcagattt tgtgggaatg atgctagctg ctgccacctc aaaaacttgt 1020 gccacaacta tagcatatcc acatgaagtt gtaagaacaa gactacgtga agagggaaca 1080 aaatacagat ctttttttca gactctatct ttgcttgttc aagaagaagg ttatgggtct 1140 ctttatcgtg gtctgacaac tcatctagtg agacagattc caaacacagc cattatgatg 1200 gccacctatg aattggtggt ttacctactc aatggatagc agcacgagga ctgctgtact 1260 gcaaaaaaag aagaccaaaa gattacagtg gaccatggga tacagaagcc agcatggcag 1320 acagaagaaa aatagtttgg gaacatgtaa ctattctaag tggaagtttt gttgtaggaa 1380 ttatagtaat cacaccacat tacttggcct ttcggtaatg tgaaaaaaaa aaaaaaacct 1440 cagagcctcc aaggaaatgc ctttagaagc actcctctct caaaattgcc attttctcta 1500 ccatgtcccc cagacacagt tgggttttgt tgatttatgg cagtcttct 1549 <210> 70 <211> 2237 <212> DNA
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 116588CB1 <400> 70 gtcactggtg ccacggggcc tcagcgcact tctgtcttaa gctcctgggc ctccttattt 60 tcccctttgc gatcgattcc agccacacct gtggatgttg ctagttactc cgcgtccgga 120 acgtggaggt cgagggactg agctgggcga gttttgtggc actcctttgc tcttcagcag 180 ctatttttgc tatgataatc ctgctgccct tcagactcaa gttaaacgag atatgcaagt 240 gaataccacg aaattcatgc tgctgtatgc ctggtattct tggcccaatg tagttttgtg 300 tttctttggt ggctttttga tagaccgagt atttggaata cgatggggca caatcatttt 360 tagctgcttt gtttgcattg gacaggttgt ttttgccctg ggtggaatat ttaatgcttt 420 ttggctgatg gaatttggaa gatttgtatt tgggattggt ggcgagtcct tagcagttgc 480 ccagaataca tatgctgtga gctggtttaa aggcaaagaa ttaaacctgg tgtttggact 540 tcaacttagc atggctagaa ttggaagtac agtaaacatg aacctcatgg gatggctgta 600 ttctaagatt gaagctttgt taggttctgc tggtcacaca accctcggga tcacacttat 660 gattgggggt gtaacgtgta ttctttcact aatctgtgcc ttggctcttg cctacttgga 720 tcagagagca gagagaatcc ttcataaaga acaaggaaaa acaggtgaag ttattaaatt 780 aactgatgta aaggacttct ccttacccct gtggcttata tttatcatct gtgtctgcta 840 ttatgttgct gtgttccctt ttattggact tgggaaagtt ttctttacag agaaatttgg 900 attttcttcc caggcagcaa gtgcaattaa cagtgttgta tatgtcatat cagctcccat 960 gtccccggtg tttgggctcc tggtggataa aacagggaag aacatcatct gggttctttg 1020 cgcagtagca gccactcttg tgtcccacat gatgctggcc tttacgatgt ggaacccttg 1080 gattgctatg tgtcttctgg gactctccta ctcattgctt gcctgtgcat tgtggccaat 1140 ggtggcattt gtagttcctg aacatcagct gggaactgca tatggcttca tgcagtccat 1200 tcagaatctt gggttggcca tcatttccat cattgctggt atgatactgg attctcgggg 1260 gtatttgttt ttggaagtgt tcttcattgc ctgtgtttct ttgtcacttt tatctgtggt 1320 cttactctat ttggtgaatc gtgcccaggg tgggaaccta aattattctg caagacaaag 1380 ggaagaaata aaattttccc atactgaatg agaagttaaa atgaatgtgt catgagaatg 1440 ggcttaacac atcgttggtt tgaaaacttc catttttaaa aatttagagt ttagtcatta 1500 gaaaaaataa tggactggaa agttatattt atatccaaat atacctattt caaagtgtat 1560 ttgtgaggcc tgttttagcc tgtgtctttt gtattgtgtg ttgctaaaga attctacttt 1620 tagtaggcta atcaacaatg aaagggttag aaaattgctg tggaacatcc aggtgaactt 1680 caggaaagac agtgaaaaat ggaaaacgtt ggagcttctg ttgagataat cttcattagg 1740 tatatatctt agggatacag ccttttcttt atcttatagc aggaaaaaaa aacttttgag 1800 ggaaatagaa gggctgcgtt acacaaaata aacaatggca ttgtcatagg ccttcctttt 1860 actagtaggg cataatgcta gggaatatgt gaagatgttt ttatgaagtc tctttctgat 1920 cacgaacaat agcttgcgct ctactctgta gttatgtgga ttgccgagca atgacccttt 1980 tcaatttctt atttctgtgt tactgaggac cctaatcact tagggatgta attttatagt 2040 ataaactttc tgtacagttt ttcttatagt ctaataagta aaaagtgtcc ttcaaattat 2100 gataattgcc tatgtacatg gataaattaa aacactgcac acggagtaaa aaaaaaaaaa 2160 aaaaaaaaaa aaaaaatgag cggccgcaag cttattccct ttagtgaggg ttaattttag 2220 cttggcactg gccgtcg 2237 <210> 71 <211> 1114 <212> DNA
<213> Homo sapiens <220>
<221> misc_feature <223> Incyte ID No: 875369CB1 <400> 71 tgggaacgga gcagccccgg gggccccctt gaggcggcga ggccgcgaag ggcgcggggc 60 tggagggtag gagagcgcgg gaaagcgccc cagacgccac tcgcggcgga cggcggccag 120 ttcccagggg tttggagccg gcccgcggcg ccatggctca cgtcggctcc cgcaagcgct 180 cgaggagtcg cagccggtcc cggggacggg ggtcggaaaa gagaaagaag aagagcagga 240 aagacacctc gaggaactgc tcggcctcca catcccaagg tcgcaaggcc agcacggccc 300 ctggggcgga ggcctcacct tctccctgca tcacagagag aagcaagcag aaggcccgga 360 ggagaacaag atccagctcc tcctcctctt cttccagttc ttctagctcc tcttcttcct 420 cctcgtcctc ctcctcttcc tccagtgatg gccggaagaa gcgggggaag tacaaggaca 480 agaggaggaa gaagaagaag aagaggaaga agctgaagaa gaagggcaag gagaaggcgg 540 aagcacagca ggtggaggct ctgccgggcc cctcgctgga ccagtggcac cgatcagctg 600 gggaggaaga ggatggccca gtcctgacgg atgagcagaa gtcccgaatc caggccatga 660 agcccatgac caaggaggag tgggatgccc ggcagagcat catccgcaag gtggtggacc 720 ctgagacggg gcgcaccagg cttattaagg gagatggcga ggtcctagag gaaatcgtaa 780 ccaaagaacg acacagagag atcaacaagc aagccacccg aggggactgc ctggccttcc 840 agatgcgagc tgggttgctt ccctgagggc ccccgctggc caaggcctgt ggacgacgct 900 ggcggcccag cctgggcagg tttcagggtg ccagtgggaa gcctgatggg tgctggtggc 960 ctttcccccg tggattggtc tctggcccag cccagtctct tctcaggggc agggggtgga 1020 ggttggggtc accggcctgc ttggcacccc catctgaaag agcagcactt ctcagctatt 1080 aaaggccccc tggatagaca aaaaaaaaaa aaaa 1114 <210> 72 <211> 998 <212> DNA
<213> Homo sapiens <220>
<221> misc_feature <223> Incyte ID No: 1325518CB1 <400> 72 gactaaaaaa gccatgtatt ctttcgtttc tctctaaaag aagaaaaata taatttaaaa 60 atacattgcg tattttctaa aacaataaat ttatagtgtt aatattcata gggtcaatca 120 aaatgaagct tctcctttgg gcctgcattg tatgtgttgc ttttgcaagg aagagacggt 180 tccccttcat tggtgaggat gacaatgacg atggtcaccc acttcatcca tctctgaata 240 ttccttatgg catacggaat ttaccacctc ctctttatta tcgcccagtg aatacagtcc 300 ccagttaccc tgggaatact tacactgaca cagggttacc ttcgtatccc tggattctaa 360 cttctcctgg attcccctat gtctatcaca tccgtggttt tcccttagct actcagttga 420 atgttcctcc tctccctcct aggggtttcc cgtttgtccc tccttcaagg tttttttcag 480 cagctgcagc acccgctgcc ccacctattg cagctgagcc tgctgcagct gcacctctta 540 cagccacacc tgtagcagct gagcctgctg caggggcccc tgttgcagct gagcctgctg 600 cagaggcacc tgttggagct gagcctgctg cagaggcacc tgttgcagct gagcctgctg 660 cagaggcacc tgttggagtg gagccagctg cagaggaacc ttcaccagct gagcctgcta 720 cagccaagcc tgctgcccca gaacctcacc cttctccctc tcttgaacag gcaaatcagt 780 gaaattctct agaagagtac catgggttca tttctatact gatgcagaaa taagtgaaat 840 ctacaaaagt tttctttctt ttccaaagac tatttcattc tgttgtattc agagtattca 900 tctcactaca ttgatttgtt tgtggtagtt tttccttgga cttaatttat attgaaaaaa 960 cattgataat taaataaata aaatagataa tttagaca 998 <210> 73 <211> 2348 <212> DNA
