Patent Publication Number: US-2004048253-A1

Title: Molecules for diagnostics and therapeutics

Description:
TECHNICAL FIELD  
       [0001] The present invention relates to human molecules and to the use of these sequences ill the diagnosis, study, prevention, and treatment of diseases associated with, as well as effects of exogenous compounds on, the expression of human molecules.  
       BACKGROUND OF THE INVENTION  
       [0002] The human genome is comprised of thousands of genes, many encoding gene products that function in the maintenance and growth of the various cells and tissues in the body. Aberrant expression or mutations in these genes and their products is the cause of, or is associated with, a variety of human diseases such as cancer and other cell proliferative disorders, autoimmune/inflammatory disorders, infections, developmental disorders, endocine disorders, metabolic disorders, neurological disorders, gastrointestinal disorders, transport disorders, and connective tissue disorders. The identification of these genes and their products is the basis of an ever-expanding effort to find markers for early detection of diseases, and targets for their prevention and treatment. Therefore, these genes and their products are useful as diagnostics and therapeutics. These genes may encode, for example, enzyme molecules, molecules associated with growth and development, biochemical pathway molecules, extracellular information transmission molecules, receptor molecules, intracellular signaling molecules, membrane transport molecules, protein modification and maintenance molecules, nucleic acid synthesis and modification molecules, adhesion molecules, antigen recognition molecules, secreted and extracellular matrix molecules, cytoskeletal molecules, ribosomal molecules, electron transfer associated molecules, transcription factor molecules, chromatin molecules, cell membrane molecules, and organelle associated molecules.  
       [0003] For example, cancer represents a type of cell proliferative disorder that affects nearly every tissue in the body. A wide variety of molecules, either aberrantly expressed or mutated, can be the cause of, or involved with, various cancers because tissue growth involves complex and ordered patterns of cell proliferation, cell differentiation, and apoptosis. Cell proliferation must be regulated to maintain both the number of cells and their spatial organization. This regulation depends upon the appropriate expression of proteins which control cell cycle progression in response to extracellular signals such as growth factors and other mitogens, and intracellular cues such as DNA damage or nutrient starvation. Molecules which directly or indirectly modulate cell cycle progression fall into several categories, including growth factors and their receptors, second messenger and signal transduction proteins, oncogene products, tumor-suppressor proteins, and mitosis-promoting factors. Aberrant expression or mutations in any of these gene products can result in cell proliferative disorders such as cancer. Oncogenes are genes generally derived from normal genes that, through abnormal expression or mutation, can effect the transformation of a normal cell to a malignant one (oncogenesis). Oncoproteins, encoded by oncogenes, can affect cell proliferation in a variety of ways and include growth factors, growth factor receptors, intracellular signal transducers, nuclear transcription factors, and cell-cycle control proteins. In contrast, tumor-suppressor genes are involved in inhibiting cell proliferation. Mutations which cause reduced function or loss of function in tumor-suppressor genes result in aberrant cell proliferation and cancer. Although many different genes and their products have been found to be associated with cell proliferative disorders such as cancer, many more may exist that are yet to be discovered.  
       [0004] DNA-based arrays can provide a simple way to explore the expression of a single polymorphic gene or a large number of genes. When the expression of a single gene is explored, DNA-based arrays are employed to detect the expression of specific gene variants. For example, a p53 tumor suppressor gene array is used to determine whether individuals are carrying mutations that predispose them to cancer. A cytocbrome p450 gene array is useful to determine whether individuals have one of a number of specific mutations that could result in increased drug metabolism, drug resistance or drug toxicity.  
       [0005] DNA-based array technology is especially relevant for the rapid screening of expression of a large number of genes. There is a growing awareness that gene expression is affected in a global fashion. A genetic predisposition, disease or therapeutic treatment may affect, directly or indirectly, the expression of a large number of genes. In some cases the interactions may be expected, such as when the genes are part of the same signaling pathway. In other cases, such as when the genes participate in separate signaling pathways, the interactions may be totally unexpected. Therefore, DNA based arrays can be used to investigate how genetic predisposition, disease, or therapeutic treatment affects the expression of a large number of genes.  
       [0006] Enzyme Molecules  
       [0007] The cellular processes of biogenesis and biodegradation involve a number of key enzyme classes including oxidoreductases, transferases, hydrolases, lyases, isomerases, and ligases. These enzyme classes are each comprised of numerous substrate-specific enzymes having precise and well regulated functions. These enzymes function by facilitating metabolic processes such as glycolysis, the tricarboxylic cycle, and fatty acid metabolism; synthesis or degradation of amino acids, steroids, phospholipids, alcohols, etc.; regulation of cell signalling, proliferation, inflamation, apoptosis, etc., and through catalyzing critical steps in DNA replication and repair, and the process of translation.  
       [0008] Oxidoreductases  
       [0009] Many pathways of biogenesis and biodegradation require oxidoreductase (dehydrogenase or reductase) activity, coupled to the reduction or oxidation of a donor or acceptor cofactor. Potential cofactors include cytochromes, oxygen, disulfide, iron-sulfur proteins, flavin adenine dinucleotide (FAD), and the nicotinamide adenine dinucleotides NAD and NADP (Newsholme, E. A and A. R. Leech (1983)  Biochemistry for the Medical Sciences , John Wiley and Sons, Chichester, U.K, pp. 779-793). Reductase activity catalyzes the transfer of electrons between substrate(s) and cofactor(s) with concurrent oxidation of the cofactor. The reverse dehydrogenase reaction catalyzes the reduction of a cofactor and consequent oxidation of the substrate. Oxidoreductase enzymes are a broad superfamily of proteins that catalyze numerous reactions in all cells of organisms ranging from bacteria to plants to humans. These reactions include metabolism of sugar, certain detoxification reactions in the liver, and the synthesis or degradation of fatty acids, amino acids, glucocorticoids, estrogens, androgens, and prostaglandins. Different family members are named according to the direction in which their reactions are typically catalyzed; thus they may be referred to as oxidoreductases, oxidases, reductases, or dehydrogenases. In addition, family members often have distinct cellular localizations, including the cytosol, the plasma membrane, mitochondrial inner or outer membrane, and peroxisomes.  
       [0010] Short-chain alcohol dehydrogenases (SCADs) are a family of dehydrogenases that only share 15% to 30% sequence identity, with similarity predominantly in the coenzyme binding domain and the substrate binding domain. In addition to the well-known role in detoxification of ethanol, SCADs are also involved in synthesis and degradation of fatty acids, steroids, and some prostaglandins, and are therefore implicated in a variety of disorders such as lipid storage disease, myopathy, SCAD deficiency, and certain genetic disorders. For example, retinol dehydrogenase is a SCAD-family member (Simon, A. et al. (1995) J. Biol. Chem. 270:1107-1112) that converts retinol to retinal, the precursor of retinoic acid. Retinoic acid, a regulator of differentiation and apoptosis, has been shown to down-regulate genes involved in cell proliferation and inflammation (Chai, X. et al. (1995) J. Biol. Chem. 270:3900-3904). In addition, retinol dehydrogenase has been linked to hereditary eye diseases such as autosomal recessive childhood-onset severe retinal dystrophy (Simon, A et al; (1996) Genomics 36:424-430).  
       [0011] Propagation of nerve impulses, modulation of cell proliferation and differentiation, induction of the immune response, and tissue homeostasis involve neurotransmitter metabolism (Weiss, B. (1991) Neurotoxicology 12:379-386; Collins, S. M. et al. (1992) Ann N.Y. Acad. Sci. 664:415424; Brown, J. K. and H. Imam (1991) J. Inherit. Metab. Dis.  14:436-458 ). Many pathways of neurotransmitter metabolism require oxidoreductase activity, coupled to reduction or oxidation of a cofactor, such as NAD + /NADH (Newsholme, E. A. and A. R. Leech (1983)  Biochemistry for the Medical Sciences , John Wiley and Sons, Chichester, U.K pp. 779-793). Degradation of catecholamies (epinephrine or norepinephrine) requires alcohol dehydrogenase (in the brain) or aldehyde dehydrogenase (in peripheral tissue). NAD + -dependent aldehyde dehydrogenase oxidizes 5-hydroxyindole-3-acetate (the product of 5-hydroxytryptamine (serotonin) metabolism) in the brain, blood platelets, liver and pulmonary endothelium (Newsholme, supra, p. 786). Other neurotransmitter degradation pathways that utilize NAD + /NADH-dependent oxidoreductase activity include those of L-DOPA (precursor of dopamine, a neuronal excitatory compound), glycine (an inhibitory neurotransmitter in the brain and spinal cord), histamine (liberated from mast cells during the inflammatory response), and taurine (an inhibitory neurotransmitter of the brain stem, spinal cord and retina) (Newsholme, supra, pp. 790, 792). Epigenetic or genetic defects in neurotransmitter metabolic pathways can result in a spectrum of disease states in different tissues including Parkinson disease and inherited myoclonus (McCance, K. L. and S. E. Huether (1994)  Pathophysiology , Mosby-Year Book, Inc., St Louis Mo., pp. 402-404; Gundlach, AL. (1990) FASEB J. 4:2761-2766).  
       [0012] Tetrahydrofolate is a derivatized glutamate molecule that acts as a carrier, providing activated is, one-carbon units to a wide variety of biosynthetic reactions, including synthesis of purines, pyrimidines, and the amino acid methionine. Tetrahydrofolate is generated by the activity of a holoenzyme complex called tetrahydrofolate synthase, which includes three enzyme activities: tetrahydrofolate dehydrogenase, tetrahydrofolate cyclohydrolase, and tetrahydrofolate synthetase. Thus, tetrahydrofolate dehydrogenase plays an important role in generating building blocks for nucleic and amino acids, crucial to proliferating cells.  
       [0013] 3-Hydroxyacyl-CoA dehydrogenase (3HACD) is involved in fatty acid metabolism. It catalyzes the reduction of 3-hydroxyacyl-CoA to 3-oxoacyl-CoA, with concomitant oxidation of NAD to NADH, in the mitochondria and peroxisomes of eukaryotic cells. In peroxisomes, 3HACD and enoyl-CoA hydratase form an enzyme complex called bifunctional enzyme, defects in which are associated with peroxisomal bifunctional enzyme deficiency. This interruption in fatty acid metabolism produces accumulation of very-long chain fatty acids, disrupting development of the brain, bone, and adrenal glands. Infants born with this deficiency typically die within 6 months (Watkins, P. et al. (1989) J. Clin. Invest. 83:771-777; Online Mendelian Inheritance in Man (OMIM) # 261-515 ). The neurodegeneration that is characteristic of Alzheimer&#39;s disease involves development of extracellular plaques in certain brain regions. A major protein component of these plaques is the peptide amyloid-β (Aβ), which is one of several cleavage products of amyloid precursor protein (APP). 3HACD has been shown to bind the Aβ peptide, and is overexpressed in neurons affected in Alzheimer&#39;s disease. In addition, an antibody against 3HACD can block the toxic effects of Aβ in a cell culture model of Alzheimer&#39;s disease (Yan, S. et al. (1997) Nature 389:689-695; OMIM, #602057).  
       [0014] Steroids, such as estrogen, testosterone, corticosterone, and others, are generated from a common precursor, cholesterol, and are interconverted into one another. A wide variety of enzymes act upon cholesterol, including a number of dehydrogenases. Steroid dehydrogenases, such as the hydroxysteroid dehydrogenases, are involved in hypertension, fertility, and cancer (Duax, W. L. and D. Ghosh (1997) Steroids 62:95-100). One such dehydrogenase is 3-oxo-5-α-steroid dehydrogenase (OASD), a microsomal membrane protein highly expressed in prostate and other androgen-responsive tissues. OASD catalyzes the conversion of testosterone into dihydrotestosterone, which is the most potent androgen. Dihydrotestosterone is essential for the formation of the male phenotype during embryogenesis, as well as for proper androgen-mediated growth of tissues such as the prostate and male genitalia. A defect in OASD that prevents the conversion of testosterone into dihydrotestosterone leads to a rare form of male pseudohermaphroditis, characterized by defective formation of the external genitalia (Andersson, S. et al. (1991) Nature 354:159-161; Labrie, F. et al. (1992) Endocrinology 131:1571-1573; OMIM #264600). Thus, OASD plays a central role in sexual differentiation and androgen physiology.  
       [0015] 17β-hydroxysteroid dehydrogenase (17βHSD6) plays an important role in the regulation of the male reproductive hormone, dihydrotestosterone (DHTT). 17βHSD6 acts to reduce levels of DHTT by oxidizing a precursor of DHTT, 3α-diol, to androsterone which is readily glucuronidated and removed from tissues. 17βHSD6 is active with both androgen and estrogen substrates when expressed in embryonic kidney 293 cells. At least five other isozymes of 17 βHSD have been identified that catalyze oxidation and/or reduction reactions in various tissues with preferences for different steroid substrates (Biswas, M. G. and D. W. Russell (1997) J. Biol. Chem. 272:15959-15966). For example, 17,HSD1 preferentially reduces estradiol and is abundant in the ovary and placenta 17βHSD2 catalyzes oxidation of androgens and is present in the endometrium and placenta. 17βHSD3 is exclusively a reductive enzyme in the testis (Geissler, W. M. et al. (1994) Nat. Genet. 7:34-39). An excess of androgens such as DHTT can contribute to certain disease states such as benign prostatic hyperplasia and prostate cancer.  
       [0016] Oxidoreductases are components of the fatty acid metabolism pathways in mitochondria and peroxisomes. The main beta-oxidation pathway degrades both saturated and unsaturated fatty acids, while the auxiliary pathway performs additional steps required for the degradation of unsaturated fatty acids. The auxiliary beta-oxidation enzyme 2,4-dienoyl-CoA reductase catalyzes the removal of even-numbered double bonds from unsaturated fatty acids prior to their entry into the main beta-oxidation pathway. The enzyme may also remove odd-numbered double bonds from unsaturated fatty acids (Koivuranta, K. T. et al. (1994) Biochem. J. 304:787-792; Smeland, T. E. et al. (1992) Proc.  
       [0017] Natl. Acad. Sci. USA 89:6673-6677). 2,4-dienoyl-CoA reductase is located in both mitochondria and peroxisomes. Inherited deficiencies in mitochondrial and peroxisomal beta-oxidation enzymes are associated with severe diseases, some of which manifest themselves soon after birth and lead to death within a few years. Defects in beta-oxidation are associated with Reye&#39;s syndrome, Zellweger syndrome, neonatal adrenoleukodystrophy, infantile Refsum&#39;s disease, acyl-CoA oxidase deficiency, and bifunctional protein deficiency (Suzuki, Y. et al. (1994) Am. J. Hum. Genet. 54:36-43; Hoefler, supra; Cotran, R. S. et al. (1994)  Robbins Pathologic Basis of Disease , W. B. Saunders Co., Philadelphia Pa., p.866). Peroxisomal beta-oxidation is impaired in cancerous tissue. Although neoplastic human breast epithelial cells have the same number of peroxisomes as do normal cells, fatty acyl-CoA oxidase activity is lower than in control tissue (el Bouhtoury, F. et al. (1992) J. Pathol. 166:27-35). Human colon carcinomas have fewer peroxisomes than normal colon tissue and have lower fatty-acyl-CoA oxidase and bifunctional enzyme (including enoyl-CoA hydratase) activities than normal tissue (Cable, S. et al. (1992) Virchows Arch. B Cell Pathol. Incl. Mol. Pathol. 62:221-226). Another important oxidoreductase is isocitrate dehydrogenase, which catalyzes the conversion of isocitrate to a-ketoglutarate, a substrate of the citric acid cycle. Isocitrate dehydrogenase can be either NAD or NADP dependent, and is found in the cytosol, mitochondria, and peroxisomes. Activity of isocitrate dehydrogenase is regulated developmentally, and by hormones, neurotransmitters, and growth factors.  
       [0018] Hydroxypyruvate reductase (HPR), a peroxisomal 2-hydroxyacid dehydrogenase in the glycolate pathway, catalyzes the conversion of hydroxypyruvate to glycerate with the oxidation of both NADH and NADPH. The reverse dehydrogenase reaction reduces NAD + and NADP + . HPR recycles nucleotides and bases back into pathways leading to the synthesis of ATP and GTP. ATP and GTP are used to produce DNA and RNA and to control various aspects of signal transduction and energy metabolism. Inhibitors of purine nucleotide biosynthesis have long been employed as antiproliferative agents to treat cancer and viral diseases. HPR also regulates biochemical synthesis of serine and cellular serine levels available for protein synthesis.  
       [0019] The mitochondrial electron transport (or respiratory) chain is a series of oxidoreductase-type enzyme complexes in the mitochondrial membrane that is responsible for the transport of electrons from NADH through a series of redox centers within these complexes to oxygen, and the coupling of this oxidation to the synthesis of ATP (oxidative phosphorylation). ATP then provides the primary source of energy for driving a cell&#39;s many energy-requiring reactions. The key complexes in the respiratory chain are NADH:ubiquinone oxidoreductase (complex I), succinate:ubiquinone oxidoreductase (complex II), cytochrome c 1 -b oxidoreductase (complex III), cytochrome c oxidase (complex IV), and ATP synthase (complex V) (Alberts, B. et al. (1994)  Molecular Biology of the Cell , Garland Publishing, Inc., New York N.Y., pp. 677-678). All of these complexes are located on the inner matrix side of the mitochondrial membrane except complex II, which is on the cytosolic side. Complex II transports electrons generated in the citric acid cycle to the respiratory chain. The electrons generated by oxidation of succinate to fumarate in the citric acid cycle are transferred through electron carriers in complex II to membrane bound ubiquinone (Q). Transcriptional regulation of these nuclear-encoded genes appears to be the predominant means for controlling the biogenesis of respiratory enzymes. Defects and altered expression of enzymes in the respiratory chain are associated with a variety of disease conditions.  