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 2060987CB1 <400> 73 gggccagccg gctcgcccgg gggccatggc agcagcggct actgcagccg agggggtccc 60 cagtcggggg cctcccgggg aagtcatcca tctgaatgtg ggaggcaaga gattcagtac 120 ctctcgccag actctcacct ggatcccaga ctccttcttc tccagtcttc tgagcggacg 180 catctcgacg ctgaaagatg agaccggagc aatcttcatc gacagggacc ctacagtctt 240 cgcccccatc ctcaacttcc tgcgcaccaa agagttggat cccaggggtg tccacggttc 300 cagcctcctc catgaagccc agttctatgg gctcactcct ctggttcgtc gcctgcagct 360 tcgagaggag ttggatcgat cttcttgtgg aaacgtcctc ttcaatggtt acctgccgcc 420 accagtgttc ccagtgaagc ggcggaaccg gcacagccta gtggggcctc agcagctagg 480 aggacggcca gcccctgtcc gacggagcaa cacgatgccc cccaaccttg gcaatgcagg 540 gctgctgggc cgaatgctgg atgagaaaac ccctccctca ccctcaggac aacctgagga 600 gccggggatg gtgcgcctgg tgtgtggaca ccataattgg atcgctgtgg cctataccca 660 gtttctagtc tgctacaggt tgaaggaagc ctctggctgg cagctggtgt tttccagccc 720 ccgcctggac tggcccatcg aacgactggc gctcacagcc cgggtgcatg gtggggcttt 780 gggtgaacat gacaagatgg tggcagcagc caccggcagc gagatcctgc tatgggctct 840 gcaggcggaa ggcggtggct ccgagatagg ggtctttcat ctgggggtgc ctgtggaggc 900 cttgttcttc gtcgggaacc agctcattgc tacaagccac acagggcgca tcggggtgtg 960 gaatgccgtc accaagcact ggcaggtcca ggaggtgcag cccatcacca gttatgacgc 1020 ggcaggctcc ttcctcctcc tgggctgcaa caacggctcc atttactacg tggatgtgca 1080 gaagttcccc ttgcgcatga aagacaacga cctccttgtc agcgagctct atcgggaccc 1140 agcggaggat ggggtcaccg ccctcagtgt ctacctcacc cccaagacca gtgacagtgg 1200 gaactggatc gagatcgcct atggcaccag ctcagggggc gtgcgggtca tcgtgcagca 1260 cccggagact gtgggctcgg ggcctcagct cttccagacc ttcactgtgc accgcagccc 1320 tgtcaccaag atcatgctgt cggagaagca cctcatctca gtctgtgccg acaacaacca 1380 cgtgcggaca tggtctgtga ctcgcttccg cggcatgatt tccacccagc ccggctccac 1440 cccactcgct tcctttaaga tcctggctct ggagtcggca gatgggcatg gcggctgcag 1500 tgctggcaat gacattggcc cctacggtga gcgggacgac cagcaagtgt tcatccagaa 1560 ggtggtgccc agtgccagcc agctcttcgt gcgtctctca tctactgggc agcgggtgtg 1620 ctccgtgcgc tccgtggacg gctcacccac gacggccttc acagtgctgg agtgcgaggg 1680 ctcccggcgg ctcggctctc ggccccggcg ctacctgctc actggccagg ccaacggcag 1740 cttggccatg tgggacctaa ccaccgccat ggacggcctc ggccaggccc ctgcaggtgg 1800 cctgacggag caagagctga tggaacagct ggaacactgt gagctggccc cgccggctcc 1860 ttcagctccc tcatggggct gtctccccag cccctcaccc cgcatctccc tcaccagcct 1920 ccactcagcc tccagcaaca cctccttgtc tggccaccgt gggagcccaa gccccccgca 1980 ggctgaggcc cggcgccgtg gtgggggcag ctttgtggaa cgctgccagg aactggtgcg 2040 gagtgggcca gacctccgac ggccacccac accagccccg tggccctcca gcggtctcgg 2100 cactcccctc acacctccca agatgaagct caatgaaact tccttttgaa caacgcagct 2160 gccatgatgc cttgggatgc cctggtcctg ggggactcag gtgcctccct gattcctgtg 2220 ggaaccccgg gttcagggcc agggcctcct tggaataaat ggttattgtt actaggtccc 2280 caccttccct cttttctgga agccaaagtc accctcccca ataaagtcct cactgccaaa 2340 aaaaaaaa 2348 <210> 74 <211> 1139 <212> DNA
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 2172064CB1 <400> 74 ctcgagctgg gatgtgtggc aggttcctgc ggcggctgct ggcggaggag agccggcgct 60 ccacccccgt ggggcgcctc ttgcttcccg tgctcctggg attccgcctt gtgctgctgg 120 ctgccagtgg gcctggagtc tatggtgatg agcagagtga attcgtgtgt cacacccagc 180 agccgggctg caaggctgcc tgcttcgatg ccttccaccc cctctccccg ctgcgttcct 240 gggtcttcca ggtcatcttg gtggctgtac ccagcgccct ctatatgggt ttcactctgt 300 atcacgtgat ctggcactgg gaattatcag gaaaggggaa ggaggaggag accctgatcc 360 agggacggga gggcaacaca gatgtcccag gggctggaag cctcaggctg ctctgggctt 420 atgtggctca gctgggggct cggcttgtcc tggagggggc agccctgggg ttgcagtacc 480 acctgtatgg gttccagatg cccagctcct ttgcatgtcg ccgagaacct tgccttggta 540 gtataacctg caatctgtcc cgcccctctg agaagaccat tttcctaaag accatgtttg 600 gagtcagcgg tttctgtctc ttgtttactt ttttggagct tgtgcttctg ggtttgggga 660 gatggtggag gacctggaag cacaaatctt cctcttctaa atacttccta acttcagaga 720 gcaccagaag acacaagaaa gcaaccgata gcctcccagt ggtggaaacc aaagagcaat 780 ttcaagaagc agttccagga agaagcttag cccaggaaaa acaaagacca gttggaccca 840 gagatgcctg agttggagat gaactttggc caactttcct catcaccata cttaaaatcc 900 tgtccaagga ggagcttatt caccattttc tatacatgtg acatatgtag cagcataacc 960 gacaactggg actgcgctgc cttgactcca cctctacata caatgactca gctaaccaga 1020 ctaataaaag ccatgtttgc accattgctc agggaggcat tgctttgggg aattattccc 1080 agtgtcctcc ttacttatcg caagtaataa aaatcccctg ggaaatcctc aaaaaaaaa 1139 <210> 75 <211> 863 <212> DNA
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 2219267CB1 <400> 75 tcagagctag cttccgccgc agcctggcac ctcaggtgcc gaccttctac ctgccctact 60 tcctggaggc cggcctggag gcggcagcct tggtcttcct gctcctgacg gcagattgct 120 gtggacgccg ccccgtgctg ctgctgggca ccatggtcac aggcctggca tccctgctgc 180 tcctcgctgg ggcccagtat ctgccaggct ggactgtgct gttcctctct gtcctggggc 240 tcctggcctc ccgggctgtg tccgcactca gcagcctctt cgcggccgag gtcttcccca 300 cggtgatcag gggggccggg ctgggcctgg tgctgggggc cgggttcctg ggccaggcag 360 ccggccccct ggacaccctg cacggccggc agggcttctt cctgcaacaa gtcgtcttcg 420 cctcccttgc tgtccttgcc ctgctgtgtg tcctgctgct gcctgagagc cgaagccggg 480 ggctgcccca gtcactgcag gacgccgacc gcctgcgccg ctccccactc ctgcggggcc 540 gcccccgcca ggaccacctg cctctgctgc cgccctccaa ctcctactgg gccggccaca 600 cccccgagca gcactagtcc tgcctggtgg ccctgggagc caggatggga ccaaagtcaa 660 ggcctggggc atggctgagt accccagacg tctggtccag ggcagacaca ttcctctcag 720 aagcccgtgt ctcagtgcag gtggagccgt ggggacagcg tgaaggtgtc tccagccagg 780 ccccaggcac tgggaggccc tgggtctccc cccagccaca cccagtaggt gtggaggata 840 aaggcttctg tggaaaaaaa aaa 863 <210> 76 <211> 1322 <212> DNA
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 2308629CB1 <400> 76 ccggggggcc ggcgcgcggg gaggccgggg cctgcaggcc cccggtacga caagatccgg 60 actccggccc ggactacgag gcgctgccgg ctggagccac tgtcaccacg cacatggtgg 120 caggcgccgt ggcagggatc ctggagcact gcgtgatgta ccccatcgac tgcgtcaaga 180 cccggatgca gagtctacag cctgacccag ctgcccgcta tcgcaatgtg ttggaggccc 240 tctggaggat tataagaacg gagggcctat ggaggcccat gagggggctg aacgtcacag 300 caacaggcgc agggcctgcc cacgcccttt attttgcctg ctacgaaaag ttaaaaaaga 360 cattgagtga tgtaatccac cctgggggca atagccatat tgccaatggt gcggccgggt 420 gtgtggcaac attacttcat gatgcagcca tgaaccctgc ggaagtggtc aagcagagga 480 tgcagatgta caactcacca taccaccggg tgacagactg tgtacgggca gtgtggcaaa 540 atgaaggggc cggggccttt taccgcagct acaccaccca gctgaccatg aacgttcctt 600 tccaagccat tcacttcatg acctatgaat tcctgcagga gcactttaac ccccagagac 660 ggtacaaccc aagctcccac gtcctctctg gagcttgcgc aggagctgta gctgccgcag 720 ccacaacccc actggacgtt tgcaaaacac tgctcaacac ccaggagtcc ttggctttga 780 actcacacat tacaggacat atcacaggca tggctagtgc cttcaggacg gtatatcaag 840 taggtggggt gaccgcctat ttccgagggg tgcaggccag agtaatttac cagatcccct 900 ccacagccat cgcatggtct gtgtatgagt tcttcaaata cctaatcact aaaaggcaag 960 aagagtggag ggctggcaag tgaagtagca ctgaacgaag ccaggggttc agatgacact 1020 gctgcatcct ggtcacattc tctgtctcct ggaatgctcc cacctcaagt ggagttagaa 1080 ggaaggtaga ggggctctcc cccaggattt tggtgttttg actaacacca gttcctgcca 1140 acctctgttg ccaccacctt tccttccagg ccctaagcac gtgcagcaaa gcacaccaca 1200 gcacctttga taacctctct ccatcctggg cctgatgacc tgctctagac tgttatagag 1260 ggataagcag ctcattcccc tggttcctaa taaaaagcct ttaaattaaa aaaaaaaaaa 1320 as 1322 <210> 77 <211> 1869 <212> DNA