       [0020] Other dehydrogenase activities using NAD as a cofactor are also important in mitochondrial function. 3-hydroxyisobutyrate dehydrogenase (3HBD), important in valine catabolism, catalyzes the NAD-dependent oxidation of 3-hydroxyisobutyrate to methylmalonate semialdehyde within mitochondria. Elevated levels of 3-hydroxyisobutyrate have been reported in a number of disease states, including ketoacidosis, methylmalonic acidemia, and other disorders associated with deficiencies in methylmalonate semialdehyde dehydrogenase (Rougraff, P. M. et al. (1989) J. Biol. Chem. 264:5899-5903).  
       [0021] Another mitochondrial dehydrogenase important in amino acid metabolism is the enzyme isovaleryl-CoA-dehydrogenase (IVD). IVD is involved in leucine metabolism and catalyzes the oxidation of isovaleryl-CoA to 3-methylcrotonyl-CoA. Human IVD is a tetrameric flavoprotein that is encoded in the nucleus and synthesized in the cytosol as a 45 kDa precursor with a mitochondrial import signal sequence. A genetic deficiency, caused by a mutation in the gene encoding IVD, results in the condition known as isovaleric acidemia. This mutation results in inefficient mitochondrial import and processing of the IVD precursor (Vockley, J. et al. (1992) J. Biol. Chem. 267:2494-2501).  
       [0022] Transferases  
       [0023] Transferases are enzymes that catalyze the transfer of molecular groups. The reaction may involve an oxidation, reduction, or cleavage of covalent bonds, and is often specific to a substrate or to particular sites on a type of substrate. Transferases participate in reactions essential to such functions as synthesis and degradation of cell components, regulation of cell functions including cell signaling, cell proliferation, inflamation, apoptosis, secretion and excretion. Transferases are involved in key steps in disease processes involving these functions. Transferases are frequently classified according to the type of group transferred. For example, methyl transferases transfer one-carbon methyl groups, amino transferases transfer nitrogenous amino groups, and similarly denominated enzymes transfer aldehyde or ketone, acyl, glycosyl, alkyl or aryl, isoprenyl, saccharyl, phosphorous-containing, sulfur-containing, or selenium-containing groups, as well as small enzymatic groups such as Coenzyme A  
       [0024] Acyl transferases include peroxisomal carnitine octanoyl transferase, which is involved in the fatty acid beta-oxidation pathway, and mitochondrial carnitine palmitoyl transferases, involved in fatty acid metabolism and transport. Choline O-acetyl transferase catalyzes the biosynthesis of the neurotransmitter acetylcholine.  
       [0025] Amino transferases play key roles in protein synthesis and degradation, and they contribute to other processes as well. For example, the amino transferase 5-aminolevulinic acid synthase catalyzes the addition of succinyl-CoA to glycine, the first step in heme biosynthesis. Other amino transferases participate in pathways important for neurological function and metabolism. For example, glutamine-phenylpyruvate amino transferase, also known as glutamine transaminase K (GTK), catalyzes several reactions with a pyridoxal phosphate cofactor. GTK catalyzes the reversible conversion of L-glutamine and phenylpyruvate to 2-oxoglutaramate and L-phenylalanine. Other amino acid substrates for GTK include L-methionine, L-histidine, and L-tyrosine. GTK also catalyzes the conversion of kynurenine to kynurenic acid, a tryptophan metabolite that is an antagonist of the N-methyl-D-aspartate (NMDA) receptor in the brain and may exert a neuromodulatory function. Alteration of the kynurenine metabolic pathway may be associated with several neurological disorders. GTK also plays a role in the metabolism of halogenated xenobiotics conjugated to glutathione, leading to nephrotoxicity in rats and neurotoxicity in humans. GTK is expressed in kidney, liver, and brain. Both human and rat GTKs contain a putative pyridoxal phosphate binding site (ExPASy ENZYME: EC 2.6.1.64; Perry, S. J. et al. (1993) Mol. Pharmacol. 43:660-665; Perry, S. et al. (1995) FEBS Lett 360:277-280; and Alberati-Giani, D. et al. (1995) J. Neurochem. 64:1448-1455). A second amino transferase associated with this pathway is kynurenine/α-aminoadipate amino transferase (AadAT. AadAT catalyzes the reversible conversion of α-aminoadipate and α-ketoglutarate to α-ketoadipate and L-glutamate during lysine metabolism. AadAT also catalyzes the transamination of kynurenine to kynurenic acid. A cytosolic AadAT is expressed in rat kidney, liver, and brain (Nakatani, Y. et al. (1970) Biochim Biophys. Acta 198:219-228; Buchli, R. et al. (1995) J. Biol. Chem. 270:29330-29335).  
       [0026] Glycosyl transferases include the mammalian UDP-glucouronosyl transferases, a family of membrane-bound microsomal enzymes catalyzing the transfer of glucouronic acid to lipophilic substrates in reactions that play important roles in detoxification and excretion of drugs, carcinogens, and other foreign substances. Another mammalian glycosyl transferase, mammalian UDP-galactose-ceramide galactosyl transferase, catalyzes the transfer of galactose to ceramide in the synthesis of galactocerebrosides in myelin membranes of the nervous system. The UDP-glycosyl transferases share a conserved signature domain of about 50 amino acid residues (PROSITE: PDOC00359, http://expasyhcuge.ch/sprotfprosite html).  
       [0027] Methyl transferases are involved in a variety of pharmacologically important processes. Nicotinamide N-methyl transferase catalyzes the N-methylation of nicotinamides and other pyridines, an important step in the cellular handling of drugs and other foreign compounds. Phenylethanolamine N-methyl transferase catalyzes the conversion of noradrenalin to adrenalin 6-O-methylguanine-DNA methyl transferase reverses DNA methylation, an important step in carcinogenesis. Uroporphyrin-III C-methyl transferase, which catalyzes the transfer of two methyl groups from S-adenosyl-L-methionine to uroporphyrinogen III, is the first specific enzyme in the biosynthesis of cobalamin, a dietary enzyme whose uptake is deficient in pernicious anemia. Protein-arginine methyl transferases catalyze the posttranslational methylation of arginine residues in proteins, resulting in the mono- and dimethylation of arginine on the guanidino group. Substrates include histones, myelin basic protein, and heterogeneous nuclear ribonucleoproteins involved in mRNA processing, splicing, and transport. Protein-arginine methyl transferase interacts with proteins upregulated by mitogens, with proteins involved in chronic lymphocytic leukemia, and with interferon, suggesting an important role for methylation in cytokine receptor signaling (Lin, W. -J. et is al. (1996) J. Biol. Chem. 271:15034-15044; Abramovich, C. et al. (1997) EMBO J. 16:260-266; and Scott, H. S. et al. (1998) Genomics  48:330-340 ).  
       [0028] Phosphotransferases catalyze the transfer of high-energy phosphate groups and are important in energy-requiring and-releasing reactions. The metabolic enzyme creatine kinase catalyzes the reversible phosphate transfer between creatine/creatine phosphate and ATP/ADP. Glycocyamine kinase catalyzes phosphate transfer from ATP to guanidoacetate, and arginine kinase catalyzes phosphate transfer from ATP to arginine. A cysteine-containing active site is conserved in this family (PROSITE: PDOC00103).  
       [0029] Prenyl transferases are heterodimers, consisting of an alpha and a beta subunit, that catalyze the transfer of an isoprenyl group. An example of a prenyl transferase is the mammalian protein farnesyl transferase. The alpha subunit of farnesyl transferase consists of 5 repeats of 34 amino acids each, with each repeat containing an invariant tryptophan (PROSITE: PDOC00703).  
       [0030] Saccharyl transferases are glycating enzymes involved in a variety of metabolic processes. Oligosacchryl transferase-48, for example, is a receptor for advanced glycation endproducts. Accumulation of these endproducts is observed in vascular complications of diabetes, macrovascular disease, renal insufficiency, and Alzheimer&#39;s disease (Thornalley, P. J. (1998) Cell Mol. Biol. (Noisy-Le-Grand) 44:1013-1023).  
       [0031] Coenzyme A (CoA) transferase catalyzes the transfer of CoA between two carboxylic acids. Succinyl CoA.3-oxoacid CoA transferase, for example, transfers CoA from succinyl-CoA to a recipient such as acetoacetate. Acetoacetate is essential to the metabolism of ketone bodies, which accumulate in tissues affected by metabolic disorders such as diabetes (PROSITE: PDOC00980).  
       [0032] Hydrolases  
       [0033] Hydrolysis is the breaking of a covalent bond in a substrate by introduction of a molecule of water. The reaction involves a nucleophilic attack by the water molecule&#39;s oxygen atom on a target bond in the substrate. The water molecule is split across the target bond, breaking the bond and generating two product molecules. Hydrolases participate in reactions essential to such functions as synthesis and degradation of cell components, and for regulation of cell functions including cell signaling, cell proliferation, inflamation, apoptosis, secretion and excretion. Hydrolases are involved in key steps in disease processes involving these functions. Hydrolytic enzymes, or hydrolases, may be grouped by substrate specificity into classes including phosphatases, peptidases, lysophospholipases, phosphodiesterases, glycosidases, and glyoxalases.  
       [0034] Phosphatases hydrolytically remove phosphate groups from proteins, an energy-providing step that regulates many cellular processes, including intracellular signaling pathways that in turn control cell growth and differentiation, cell-cell contact, the cell cycle, and oncogenesis.  
       [0035] Lysophospholipases (LPLs) regulate intracellular lipids by catalyzing the hydrolysis of ester bonds to remove an acyl group, a key step in lipid degradation. Small LPL isoforms, approximately 15-30 kD, function as hydrolases; larger isoforms function both as hydrolases and transacylases. A particular substrate for LPLs, lysophosphatidylcholine, causes lysis of cell membranes. LPL activity is regulated by signaling molecules important in numerous pathways, including the inflammatory response.  
       [0036] Peptidases, also called proteases, cleave peptide bonds that form the backbone of peptide or protein chains. Proteolytic processing is essential to cell growth, differentiation, remodeling, and homeostasis as well as inflammation and immune response. Since typical protein half-lives range from hours to a few days, peptidases are continually cleaving precursor proteins to their active form, removing signal sequences from targeted proteins, and degrading aged or defective proteins. Peptidases function in bacterial, parasitic, and viral invasion and replication within a host. Examples of peptidases include trypsin and chymotrypsin (components of the complement cascade and the blood-clotting cascade) lysosomal cathepsins, calpains, pepsin, renin, and chymosin (Beynon, R. J. and J. S. Bond (1994)  Proteolytic Enzymes: A Practical Approach , Oxford University Press, New York N.Y., pp. 1-5).  
       [0037] The phosphodiesterases catalyze the hydrolysis of one of the two ester bonds in a phosphodiester compound. Phosphodiesterases are therefore crucial to a variety of cellular processes. Phosphodiesterases include DNA and RNA endo- and exo-nucleases, which are essential to cell growth and replication as well as protein synthesis. Another phosphodiesterase is acid sphingomyelinase, which hydrolyzes the membrane phosphoilpid sphingomyelin to ceramide and phosphorylcholine. Phosphorylcholine is used in the synthesis of phosphatidylcholine, which is involved in numerous intracellular signaling pathways. Ceramide is an essential precursor for the generation of gangliosides, membrane lipids found in high concentration in neural tissue. Defective acid sphingomyelinase phosphodiesterase leads to a build-up of sphingomyelin molecules in lysosomes, resulting in Niemann-Pick disease.  
       [0038] Glycosidases catalyze the cleavage of hemiacetyl bonds of glycosides, which are compounds that contain one or more sugar. Mammalian lactase-phlorizin hydrolase, for example, is an intestinal enzyme that splits lactose. Mammalian beta-galactosidase removes the terminal galactose from gangliosides, glycoproteins, and glycosaminoglycans, and deficiency of this enzyme is associated with a gangliosidosis known as Morquio disease type B. Vertebrate lysosomal alpha-glucosidase, which hydrolyzes glycogen, maltose, and isomaltose, and vertebrate intestinal sucrase-isomaltase, which hydrolyzes sucrose, maltose, and isomaltose, are widely distributed members of this family with highly conserved sequences at their active sites.  
       [0039] The glyoxylase system is involved in gluconeogenesis, the production of glucose from storage compounds in the body. It consists of glyoxylase I, which catalyzes the formation of S-D-lactoylglutathione from methyglyoxal, a side product of triose-phosphate energy metabolism, and glyoxylase II, which hydrolyzes S-D-lactoylglutathione to D-lactic acid and reduced glutathione. Glyoxylases are involved in hyperglycemia, non-insulin-dependent diabetes mellitus, the detoxification of bacterial toxins, and in the control of cell proliferation and microtubule assembly.  
       [0040] Lyases  
       [0041] Lyases are a class of enzymes that catalyze the cleavage of C—C, C—O, C—N, C—S, C-(halide), P—O or other bonds without hydrolysis or oxidation to form two molecules, at least one of which contains a double bond (Stryer, L. (1995)  Biochemistry  W. H. Freeman and Co. New York, N.Y. p.620). Lyases are critical components of cellular biochemistry with roles in metabolic energy production including fatty acid metabolism, as well as other diverse enzymatic processes. Further classification of lyases reflects the type of bond cleaved as well as the nature of the cleaved group.  
       [0042] The group of C—C lyases include carboxyl-lyases (decarboxylases), aldehyde-lyases (aldolases), oxo-acid-lyases and others. The C—O lyase group includes hydro-lyases, lyases acting on polysaccharides and other lyases. The C—N lyase group includes ammonia-lyases, amidine-lyases, amine-lyases (deaminases) and other lyases.  
       [0043] Proper regulation of lyases is critical to normal physiology. For example, mutation induced deficiencies in the uroporphyrinogen decarboxylase can lead to photosensitive cutaneous lesions in the genetically-linked disorder familial porphyria cutanea tarda (Mendez, M. et al. (1998) Am. J. Genet. 63:1363-1375). It has also been shown that adenosine deaminase (ADA) deficiency stems from genetic mutations in the ADA gene, resulting in the disorder severe combined immunodeficiency disease (SCID) (Hershfield, M. S. (1998) Semin. Hematol. 35:291-298).  
       [0044] Isomerases  
       [0045] Isomerases are a class of enzymes that catalyze geometric or structural changes within a molecule to form a single product. This class includes racemases and epimerases, cis-trans-isomerases, intramolecular oxidoreductases, intramolecular transferases (mutases) and intramolecular lyases. Isomerases are critical components of cellular biochemistry with roles in metabolic energy production including glycolysis, as well as other diverse enzymatic processes (Stryer, L. (1995)  Biochemistry , W. H. Freeman and Co., New York N.Y., pp.483-507).  
       [0046] Racemases are a subset of isomerases that catalyze inversion of a molecules configuration around the asymmetric carbon atom in a substrate having a single center of asymmetry, thereby interconverting two racemers. Epimerases are another subset of isomerases that catalyze inversion of configuration around an asymmetric carbon atom in a substrate with more than one center of symmetry, thereby interconverting two epimers. Racemases and epimerases can act on amino acids and derivatives, hydroxy acids and derivatives, as well as carbohydrates and derivatives. The interconversion of UDP-galactose and UDP-glucose is catalyzed by UDP-galactose4′-epimerase. Proper regulation and function of this epimerase is essential to the synthesis of glycoproteins and glycolipids. Elevated blood galactose levels have been correlated with UDP-galactose4′-epimerase deficiency in screening programs of infants (Gitzelmann, R. (1972) Helv. Paediat. Acta 27:125-130).  
       [0047] Oxidoreductases can be isomerases as well. Oxidoreductases catalyze the reversible transfer of electrons from a substrate that becomes oxidized to a substrate that becomes reduced. This class of enzymes includes dehydrogenases, hydroxylases, oxidases, oxygenases, peroxidases, and reductases. Proper maintenance of oxidoreductase levels is physiologically important. For example, genetically-linked deficiencies in lipoamide dehydrogenase can result in lactic acidosis (Robinson, B. H. et al. (1977) Pediat. Res. 11:1198-1202).  
       [0048] Another subgroup of isomerases are the transferases (or mutases). Transferases transfer a chemical group from one compound (the donor) to another compound (the acceptor). The types of groups transferred by these enzymes include acyl groups, amino groups, phosphate groups (phosphotransferases or phosphomutases), and others. The transferase carnitine palmitoyltransferase is an important component of fatty acid metabolism. Genetically-linked deficiencies in this transferase can lead to myopathy (Scriver, C. R. et al. (1995)  The Metabolic and Molecular Basis of Inherited Disease , McGraw-Hill, New York N.Y., pp.1501-1533).  
       [0049] Yet another subgroup of isomerases are the topoisomersases. Topoisomerases are enzymes that affect the topological state of DNA. For example, defects in topoisomerases or their regulation can affect normal physiology. Reduced levels of topoisomerase II have been correlated with some of the DNA processing defects associated with the disorder ataxia-telangiectasia (Singh, S. P. et al. (1988) Nucleic Acids Res. 16:3919-3929).  
       [0050] Ligases  
       [0051] Ligases catalyze the formation of a bond between two substrate molecules. The process involves the hydrolysis of a pyrophosphate bond in ATP or a similar energy donor. Ligases are classified based on the nature of the type of bond they form, which can include carbon-oxygen, carbon-sulfur, carbon-nitrogen, carbon-carbon and phosphoric ester bonds.  
       [0052] Ligases forming carbon-oxygen bonds include the aminoacyl-transfer RNA (tRNA) synthetases which are important RNA-associated enzymes with roles in translation. Protein biosynthesis depends on each amino acid forming a linkage with the appropriate tRNA. The aminoacyl-tRNA synthetases are responsible for the activation and correct attachment of an ammo acid with its cognate tRNA. The 20 aminoacyl-tRNA synthetase enzymes can be divided into two structural classes, and each class is characterized by a distinctive topology of the catalytic domain.  
       [0053] Class I enzymes contain a catalytic domain based on the nucleotide-binding Rossman fold. Class II enzymes contain a central catalytic domain, which consists of a seven-stranded antiparallel β-sheet motif, as well as N- and C-terminal regulatory domains. Class II enzymes are separated into two groups based on the heterodimeric or homodimeric structure of the enzyme; the latter group is further subdivided by the structure of the N- and C-terminal regulatory domains (Hartlein, M. and S. Cusack (1995) J. Mol. Evol. 40:519-530). Autoantibodies against aminoacyl-tRNAs are generated by patients with dermatomyositis and polymyositis, and correlate strongly with complicating interstitial lung disease (ILD). These antibodies appear to be generated in response to viral infection, and coxsackie virus has been used to induce experimental viral myositis in animals.  
       [0054] Ligases forming carbon-sulfur bonds (Acid-thiol ligases) mediate a large number of cellular biosynthetic intermediary metabolism processes involve intermolecular transfer of carbon atom-containing substrates (carbon substrates). Examples of such reactions include the tricarboxylic acid cycle, synthesis of fatty acids and long-chain phospholipids, synthesis of alcohols and aldehydes, synthesis of intermediary metabolites, and reactions involved in the amino acid degradation pathways. Some of these reactions require input of energy, usually in the form of conversion of ATP to either ADP or AMP and pyrophosphate.  
       [0055] In many cases, a carbon substrate is derived from a small molecule containing at least two carbon atoms. The carbon substrate is often covalently bound to a larger molecule which acts as a carbon substrate carrier molecule within the cell. In the biosynthetic mechanisms described above, the carrier molecule is coenzyme A. Coenzyme A (CoA) is structurally related to derivatives of the nucleotide ADP and consists of 4′-phosphopantetheine linked via a phosphodiester bond to the alpha phosphate group of adenosine 3′,5′-bisphosphate. The terminal thiol group of 4′-phosphopantetheine acts as the site for carbon substrate bond formation. The predominant carbon substrates which utilize CoA as a carrier molecule during biosynthesis and intermediary metabolism in the cell are acetyl, succinyl, and propionyl moieties, collectively referred to as acyl groups. Other carbon substrates include enoyl lipid, which acts as a fatty acid oxidation intermediate, and carnitine, which acts as an acetyl-CoA flux regulator/mitochondrial acyl group transfer protein. Acyl-CoA and acetyl-CoA are synthesized in the cell by acyl-CoA synthetase and acetyl-CoA synthetase, respectively.  
       [0056] Activation of fatty acids is mediated by at least three forms of acyl-CoA synthetase activity: i) acetyl-CoA synthetase, which activates acetate and several other low molecular weight carboxylic acids and is found in muscle mitochondria and the cytosol of other tissues; ii) medium-chain acyl-CoA synthetase, which activates fatty acids containing between four and eleven carbon atoms (predominantly from dietary sources), and is present only in liver mitochondria; and iii) acyl CoA synthetase, which is specific for long chain fatty acids with between six and twenty carbon atoms, and is found in microsomes and the mitochondria. Proteins associated with acyl-CoA synthetase activity have been identified from many sources including bacteria, yeast, plants, mouse, and man. The activity of acyl-CoA synthetase may be modulated by phosphorylation of the enzyme by cAMP-dependent protein kinase.  
       [0057] Ligases forming carbon-nitrogen bonds include amide synthases such as glutamine synthetase (glutamate-ammonia ligase) that catalyzes the amination of glutamic acid to glutamine by ammonia using the energy of ATP hydrolysis. Glutamine is the primary source for the amino group in various amide transfer reactions involved in de novo pyrimidine nucleotide synthesis and in purine and pyrimidine ribonucleotide interconversions. Overexpression of glutamine synthetase has been observed in primary liver cancer (Christa, L. et al. (1994) Gastroent. 106:1312-1320).  
       [0058] Acid-amino-acid ligases (peptide synthases) are represented by the ubiquitin proteases which are associated with the ubiquitin conjugation system (UCS), a major pathway for the degradation of cellular proteins in eukaryotic cells and some bacteria. The UCS mediates the elimination of abnormal proteins and regulates the half-lives of important regulatory proteins that control cellular processes such as gene transcription and cell cycle progression. In the UCS pathway, proteins targeted for degradation are conjugated to a ubiquitin (Ub), a small heat stable protein. Ub is first activated by a ubiquitin-activating enzyme (E1), and then transferred to one of several Ub-conjugating enzymes (E2). E2 then links the Ub molecule through its C-terminal glycine to an internal lysine (acceptor lysine) of a target protein. The ubiquitinated protein is then recognized and degraded by proteasome, a large, multisubunit proteolytic enzyme complex, and ubiquitin is released for reutilization by ubiquitin protease. The UCS is implicated in the degradation of mitotic cyclic kinases, oncoproteins, tumor suppressor genes such as p53, viral proteins, cell surface receptors associated with signal transduction, transcriptional regulators, and mutated or damaged proteins (Ciechanover, A (1994) Cell 79:13-21). A murine proto-oncogene, Unp, encodes a nuclear ubiquitin protease whose overexpression leads to oncogenic transformation of NIH3T3 cells, and the human homolog of this gene is consistently elevated in small cell tumors and adenocarcinomas of the lung (Gray, D. A. (1995) Oncogene 10:2179-2183).  
       [0059] Cyclo-ligases and other carbon-nitrogen ligases comprise various enzymes and enzyme complexes that participate in the de novo pathways to purine and pyrimidine biosynthesis. Because these pathways are critical to the synthesis of nucleotides for replication of both RNA and DNA, many of these enzymes have been the targets of clinical agents for the treatment of cell proliferative disorders such as cancer and infectious diseases.  
       [0060] Purine biosynthesis occurs de novo from the amino acids glycine and glutamine, and other small molecules. Three of the key reactions in this process are catalyzed by a trifunctional enzyme composed of glycinamide-ribonucleotide synthetase (GARS), aminoimidazole ribonucleotide synthetase (AIRS), and glycinamide ribonucleotide transformylase (GART). Together these three enzymes combine ribosylamine phosphate with glycine to yield phosphoribosyl aminoimidazole, a precursor to both adenylate and guanylate nucleotides. This trifunctional protein has been implicated in the pathology of Downs syndrome (Aimi, J. et al. (1990) Nucleic Acid Res. 18:6665-6672). Adenylosuccinate synthetase catalyzes a later step in purine biosynthesis that converts inosinic acid to adenylosuccinate, a key step on the path to ATP synthesis. This enzyme is also similar to another carbon-nitrogen ligase, argininosuccinate synthetase, that catalyzes a similar reaction in the urea cycle (Powell, S. M. et al. (1992) FEBS Lett. 303:4-10).  
       [0061] Like the de novo biosynthesis of purines, de novo synthesis of the pyrimidine nucleotides uridylate and cytidylate also arises from a common precursor, in this instance the nucleotide orotidylate derived from orotate and phosphoribosyl pyrophosphate (PPRP). Again a trifunctional enzyme comprising three carbon-nitrogen ligases plays a key role in the process. In this case the enzymes aspartate transcarbamylase (ATCase), carbamyl phosphate synthetase II, and dihydroorotase (DHOase) are encoded by a single gene called CAD. Together these three enzymes combine the initial reactants in pyrimidine biosynthesis, glutamine, CO 2 , and ATP to form dihydroorotate, the precursor to orotate and orotidylate (Iwahana, H. et al. (1996) Biochem. Biophys. Res. Commun. 219:249-255). Further steps then lead to the synthesis of uridine nucleotides from orotidylate. Cytidine nucleotides are derived from uridine-5′-triphosphate (UTP) by the amidation of UTP using glutamine as the amino donor and the enzyme CTP synthetase. Regulatory mutations in the human CTP synthetase are believed to confer multi-drug resistance to agents widely used in cancer therapy (Yamauchi, M. et al. (1990) EMBO J. 9:2095-2099).  
       [0062] Ligases forming carbon-carbon bonds include the carboxylases acetyl-CoA carboxylase and pyruvate carboxylase. Acetyl-CoA carboxylase catalyzes the carboxylation of acetyl-CoA from CO 2  and H 2 O using the energy of ATP hydrolysis. Acetyl-CoA carboxylase is the rate-limiting step in the biogenesis of long-chain fatty acids. Two isoforms of acetyl-CoA carboxylase, types I and types II, are expressed in human in a tissue-specific manner (Ha, J. et al. (1994) Eur. J. Biochem. 219:297-306): Pyruvate carboxylase is a nuclear-encoded mitochondrial enzyme that catalyzes the conversion of pyruvate to oxaloacetate, a key intermediate in the citric acid cycle.  
       [0063] Ligases forming phosphoric ester bonds include the DNA ligases involved in both DNA replication and repair. DNA ligases seal phosphodiester bonds between two adjacent nucleotides in a DNA chain using the energy from ATP hydrolysis to first activate the free 5′-phosphate of one nucleotide and then react it with the 3′-OH group of the adjacent nucleotide. This resealing reaction is used in both DNA replication to join small DNA fragments called Okazaki fragments that are transiently formed in the process of replicating new DNA, and in DNA repair. DNA repair is the process by which accidental base changes, such as those produced by oxidative damage, hydrolytic attack, or uncontrolled methylation of DNA, are corrected before replication or transcription of the DNA can occur. Bloom&#39;s syndrome is an inherited human disease in which individuals are partially deficient in DNA ligation and consequently have an increased incidence of cancer (Alberts, B. et al. (1994)  The Molecular Biology of the Cell , Garland Publishing Inc., New York N.Y., p. 247).  
       [0064] Molecules Associated with Growth and Development  
       [0065] Human growth and development requires the spatial and temporal regulation of cell differentiation, cell proliferation, and apoptosis. These processes coordinately control reproduction, aging, embryogenesis, morphogenesis, organogenesis, and tissue repair and maintenance. At the cellular level, growth and development is governed by the cell&#39;s decision to enter into or exit from the cell division cycle and by the cell&#39;s commitment to a terminally differentiated state. These decisions are made by the cell in response to extracellular signals and other environmental cues it receives. The following discussion focuses on the molecular mechanisms of cell division, reproduction, cell differentiation and proliferation, apoptosis, and aging.  
       [0066] Cell Division  
       [0067] Cell division is the fundamental process by which all living things grow and reproduce. In unicellular organisms such as yeast and bacteria, each cell division doubles the number of organisms, while in multicellular species many rounds of cell division are required to replace cells lost by wear or by programmed cell death, and for cell differentiation to produce a new tissue or organ Details of the cell division cycle may vary, but the basic process consists of three principle events. The first event, interphase, involves preparations for cell division, replication of the DNA, and production of essential proteins. In the second event, mitosis, the nuclear material is divided and separates to opposite sides of the cell. The final event, cytokinesis, is division and fission of the cell cytoplasm. The sequence and timing of cell cycle transitions is under the control of the cell cycle regulation system which controls the process by positive or negative regulatory circuits at various check points.  
       [0068] Regulated progression of the cell cycle depends on the integration of growth control pathways with the basic cell cycle machinery. Cell cycle regulators have been identified by selecting for human and yeast cDNAs that block or activate cell cycle arrest signals in the yeast mating pheromone pathway when they are overexpressed. Known regulators include human CPR (cell cycle progression restoration) genes, such as CPR8 and CPR2, and yeast CDC (cell division control) genes, including CDC91, that block the arrest signals. The CPR genes express a variety of proteins including cyclins, tumor suppressor binding proteins, chaperones, transcription factors, translation factors, and RNA-binding proteins (Edwards, M. C. et al.(1997) Genetics 147:1063-1076).  
       [0069] Several cell cycle transitions, including the entry and exit of a cell from mitosis, are dependent upon the activation and inhibition of cyclin-dependent kinases (Cdks). The Cdks are composed of a kinase subunit, Cdk, and an activating subunit, cyclin, in a complex that is subject to many levels of regulation. There appears to be a single Cdk in  Saccharomyces cerevisiae  and  Saccharomyces pombe  whereas mammals have a variety of specialized Cdks. Cyclins act by binding to and activating cyclin-dependent protein kinases which then phosphorylate and activate selected proteins involved in the mitotic process. The Cdk-cyclin complex is both positively and negatively regulated by phosphorylation, and by targeted degradation involving molecules such as CDC4 and CDC53. In addition, Cdks are further regulated by binding to inhibitors and other proteins such as Suc1 that modify their specificity or accessibility to regulators (Patra, D. and W. G. Dunphy (1996) Genes Dev. 10:1503-1515; and Mathias, N. et al. (1996) Mol. Cell Biol. 16:66346643).  
       [0070] Reproduction  
       [0071] The male and female reproductive systems are complex and involve many aspects of growth and development. The anatomy and physiology of the male and female reproductive systems are reviewed in (Guyton, A. C. (1991)  Textbook of Medical Physiology , W. B. Saunders Co., Philadelphia Pa., pp. 899-928).  
       [0072] The male reproductive system includes the process of spermatogenesis, in which the sperm are formed, and male reproductive functions are regulated by various hormones and their effects on accessory sexual organs, cellular metabolism, growth, and other bodily functions.  
       [0073] Spermatogenesis begins at puberty as a result of stimulation by gonadotropic hormones released from the anterior pituitary. Immature sperm (spermatogonia) undergo several mitotic cell divisions before undergoing meiosis and full maturation. The testes secrete several male sex hormones, the most abundant being testosterone, that is essential for growth and division of the immature sperm, and for the masculine characteristics of the male body. Three other male sex hormones, gonadotropin-releasing hormone (GnRH), luteinizing hormone (LH), and follicle-stimulating hormone (FSH) control sexual function.  
       [0074] The uterus, ovaries, fallopian tubes, vagina, and breasts comprise the female reproductive system. The ovaries and uterus are the source of ova and the location of fetal development, respectively. The fallopian tubes and vagina are accessory organs attached to the top and bottom of the uterus, respectively. Both the uterus and ovaries have additional roles in the development and loss of reproductive capability during a female&#39;s lifetime. The primary role of the breasts is lactation Multiple endocrine signals from the ovaries, uterus, pituitary, hypothalamus, adrenal glands, and other tissues coordinate reproduction and lactation. These signals vary during the monthly menstruation cycle and during the female&#39;s lifetime. Similarly, the sensitivity of reproductive organs to these endocrine signals varies during the female&#39;s lifetime.  
       [0075] A combination of positive and negative feedback to the ovaries, pituitary and hypothalamus glands controls physiologic changes during the monthly ovulation and endometrial cycles. The anterior pituitary secretes two major gonadotropin hormones, follicle-stimulating hormone (FSH) and luteinizing hormone (LH), regulated by negative feedback of steroids, most notably by ovarian estradiol. If fertilization does not occur, estrogen and progesterone levels decrease. This sudden reduction of the ovarian hormones leads to menstruation, the desquamation of the endometrium.  
       [0076] Hormones further govern an the steps of pregnancy, parturition, lactation, and menopause. During pregnancy large quantities of human chorionic gonadotropin (hCG), estrogens, progesterone, and human chorionic somatomammotropin (hCS) are formed by the placenta. hCG, a glycoprotein similar to luteinizing hormone, stimulates the corpus luteum to continue producing more progesterone and estrogens, rather than to involute as occurs if the ovum is not fertilized hCS is similar to growth hormone and is crucial for fetal nutrition.  
       [0077] The female breast also matures during pregnancy. Large amounts of estrogen secreted by the placenta trigger growth and branching of the breast milk ductal system while lactation is initiated by the secretion of prolactin by the pituitary gland.  
       [0078] Parturition involves several hormonal changes that increase uterine contractility toward the end of pregnancy, as follows. The levels of estrogens increase more than those of progesterone. Oxytocin is secreted by the neurohypophysis. Concomitantly, uterine sensitivity to oxytocin increases. The fetus itself secretes oxytocin, cortisol (from adrenal glands), and prostaglandins.  
       [0079] Menopause occurs when most of the ovarian follicles have degenerated. The ovary then produces less estradiol, reducing the negative feedback on the pituitary and hypothalamus glands. Mean levels of circulating FSH and LH increase, even as ovulatory cycles continue. Therefore, the ovary is less responsive to gonadotropins, and there is an increase in the time between menstrual cycles.  
       [0080] Consequently, menstrual bleeding ceases and reproductive capability ends.  
       [0081] Cell Differentiation and Proliferation  
       [0082] Tissue growth involves complex and ordered patterns of cell proliferation, cell differentiation, and apoptosis. Cell proliferation must be regulated to maintain both the number of cells and their spatial organization. This regulation depends upon the appropriate expression of proteins which control cell cycle progression in response to extracellular signals, such as growth factors and other mitogens, and intracellular cues, such as DNA damage or nutrient starvation. Molecules which directly or indirectly modulate cell cycle progression fall into several categories, including growth factors and their receptors, second messenger and signal transduction proteins, oncogene products, tumor-suppressor proteins, and mitosis-promoting factors.  
       [0083] Growth factors were originally described as serum factors required to promote cell proliferation. Most growth factors are large, secreted polypeptides that act on cells in their local environment. Growth factors bind to and activate specific cell surface receptors and initiate intracellular signal transduction cascades. Many growth factor receptors are classified as receptor tyrosine kinases which undergo autophosphorylation upon ligand binding. Autophosphorylation enables the receptor to interact with signal transduction proteins characterized by the presence of SH2 or SH3 domains (Src homology regions 2 or 3). These proteins then modulate the activity state of small G-proteins, such as Ras, Rab, and Rho, along with GTPase activating proteins (GAPs), guanine nucleotide releasing proteins (GNRPs), and other guanine nucleotide exchange factors. Small G proteins act as molecular switches that activate other downstream events, such as mitogen-activated protein kinase (MAP kinase) cascades. MAP kinases ultimately activate transcription of mitosis-promoting genes.  