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 2660038CB1 <400> 77 cagggacgct cattcctggc cagagtcctg acttctttct cggatcgaga tctcgttgct 60 ggctcgagga aatcaccggc tcttctcccg gatccttttc ttctctcact tgctggcttt 120 cttcttcctg cctccctggc ttccatctcc caaccccgcg attccctcct ctactcccgt 180 gctcccgtcg cccgccatcc tgagccatcc cacctgcaac cttctgtctt ttgcccctcc 240 ttgacctcag agggtcctgc cttaagcttc tcaccagaat ctcctagatt tctatctctt 300 ccctgcttgc cagctcttga ctcccaaatt ccagctgacg tttgaccact tgacatttga 360 ccctgacacc cttgactgca aatctaaatt cttatcttct gcaacctgta ctgctgaaac 420 aggccttccc cctgtcttcc aaccccgctt tctgacaccc atttttactt tcttactctt 480 gggtcagttc ctgctacagc tatcccacca tggacttctt gatgagtggc ctggcagcct 540 gcggggcctg tgtattcacc aatcccctgg aggtggtgaa gaccaggatg cagttgcaag 600 gagaactgca ggcccctggc acataccagc ggcactaccg aaatgtcttc catgccttca 660 tcaccatcgg caaggtggat ggccttgctg ccctgcagaa aggcctggcc cccgccctct 720 tgtaccagtt cctgatgaat ggcatccgac tgggcaccta tgggctggct gaggctgggg 780 gctacctgca cacagccgaa gcgacccaca gtcctgcccg cagcgcagca gctggggcca 840 tggctggggt catgggagcc tacttgggga gccccatcta catggtgaag acacacctgc 900 aggcacaggc agcctcagaa attgctgtag ggcaccagta taagcatcag ggcatgtttc 960 aggcgctaac cgagattggc cagaaacatg gtctggtggg gttatggcgt ggggctctgg 1020 gcggcctgcc ccgagttatc gtcggttcct ccacccagct gtgcaccttc tcatccacca 1080 aggacctcct gagccagtgg gagatctttc ctccccagag ctggaagttg gcgctggtgg 1140 ctgccatgat gagtggcatt gcagttgtct tggccatggc accctttgat gtggcctgca 1200 caaggctcta caaccagccc acagatgcac agggcaaggg cctcatgtac cgggggatac 1260 tggacgctct gctgcagaca gctcggaccg agggcatttt tggcatgtac aagggtatag 1320 gtgcctccta cttccgcctc ggcccccaca ccatcctctc cctcttcttc tgggaccagc 1380 tgcgctccct ctactacaca gacactaaat aacagccgct ttcccagtct ccaccaaatg 1440 agcactcctt ggccacttgt gcctccacca ctatgtcctg gtgactactg attaggtgac 1500 ctttcatcca tccatggggg acagccaacc ccactcccca tctgttctca gggttgaatc 1560 actacaagag atgagtttcc cttctttcct tgggtgttgc tttaaacctt ccctacccat 1620 tccctgggta actcacaccc ctctctcagg gctgaacgag tcatcccaaa gtgtatttcc 1680 tcccactcac cactgccacc cttgagtccc tcctgctccc atgcacagtt ttaaactcct 1740 ccctccaaaa ccaaagggaa tcgagagacc caattcccag gcgtctggga cccaggtgtc 1800 ctgttagatt caaaggcaca gagattatat tgattataaa gcaagtttat tctgaaaaaa 1860 aaaaaaaaa 1869 <210> 78 <211> 1881 <212> DNA
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 2670745CB1 <400> 78 gaagaaccga gcttggctgt gtttatctcg ttggggacta aggcgtcggt tggcgcgcaa 60 cgggttctag gctgcaggca gctcgaggac ccgcggcccc gccccggctc ggcctggcag 120 atagcagagg cagcaggccg tgccgggggg gcatgttgct gtaaccagtg gcccagggga 180 tgttacggtg gacagtgcac ctggagggcg ggccccgcag ggtgaaccat gctgcagtgg 240 ctgtcgggca tcgggtatac tccttcgggg gttactgctc tggtgaagac tatgagacac 300 tgcgtcagat agatgtgcac attttcaatg cagtgtcctt gcgttggaca aagctgcccc 360 cggtgaagtc tgccatccgt gggcaagctc ctgtggtacc ctacatgcgc tatggacact 420 caaccgtcct catcgacgac acagtcctcc tttggggcgg gcggaatgac accgaagggg 480 cctgcaatgt gctctatgcc tttgacgtca atacgcacaa gtggttcaca ccccgagtgt 540 cagggacagt tcctggggcc cgggatggac attcagcctg tgtcctaggc aagatcatgt 600 acatttttgg gggctacgag cagcaggcgg actgtttttc caatgacatt cacaagctag 660 ataccagcac catgacatgg actcttatct gtacaaaggg cagccctgca cgctggaggg 720 acttccactc agccacaatg ctgggaagtc acatgtatgt ctttgggggc cgtgccgacc 780 gctttgggcc attccattcc aacaatgaga tttactgcaa ccgcattcga gtctttgaca 840 ccagaactga ggcttggctg gactgtcccc cgactccagt gctgcctgag gggcgccgga 900 gccactcggc ctttggctac aatggggagc tgtacatctt tggtggttat aatgcaaggc 960 tgaaccggca cttccatgac ctctggaagt ttaatcctgt gtcctttacc tggaaaaaga 1020 ttgaaccgaa ggggaagggg ccatgtcccc gccggcgcca gtgctgctgt attgttggtg 1080 acaagattgt cctctttggg ggtaccagtc catctcctga ggaaggcctg ggagatgaat 1140 ttgaccttat agatcattct gacttacaca ttttggactt tagccctagt ctgaagactc 1200 tgtgcaaact ggccgtgatt cagtataacc tagaccagtc ctgtttgcct catgatatca 1260 ggtgggagct gaatgccatg accaccaaca gcaatatcag tcgccccatc gtctcctccc 1320 atgggtagga ggaagtttct gccacctccc ctcctgagcc tgctgtcatc ttcactgccc 1380 ctgcccatct gtcacccacc tgctcctttg acccctggac ttggtatacc tccatgtgga 1440 gttgttgggc gagaggtgtt ctctgtgctg tgaattcagt ggggagctgt agcggggtgg 1500 gggctaggtt cctcccccct tgggccgagg gccccttccc cttggtgctc tgtccccatc 1560 cacctccttt cagctgctcc tgggcctcag ctctgcccag ggccagccag gttctgctgg 1620 gaagggaagg gaatggggag aagggagaag caagcagtgt ctgagcctca ggagcttccc 1680 cctccccctt tgcctatccc ctcccctctg cttgagcctt gagccttgac tgggagctga 1740 aaggagttgc agctgttggc atgagacctc cttctccccg tcttggggag gtggggacca 1800 gcagataaat cccacccttc cttgagctgt cgctgtactc tgaagttcag ccagctcaga 1860 ttttataaaa attaattaaa a 1881 <210> 79 <211> 2004 <212> DNA
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 2676443CB1 <400> 79 aatcgcaggg ggcgtggcct agcggtcccg cccccggggc gcgcgcgcgc attggctgtg 60 cggggtgcgg gcgcgcgggc ggcgctttga accgggcgcg gggcgcgggg cgcggggcgc 120 tgcggccggt acacgccggg gtagggccgg ggtcgggttg tggtcgggcc gggattgggc 180 tctcctgggc catggcagcc gaggcgcgcg tgtcgcgctg gtacttcggg gggctggcct 240 cctgcggggc cgcctgctgc acgcacccgc tggacctgct caaggtgcat ctgcagacgc 300 agcaggaggt gaagctgcgc atgacgggca tggcgctgcg ggtggtgcgt accgacggca 360 tcctggcact ctacagcggc ctgagcgcct cgctgtgcag acagatgacc tactccctga 420 ctcggttcgc catctacgag actgtgcggg accgcgtggc caagggcagc caggggcctc 480 tccccttcca cgagaaggtg ttgctgggct ccgtcagcgg tttagctgga ggcttcgtgg 540 ggacgcccgc agacttggtc aacgtcagga tgcagaacga cgtgaagctg ccccagggtc 600 agcggcgcaa ctacgcccat gcgctggatg gcctgtaccg cgtagctcgt gaagagggtc 660 tcaggagact gttctcgggt gcaaccatgg catccagccg aggggcctta gtcactgtgg 720 gccagctgtc ctgctacgac caggccaagc agctggtcct tagcaccggg tacctctctg 780 acaacatctt cactcacttt gtcgccagct ttattgcagg tggatgtgcc acgttcctgt 840 gccagcccct ggatgtgctg aagactcgcc tgatgaactc caagggggag tatcagggcg 900 ttttccactg cgccgtggag acagcgaagc tcgggcctct ggccttttac aagggcctcg 960 tcccagctgg catccgcctc atcccccaca ccgtgctcac ttttgtgttt ctggaacagc 1020 tacgcaaaaa ctttggcatc aaagtgccat cctgaccagc cgtgggaatg gctgggctgc 1080 caggccagac acgctaggtt cttccaaaga gtcccaagcc cagcacctgc tcctggggcc 1140 acgacctccc tggccgtggc