       [0084] In addition to growth factors, small signaling peptides and hormones also influence cell proliferation. These molecules bind primarily to another class of receptor, the trimeric G-protein coupled receptor (GPCR), found predominantly on the surface of immune, neuronal and neuroendocrine cells. Upon ligand binding, the GPCR activates a trimeric G protein which in turn triggers increased levels of intracellular second messengers such as phospholipase C, Ca2+, and cyclic AMP. Most GPCR-mediated signaling pathways indirectly promote cell proliferation by causing the secretion or breakdown of other signaling molecules that have direct mitogenic effects. These signaling cascades often involve activation of kinases and phosphatases. Some growth factors, such as some members of the transforming growth factor beta (TGF-β) family, act on some cells to stimulate cell proliferation and on other cells to inhibit it Growth factors may also stimulate a cell at one concentration and inhibit the same cell at another concentration. Most growth factors also have a multitude of other actions besides the regulation of cell growth and division: they can control the proliferation, survival, differentiation, migration, or function of cells depending on the circumstance. For example, the tumor necrosis factor/nerve growth factor (TNF/NGF) family can activate or inhibit cell death, as well as regulate proliferation and differentiation. The cell response depends on the type of cell, its stage of differentiation and transformation status, which surface receptors are stimulated, and the types of stimuli acting on the cell (Smith, A. et al. (1994) Cell 76:959-962; and Nocentini, G. et al. (1997) Proc. Natl. Acad. Sci. USA 94:6216-6221).  
       [0085] Neighboring cells in a tissue compete for growth factors, and when provided with “ited” quantities in a perfused system win grow to even higher cell densities before reaching density-dependent inhibition of cell division. Cells often demonstrate an anchorage dependence of cell division as well. This anchorage dependence may be associated with the formation of focal contacts linking the cytoskeleton with the extracellular matrix (ECM). The expression of ECM components can be stimulated by growth factors. For example, TGF-5 stimulates fibroblasts to produce a variety of ECM proteins, including fibronectin, collagen, and tenascin (Pearson, C. A. et al. (1988) EMBO J. 7:2677-2981). In fact, for some cell types specific ECM molecules, such as laminin or fibronectin, may act as growth factors. Tenascin-C and -R, expressed in developing and lesioned neural tissue, provide stimulatory/anti-adhesive or inhibitory properties, respectively, for axonal growth (Faissner, A (1997) Cell Tissue Res. 290:331-341).  
       [0086] Cancers are associated with the activation of oncogenes which are derived from normal cellular genes. These oncogenes encode oncoproteins which convert normal cells into malignant cells. Some oncoproteins are mutant isoforms of the normal protein, and other oncoproteins are abnormally expressed with respect to location or amount of expression The latter category of oncoprotein causes cancer by altering transcriptional control of cell proliferation. Five classes of oncoproteins are known to affect cell cycle controls. These classes include growth factors, growth factor receptors, intracellular signal transducers, nuclear transcription factors, and cell-cycle control proteins. Viral oncogenes are integrated into the human genome after infection of human cells by certain viruses. Examples of viral oncogenes include v-src, v-abl, and v-fps.  
       [0087] Many oncogenes have been identified and characterized These include v-src, erbA, erbB, her-2, mutated G S , src, abl, ras, crk, jun, fos, myc, and mutated tumor-suppressor genes such as RB, p53, mdm2, Cip1, p16, and cyclin D. Transformation of normal genes to oncogenes may also occur by chromosomal translocation. The Philadelphia chromosome, characteristic of chronic myeloid leukemia and a subset of acute lymphoblastic leukemias, results from a reciprocal translocation between chromosomes 9 and 22 that moves a truncated portion of the proto-oncogene c-abl to the breakpoint cluster region (bcr) on chromosome 22.  
       [0088] Tumor-suppressor genes are involved in regulating cell proliferation. Mutations which cause reduced or loss of function in tumor-suppressor genes result in uncontrolled cell proliferation. For example, the retinoblastoma gene product (RB), in a non-phosphorylated state, binds several early-response genes and suppresses their transcription, thus blocking cell division. Phosphorylation of RB causes it to dissociate from the genes, releasing the suppression, and allowing cell division to proceed.  
       [0089] Apoptosis  
       [0090] Apoptosis is the genetically controlled process by which unneeded or defective cells undergo programmed cell death. Selective elimination of cells is as important for morphogenesis and tissue remodeling as is cell proliferation and differentiation. Lack of apoptosis may result in hyperplasia and other disorders associated with increased cell proliferation. Apoptosis is also a critical component of the immune response. Immune cells such as cytotoxic T-cells and natural killer cells prevent the spread of disease by inducing apoptosis in tumor cells and virus-infected cells. In addition, immune cells that fail to distinguish self molecules from foreign molecules must be eliminated by apoptosis to avoid an autoimmune response.  
       [0091] Apoptotic cells undergo distinct morphological changes. Hallmarks of apoptosis include cell shrinkage, nuclear and cytoplasmic condensation, and alterations in plasma membrane topology. Biochemically, apoptotic cells are characterized by increased intracellular calcium concentration, fragmentation of chromosomal DNA, and expression of novel cell surface components.  
       [0092] The molecular mechanisms of apoptosis are highly conserved, and many of the key protein regulators and effectors of apoptosis have been identified. Apoptosis generally proceeds in response to a signal which is transduced intracellularly and results in altered patterns of gene expression and protein activity. Signaling molecules such as hormones and cytokines are known both to stimulate and to inhibit apoptosis through interactions with cell surface receptors. Transcription factors also play an important role in the onset of apoptosis. A number of downstream effector molecules, particularly proteases such as the cysteine proteases called caspases, have been implicated in the degradation of cellular components and the proteolytic activation of other apoptotic effectors.  
       [0093] Aging and Senescence  
       [0094] Studies of the aging process or senescence have shown a member of characteristic cellular and molecular changes (Fauci et al. (1998)  Harrison&#39;s Principles of Internal Medicine , McGraw-Hill, New York N.Y., p.37). These characteristics include increases in chromosome structural abnormalities, DNA cross-linking, incidence of single-stranded breaks in DNA, losses in DNA methylation, and degradation of telomere regions. In addition to these DNA changes, post-translational alterations of proteins increase including, deamidation, oxidation, cross-linking, and nonenzymatic glycation. Still further molecular changes occur in the mitochondria of aging cells through deterioration of structure. These changes eventually contribute to decreased function in every organ of the body.  
       [0095] Biochemical Pathway Molecules  
       [0096] Biochemical pathways are responsible for regulating metabolism, growth and development, protein secretion and trafficking, environmental responses, and ecological interactions including immune response and response to parasites.  
       [0097] DNA Replication  
       [0098] Deoxyribonucleic acid (DNA), the genetic material, is found in both the nucleus and mitochondria of human cells. The bulk of human DNA is nuclear, in the form of linear chromosomes, while mitochondrial DNA is circular. DNA replication begins at specific sites called origins of replication. Bidirectional synthesis occurs from the origin via two growing forks that move in opposite directions. Replication is semi-conservative, with each daughter duplex containing one old strand and its newly synthesized complementary partner. Proteins involved in DNA replication include DNA polymerases, DNA primase, telomerase, DNA helicase, topoisomerases, DNA ligases, replication factors, and DNA-binding proteins.  
       [0099] DNA Recombination and Repair  
       [0100] Cells are constantly faced with replication errors and environmental assault (such as ultraviolet irradiation) that can produce DNA damage. Damage to DNA consists of any change that modifies the structure of the molecule. Changes to DNA can be divided into two general classes, single base changes and structural distortions. Any damage to DNA can produce a mutation, and the mutation may produce a disorder, such as cancer.  
       [0101] Changes in DNA are recognized by repair systems within the cell. These repair systems act to correct the damage and thus prevent any deleterious affects of a mutational event Repair systems can be divided into three general types, direct repair, excision repair, and retrieval systems. Proteins involved in DNA repair include DNA polymerase, excision repair proteins, excision and cross link repair proteins, recombination and repair proteins, RAD51 proteins, and BLN and WRN proteins that are homologs of RecQ helicase. When the repair systems are eliminated, cells become exceedingly sensitive to environmental mutagens, such as ultraviolet irradiation Patients with disorders associated with a loss in DNA repair systems often exhibit a high sensitivity to environmental mutagens. Examples of such disorders include xeroderma pigmentosum (XP), Bloom&#39;s syndrome (BS), and Werner&#39;s syndrome (WS) (Yamagata, K et al. (1998) Proc. Natl. Acad. Sci. USA 95:8733-8738), ataxia telangiectasia, Cockayne&#39;s syndrome, and Fanconi&#39;s anemia.  
       [0102] Recombination is the process whereby new DNA sequences are generated by the movements of large pieces of DNA. In homologous recombination, which occurs during meiosis and DNA repair, parent DNA duplexes align at regions of sequence similarity, and new DNA molecules form by the breakage and joining of homologous segments. Proteins involved include RAD51 recombinase. In site-specific recombination, two specific but not necessarily homologous DNA sequences are exchanged. In the immune system this process generates a diverse collection of anitibody and T cell receptor genes. Proteins involved in site-specific recombination in the immune system include recombination activating genes 1 and 2 (RAG1 and RAG2). A defect in immune system site-specific recombination causes severe combined immunodeficiency disease in mice.  
       [0103] RNA Metabolism  
       [0104] Ribonucleic acid (RNA) is a linear single-stranded polymer of four nucleotides, ATP, CTP, UTP, and GTP. In most organisms, RNA is transcribed as a copy of DNA, the genetic material of the organism. In retroviruses RNA rather than DNA serves as the genetic material. RNA copies of the genetic material encode proteins or serve various structural, catalytic, or regulatory roles in organisms. RNA is classified according to its cellular localization and function Messenger RNAs (mRNAs) encode polypeptides. Ribosomal RNAs (rRNAs) are assembled, along with ribosomal proteins, into ribosomes, which are cytoplasmic particles that translate mRNA into polypeptides. Transfer RNAs (tRNAs) are cytosolic adaptor molecules that function in mRNA translation by recognizing both an mRNA codon and the amino acid that matches that codon. Heterogeneous nuclear RNAs (hnRNAs) include mRNA precursors and other nuclear RNAs of various sizes. Small nuclear RNAs (snRNAs) are a part of the nuclear spliceosome complex that removes intervening, non-coding sequences (introns) and rejoins exons in pre-mRNAs.  
       [0105] RNA Transcription  
       [0106] The transcription process synthesizes an RNA copy of DNA. Proteins involved include multi-subunit RNA polymerases, transcription factors IIA, IIB, IID, IIE, IIF, IIH, and IIJ. Many transcription factors incorporate DNA-binding structural motifs which comprise either α-helices or β-sheets that bind to the major groove of DNA. Four well-characterized structural motifs are helix-turn-helix, zinc finger, leucine zipper, and helix-loop-helix.  
       [0107] RNA Processing  
       [0108] Various proteins are necessary for processing of transcribed RNAs in the nucleus. Pre-mRNA processing steps include capping at the 5′ end with methylguanosine, polyadenylating the 3′ end, and splicing to remove introns. The spliceosomal complex is comprised of five small nuclear ribonucleoprotein particles (snRNPs) designated U1, U2, U4, U5, and U6. Each snRNP contains a single species of snRNA and about ten proteins. The RNA components of some snRNPs recognize and base-pair with intron consensus sequences. The protein components mediate spliceosome assembly and the splicing reaction. Autoantibodies to snRNP proteins are found in the blood of patients with systemic lupus erythematosus (Stryer, L. (1995)  Biochemistry  W. H. Freeman and Company, New York N.Y., p. 863).  
       [0109] Heterogeneous nuclear ribonucleoproteins (hnRNPs) have been identified that have roles in splicing, exporting of the mature RNAs to the cytoplasm, and mRNA translation (Biamonti, G. et al. (1998) Clin Exp. Rheumatol. 16:317-326). Some examples of hnRNPs include the yeast proteins Hrplp, involved in cleavage and polyadenylation at the 3′ end of the RNA; Cbp80p, involved in capping the 5′ end of the RNA; and Npl3p, a homolog of mammalian hnRNP A1, involved in export of mRNA from the nucleus (Shen, E. C. et al. (1998) Genes Dev. 12:679-691). HnRNPs have been shown to be important targets of the autoimmune response in rheumatic diseases (Biamonti, supra).  
       [0110] Many snRNP proteins, and alternative splicing factors are characterized by an RNA recognition motif (RRM). (Reviewed in Birney, E. et al. (1993) Nucleic Acids Res. 21:5803-5816.) The RRM is about 80 amino acids in length and forms four β-strands and two α-helices arranged in an a/0 sandwich. The RRM contains a core RNP-1 octapeptide motif along with surrounding conserved sequences.  
       [0111] RNA Stability and Degradation  
       [0112] RNA helicases alter and regulate RNA conformation and secondary structure by using energy derived from ATP hydrolysis to destabilize and unwind RNA duplexes. The most well-characterized and ubiquitous family of RNA helicases is the DEAD-box family, so named for the conserved B-type ATP-binding motif which is diagnostic of proteins in this family. Over 40 DEAD-box helicases have been identified in organisms as diverse as bacteria, insects, yeast, amphibians, mammals, and plants. DEAD-box helicases function in diverse processes such as translation initiation, splicing, ribosome assembly, and RNA editing, transport, and stability. Some DEAD-box helicases play tissue- and stage-specific roles in spermatogenesis and embryogenesis. (Reviewed in Linder, P. et al. (1989) Nature 337:121-122.)  
       [0113] Overexpression of the DEAD-box 1 protein (DDX1) may play a role in the progression of neuroblastoma (Nb) and retinoblastoma (Rb) tumors. Other DEAD-box helicases have been implicated either directly or indirectly in ultraviolet light-induced tumors, B cell lymphoma, and myeloid malignancies. (Reviewed in Godbout, R. et al. (1998) J. Biol. Chem. 273:21161-21168.)  
       [0114] Ribonucleases (RNases) catalyze the hydrolysis of phosphodiester bonds in RNA chains, thus cleaving the RNA. For example, RNase P is a ribonucleoprotein enzyme which cleaves the 5′ end of pre-tRNAs as part of their maturation process. RNase H digests the RNA strand of an RNA/DNA hybrid. Such hybrids occur in cells invaded by retroviruses, and RNase H is an important enzyme in the retroviral replication cycle. RNase H domains are often found as a domain associated with reverse transcriptases. RNase activity in serum and cell extracts is elevated in a variety of cancers and infectious diseases (Schein, C. H. (1997) Nat. Biotechnol. 15:529-536). Regulation of RNase activity is being investigated as a means to control tumor angiogenesis, allergic reactions, viral infection and replication, and fungal infections.  
       [0115] Protein Translation  
       [0116] The eukaryotic ribosome is composed of a 60S (large) subunit and a 40S (small) subunit, which together form the 80S ribosome. In addition to the 18S, 28S, 5S, and 5.8S rRNAs, the ribosome also contains more than fifty proteins. The ribosomal proteins have a prefix which denotes the subunit to which they belong, either L (large) or S (small). Three important sites are identified on the ribosome. The aminoacyl-tRNA site (A site) is where charged tRNAs (with the exception of the initiator-tRNA) bind on arrival at the ribosome. The peptidyl-tRNA site (P site) is where new peptide bonds are formed, as well as where the initiator tRNA binds. The exit site (E site) is where deacylated tRNAs bind prior to their release from the ribosome. (Translation is reviewed in Stryer, L. (1995)  Biochemistry , W. H. Freeman and Company, New York N.Y., pp. 875-908; and Lodish, H. et al. (1995)  Molecular Cell Biology , Scientific American Books, New York N.Y., pp. 119-138.)  
       [0117] tRNA Charging  
       [0118] Protein biosynthesis depends on each amino acid forming a linkage with the appropriate tRNA The aminoacyl-tRNA synthetases are responsible for the activation and correct attachment of an amino acid with its cognate tRNA The 20 aminoacyl-tRNA synthetase enzymes can be divided into two structural classes, Class I and Class II. Autoantibodies against aminoacyl-tRNAs are generated by patients with dermatomyositis and polymyositis, and correlate strongly with complicating interstitial lung disease (ILD). These antibodies appear to be generated in response to viral infection, and coxsackie virus has been used to induce experimental viral myositis in animals.  
       [0119] Translation Initiation  
       [0120] Initiation of translation can be divided into three stages. The first stage brings an initiator transfer RNA (Met-tRNA) together with the 40S ribosomal subunit to form the 43S preinitiation complex. The second stage binds the 43S preinitiation complex to the mRNA, followed by migration of the complex to the correct AUG initiation codon. The third stage brings the 60S ribosomal subunit to the 40S subunit to generate an 80S ribosome at the initiation codon. Regulation of translation primarily involves the first and second stage in the initiation process (Pain, V. M. (1996) Eur. J. Biochem. 236:747-771).  
       [0121] Several initiation factors, many of which contain multiple subunits, are involved in bringing an initiator tRNA and 40S ribosomal subunit together. eIF2, a guanine nucleotide binding protein, recruits the initiator tRNA to the 40S ribosomal subunit. Only when eIF2 is bound to GTP does it associate with the initiator tRNA eIF2B, a guanine nucleotide exchange protein, is responsible for converting eIF2 from the GDP-bound inactive form to the GTP-bound active form. Two other factors, eIF1A and eIF3 bind and stabilize the 40S subunit by interacting with 18S ribosomal RNA and specific ribosomal structural proteins. eIF3 is also involved in association of the 40S ribosomal subunit with mRNA. The Met-tRNA f , eIF1A, eIF3, and 40S ribosomal subunit together make up the 43S preinitiation complex (Pain, supra).  
       [0122] Additional factors are required for binding of the 43S preinitiation complex to an mRNA molecule, and the process is regulated at several levels. eIF4F is a complex consisting of three proteins: eIF4E, eIF4A, and eIF4G. eIF4E recognizes and binds to the mRNA 5′-terminal m 7 GTP cap, eIF4A is a bidirectional RNA-dependent helicase, and eIF4G is a scaffolding polypeptide. eIF4G has three binding domains. The N-terminal third of eIF4G interacts with eIF4E, the central third interacts with eIF4A, and the C-terminal third interacts with eIF3 bound to the 43S preinitiation complex. Thus, eIF4G acts as a bridge between the 40S ribosomal subunit and the mRNA (Hentze, M. W. (1997) Science 275:500-501).  