cacccatcct ccgcagcagg cccctgctgt ccccccacct 1200 gctggctgag ctcctcctgg cctcgtcccc tctcagctgt agctgcacca cccccgctct 1260 ggctaccagg ctctcccggc tgggcactgc gtggccttgc ccctctcccg ctggcagctc 1320 ctcaggggaa caggggctac cagaggctga tttctcccct ctcctgggcc aggggagggg 1380 tattatccct gcctcctgcc cccgatgccc aaagcagcat cttccagcac tttccatcga 1440 ggacttgggt ggcagagtgt gggtgcagcc tggctgttgc tcacccaagt gctagctctg 1500 cacttcgtgt ctgctgagag caaccagacc ttccatgtcc tcgggcagct gcaactcccc 1560 gcgagacccc gcagctgggt gggatgaaca agcaacgcag accacaagcg agtgcctggg 1620 agggagtggc ccagggtggt tctggagcca ttgtgggtga gggtcgaggg ccaccgaggt 1680 cccgcgcacc gctgcctgcc ctgcagtggc tttaacagtt agttttgcca aagcctctcc 1740 actcaccagc aggcggtctc tgtcttcagg gattgtgcct gcgtccctcg ggcacctggg 1800 cccccccgct tggctccctg ggggaatggc ccaggcgggc cgcggttcct ccttagggcc 1860 ttctccccga caaggagtcc gacggggcgg atgctgcatc ctctgcctcc ctggtcgctg 1920 ggcttcaccc cacctgggaa gggcagtgtg ctctgtgggg gctgcaatca ataaatgccg 1980 ggagctgcca aaaaaaaaaa aaaa 2004 <210> 80 <211> 3555 <212> DNA
<213> Homo sapiens <220>
<221> misc_feature <223> Incyte ID No: 3295764CB1 <400> 80 gaccgtgggc gagtccagaa cgtcctggcc ttacagggag aaggcgtcac tcgcggttac 60 aagtgcctga ccctcactcc agttggcgga ggaggagaag gaaggggccg ggccgggtcc 120 cctcccctcg cgccccggat ggatgtgccc ggcccggtgt ctcggcgggc ggcggcggcg 180 gcggccactg tgctcctgcg gaccgctcgg gtccgtcgcg aatgctggtt cttgccgacc 240 gcgctgctct gcgcctacgg cttcttcgcc agcctcaggc cgtccgagcc cttcctgacc 300 ccgtacctgc tggggccgga caagaacctg accgagaggg aggtcttcaa tgaaatttat 360 ccagtatgga cttactctta cctggtgcta ctgtttcctg tgttccttgc cacagactac 420 ctccgttata aacctgttgt tctactgcag gggctcagcc ttattgttac atggtttatg 480 ctgctctatg cccagggact gctggccatt caatttctag aattttttta tggcatcgcc 540 acagccactg aaattgccta ttactcttat atctacagtg tggtggacct gggcatgtac 600 cagaaagtca caagttactg tcgaagtgcc actttggtgg gctttacagt gggctctgtc 660 ctagggcaaa tccttgtctc agtggcaggc tggtcgctgt tcagcctgaa tgtcatctct 720 cttacctgtg tttcagtggc ttttgctgtg gcctggtttt tacctatgcc acagaagagc 780 ctcttctttc accacattcc ttctacctgc cagagagtga atggcatcaa ggtacaaaat 840 ggtggcattg ttactgacac cccagcttct aaccaccttc ctggctggga ggacattgag 900 tcaaaaatcc ctctaaatat ggaggagcct cccgtggagg aaccggaacc caagccagac 960 cgtctccttg tattgaaagt actatggaat gatttcctga tgtgctactc ctctcgccct 1020 cttctctgct ggtctgtgtg gtgggccctc tctacctgtg gctattttca agttgtgaac 1080 tacacacagg gcctgtggga gaaagtgatg ccttctcgct atgctgctat ctataatggt 1140 ggcgtggagg ccgtttcaac cttactgggt gctgttgctg tgtttgcagt tggttatata 1200 aaaatatcct ggtcaacttg gggagaaatg acattatctc tcttttctct cctgattgct 1260 gctgcagtgt atatcatgga cactgtgggt aacatttggg tgtgctatgc atcctatgtt 1320 gtcttcagaa tcatctacat gttactcatc acgatagcaa cttttcaaat tgctgcaaac 1380 ctcagcatgg aacgctatgc cctagtattt ggtgtaaata ccttcattgc cctggcactg 1440 cagacgctgc tcactctaat tgtggtagat gccagtggcc ttggattaga aattaccact 1500 cagtttttga tctatgccag ttattttgca ctcatcgctg tggttttcct ggccagtggt 1560 gcagtcagtg ttatgaagaa atgtagaaag ctggaagatc cacaatcaag ttctcaagta 1620 accacttcat aatatactgc tgaagggctt cttcttatag caagaactct gcacagcaac 1680 tgcctggatg tatttgattt tttaaagcgt agacatatat ttatgaatgt gcatttcttg 1740 acttcacagc agccacttga ctaatacctt gtgttccggg aataacatga tactattcag 1800 aggagccaga agtaaagttt atttcatgga ttatttatga gagctaattt aaggatgact 1860 ttttttctga ttcaaaagtg aacttgattt taaaaaccag tcaagagcaa tcaaagcagc 1920 acatggtgtt gtatacttca ttagcaagtg agtttggtgt tttataggtc acatatgtct 1980 gtatctactt agccagatgc ttggcctggt gggaccaggg ctccacagag gccacaaaat 2040 gttgcaagtc atgatggatg gaaatatgtt ctaacagcat ctgcctctat tcaatttaat 2100 tcttatttct gtgttactca tgtacattgg tctttctaca tagttattct atcactggca 2160 atatttgttc tggtttagtg ttctgtattt taaggtgtac gtatcatttc taattttaag 2220 ttattttaaa aaaattcatc atatgaatgt tcttggttcc cattgtgacg attatttatt 2280 tctgtaaaat ttgtttagaa gtacgttttt gcattattca tatgcttccc agagaagctc 2340 atttagttag aaaataaggc aagttttgaa gcctgctaaa tgaagagact taagaaagct 2400 taaggtacgc ttgcttgtct ttaaatcttc aatatgaagg actattaatt ccaagattaa 2460 aagttcatat ataggctaaa gatgtaacta ggccatttgt atttgtattc ccttttattt 2520 ccaaaataaa atgaaaaatc tttttttaat aatttcatcc ctatttatag tttttatatt 2580 aatttgtttt tcttatccaa gtaaagatgt caataggaat tgcattagtc caaggccttt 2640 ttcataaact gagcctcttt tcaattattt caatgggaca ggaactagga tagatgtgat 2700 tcctgcattt ttttacctta aatctgcctt tgtttctaaa ggtagatcat cttgaatatt 2760 tgcttaaaat tgctagtgat ttcattacca agttacttga aaaaatgttc tatatgcatt 2820 taattctgaa atcagtctac caaggggctg ctagtatatg tcagacatga aaactatttt 2880 aaagctgact ttgttgcctt atcttgaaaa gaatctagat aggtgctttt aactggggta 2940 ttaacttttt tagaatgaca cagctgaaca gtgttaataa tagtgtgtca agattgcaaa 3000 gtcgacatac tcatttggtt taagcaggaa tcctagaagc aaatggatgg ggataagaat 3060 aggtcatttt ctattcacca tcctttacta ttaagggaaa ggaaaagaac actagctaag 3120 gaagggaaag ggaagtgata tcataaaagt agcaaccttc attttacatt ctgtctgttt 3180 ttcttttttt gctttgtttt gtttgtgcta atttgggaat tgtgtactcc gaaacaagta 3240 gaaaagtgct gtttgaggga ttttattaaa tcttttttta atggaatgtg gtacaaattg 3300 ttcatgttac caaagcaata tttccctgga atttaattca aagtttgtgg catacaacct 3360 gagccttttc ttatataaga caagaatatg ttcacatctt ggtatgtggc catatttata 3420 gaatgctgaa ctcaatgtgc aagttgtact gtatgcagtt ttgtaaataa gtgaaaataa 3480 tttgttgtac tttttattca attctgtata gattataaaa ttatttttat taaataaata 3540 ttttacagta tattt 3555 <210> 81 <211> 1293 <212> DNA
<213> Homo sapiens <220>
<221> misc_feature <223> Incyte ID No: 3438320CB1 <400> 81 atatgcccca cgcgggactc atactacgtt tcccgtgaac acgtgcagtc caaaccccgc 60 ccctgatatt tatctcagtg gacggtggcc ggaaaaggac aatggtttcc atgtcagcgg 120 ataaacgctc tcccctcggc tcccggacgc gacggaggtc gtagtagtag tgagtacgtg 180 ctgaggagca aaggagtaac caagagatcc agtgaccgac agagcaagag ccatgccgcg 240 ccggggcctg gtggctgggc cagacttgga gtattttcag cgtcgctatt tcacgccggc 300 ggaggtggcc caacataaca ggcccgaaga cctctgggta tcttacctgg gacgcgtgta 360 cgacctaacg tcattggcac aggaatacaa ggggaacctg ctgctgaaac ccatcgtgga 420 agttgcaggc caggatatca gccactggtt tgatccaaag accagagaca tccgcaagca 480 catagatccg ctgaccggct gcctgaggta ctgcaccccg cggggccgct ttgtgcacgt 540 tccgcctcag ctgccctgtt cggactgggc caacgatttt gggaagccct ggtggcaggg 600 gtcgtattat gaggtggggc ggctgtctgc caagacccgg agcatccgca tcattaacac 660 gctcacgtcg caggagcaca cactggaggt gggggttctg gagtccatat gggaaatcct 720 acaccgctat ctcccctata actcacatgc tgccagctac acgtggaaat atgaagggaa 780 gaacctgaac atggatttta ccctggaaga gaatgggatc cgggatgagg aggaagaatt 840 tgactatctc agtatggacg gtacacttca cacacctgca atacttctgt acttcaatga 900 tgatctcacg gagttgtagg caaggagatg tacactcgtg tagactcaag acgtatttcg 960 agtttggctt tttctgtgcc ttgaggaaaa gtggtggggc cgaggggtgc ctggacccag 1020 atctccactc ctctccagga gctagcctgt gcccttctga agtgtaaagg ccctattccc 1080 tgccttcatt acagtttgct ctgagaaaat tagtgaatta atctttggga atgatacaag 1140 aagatcaagt accttggttt agggagatgt agaagaggat agtcagagtt caggcagaac 1200 tgtttgatag ttaagagaga gtagttctac aggggtgagg gatggaagga cttttttggc 1260 aatgatggaa atgagatgtc tgcaggagat ggg 1293 <210> 82 <211> 1489 <212> DNA
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 3986488CB1 <400> 82 cggggctggc gggcggcgct cttctacggg acctgctcct tcctcatcgt gcttgtcaac 60 aaggcgctgc tgaccaccta cggtttcccg tcaccaattt tccttggaat tggacagatg 120 gcagccacca taatgatact atatgtgtcc aagctaaaca aaatcattca cttccctgat 180 tttgataaga aaattcctgt aaagctgttt cctctgcctc tcctctacgt tggaaaccac 240 ataagtggat tatcaagcac aagtaaatta agcctaccga tgttcaccgt gctcaggaaa 300 ttcaccattc cacttacctt acttctggaa accatcatac ttgggaagca gtattcactc 360.
aacatcatcc tcagtgtctt tgccattatt ctcggggctt tcatagcagc tgggtctgac 420 cttgctttta acttagaagg ctatattttt gtattcctga atgatatctt cacagcagca 480 aatggagttt ataccaaaca gaaaatggac ccaaaggagc tagggaaata cggagtactt 540 ttctacaatg cctgcttcat gattatccca actcttatta ttagtgtctc cactggagac 600 ctgcaacagg ctactgaatt caaccaatgg aagaatgttg tgtttatcct acagtttctt 660 ctttcctgtt ttttggggtt tctgctgatg tactccacgg ttctgtgcag ctattacaat 720 tcagccctga cgacagcagt ggttggagcc atcaagaatg tatccgttgc ctacattggg 780 atattaatcg gtggagacta cattttctct ttgttaaact ttgtagggtt aaatatttgc 840 atggcagggg gcttgagata ttccttttta acactgagca gccagttaaa acctaaacct 900 gtgggtgaag aaaacatctg tttggatttg aagagctaaa gagtctgcag caggattgga 960 gactgacttg tgactgcggg ctgggggggc attcccagta ggaatgtgaa gccagaggtt 1020 tcggattcgt gacatccacc ccctgggcaa gtgagagcat ctgcaaaatg caaagagaac 1080 tacctcatat gcaggatgag ccaatggcag tctcaagaaa tgtactcggg cgacacctta 1140 cctgtggaaa gcaaatcttt tcaaaataag ccactgggac tcggtaggtg gagccccagc 1200 tgctcttcta gggacctatg gggccttcgt ggcatctctg tgctgtgtgc tggggaggag 1260 gttgatgtaa tggtgactct tttctgatca gcaccttggc cgtgattccc aaggtcccag 1320 ccaaagcaaa gggccagttg tttcagttta aacagacatg tctttagtct aataaaatta 1380 gttaactgcc agtaaagtta tttgttagct ttgatgaaag ctatgttggt atctttccct 1440 aatcatcaaa gtaaataaaa aatcatttct atgtaaaaaa aaaaaaaaa 1489 <210> 83 <211> 927 <212> DNA
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 4378816CB1 <400> 83 ctccctgctt tcctctgccg catggtcctg ggccgttggc gtcggaagcc tgaagcatgg 60 gcgctgagtg ggagctgggg gccgaggctg gcggttcgct gctgctgtgc gccgcgctgc 120 tggcggcggg ctgcgccctg ggcctgcgcc tgggccgcgg gcagggggcg gcggaccgcg 180 gggcgctcat ctggctctgc tacgacgcgc tggtgcactt cgcgctggaa ggcccttttg 240 tctacttgtc tttagtagga aacgttgcaa attccgatgg cttgattgct tctttatgga 300 aagaatatgg caaagctgat gcaagatggg tttattttga tccaaccatt gtgtctgtgg 360 aaattctgac cgtcgccctg gatgggtctc tggcattgtt cctcatttat gccatagtca 420 aagaaaaata ttaccggcat ttcctgcaga tcaccctgtg cgtgtgcgag ctgtatggct 480 gctggatgac cttcctccca gagtggctca ccagaagccc caacctcaac accagcaact 540 ggctgtactg ttggctttac ctgttttttt ttaacggtgt gtgggttctg atcccaggac 600 tgctactgtg gcagtcatgg ctagaactca agaaaatgca tcagaaagaa accagttcag 660 tgaagaagtt tcagtgaact ttcaaaacca taaacaccat tatctaactt catgaaccag 720 aatgaatcaa atctttttgt ttggccaaaa tgtaatacat tccagtctac actttgtttt 780 tgtattgttg ctcctgaaca acctgtttca aattggtttt aaggcgacca gttttcgttg 840 tattgttgtt caattaaatg gtgatatagg gaaaagagaa caaatttgaa tttgtaataa 900 taaaatgttt aattatacaa aaaaaaa 927 <210> 84 <211> 970 <212> DNA
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 4797137CB1 <400> 84 ggatgcagca gagaggagca gctggaagcc gtggctgcgc tctcttccct ctgctgggcg 60 tcctgttctt ccagggtgtt tatatcgtct tttccttgga gattcgtgca gatgcccatg 120 tccgaggtta tgttggagaa aagatcaagt tgaaatgcac tttcaagtca acttcagatg 180 tcactgacaa gcttactata gactggacat atcgccctcc cagcagcagc cacacagtat 240 caatatttca ttatcagtct ttccagtacc caaccacagc aggcacattt cgggatcgga 300 tttcctgggt tggaaatgta tacaaagggg atgcatctat aagtataagc aaccctacca 360 taaaggacaa tgggacattc agctgtgctg tgaagaatcc cccagatgtg caccataata 420 ttcccatgac agagctaaca gtcacagaaa ggggttttgg caccatgctt tcctctgtgg 480 cccttctttc catccttgtc tttgtgccct cagccgtggt ggttgctctg ctgctggtga 540 gaatggggag gaaggctgct gggctgaaga agaggagcag gtctggctat aagaagtcat 600 ctattgaggt ttccgatgac actgatcagg aggaggaaga ggcgtgtatg gcgaggcttt 660 gtgtccgttg cgctgagtgc ctggattcag actatgaaga gacatattga tgaaagtctg 720 tatgacacaa gaagagtcac ctaaagacag gaaacatccc attccactgg cagctaaagc 780 ctgtcagaga aagtggagct ggcctggacc atagcgatgg acaatcctgg agatcatcag 840 taaagacttt aggaaccact tatttattga ataaatgttc ttgttgtatt tataaactgt 900 tcaggaagtc tcataagaga ctcatgactt cccctttcaa tgaattatgc tgtaattgaa 960 tgaagaaatc 970 <210> 85 <211> 594 <212> DNA
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 5470806CB1 <400> 85 gacaggatgg cttcccttcg cctgttcctc ctctgcctcg ctgtactgat atttgcgtct 60 gaagctggcc ctgggggtgc tggagaatcc aagtgtcctc tgatggtcaa agtcctggat 120 gctgtccgag gcagccctgc tgttgatgtg gccgtgaaag tgttcaaaaa gactgcagac 180 ggaagctggg agccgtttgc ctctgggaag accgccgagt ctggagagct gcacgggctc 240 accacagatg agaagttcac ggaaggggtg tacagggtag aactggacac caaatcgtac 300 tggaaggctc ttggcatttc cccattccat gaatacgcag aggtggtttt cacagccaat 360 gactctggtc atcgccacta caccatcgca gccctgctca gcccgtactc ctacagcacc 420 actgctgtcg tcagtaaccc ccagaactga gggacccagc ccagtaggac caggatcttg 480 ccaaagcagt agcttcccat ttgtactgaa acagtgttct tgctctataa accgtgttag 540 caactcggga agatgccgtg aaacgatctt attaaaccac ctgtgatgcc aaaa 594 <210> 86 <211> 618 <212> DNA
<213> Homo Sapiens <220>
<221> misc_feature <223> Incyte ID No: 5473242CB1 <400> 86 gtgttgactc gcaacctcag gaacagacac catggtgcac ctaactgatg ctgagaaggc 60 tactgttaat ggcctgtggg gaaaggtgaa ccctgttgaa attggcgctg agtcccttgc 120 cagtctgctg attgtctacc cttggaccca gaggtacttt tctaaatttg gggacctgtc 180 ctctgtctct gctatcatgg gtaaccccca ggtgaaggcc catggcgaaa aggtgataaa 240 cgccttcgat gatggcctga aacacttgga caacctcaag ggcacctttg ccagcctcag 300 tgaactccac tgtgacaagc tgcatgtgga tcctgagaac ttcaggctcc tgggcaatat 360 gattgtgatt atgatgggcc accacctggg caaggaattc accccgagtg cacaggctgc 420 cttccagaag gtggtggctg gagtggccag tgccctggct cacaagtacc actaaacctc 480 ttttcctgct cttgtctttg tgcaatggtc aattgttccc aagagagctt ctgtcagttg 540 ttgtcaaaat gacaaagacc tttgaaaatc tgtcctacta attaaagcat ttggttcaag 600 tgttctgttg agataccc 618