       [0123] The ability of eIF4F to initiate binding of the 43S preinitiation complex is regulated by structural features of the mRNA The mRNA molecule has an untranslated region (UM) between the 5, cap and the AUG start codon. In some mRNAs this region forms secondary structures that impede binding of the 43S preinitiation complex. The helicase activity of eIF4A is thought to function in removing this secondary structure to facilitate binding of the 43S preinitiation complex (Pain, supra).  
       [0124] Translation Elongation  
       [0125] Elongation is the process whereby additional amino acids are joined to the initiator methionine to form the complete polypeptide chain. The elongation factors EF1α, EF1βγ, and EF2 are involved in elongating the polypeptide chain following initiation. EF1α is a GTP-binding protein. InEFla&#39;s GTP-bound form, it brings an aminoacyl-tRNA to the ribosome&#39;s A site. The amino acid attached to the newly arrived aminoacyl-tRNA forms a peptide bond with the initiator methionine. The GTP on EF1α is hydrolyzed to GDP, and EF1α-GDP dissociates from the ribosome. EF1βγ binds EF1α-GDP and induces the dissociation of GDP from EF1α, allowing EF1α to bind GTP and a new cycle to begun  
       [0126] As subsequent aminoacyl-tRNAs are brought to the ribosome, EF-G, another GTP-binding protein, catalyzes the translocation of tRNAs from the A site to he P site and finally to the E site of the ribosome. This allows the processivity of translation.  
       [0127] Translation Termination  
       [0128] The release factor eRF carries out termination of translation. eRF recognizes stop codons in the mRNA, leading to the release of the polypeptide chain from the ribosome.  
       [0129] Post-Translational Pathways  
       [0130] Proteins may be modified after translation by the addition of phosphate, sugar, prenyl, fatty acid, and other chemical groups. These modifications are often required for proper protein activity. Enzymes involved in post-translational modification include kinases, phosphatases, glycosyltransferases, and prenyltransferases. The conformation of proteins may also be modified after translation by the introduction and rearrangement of disulfide bonds (rearrangement catalyzed by protein disulfide isomerase), the isomerization of proline sidechains by prolyl isomerase, and by interactions with molecular chaperone proteins.  
       [0131] Proteins may also be cleaved by proteases. Such cleavage may result in activation, inactivation, or complete degradation of the protein. Proteases include serine proteases, cysteine proteases, aspartic proteases, and metalloproteases. Signal peptidase in the endoplasmic reticulum (ER) lumen cleaves the signal peptide from membrane or secretory proteins that are imported into the ER. Ubiquitin proteases are associated with the ubiquitin conjugation system (UCS), a major pathway for the degradation of cellular proteins in eukaryotic cells and some bacteria. The UCS mediates the elimination of abnormal proteins and regulates the half-lives of important regulatory proteins that control cellular processes such as gene transcription and cell cycle progression. In-the UCS pathway, proteins targeted for degradation are conjugated to a ubiquitin, a small heat stable protein. Proteins involved in the UCS include ubiquitin-activating enzyme, ubiquitin-conjugating enzymes, ubiquitin-ligases, and ubiquitin C-terminal hydrolases. The ubiquitinated protein is then recognized and degraded by the proteasome, a large, multisubunit proteolytic enzyme complex, and ubiquitin is released for reutilization by ubiquitin protease.  
       [0132] Lipid Metabolism  
       [0133] Lipids are water-insoluble, oily or greasy substances that are soluble in nonpolar solvents such as chloroform or ether. Neutral fats (triacylglycerols) serve as major fuels and energy stores. Polar lipids, such as phospholipids, sphingolipids, glycolipids, and cholesterol, are key structural components of cell membranes.  
       [0134] Lipid metabolism is involved in human diseases and disorders. In the arterial disease atherosclerosis, fatty lesions form on the inside of the arterial wall. These lesions promote the loss of arterial flexibility and the formation of blood clots (Guyton, A. C.  Textbook of Medical Physiology  (1991) W. B. Saunders Company, Philadelphia Pa., pp.760-763). In Tay-Sachs disease, the GM 2  ganglioside (a sphingolipid) accumulates in lysosomes of the central nervous system due to a lack of the enzyme N-acetylhexosaminidase. Patients suffer nervous system degeneration leading to early death (Fauci, AS. et al. (1998)  Harrison&#39;s Principles of Internal Medicine  McGraw-Hill, New York N.Y., p. 2171). The Niemann-Pick diseases are caused by defects in lipid metabolism. Niemann-Pick diseases types A and B are caused by accumulation of sphingomyelin (a sphingolipid) and other lipids in the central nervous system due to a defect in the enzyme sphingomyelinase, leading to neurodegeneration and lung disease. Niemann-Pick disease type C results from a defect in cholesterol transport, leading to the accumulation of sphingomyelin and cholesterol in lysosomes and a secondary reduction in sphingomyelinase activity. Neurological symptoms such as grand mal seizures, ataxia, and loss of previously learned speech, manifest 1-2 years after birth. A mutation in the NPC protein, which contains a putative cholesterol-sensing domain, was found in a mouse model of Niemann-Pick disease type C (Fauci, supra, p. 2175; Loftus, S. K et al. (1997) Science 277:232-235). (Lipid metabolism is reviewed in Stryer, L. (1995)  Biochemistry , W. H. Freeman and Company, New York N.Y.; Lehninger, A (1982)  Principles of Biochemistry  Worth Publishers, Inc., New York N.Y.; and ExPASy “Biochemical Pathways” index of Boehringer Mannheim World Wide Web site.)  
       [0135] Fatty Acid Synthesis  
       [0136] Fatty acids are long-chain organic acids with a single carboxyl group and a long non-polar hydrocarbon tail. Long-chain fatty acids are essential components of glycolipids, phospholipids; and cholesterol, which are building blocks for biological membranes, and of triglycerides, which are biological fuel molecules. Long-chain fatty acids are also substrates for eicosanoid production, and are important in the functional modification of certain complex carbohydrates and proteins. 16-carbon and 18-carbon fatty acids are the most common.  
       [0137] Fatty acid synthesis occurs in the cytoplasm. In the first step, acetyl-Coenzyme A (CoA) carboxylase (ACC) synthesizes malonyl-CoA from acetyl-CoA and bicarbonate. The enzymes which catalyze the remaining reactions are covalently linked into a single polypeptide chain, referred to as the multifunctional enzyme fatty acid synthase (FAS). FAS catalyzes the synthesis of palmitate from acetyl-CoA and malonyl-CoA FAS contains acetyl transferase, malonyl transferase, β-ketoacetyl synthase, acyl carrier protein, β-ketoacyl reductase, dehydratase, enoyl reductase, and thioesterase activities. The final product of the FAS reaction is the 16 carbon fatty acid palmitate. Further elongation, as well as unsaturation, of palmitate by accessory enzymes of the ER produces the variety of long chain fatty acids required by the individual cell. These enzymes include a NADH-cytocbrome b 5  reductase, cytochrome b 5 , and a desaturase.  
       [0138] Phospholipid and Triacylglcerol Synthesis  
       [0139] Triacylglycerols, also known as triglycerides and neutral fats, are major energy stores in animals. Triacylglycerols are esters of glycerol with three fatty acid chains. Glycerol-3-phosphate is produced from dihydroxyacetone phosphate by the enzyme glycerol phosphate dehydrogenase or from glycerol by glycerol kinase. Fatty acid-CoA&#39;s are produced from fatty acids by fatty acyl-CoA synthetases. Glyercol-3-phosphate is acylated with two fatty acyl-CoA&#39;s by the enzyme glycerol phosphate acyltransferase to give phosphatidate. Phosphatidate phosphatase converts phosphatidate to diacylglycerol, which is subsequently acylated to a triacylglyercol by the enzyme diglyceride acyltransferase. Phosphatidate phosphatase and diglyceride acyltransferase form a triacylglyerol synthease complex bound to the ER membrane.  
       [0140] A major class of phospholipids are the phosphoglycerides, which are composed of a glycerol backbone, two fatty acid chains, and a phosphorylated alcohol. Phosphoglycerides are components of cell membranes. Principal phosphoglycerides are phosphatidyl choline, phosphatidyl ethanolamine, phosphatidyl serine, phosphatidyl inositol, and diphosphatidyl glycerol. Many enzymes involved in phosphoglyceride synthesis are associated with membranes (Meyers, R. A. (1995)  Molecular Biology and Biotechnology , VCH Publishers Inc., New York N.Y., pp. 494-501). Phosphatidate is converted to CDP-diacylglycerol by the enzyme phosphatidate cytidylyltransferase (ExPASy ENZYME EC 2.7.7.41). Transfer of the diacylglycerol group from CDP-diacylglycerol to serine to yield phosphatidyl serine, or to inositol to yield phosphatidyl inositol, is catalyzed by the enzymes CDP-diacylglycerol-serine O-phosphatidyltransferase and CDP-diacylglycerol-inositol 3-phosphatidyltransferase, respectively (ExPASy ENZYME EC 2.7.8.8; ExPASy ENZYME EC 2.7.8.11). The enzyme phosphatidyl serine decarboxylase catalyzes the conversion of phosphatidyl serine to phosphatidyl ethanolamine, using a pyruvate cofactor (Voelker, D. R. (1997) Biochim. Biophys. Acta 1348:236-244).  
       [0141] Phosphatidyl choline is formed using diet-derived choline by the reaction of CDP-choline with 1,2-diacylglycerol, catalyzed by diacylglycerol cholinephosphotransferase (ExPASy ENZYME 2.7.8.2).  
       [0142] Sterol, Steroid, and Isoprenoid Metabolism  
       [0143] Cholesterol, composed of four fused hydrocarbon rings with an alcohol at one end, moderates the fluidity of membranes in which it is incorporated. In addition, cholesterol is used in the synthesis of steroid hormones such as cortisol, progesterone, estrogen, and testosterone. Bile salts derived from cholesterol facilitate the digestion of lipids. Cholesterol in the skin forms a barrier that prevents excess water evaporation from the body. Farnesyl and geranylgeranyl groups, which are derived from cholesterol biosynthesis intermediates, are post-translationally added to signal transduction proteins such as ras and protein-targeting proteins such as rab. These modifications are important for the activities of these proteins (Guyton, supra; Stryer, supra, pp. 279-280, 691-702, 934).  
       [0144] Mammals obtain cholesterol derived from both de novo biosynthesis and the diet. The liver is the major site of cholesterol biosynthesis in mammals. Two acetyl-CoA molecules initially condense to form acetoacetyl-CoA, catalyzed by a thiolase. Acetoacetyl-CoA condenses with a third acetyl-CoA to form hydroxymethylglutaryl-CoA (HMG-CoA), catalyzed by HMG-CoA synthase. Conversion of HMG-COA to cholesterol is accomplished via a series of enzymatic steps known as the mevalonate pathway. The rate-limiting step is the conversion of HMG-CoA to mevalonate by HMG-CoA reductase. The drug lovastatin, a potent inhibitor of HMG-CoA reductase, is given to patients to reduce their serum cholesterol levels. Other mevalonate pathway enzymes include mevalonate kinase, phosphomevalonate kinase, diphosphomevalonate decarboxylase, isopentenyldiphosphate isomerase, dimethylallyl transferase, geranyl transferase, farnesyl-diphosphate farnesyltransferase, squalene monooxygenase, lanosterol synthase, lathosterol oxidase, and 7-dehydrocholesterol reductase.  
       [0145] Cholesterol is used in the synthesis of steroid hormones such as cortisol, progesterone, aldosterone, estrogen, and testosterone. First, cholesterol is converted to pregnenolone by cholesterol monooxygenases. The other steroid hormones are synthesized from pregnenolone by a series of enzyme-catalyzed reactions including oxidations, isomerizations, hydroxylations, reductions, and demethylations. Examples of these enzymes include steroid Δ-isomerase, 3β-hydroxy-Δ 5 -steroid dehydrogenase, steroid 21-monooxygenase, steroid 19-hydroxylase, and 3β-hydroxysteroid dehydrogenase. Cholesterol is also the precursor to vitamin D.  
       [0146] Numerous compounds contain 5-carbon isoprene units derived from the mevalonate pathway intermediate isopentenyl pyrophosphate. Isoprenoid groups are found in vitamin K, ubiquinone, retinal, dolichol phosphate (a carrier of oligosaccharides needed for N-lined glycosylation), and farnesyl and geranylgeranyl groups that modify proteins. Enzymes involved include farnesyl transferase, polyprenyl transferases, dolichyl phosphatase, and dolichyl kinase.  
       [0147] Sphingolipid Metabolism  
       [0148] Sphingolipids are an important class of membrane lipids that contain sphingosine, a long chain amino alcohol. They are composed of one long-chain fatty acid, one polar head alcohol, and sphingosine or sphingosine derivative. The three classes of sphingolipids are sphingomyelins, cerebrosides, and gangliosides. Sphingomyelins, which contain phosphocholine or phosphoethanolamine as their head group, are abundant in the myelin sheath surrounding nerve cells. Galactocerebrosides, which contain a glucose or galactose head group, are characteristic of the brain. Other cerebrosides are found in nonneural tissues. Gangliosides, whose head groups contain multiple sugar units, are abundant in the brain, but are also found in nonneural tissues.  
       [0149] Sphingolipids are built on a sphingosine backbone. Sphingosine is acylated to ceramide by the enzyme sphingosine acetyltransferase. Ceramide and phosphatidyl choline are converted to sphingomyelin by the enzyme ceramide choline phosphotransferase. Cerebrosides are synthesized by the linkage of glucose or galactose to ceramide by a transferase. Sequential addition of sugar residues to ceramide by transferase enzymes yields gangliosides.  
       [0150] Eicosanoid Metabolism  
       [0151] Eicosanoids, including prostaglandins, prostacyclin, thromboxanes, and leukotrienes, are 20-carbon molecules derived from fatty acids. Eicosanoids are signaling molecules which have roles in pain, fever, and inflammation. The precursor of all eicosanoids is arachidonate, which is generated from phospholipids by phospholipase A 2  and from diacylglycerols by diacylglycerol lipase. Leukotrienes are produced from arachidonate by the action of lipoxygenases. Prostaglandin synthase, reductases, and isomerases are responsible for the synthesis of the prostaglandins. Prostaglandins have roles in inflammation, blood flow, ion transport, synaptic transmission, and sleep. Prostacyclin and the thromboxanes are derived from a precursor prostaglandin by the action of prostacyclin synthase and thromboxane synthases, respectively.  
       [0152] Ketone Body Metabolism  
       [0153] Pairs of acetyl-CoA molecules derived from fatty acid oxidation in the liver can condense to form acetoacetyl-CoA, which subsequently forms acetoacetate, D-3-hydroxybutyrate, and acetone. These three products are known as ketone bodies. Enzymes involved in ketone body metabolism include HMG-COA synthetase, HMG-CoA cleavage enzyme, D-3-hydroxybutyrate dehydrogenase, acetoacetate decarboxylase, and 3-ketoacyl-CoA transferase. Ketone bodies are a normal fuel supply of the heart and renal cortex. Acetoacetate produced by the liver is transported to cells where the acetoacetate is converted back to acetyl-CoA and enters the citric acid cycle. In times of starvation, ketone bodies produced from stored triacylglyerols become an important fuel source, especially for the brain Abnormally high levels of ketone bodies are observed in diabetics. Diabetic coma can result if ketone body levels become too great  
       [0154] Lipid Mobilization  
       [0155] Within cells, fatty acids are transported by cytoplasmic fatty acid binding proteins (Online Mendelian Inheritance in Man (OMIM)*134650 Fatty Acid-Binding Protein 1, Liver; FABP1). Diazepam binding inhibitor (DBI), also known as endozepine and acyl CoA-binding protein, is an endogenous γ-aminobutyric acid (GABA) receptor ligand which is thought to down-regulate the effects of GABA DBI binds medium- and long-chain acyl-CoA esters with very high affinity and may function as an intracellular carrier of acyl-CoA esters (OMIM*125950 Diazepam Binding inhibitor; DBI; PROSITE PDOC00686 Acyl-CoA-binding protein signature).  
       [0156] Fat stored in liver and adipose triglycerides may be released by hydrolysis and transported in the blood Free fatty acids are transported in the blood by albumin Triacylglycerols and cholesterol esters in the blood are transported in lipoprotein particles. The particles consist of a core of hydrophobic lipids surrounded by a shell of polar lipids and apolipoproteins. The protein components serve in the solubilization of hydrophobic lipids and also contain cell-targeting signals. Lipoproteins include chylomicrons, chylomicron remnants, very-low-density lipoproteins (VLDL), intermediate-density lipoproteins (IDL), low-density lipoproteins (LDL), and high-density lipoproteins (DL). There is a strong inverse correlation between the levels of plasma HDL and risk of premature coronary heart disease.  
       [0157] Triacylglycerols in chylomicrons and VLDL are hydrolyzed by lipoprotein lipases that line blood vessels in muscle and other tissues that use fatty acids. Cell surface LDL receptors bind LDL particles which are then internalized by endocytosis. Absence of the LDL receptor, the cause of the disease familial hypercholesterolemia, leads to increased plasma cholesterol levels and ultimately to atherosclerosis. Plasma cholesteryl ester transfer protein mediates the transfer of cholesteryl esters from HDL to apolipoprotein B-containing lipoproteins. Cholesteryl ester transfer protein is important in the reverse cholesterol transport system and may play a role in atherosclerosis (Yamashita, S. et al. (1997) Curr. Opin. Lipidol. 8:101-110). Macrophage scavenger receptors, which bind and internalize modified lipoproteins, play a role in lipid transport and may contribute to atherosclerosis (Greaves, D. R. et al. (1998) Curr. Opin. Lipidol. 9:425-432).  
       [0158] Proteins involved in cholesterol uptake and biosynthesis are tightly regulated in response to cellular cholesterol levels. The sterol regulatory element binding protein (SREBP) is a sterol-responsive transcription factor. Under normal cholesterol conditions, SREBP resides in the ER membrane. When cholesterol levels are low, a regulated cleavage of SREBP occurs which releases the extracellular domain of the protein. This cleaved domain is then transported to the nucleus where it activates the transcription of the LDL receptor gene, and genes encoding enzymes of cholesterol synthesis, by binding the sterol regulatory element (SRE) upstream of the genes (Yang, J. et al. (1995) J. Biol. Chem. 270:12152-12161). Regulation of cholesterol uptake and biosynthesis also occurs via the oxysterol-binding protein (OSBP). OSBP is a high-affinity intracellular receptor for a variety of oxysterols that down-regulate cholesterol synthesis and stimulate cholesterol esterification (Lagace, T. A et al. (1997) Biochem. J. 326:205-213).  
       [0159] Beta-oxidation  
       [0160] Mitochondrial and peroxisomal beta-oxidation enzymes degrade saturated and unsaturated fatty acids by sequential removal of two-carbon units from CoA-activated fatty acids. The main beta-oxidation pathway degrades both saturated and unsaturated fatty acids while the auxiliary pathway performs additional steps required for the degradation of unsaturated fatty acids.  
       [0161] The pathways of mitochondrial and peroxisomal beta-oxidation use similar enzymes, but have different substrate specificities and functions. Mitochondria oxidize short-, medium-, and long-chain fatty acids to produce energy for cells. Mitochondrial beta-oxidation is a major energy source for cardiac and skeletal muscle. In liver, it provides ketone bodies to the peripheral circulation when glucose levels are low as in starvation, endurance exercise, and diabetes (Eaton, S. et al. (1996) Biochem. J. 320:345-357). Peroxisomes oxidize medium-, long-, and very-long-chain fatty acids, dicarboxylic fatty acids, branched fatty acids, prostaglandins, xenobiotics, and bile acid intermediates. The chief roles of peroxisomal beta-oxidation are to shorten toxic lipophilic carboxylic acids to facilitate their excretion and to shorten very-long-chain fatty acids prior to mitochondrial beta-oxidation (Mannaerts, G. P. and P. P. van Veldhoven (1993) Biochimie 75:147-158).  
       [0162] Enzymes involved in beta-oxidation include acyl CoA synthetase, carnitine acyltransferase, acyl CoA dehydrogenases, enoyl CoA hydratases, L-3-hydroxyacyl CoA dehydrogenase, β-ketothiolase, 2,4-dienoyl CoA reductase, and isomerase.  
       [0163] Lipid Cleavage and Degradation  
       [0164] Triglycerides are hydrolyzed to fatty acids and glycerol by lipases. Lysophospholipases (LPLs) are widely distributed enzymes that metabolize intracellular lipids, and occur in numerous isoforms. Small isoforms, approximately 15-30 kD, function as hydrolases; large isoforms, those exceeding 60 kD, function both as hydrolases and transacylases. A particular substrate for LPLS, lysophosphatidylcholine, causes lysis of cell membranes when it is formed or imported into a cell. LPLs are regulated by lipid factors including acylcarnitine, arachidonic acid, and phosphatidic acid. These lipid factors are signaling molecules important in numerous pathways, including the inflammatory response. (Anderson, R. et al. (1994) Toxicol. Appl. Pharmacol. 125:176-183; Selle, H. et al. (1993); Eur. J. Biochem.  212:411-416 .)  
       [0165] The secretory phospholipase A 2  (PLA2) superfamily comprises a number of heterogeneous enzymes whose common feature is to hydrolyze the sn-2 fatty acid acyl ester bond of phosphoglycerides. Hydrolysis of the glycerophospholipids releases free fatty acids and lysophospholipids. PLA2 activity generates precursors for the biosynthesis of biologically active lipids, hydroxy fatty acids, and platelet-activating factor. PLA2 hydrolysis of the sn-2 ester bond in phospholipids generates free fatty acids, such as arachidonic acid and lysophospholipids.  
       [0166] Carbon and Carbohydrate Metabolism Carbohydrates, including sugars or saccharides, starch, and cellulose, are aldehyde or ketone compounds with multiple hydroxyl groups. The importance of carbohydrate metabolism is demonstrated by the sensitive regulatory system in place for maintenance of blood glucose levels. Two pancreatic hormones, insulin and glucagon, promote increased glucose uptake and storage by cells, and increased glucose release from cells, respectively. Carbohydrates have three important roles in mammalian cells. First, carbohydrates are used as energy stores, fuels, and metabolic intermediates. Carbohydrates are broken down to form energy in glycolysis and are stored as glycogen for later use. Second, the sugars deoxyribose and ribose form part of the structural support of DNA and RNA, respectively. Third, carbohydrate modifications are added to secreted and membrane proteins and lipids as they traverse the secretory pathway. Cell surface carbohydrate-containing macromolecules, including glycoproteins, glycolipids, and transmembrane proteoglycans, mediate adhesion with other cells and with components of the extracellular matrix. The extracellular matrix is comprised of diverse glycoproteins, glycosaminoglycans (GAGs), and carbohydrate-binding proteins which are secreted from the cell and assembled into an organized meshwork in close association with the cell surface. The interaction of the cell with the surrounding matrix profoundly influences cell shape, strength, flexibility, motility, and adhesion. These dynamic properties are intimately associated with signal transduction pathways controlling cell proliferation and differentiation, tissue construction, and embryonic development.  
       [0167] Carbohydrate metabolism is altered in several disorders including diabetes mellitus, hyperglycemia, hypoglycemia, galactosemia, galactokinase deficiency, and UDP-galactose4-epimerase deficiency (Fauci, A. S. et al. (1998)  Harrison&#39;s Principles of Internal Medicine , McGraw-Hill New York N.Y., pp. 2208-2209). Altered carbohydrate metabolism is associated with cancer. Reduced GAG and proteoglycan expression is associated with human lung carcinomas (Nackaerts, K. et al. (1997) Int. J. Cancer 74:335-345). The carbohydrate determinants sialyl Lewis A and sialyl Lewis X are frequently expressed on human cancer cells (Kannagi, R. (1997) Glycoconj. J. 14:577-584). Alterations of the N-linked carbohydrate core structure of cell surface glycoproteins are linked to colon and pancreatic cancers (Schwarz, R. E. et al. (1996) Cancer Lett. 107:285-291). Reduced expression of the Sda blood group carbohydrate structure in cell surface glycolipids and glycoproteins is observed in gastrointestinal cancer (Dohi, T. et al. (1996) Int. J. Cancer 67:626-663). (Carbon and carbohydrate metabolism is reviewed in Stryer, L. (1995)  Biochemistry  W. H. Freeman and Company, New York N.Y.; Lehninger, A. L. (1982)  Principles of Biochemistry  Worth Publishers Inc., New York N.Y.; and Lodish, H. et al. (1995)  Molecular Cell Biology  Scientific American Books, New York N.Y.)  
       [0168] Glycolysis  
       [0169] Enzymes of the glycolytic pathway convert the sugar glucose to pyruvate while simultaneously producing ATP. The pathway also provides building blocks for the synthesis of cellular components such as long-chain fatty acids. After glycolysis, pyrvuate is converted to acetyl-Coenzyme A, which, in aerobic organisms, enters the citric acid cycle. Glycolytic enzymes include hexokinase, phosphoglucose isomerase, phosphofructokinase, aldolase, triose phosphate isomerase, glyceraldehyde 3-phosphate dehydrogenase, phosphoglycerate kinase, phosphoglyceromutase, enolase, and pyruvate kinase. Of these, phosphofructokinase, hexokinase, and pyruvate kinase are important in regulating the rate of glycolysis.  
       [0170] Gluconeogenesis  
       [0171] Gluconeogenesis is the synthesis of glucose from noncarbohydrate precursors such as lactate and amino acids. The pathway, which functions mainly in times of starvation and intense exercise, occurs mostly in the liver and kidney. Responsible enzymes include pyruvate carboxylase, phosphoenolpyruvate carboxykinase, fructose 1,6-bisphosphatase, and glucose-6-phosphatase.  
       [0172] Pentose Phosohate Pathway  
       [0173] Pentose phosphate pathway enzymes are responsible for generating the reducing agent NADPH, while at the same time oxidizing glucose-6-phosphate to ribose-5-phosphate. Ribose-5-phosphate and its derivatives become part of important biological molecules such as ATP, Coenzyme A, NAD + , FAD, RNA, and DNA The pentose phosphate pathway has both oxidative and non-oxidative branches. The oxidative branch steps, which are catalyzed by the enzymes glucose-6-phosphate dehydrogenase, lactonase, and 6-phosphogluconate dehydrogenase, convert glucose-6-phosphate and NADP +  to ribulose-6-phosphate and NADPH. The non-oxidative branch steps, which are catalyzed by the enzymes phosphopentose isomerase, phosphopentose epimerase, transketolase, and transaldolase, allow the interconversion of three-, four-, five-, six-, and seven-carbon sugars.  
       [0174] Glucouronate Metabolism  
       [0175] Glucuronate is a monosaccharide which, in the form of D-glucuronic acid, is found in the GAGs chondroitin and dermatan. D-glucuronic acid is also important in the detoxification and excretion of foreign organic compounds such as phenol. Enzymes involved in glucuronate metabolism include UDP-glucose dehydrogenase and glucuronate reductase.  
       [0176] Disaccharide Metabolism  
       [0177] Disaccharides must be hydrolyzed to monosaccharides to be digested. Lactose, a disaccharide found in mil, is hydrolyzed to galactose and glucose by the enzyme lactase. Maltose is derived from plant starch and is hydrolyzed to glucose by the enzyme maltase. Sucrose is derived from plants and is hydrolyzed to glucose and fructose by the enzyme sucrase. Trehalose, a disaccharide found mainly in insects and mushrooms, is hydrolyzed to glucose by the enzyme trehalase (OMIM*275360 Trehalase; Ruf, J. et al. (1990) J. Biol. Chem. 265:1503415039). Lactase, maltase, sucrase, and trehalase are bound to mucosal cells lining the small intestine, where they participate in the digestion of dietary disaccharides. The enzyme lactose synthetase, composed of the catalytic subunit galactosyltransferase and the modifier subunit α-lactalbumin, converts UDP-galactose and glucose to lactose in the mammary glands.  
       [0178] Glycogen, Starch, and Chitin Metabolism  
       [0179] Glycogen is the storage form of carbohydrates in mammals. Mobilization of glycogen maintains glucose levels between meals and during muscular activity. Glycogen is stored mainly in the liver and in skeletal muscle in the form of cytoplasmic granules. These granules contain enzymes that catalyze the synthesis and degradation of glycogen, as well as enzymes that regulate these processes. Enzymes that catalyze the degradation of glycogen include glycogen phosphorylase, a transferase, α-1,6-glucosidase, and phosphoglucomutase. Enzymes that catalyze the synthesis of glycogen include UDP-glucose pyrophosphorylase, glycogen synthetase, a branching enzyme, and nucleoside diphosphokinase. The enzymes of glycogen synthesis and degradation are tightly regulated by the hormones insulin, glucagon, and epinephrine. Starch, a plant-derived polysaccharide, is hydrolyzed to maltose, maltotriose, and α-dextrin by α-amylase, an enzyme secreted by the salivary glands and pancreas. Chitin is a polysaccharide found in insects and crustacea. A chitotriosidase is secreted by macrophages and may play a role in the degradation of chitin-containing pathogens (Boot, R. G. et al. (1995) J. Biol. Chem. 270:26252-26256).  
       [0180] Peptidoglycans and Glycosaminoglycans  
       [0181] Glycosaminoglycans (GAGs) are anionic linear unbranched polysaccharides composed of repetitive disaccharide units. These repetitive units contain a derivative of an amino sugar, either glucosamine or galactosamine. GAGs exist free or as part of proteoglycans, large molecules composed of a core protein attached to one or more GAGs. GAGs are found on the cell surface, inside cells, and in the extracellular matrix. Changes in GAG levels are associated with several autoimmune diseases including autoimmune thyroid disease, autoimmune diabetes mellitus, and systemic lupus erythematosus (Hansen, C. et al. (1996) Clin. Exp. Rheum 14 (Suppl. 15):S59-S67). GAGs include chondroitin sulfate, keratan sulfate, heparin, heparan sulfate, dermatan sulfate, and hyaluronan  
       [0182] The GAG hyaluronan (HA) is found in the extracellular matrix of many cells, especially in soft connective tissues, and is abundant in synovial fluid (Pitsillides, A. A. et al. (1993) Int. J. Exp. Pathol. 74:27-34). HA seems to play important roles in cell regulation, development, and differentiation (Laurent, T. C. and J. R. Fraser (1992) FASEB J. 6:2397-2404). Hyaluronidase is an enzyme that degrades HA to oligosaccharides. Hyaluronidases may function in cell adhesion, infection, angiogenesis, signal transduction, reproduction, cancer, and inflammation.  
       [0183] Proteoglycans, also known as peptidoglycans, are found in the extracellular matrix of connective tissues such as cartilage and are essential for distributing the load in weight-bearing joints. Cell-surface-attached proteoglycans anchor cells to the extracellular matrix. Both extracellular and cell-surface proteoglycans bind growth factors, facilitating their binding to cell-surface receptors and subsequent triggering of signal transduction pathways.  
       [0184] Amino Acid and Nitrogen Metabolism  
       [0185] NH 4   +  is assimilated into amino acids by the actions of two enzymes, glutamate dehydrogenase and glutamine synthetase. The carbon skeletons of amino acids come from the intermediates of glycolysis, the pentose phosphate pathway, or the citric acid cycle. Of the twenty amino acids used in proteins, humans can synthesize only thirteen (nonessential amino acids). The remaining nine must come from the diet (essential amino acids). Enzymes involved in nonessential amino acid biosynthesis include glutamate kinase dehydrogenase, pyrroline carboxylate reductase, asparagine synthetase, phenylalanine oxygenase, methionine adenosyltransferase, adenosylhomocysteinase, cystathionine β-synthase, cystathionine γ-lyase, phosphoglycerate dehydrogenase, phosphoserine transaminase, phosphoserine phosphatase, serine hydroxylmethyltransferase, and glycine synthase.  
       [0186] Metabolism of amino acids takes place almost entirely in the liver, where the amino group is removed by aminotransferases (transaminases), for example, alanine aminotransferase. The amino group is transferred to α-ketoglutarate to form glutamate. Glutamate dehydrogenase converts glutamate to NH 4   +  and α-ketoglutarate. NH 4   +  is converted to urea by the urea cycle which is catalyzed by the enzymes arginase, ornithine transcarbamoylase, arginosuccinate synthetase, and arginosuccinase. Carbamoyl phosphate synthetase is also involved in urea formation. Enzymes involved in the metabolism of the carbon skeleton of amino acids include serine dehydratase, asparaginase, glutaminase, propionyl CoA carboxylase, methylmalonyl CoA mutase, branched-chain α-keto dehydrogenase complex, isovaleryl CoA dehydrogenase, β-methylcrotonyl CoA carboxylase, phenylalanine hydroxylase, p-hydroxylphenylpyruvate hydroxylase, and homogentisate oxidase.  
       [0187] Polyamines, which include spermidine, putrescine, and spermine, bind tightly to nucleic acids and are abundant in rapidly proliferating cells. Enzymes involved in polyamine synthesis include ornithine decarboxylase.  
       [0188] Diseases involved in amino acid and nitrogen metabolism include hyperammonemia, carbamoyl phosphate synthetase deficiency, urea cycle enzyme deficiencies, methylmalonic aciduria, maple syrup disease, alcaptonuria, and phenylketonuria.  
       [0189] Energy Metabolism  
       [0190] Cells derive energy from metabolism of ingested compounds that may be roughly categorized as carbohydrates, fats, or proteins. Energy is also stored in polymers such as triglycerides (fats) and glycogen (carbohydrates). Metabolism proceeds along separate reaction pathways connected by key intermediates such as acetyl coenzyme A (acetyl-CoA). Metabolic pathways feature anaerobic and aerobic degradation, coupled with the energy-requiring reactions such as phosphorylation of adenosine diphosphate (ADP) to the triphosphate (ATP) or analogous phosphorylations of guanosine (GDP/GTP), uridine (UDP/UTP), or cytidine (CDP/CTP). Subsequent dephosphorylation of the triphosphate drives reactions needed for cell maintenance, growth, and proliferation.  
       [0191] Digestive enzymes convert carbohydrates and sugars to glucose; fructose and galactose are converted in the liver to glucose. Enzymes involved in these conversions include galactose-1-phosphate uridyl transferase and UDP-galactose-4 epimerase. In the cytoplasm, glycolysis converts glucose to pyruvate in a series of reactions coupled to ATP synthesis.  
       [0192] Pyruvate is transported into the mitochondria and converted to acetyl-CoA for oxidation via the citric acid cycle, involving pyruvate dehydrogenase components, dihydrolipoyl transacetylase, and dihydrolipoyl dehydrogenase. Enzymes involved in the citric acid cycle include: citrate synthetase, aconitases, isocitrate dehydrogenase, alpha-ketoglutarate dehydrogenase complex including transsuccinylases, succinyl CoA synthetase, succinate dehydrogenase, fumarases, and malate dehydrogenase. Acetyl CoA is oxidized to CO 2  with concomitant formation of NADH, FADH 2 , and GTP. In oxidative phosphorylation, the transport of electrons from NADH and FADH 2  to oxygen by dehydrogenases is coupled to the synthesis of ATP from ADP and P i  by the F 0 F 1  ATPase complex in the mitochondrial inner membrane. Enzyme complexes responsible for electron transport and ATP synthesis include the F 0 F 1  ATPase complex, ubiquinone(CoQ)-cytochrome c reductase, ubiquinone reductase, cytochrome b, cytochrome c 1 , FeS protein, and cytochrome c oxidase.  
       [0193] Triglycerides are hydrolyzed to fatty acids and glycerol by lipases. Glycerol is then is phosphorylated to glycerol-3-phosphate by glycerol kinase and glycerol phosphate dehydrogenase, and degraded by the glycolysis. Fatty acids are transported into the mitochondria as fatty acyl-carnitine esters and undergo oxidative degradation.  