Claims (33)

What is claimed is:

1. An isolated polypeptide comprising an amino acid sequence selected from the group consisting of:
a) an amino acid sequence selected from the group consisting of SEQ ID NO:1.
SEQ ID
NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID
NO:9, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID
NO:16, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:21, SEQ ID
NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:26, SEQ ID NO:27, SEQ ID
NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:33, SEQ ID
NO:34, SEQ ID NO:35, SEQ ID NO:36, SEQ ID NO:38, SEQ ID NO:39, SEQ ID NO:41, SEQ ID
NO:42, and SEQ ID NO:43, b) a naturally occurring amino acid sequence having at least 90% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NO:1, SEQ ID
NO:2, SEQ ID
NO:3, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID
NO:10, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID
NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:22, SEQ ID
NO:23, SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:26, SEQ ID NO:27, SEQ ID NO:28, SEQ ID
NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO:34, SEQ ID
NO:35, SEQ ID NO:36, SEQ ID NO:38, SEQ ID NO:39, SEQ ID NO:41, SEQ ID NO:42, and SEQ
ID
NO:43, c) a biologically active fragment of an amino acid sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:6, SEQ ID
NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:13, SEQ
ID
NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID
NO:20, SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:25, SEQ ID
NO:26, SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID
NO:32, SEQ ID NO:33, SEQ ID NO:34, SEQ ID NO:35, SEQ ID NO:36, SEQ ID NO:38, SEQ ID
NO:39, SEQ ID NO:41, SEQ ID NO:42, and SEQ ID NO:43, and d) an immunogenic fragment of an amino acid sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:6, SEQ ID
NO:7, SEQ
ID NO:8, SEQ ID NO:9, SEQ ID NO:1O, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID
NO:15, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID
NO:21, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:26, SEQ ID
NO:27, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID
NO:33, SEQ ID NO:34, SEQ ID NO:35, SEQ ID NO:36, SEQ ID NO:38, SEQ ID NO:39, SEQ ID

NO:41, SEQ ID NO:42, and SEQ ID NO:43.

2. An isolated polypeptide of claim 1 selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID
NO:9, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID
NO:16, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:21, SEQ ID
NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:26, SEQ ID NO:27, SEQ ID
NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:33, SEQ ID
NO:34, SEQ ID NO:35, SEQ ID NO:36, SEQ ID NO:38, SEQ ID NO:39, SEQ ID NO:41, SEQ ID
NO:42, and SEQ ID NO:43.

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 selected from the group consisting of SEQ ID
NO:44, SEQ ID NO:45, SEQ ID NO:46, SEQ ID NO:47, SEQ ID NO:49, SEQ ID NO:50, SEQ ID
NO:51, SEQ ID NO:52, SEQ ID NO:53, SEQ ID NO:55, SEQ ID NO:56, SEQ ID NO:57, SEQ ID
NO:58, SEQ ID NO:59, SEQ ID NO:60, SEQ ID NO:61, SEQ ID NO:62, SEQ ID NO:63, SEQ ID
NO:64, SEQ ID NO:65, SEQ ID NO:66, SEQ ID NO:67, SEQ ID NO:68, SEQ ID NO:69, SEQ ID
NO:70, SEQ ID NO:71, SEQ ID NO:72, SEQ ID NO:73, SEQ ID NO:74, SEQ ID NO:75, SEQ ID
NO:76, SEQ ID NO:77, SEQ ID NO:78, SEQ ID NO:79, SEQ ID NO:81, SEQ ID NO:82, SEQ ID
NO:84, SEQ ID NO:85, SEQ ID NO:86.