       [0194] In addition to metabolic disorders such as diabetes and obesity, disorders of energy metabolism are associated with cancers (Dorward, A. et al. (1997) J. Bioenerg. Biomembr. 29:385-392), autism (Lombard, J. (1998) Med. Hypotheses 50:497-500), neurodegenerative disorders (Alexi, T. et al. (1998) Neuroreport 9:R57-64), and neuromuscular disorders (DiMauro, S. et al. (1998) Biochim. Biophys. Acta 1366:199-210). The myocardium is heavily dependent on oxidative metabolism, so metabolic dysfunction often leads to heart disease (DiMauro, S. and M. Hirano (1998) Curr. Opin. Cardiol. 13:190-197).  
       [0195] For a review of energy metabolism enzymes and intermediates, see Stryer, L. et al. (1995)  Biochemistry , W. H. Freeman and Co., San Francisco Calif., pp.443-652. For a review of energy metabolism regulation, see Lodish, H. et al. (1995)  Molecular Cell Biology , Scientific American Books, New York N.Y., pp. 744-770.  
       [0196] Cofactor Metabolism  
       [0197] Cofactors, including coenzymes and prosthetic groups, are small molecular weight inorganic or organic compounds that are required for the action of an enzyme. Many cofactors contain vitamins as a component. Cofactors include thiamine pyrophosphate, flavin adenine dinucleotide, flavin mononucleotide, nicotinamide adenine dinucleotide, pyridoxal phosphate, coenzyme A, tetrahydrofolate, lipoamide, and heme. The vitamins biotin and cobalamin are associated with enzymes as well. Heme, a prosthetic group found in myoglobin and hemoglobin, consists of protoporphyrin group bound to iron. Porphyrin groups contain four substituted pyrroles covalently joined in a ring, often with a bound metal atom. Enzymes involved in porphyrin synthesis include δ-aminolevulinate synthase, δ-aminolevulinate dehydrase, porphobilinogen deaminase, and cosynthase. Deficiencies in heme formation cause porphyrias. Heme is broken down as a part of erythrocyte turnover. Enzymes involved in heme degradation include heme oxygenase and biliverdin reductase.  
       [0198] Iron is a required cofactor for many enzymes. Besides the heme-containing enzymes, iron is found in iron-sulfur clusters in proteins including aconitase, succinate dehydrogenase, and NADH-Q reductase. Iron is transported in the blood by the protein transferrin Binding of transferrin to the transferrin receptor on cell surfaces allows uptake by receptor mediated endocytosis. Cytosolic iron is bound to ferritin protein.  
       [0199] A molybdenum-containing cofactor (molybdopterin) is found in enzymes including sulfite oxidase, xanthine dehydrogenase, and aldehyde oxidase. Molybdopterin biosynthesis is performed by two molybdenum cofactor synthesizing enzymes. Deficiencies in these enzymes cause mental retardation and lens dislocation. Other diseases caused by defects in cofactor metabolism include pernicious anemia and methylmalonic aciduria.  
       [0200] Secretion and Trafficking  
       [0201] Eukaryotic cells are bound by a lipid bilayer membrane and subdivided into functionally distinct, membrane bound compartments. The membranes maintain the essential differences between the cytosol, the extracelluar environment, and the lumenal space of each intracellular organelle. As lipid membranes are highly impermeable to most polar molecules, transport of essential nutrients, metabolic waste products, cell signaling molecules, macromolecules and proteins across lipid membranes and between organelles must be mediated by a variety of transport-associated molecules.  
       [0202] Protein Trafficking  
       [0203] In eukaryotes, some proteins are synthesized on ER-bound ribosomes, co-translationally imported into the ER, delivered from the ER to the Golgi complex for post-translational processing and sorting, and transported from the Golgi to specific intracellular and extracellular destinations. All cells possess a constitutive transport process which maintains homeostasis between the cell and its environment. In many differentiated cell types, the basic machinery is modified to carry out specific transport functions. For example, in endocrine glands, hormones and other secreted proteins are packaged into secretory granules for regulated exocytosis to the cell exterior. In macrophage, foreign extracellular material is engulfed (phagocytosis) and delivered to lysosomes for degradation. In fat and muscle cells, glucose transporters are stored in vesicles which fuse with the plasma membrane only in response to insulin stimulation.  
       [0204] The Secretory Pathway  
       [0205] Synthesis of most integral membrane proteins, secreted proteins, and proteins destined for the lumen of a particular organelle occurs on ER-bound ribosomes. These proteins are co-translationally imported into the ER. The proteins leave the ER via membrane-bound vesicles which bud off the ER at specific sites and fuse with each other (homotypic fusion) to form the ER-Golgi Intermediate Compartment (ERGIC). The ERGIC matures progressively through the cis, medial, and trans cisternal stacks of the Golgi, modifying the enzyme composition by retrograde transport of specific Golgi enzymes. In this way, proteins moving through the Golgi undergo post-translational modification, such as glycosylation. The final Golgi compartment is the Trans-Golgi Network (TGN), where both membrane and lumenal proteins are sorted for their final destination. Transport vesicles destined for intracellular compartments, such as the lysosome, bud off the TGN. What remains is a secretory vesicle which contains proteins destined for the plasma membrane, such as receptors, adhesion molecules, and ion channels, and secretory proteins, such as hormones, neurotransmitters, and digestive enzymes. Secretory vesicles eventually fuse with the plasma membrane (Glick, B. S. and V. Malhotra (1998) Cell 95:883-889).  
       [0206] The secretory process can be constitutive or regulated. Most cells have a constitutive pathway for secretion, whereby vesicles derived from maturation of the TGN require no specific signal to fuse with the plasma membrane. In many cells, such as endocrine cells, digestive cells, and neurons, vesicle pools derived from the TGN collect in the cytoplasm and do not fuse with the plasma membrane until they are directed to&#39;by a specific signal.  
       [0207] Endocytosis  
       [0208] Endocytosis, wherein cells internalize material from the extracellular environment, is essential for transmission of neuronal, metabolic, and proliferative signals; uptake of many essential nutrients; and defense against invading organisms. Most cells exhibit two forms of endocytosis. The first, phagocytosis, is an actin-driven process exemplified in macrophage and neutrophils. Material to be endocytosed contacts numerous cell surface receptors which stimulate the plasma membrane to extend and surround the particle, enclosing it in a membrane-bound phagosome. In the mammalian immune system, IgG-coated particles bind Fc receptors on the surface of phagocytic leukocytes. Activation of the Fc receptors initiates a signal cascade involving src-family cytosolic kinases and the monomeric GTP-binding (G) protein Rho. The resulting actin reorganization leads to phagocytosis of the particle. This process is an important component of the humoral immune response, allowing the processing and presentation of bacterial-derived peptides to antigen-specific T-lymphocytes.  
       [0209] The second form of endocytosis, pinocytosis, is a more generalized uptake of material from the external milieu. Like phagocytosis, pinocytosis is activated by ligand binding to cell surface receptors. Activation of individual receptors stimulates an internal response that includes coalescence of the receptor-ligand complexes and formation of clathrin-coated pits. Invagination of the plasma membrane at clathrin-coated pits produces an endocytic vesicle within the cell cytoplasm. These vesicles undergo homotypic fusion to form an early endosomal (EE) compartment. The tubulovesicular EE serves as a sorting site for incoming material. ATP-driven proton pumps in the EE membrane lowers the pH of the EE lumen (pH 6.3-6.8). The acidic environment causes many ligands to dissociate from their receptors. The receptors, along with membrane and other integral membrane proteins, are recycled back to the plasma membrane by budding off the tubular extensions of the EE in recycling vesicles (RV). This selective removal of recycled components produces a carrier vesicle containing ligand and other material from the external environment. The carrier vesicle fuses with TGN-derived vesicles which contain hydrolytic enzymes. The acidic environment of the resulting late endosome (LE) activates the hydrolytic enzymes which degrade the ligands and other material. As digestion takes place, the LE fuses with the lysosome where digestion is completed (Mellman, I. (1996) Annu. Rev. Cell Dev. Biol. 12:575-625).  
       [0210] Recycling vesicles may return directly to the plasma membrane. Receptors internalized and returned directly to the plasma membrane have a turnover rate of 2-3 minutes. Some RVs undergo microtubule-directed relocation to a perinuclear site, from which they then return to the plasma membrane. Receptors following this route have a turnover rate of 5-10 minutes. Still other RVs are retained within the cell until an appropriate signal is received (Mellman, supra; and James, D. E. et al. (1994) Trends Cell Biol. 4:120-126).  
       [0211] Vesicle Formation  
       [0212] Several steps in the transit of material along the secretory and endocytic pathways require the formation of transport vesicles. Specifically, vesicles form at the transitional endoplasmic reticulum (tER), the rim of Golgi cisternae, the face of the Trans-Golgi Network (TGN), the plasma membrane (PM), and tubular extensions of the endosomes. The process begins with the budding of a vesicle out of the donor membrane. The membrane-bound vesicle contains proteins to be transported and is surrounded by a protective coat made up of protein subunits recruited from the cytosol. The initial budding and coating processes are controlled by a cytosolic ras-like GTP-binding protein, ADP-ribosylating factor (Arf), and adapter proteins (AP). Different isoforms of both Arf and AP are involved at different sites of budding. Another small G-protein, dynamin, forms a ring complex around the neck of the forming vesicle and may provide the mechanochemical force to accomplish the final step of the budding process. The coated vesicle complex is then transported through the cytosol. During the transport process, Arf-bound GTP is hydrolyzed to GDP and the coat dissociates from the transport vesicle (West, M. A. et al. (1997) J. Cell Biol. 138:1239-1254). Two different classes of coat protein have also been identified Clathrin coats form on the TGN and PM surfaces, whereas coatomer or COP coats form on the ER and Golgi. COP coats can further be distinguished as COPI, involved in retrograde traffic through the Golgi and from the Golgi to the ER, and COPII, involved in anterograde traffic from the ER to the Golgi (Mellman, supra). The COP coat consists of two major components, a G-protein (Arf or Sar) and coat protomer (coatomer). Coatomer is an equimolar complex of seven proteins, termed alpha-, beta-, beta&#39;-, gamma-, delta-, epsilon- and zeta-COP. (Harter, C. and F. T. Wieland (1998) Proc. Natl. Acad. Sci. USA 95:11649-11654.)  
       [0213] Membrane Fusion  
       [0214] Transport vesicles undergo homotypic or heterotypic fusion in the secretory and endocytotic pathways. Molecules required for appropriate targeting and fusion of vesicles with their target membrane include proteins incorporated in the vesicle membrane, the target membrane, and proteins recruited from the cytosol. During budding of the vesicle from the donor compartment, an integral membrane protein, VAMP (vesicle-associated membrane protein) is incorporated into the vesicle. Soon after the vesicle uncoats, a cytosolic prenylated GTP-binding protein, Rab (a member of the Ras superfamily), is inserted into the vesicle membrane. GTP-bound Rab proteins are directed into nascent transport vesicles where they interact with VAMP. Following vesicle transport, GTPase activating proteins (GAPs) in the target membrane convert Rab proteins to the GDP-bound form A cytosolic protein, guanine-nucleotide dissociation inhibitor (GDI) helps return GDP-bound Rab proteins to their membrane of origin. Several Rab isoforms have been identified and appear to associate with specific compartments within the cell. Rab proteins appear to play a role in mediating the function of a viral gene, Rev, which is essential for replication of HIV-1, the virus responsible for AIDS (Flavell, R. A. et al. (1996) Proc. Natl. Acad. Sci. USA  93:4421-4424 ).  
       [0215] Docking of the transport vesicle with the target membrane involves the formation of a complex between the vesicle SNAP receptor (v-SNARE), target membrane (t-) SNAREs, and certain other membrane and cytosolic proteins. Many of these other proteins have been identified although their exact functions in the docking complex remain uncertain (Tellam, J. T. et al. (1995) J. Biol. Chem. 270:5857-5863; and Hata, Y. and T. C. Sudhof (1995) J. Biol. Chem. 270:13022-13028). N-ethylmaleimide sensitive factor (NSF) and soluble NSF-attachment protein (α-SNAP and, β-SNAP) are two such proteins that are conserved from yeast to man and function in most intracellular membrane fusion reactions. Sec1 represents a family of yeast proteins that function at many different stages in the secretory pathway including membrane fusion Recently, mammalian homologs of Sec1, called Munc-18 proteins, have been identified (Katagiri, H. et al. (1995) J. Biol. Chem 270:4963-4966; Hata et al. supra).  
       [0216] The SNARE complex involves three SNARE molecules, one in the vesicular membrane and two in the target membrane. Synaptotagmin is an integral membrane protein in the synaptic vesicle which associates with the t-SNARE syntaxin in the docking complex. Synaptotagmin binds calcium in a complex with negatively charged phospholipids, which allows the cytosolic SNAP protein to displace synaptotagmin from syntaxin and fusion to occur. Thus, synaptotagmin is a negative regulator of fusion in the neuron (Littleton, J. T. et al. (1993) Cell 74:1125-1134). The most abundant membrane protein of synaptic vesicles appears to be the glycoprotein synaptophysin, a 38 kDa protein with four transmembrane domains.  
       [0217] Specificity between a vesicle and its target is derived from the v-SNARE, t-SNAREs, and associated proteins involved. Different isoforms of SNAREs and Rabs show distinct cellular and subcellular distributions. VAMP-1/synaptobrevin, membrane-anchored synaptosome-associated protein of 25 kDa (SNAP-25), syntaxin-1, Rab3A, Rab15, and Rab23 are predominantly expressed in the brain and nervous system. Different syntaxin, VAMP, and Rab proteins are associated with distinct subcellular compartments and their vesicular carriers.  
       [0218] Nuclear Transport  
       [0219] Transport of proteins and RNA between the nucleus and the cytoplasm occurs through nuclear pore complexes (NPCs). NPC-mediated transport occurs in both directions through the nuclear envelope. All nuclear proteins are imported from the cytoplasm, their site of synthesis. tRNA and mRNA are exported from the nucleus, their site of synthesis, to the cytoplasm, their site of function. Processing of small nuclear RNAs involves export into the cytoplasm, assembly with proteins and modifications such as hypermethylation to produce small nuclear ribonuclear proteins (s RNs), and subsequent import of the snRNPs back into the nucleus. The assembly of ribosomes requires the initial import of ribosomal proteins from the cytoplasm, their incorporation with RNA into ribosomal subunits, and export back to the cytoplasm. (Görlich, D. and I. W. Mattaj (1996) Science 271:1513-1518.)  
       [0220] The transport of proteins and mRNAs across the NPC is selective, dependent on nuclear localization signals, and generally requires association with nuclear transport factors. Nuclear localization signals (NLS) consist of short stretches of amino acids enriched in basic residues. NLS 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). NTF2 binds the GDP-bound form of Ran and to multiple proteins of the nuclear pore complex containing FXFG repeat motifs, such as p62. (Paschal, B. et al. (1997) J. Biol. Chem.  272:21534-21539 ; and Wong, D. H. et al. (1997) Mol. Cell Biol. 17:3755-3767). Some proteins are dissociated before nuclear mRNAs are transported across the NPC while others are dissociated shortly after nuclear mRNA transport across the NPC and are reimported into the nucleus.  
       [0221] Disease Correlation  
       [0222] The etiology of numerous human diseases and disorders can be attributed to defects in the transport or secretion of proteins. For example, abnormal hormonal secretion is linked to disorders such as diabetes insipidus (vasopressin), hyper- and hypoglycemia (insulin, glucagon), Grave&#39;s disease and goiter (thyroid hormone), and Cushing&#39;s and Addison&#39;s diseases (adrenocorticotropic hormone, ACTH). Moreover, cancer cells secrete excessive amounts of hormones or other biologically active peptides. Disorders related to excessive secretion of biologically active peptides by tumor cells include fasting hypoglycemia due to increased insulin secretion from insulinoma-islet cell tumors; hypertension due to increased epinephrine and norepinephrine secreted from pheochromocytomas of the adrenal medulla and sympathetic paraganglia; and carcinoid syndrome, which is characterized by abdominal cramps, diarrhea, and valvular heart disease caused by excessive amounts of vasoactive substances such as serotonin, bradykinin, histamine, prostaglandins, and polypeptide hormones, secreted from intestinal tumors. Biologically active peptides that are ectopically synthesized in and secreted from tumor cells include ACTH and vasopressin (lung and pancreatic cancers); parathyroid hormone (lung and bladder cancers); calcitonin (lung and breast cancers); and thyroid-stimulating hormone (medullary thyroid carcinoma). Such peptides may be useful as diagnostic markers for tumorigenesis(Schwartz, M. Z. (1997) Semin. Pediatr. Surg. 3:141-146; and Said, S. I. and G. R. Faloona (1975) N. Engl. J. Med. 293:155-160).  
       [0223] Defective nuclear transport may play a role in cancer. The BRCA1 protein contains three potential NLSs which interact with importin alpha, and is transported into the nucleus by the importin/NPC pathway. In breast cancer cells the BRCA1 protein is aberrantly localized in the cytoplasm. The mislocation of the BRCA1 protein in breast cancer cells may be due to a defect in the NPC nuclear import pathway (Chen, C. F. et al. (1996) J. Biol. Chem. 271:32863-32868).  
       [0224] It has been suggested that in some breast cancers, the tumor-suppressing activity of p53 is inactivated by the sequestration of the protein in the cytoplasm, away from its site of action in the cell nucleus. Cytoplasmic wild-type p53 was also found inhuman cervical carcinoma cell lines. (Moll, U. M. et al. (1992) Proc. Natl. Acad. Sci. USA 89:7262-7266; and Liang, X. H. et al. (1993) Oncogene 8:2645-2652.)  
       [0225] Environmental Responses  
       [0226] Organisms respond to the environment by a number of pathways. Heat shock proteins, including hsp 70, hsp60, hsp90, and hsp 40, assist organisms in coping with heat damage to cellular proteins.  