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 for producing a polypeptide of claim I, 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. An isolated antibody which specifically binds to a polypeptide of claim 1.

11. An isolated polynucleotide comprising a polynucleotide sequence selected from the group consisting of:
a) a polynucleotide sequence selected from the group consisting of SEQ ID
NO:44, SEQ ID
NO:45, SEQ ID NO:46, SEQ ID NO:47, SEQ ID NO:49, SEQ ID NO:50, SEQ ID NO:51, SEQ ID
NO:52, SEQ ID NO:53, SEQ ID NO:55, SEQ ID NO:56, SEQ ID NO:57, SEQ ID NO:58, SEQ ID
NO:59, SEQ ID NO:60, SEQ ID NO:61, SEQ ID NO:62, SEQ ID NO:63, SEQ ID NO:64, SEQ ID
NO:65, SEQ ID NO:66, SEQ ID NO:67, SEQ ID NO:68, SEQ ID NO:69, SEQ ID NO:70, SEQ ID
NO:71, SEQ ID NO:72, SEQ ID NO:73, SEQ ID NO:74, SEQ ID NO:75, SEQ ID NO:76, SEQ ID
NO:77, SEQ ID NO:78, SEQ ID NO:79, SEQ ID NO:81, SEQ ID NO:82, SEQ ID NO:84, SEQ ID
NO:85, SEQ ID NO:86, b) a naturally occurring polynucleotide sequence having at least 70% sequence identity to a polynucleotide sequence selected from the group consisting of SEQ ID NO:44, SEQ ID NO:45, SEQ
ID NO:46, SEQ ID NO:47, SEQ ID NO:49, SEQ ID NO:50, SEQ ID NO:51, SEQ ID
NO:52, SEQ ID
NO:53, SEQ ID NO:55, SEQ ID NO:56, SEQ ID NO:57, SEQ ID NO:58, SEQ ID NO:59, SEQ ID
NO:60, SEQ ID NO:61, SEQ ID NO:62, SEQ ID NO:63, SEQ ID NO:64, SEQ ID NO:65, SEQ ID
NO:66, SEQ ID NO:67, SEQ ID NO:68, SEQ ID NO:69, SEQ ID NO:70, SEQ ID NO:71, SEQ ID
NO:72, SEQ ID NO:73, SEQ ID NO:74, SEQ ID NO:75, SEQ ID NO:76, SEQ ID NO:77, SEQ ID
NO:78, SEQ ID NO:79, SEQ ID NO:81, SEQ ID NO:82, SEQ ID NO:84, SEQ ID NO:85, SEQ ID
NO:86, c) a polynucleotide sequence complementary to a), d) a polynucleotide sequence complementary to b), and e) an RNA equivalent of a)-d).

12. An isolated polynucleotide comprising at least 60 contiguous nucleotides of a polynucleotide of claim 11.

13. A method for detecting a target polynucleotide in a sample, said target polynucleotide having a sequence of a polynucleotide of claim 11, 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.

14. A method of claim 13, wherein the probe comprises at least 60 contiguous nucleotides.

15. A method for detecting a target polynucleotide in a sample, said target polynucleotide having a sequence of a polynucleotide of claim 11, 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.

16. A pharmaceutical composition comprising an effective amount of a polypeptide of claim 1 and a pharmaceutically acceptable excipient.

17. A pharmaceutical composition of claim 16, wherein the polypeptide comprises an amino acid sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID
NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:17, SEQ ID
NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:23, SEQ ID
NO:24, SEQ ID NO:25, SEQ ID NO:26, SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:29, SEQ ID
NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO:34, SEQ ID NO:35, SEQ ID
NO:36, SEQ ID NO:38, SEQ ID NO:39, SEQ ID NO:41, SEQ ID NO:42, and SEQ ID
NO:43.

18. A method for treating a disease or condition associated with decreased expression of functional TPPT, comprising administering to a patient in need of such treatment the pharmaceutical composition of claim 16.

19. A method for 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.

20. A pharmaceutical composition comprising an agonist compound identified by a method of claim 19 and a pharmaceutically acceptable excipient.

21. A method for treating a disease or condition associated with decreased expression of functional TPPT, comprising administering to a patient in need of such treatment a pharmaceutical composition of claim 20.

22. A method for 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.

23. A pharmaceutical composition comprising an antagonist compound identified by a method of claim 22 and a pharmaceutically acceptable excipient.

24. A method for treating a disease or condition associated with overexpression of functional TPPT, comprising administering to a patient in need of such treatment a pharmaceutical composition of claim 23.

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

26. A method of screening for a compound that modulates the activity of the polypeptide of claim 1, said 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.

27. A method for 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, and b) detecting altered expression of the target polynucleotide.

28. An isolated polynucleotide comprising a polynucleotide sequence of SEQ ID
NO:83.

29. A recombinant polynucleotide comprising a promoter sequence operably linked to a polynucleotide of claim 28.

30. A cell transformed with a recombinant polynucleotide of claim 29.

31. A transgenic organism comprising a recombinant polynucleotide of claim 29.

32. A method for producing a polypeptide comprising an amino acid sequence of SEQ ID
NO:40, the method comprising:
a) culturing the cell of claim 30 under conditions suitable for expression of the polypeptide, and b) recovering the polypeptide so expressed.

33. A method for screening a compound for effectiveness in altering expression of a target polynucleotide, wherein said target polynucleotide comprises a sequence of claim 28, the method comprising:
a) exposing a sample comprising the target polynucleotide to a compound, and b) detecting altered expression of the target polynucleotide.