       [0227] Aquaporins (AQP) are channels that transport water and, in some cases, nonionic small solutes such as urea and glycerol. Water movement is important for a number of physiological processes including renal fluid filtration, aqueous humor generation in the eye, cerebrospinal fluid production in the brain, and appropriate hydration of the lung. Aquaporins are members of the major intrinsic protein (MIP) family of membrane transporters (King, L. S. and P. Agre (1996) Annu. Rev. Physiol. 58:619-648; Ishibashi, K. et al. (1997) J. Biol. Chem. 272:20782-20786). The study of aquaporins may have relevance to understanding edema formation and fluid balance in both normal physiology and disease states (King, supra). Mutations in AQP2 cause autosomal recessive nephrogenic diabetes insipidus (OMIM*107777 Aquaporin 2; AQP2). Reduced AQP4 expression in skletal muscle may be associated with Duchenne muscular dystrophy (Frigeri, A. et al. (1998) J. Clin. Invest. 102:695-703). Mutations in AQPO cause autosomal dominant cataracts in the mouse (OMIM *154050 Major Intrinsic Protein of Lens Fiber; MIP).  
       [0228] The metallothioneins (M Is) are a group of small (61 amino acids), cysteine-rich proteins that bind heavy metals such as cadmium, zinc, mercury, lead, and copper and are thought to play a role in metal detoxification or the metabolism and homeostasis of metals. Arsenite-resistance proteins have been identified in hamsters that are resistant to toxic levels of arsenite (Rossman, T. G. et al. (1997) Mutat. Res. 386:307-314).  
       [0229] Humans respond to light and odors by specific protein pathways. Proteins involved in light perception include rhodopsin, transducin, and cGMP phosphodiesterase. Proteins involved in odor perception include multiple olfactory receptors. Other proteins are important inhuman Circadian rhythms and responses to wounds.  
       [0230] Immunity and Host Defense  
       [0231] All vertebrates have developed sophisticated and complex immune systems that provide protection from viral, bacterial, fungal and parasitic infections. Included in these systems are the processes of humoral immunity, the complement cascade and the inflammatory response (Paul, W. E. (1993)  Fundamental Immunology , Raven Press, Ltd., New York N.Y., pp.1-20).  
       [0232] The cellular components of the humoral immune system include six different types of leukocytes: monocytes, lymphocytes, polymorphonuclear granulocytes (consisting of neutrophils, eosinophils, and basophils) and plasma cells. Additionally, fragments of megakaryocytes, a seventh type of white blood cell in the bone marrow, occur in large numbers in the blood as platelets.  
       [0233] Leukocytes are formed from two stem cell lineages in bone marrow. The myeloid stem cell line produces granulocytes and monocytes and, the lymphoid stem cell produces lymphocytes. Lymphoid cells travel to the thymus, spleen and lymph nodes, where they mature and differentiate into lymphocytes. Leukocytes are responsible for defending the body against invading pathogens.  
       [0234] Neutrophils and monocytes attack invading bacteria, viruses, and other pathogens and destroy them by phagocytosis. Monocytes enter tissues and differentiate into macrophages which are extremely phagocytic. Lymphocytes and plasma cells are a part of the immune system which recognizes specific foreign molecules and organisms and inactivates them, as well as signals other cells to attack the invaders.  
       [0235] Granulocytes and monocytes are formed and stored in the bone marrow until needed. Megakaryocytes are produced in bone marrow, where they fragment into platelets and are released into the bloodstream. The main function of platelets is to activate the blood clotting mechanism. Lymphocytes and plasma cells are produced in various lymphogenous organs, including the lymph nodes, spleen, thymus, and tonsils.  
       [0236] Both neutrophils and macrophages exhibit chemotaxis towards sites of inflammation. Tissue inflammation in response to pathogen invasion results in production of chemo-attractants for leukocytes, such as endotoxins or other bacterial products, prostaglandins, and products of leukocytes or platelets.  
       [0237] Basophils participate in the release of the chemicals involved in the inflammatory process. The main function of basophils is secretion of these chemicals to such a degree that they have been referred to as “unicellular endocrine glands.” A distinct aspect of basophilic secretion is that the contents of granules go directly into the extracellular environment, not into vacuoles as occurs with neutrophils, eosinophils and monocytes. Basophils have receptors for the Fc fragment of immunoglobulin E (IgE) that are not present on other leukocytes. Crosslinking of membrane IgE with anti-IgE or other ligands triggers degranulation.  
       [0238] Eosinophils are bi- or multi-nucleated white blood cells which contain eosinophilic granules. Their plasma membrane is characterized by Ig receptors, particularly IgG and IgE. Generally, eosinophils are stored in the bone marrow until recruited for use at a site of inflammation or invasion. They have specific functions in parasitic infections and allergic reactions, and are thought to detoxify some of the substances released by mast cells and basophils which cause inflammation. Additionally, they phagocytize antigen-antibody complexes and further help prevent spread of the inflammation.  
       [0239] Macrophages are monocytes that have left the blood stream to settle in tissue. Once monocytes have migrated into tissues, they do not reenter the bloodstream. The mononuclear phagocyte system is comprised of precursor cells in the bone marrow, monocytes in circulation, and macrophages in tissues. The system is capable of very fast and extensive phagocytosis. A macrophage may phagocytize over 100 bacteria, digest them and extrude residues, and then survive for many more months. Macrophages are also capable of ingesting large particles, including red blood cells and malarial parasites. They increase several-fold in size and transform into macrophages that are characteristic of the tissue they have entered, surviving in tissues for several months.  
       [0240] Mononuclear phagocytes are essential in defending the body against invasion by foreign pathogens, particularly intracellular microorganisms such as  M. tuberculosis , listeria, leishmania and toxoplasma. Macrophages can also control the growth of tumorous cells, via both phagocytosis and secretion of hydrolytic enzymes. Another important function of macrophages is that of processing antigen and presenting them in a biochemically modified form to lymphocytes.  
       [0241] The immune system responds to invading microorganisms in two major ways: antibody production and cell mediated responses. Antibodies are immunoglobulin proteins produced by B-lymphocytes which bind to specific antigens and cause inactivation or promote destruction of the antigen by other cells. Cell-mediated immune responses involve T-lymphocytes (T cells) that react with foreign antigen on the surface of infected host cells. Depending on the type of T cell, the infected cell is either killed or signals are secreted which activate macrophages and other cells to destroy the infected cell (Paul, supra).  
       [0242] T-lymphocytes originate in the bone marrow or liver in fetuses. Precursor cells migrate via the blood to the thymus, where they are processed to mature into T-lymphocytes. This processing is crucial because of positive and negative selection of T cells that will react with foreign antigen and not with self molecules. After processing, T cells continuously circulate in the blood and secondary lymphoid tissues, such as lymph nodes, spleen, certain epithelium-associated tissues in the gastrointestinal tract, respiratory tract and skin. When T-lymphocytes are presented with the complementary antigen, they are stimulated to proliferate and release large numbers of activated T cells into the lymph system and the blood system. These activated T cells can survive and circulate for several days. At the same time, T memory cells are created, which remain in the lymphoid tissue for months or years. Upon subsequent exposure to that specific antigen, these memory cells will respond more rapidly and with a stronger response than induced by the original antigen. This creates an “immunological memory” that can provide immunity for years.  
       [0243] There are two major types of T cells: cytotoxic T cells destroy infected host cells, and helper T cells activate other white blood cells via chemical signals. One class of helper cell, T H 1, activates macrophages to destroy ingested microorganisms, while another, T H 2, stimulates the production of antibodies by B cells.  
       [0244] Cytotoxic T cells directly attack the infected target cell. In virus-infected cells, peptides derived from viral proteins are generated by the proteasome. These peptides are transported into the ER by the transporter associated with antigen processing (TAP) (Pamer, E. and P. Cresswell (1998) Annu. Rev. Immunol. 16:323-358). Once inside the ER, the peptides bind MHC I chains, and the peptide/MHC I complex is transported to the cell surface. Receptors on the surface of T cells bind to antigen presented on cell surface MHC molecules. Once activated by binding to antigen, T cells secrete γ-interferon, a signal molecule that induces the expression of genes necessary for presenting viral (or other) antigens to cytotoxic T cells. Cytotoxic T cells kill the infected cell by stimulating programmed cell death.  
       [0245] Helper T cells constitute up to 75% of the total T cell population. They regulate the immune functions by producing a variety of lymphokines that act on other cells in the immune system and on bone marrow. Among these lymphokines are: interleukins-2,3,4,5,6; granulocyte-monocyte colony stimulating factor, and γ-interferon.  
       [0246] Helper T cells are required for most B cells to respond to antigen. When an activated helper cell contacts a B cell its centrosome and Golgi apparatus become oriented toward the B cell, aiding the directing of signal molecules, such as transmembrane-bound protein called CD40 ligand, onto the B cell surface to interact with the CD40 transmembrane protein Secreted signals also help B cells to proliferate and mature and, in some cases, to switch the class of antibody being produced.  
       [0247] B-lymphocytes (B cells) produce antibodies which react with specific antigenic proteins presented by pathogens. Once activated, B cells become filled with extensive rough endoplasmic reticulum and are known as plasma cells. As with T cells, interaction of B cells with antigen stimulates proliferation of only those B cells which produce antibody specific to that antigen. There are five classes of antibodies, known as immunoglobulins, which together comprise about 20% of total plasma protein. Each class mediates a characteristic biological response after antigen binding. Upon activation by specific antigen B cells switch from making membrane-bound antibody to secretion of that antibody.  
       [0248] Antibodies, or immunoglobulins (g), are the founding members of the Ig superfamily and the central components of the humoral immune response. Antibodies are either expressed on the surface of B cells or secreted by B cells into the circulation. Antibodies bind and neutralize blood-borne foreign antigens. The prototypical antibody is a tetramer consisting of two identical heavy polypeptide chains (H-chains) and two identical light polypeptide chains (L-chains) interlinked by disulfide bonds. This arrangement confers the characteristic Y-shape to antibody molecules. Antibodies are classified based on their H-chain composition. The five antibody classes, IgA, IgD, IgE, IgG and IgM, are defined by the α, δ, ε, γ, and μ H-chain types. There are two types of L-chains, κ and λ, either of which may associate as a pair with any H-chain pair. IgG, the most common class of antibody found in the circulation, is tetrameric, while the other classes of antibodies are generally variants or multimers of this basic structure.  
       [0249] H-chains and L-chains each contain an N-terminal variable region and a C-terminal constant region. Both H-chains and L-chains contain repeated Ig domains. For example, a typical H-chain contains four Ig domains, three of which occur within the constant region and one of which occurs within the variable region and contributes to the formation of the antigen recognition site. Likewise, a typical L-chain contains two Ig domains, one of which occurs within the constant region and one of which occurs within the variable region. In addition, H chains such as μ have been shown to associate with other polypeptides during differentiation of the B cell.  
       [0250] Antibodies can be described in terms of their two main functional domains. Antigen recognition is mediated by the Fab (antigen binding fragment) region of the antibody, while effector functions are mediated by the Fc (crystallizable fragment) region. Binding of antibody to an antigen, such as a bacterium, triggers the destruction of the antigen by phagocytic white blood cells such as macrophages and neutrophils. These cells express surface receptors that specifically bind to the antibody Fc region and allow the phagocytic cells to engulf, ingest, and degrade the antibody-bound antigen. The Fc receptors expressed by phagocytic cells are single-pass transmembrane glycoproteins of about 300 to 400 amino acids (Sears, D. W. et al. (1990) J. Immunol. 144:371-378). The extracellular portion of the Fc receptor typically contains two or three Ig domains.  
       [0251] Diseases which cause over- or under-abundance of any one type of leukocyte usually result in the entire immune defense system becoming involved. A well-known autoimmune disease is AIDS (Acquired Immunodeficiency Syndrome) where the number of helper T cells is depleted, leaving the patient susceptible to infection by microorganisms and parasites. Another widespread medical condition attributable to the immune system is that of allergic reactions to certain antigens. Allergic reactions include: hay fever, asthma, anaphylaxis, and urticaria (hives). Leukemias are an excess production of white blood cells, to the point where a major portion of the body&#39;s metabolic resources are directed solely at proliferation of white blood cells, leaving other tissues to starve. Leukopenia or agranulocytosis occurs when the bone marrow stops producing white blood cells. This leaves the body unprotected against foreign microorganisms, including those which normally inhabit skin, mucous membranes, and gastrointestinal tract. If all white blood cell production stops completely, infection will occur within two days and death may follow only 1 to 4 days later.  
       [0252] Impaired phagocytosis occurs in several diseases, including monocytic leukemia, systemic lupus, and granulomatous disease. In such a situation, macrophages can phagocytize normally, but the enveloped organism is not killed. A defect in the plasma membrane enzyme which converts oxygen to lethally reactive forms results in abscess formation in liver, lungs, spleen, lymph nodes, and beneath the skin. Eosinophilia is an excess of eosinophils commonly observed in patients with allergies (hay fever, asthma), allergic reactions to drugs, rheumatoid arthritis, and cancers (Hodgkin&#39;s disease, lung, and liver cancer) (Isselbacher, K. J. et al. (1994)  Harrison&#39;s Principles of Internal Medicine , McGraw-Hill, Inc., New York N.Y.).  
       [0253] Host defense is further augmented by the complement system. The complement system serves as an effector system and is involved in infectious agent recognition. It can function as an independent immune network or in conjunction with other humoral immune responses. The complement system is comprised of numerous plasma and membrane proteins that act in a cascade of reaction sequences whereby one component activates the next. The result is a rapid and amplified response to infection through either an inflammatory response or increased phagocytosis.  
       [0254] The complement system has more than 30 protein components which can be divided into functional groupings including modified serine proteases, membrane-binding proteins and regulators of complement activation. Activation occurs through two different pathways the classical and the alternative. Both pathways serve to destroy infectious agents through distinct triggering mechanisms that eventually merge with the involvement of the component C3.  
       [0255] The classical pathway requires antibody binding to infectious agent antigens. The antibodies serve to define the target and initiate the complement system cascade, culminating in the destruction of the infectious agent. In this pathway, since the antibody guides initiation of the process, the complement can be seen as an effector arm of the humoral immune system.  
       [0256] The alternative pathway of the complement system does not require the presence of pre-existing antibodies for targeting infectious agent destruction. Rather, this pathway, through low levels of an activated component, remains constantly primed and provides surveillance in the non-immune host to enable targeting and destruction of infectious agents. In this case foreign material triggers the cascade, thereby facilitating phagocytosis or lysis (Paul, supra, pp.918-919).  
       [0257] Another important component of host defense is the process of inflammation. Inflammatory responses are divided into four categories on the basis of pathology and include allergic inflammation, cytotoxic antibody mediated inflammation, immune complex mediated inflammation and monocyte mediated inflammation. Inflammation manifests as a combination of each of these forms with one predominating.  
       [0258] Allergic acute inflammation is observed in individuals wherein specific antigens stimulate IgE antibody production. Mast cells and basophils are subsequently activated by the attachment of antigen-IgE complexes, resulting in the release of cytoplasmic granule contents such as histamine. The products of activated mast cells can increase vascular permeability and constrict the smooth muscle of breathing passages, resulting in anaphylaxis or asthma. Acute inflammation is also mediated by cytotoxic antibodies and can result in the destruction of tissue through the binding of complement-fixing antibodies to cells. The responsible antibodies are of the IgG or IgM types. Resultant clinical disorders include autoimmune hemolytic anemia and thrombocytopenia as associated with systemic lupus erythematosis.  
       [0259] Immune complex mediated acute inflammation involves the IgG or IgM antibody types which combine with antigen to activate the complement cascade. When such immune complexes bind to neutrophils and macrophages they activate the respiratory burst to form protein- and vessel-damaging agents such as hydrogen peroxide, hydroxyl radical, hypochlorous acid, and chloramines. Clinical manifestations include rheumatoid arthritis and systemic lupus erythematosus.  
       [0260] In chronic inflammation or delayed-type hypersensitivity, macrophages are activated and process antigen for presentation to T cells that subsequently produce lymphokines and monokines. This type of inflammatory response is likely important for defense against intracellular parasites and certain viruses. Clinical associations include, granulomatous disease, tuberculosis, leprosy, and sarcoidosis (Paul, W. E., supra, pp.1017-1018).  
       [0261] Extracellular Information Transmission Molecules  
       [0262] Intercellular communication is essential for the growth and survival of multicellular organisms, and in particular, for the function of the endocrine, nervous, and immune systems. In addition, intercellular communication is critical for developmental processes such as tissue construction and organogenesis, in which cell proliferation, cell differentiation, and morphogenesis must be spatially and temporally regulated in a precise and coordinated manner. Cells communicate with one another through the secretion and uptake of diverse types of signaling molecules such as hormones, growth factors, neuropeptides, and cytokines.  
       [0263] Hormones  
       [0264] Hormones are signaling molecules that coordinately regulate basic physiological processes from embryogenesis throughout adulthood. These processes include metabolism, respiration, reproduction, excretion, fetal tissue differentiation and organogenesis, growth and development, homeostasis, and the stress response. Hormonal secretions and the nervous system are tightly integrated and interdependent. Hormones are secreted by endocrine glands, primarily the hypothalamus and pituitary, the thyroid and parathyroid, the pancreas, the adrenal glands, and the ovaries and testes.  
       [0265] The secretion of hormones into the circulation is tightly controlled Hormones are often secreted in diurnal, pulsatile, and cyclic patterns. Hormone secretion is regulated by perturbations in blood biochemistry, by other upstream-acting hormones, by neural impulses, and by negative feedback loops. Blood hormone concentrations are constantly monitored and adjusted to maintain opal, steady-state levels. Once secreted, hormones act only on those target cells that express specific receptors.  
       [0266] Most disorders of the endocrine system are caused by either hyposecretion or hypersecretion of hormones. Hyposecretion often occurs when a hormone&#39;s gland of origin is damaged or otherwise impaired. Hypersecretion often results from the proliferation of tumors derived from hormone-secreting cells. Inappropriate hormone levels may also be caused by defects in regulatory feedback loops or in the processing of hormone precursors. Endocrine malfunction may also occur when the target cell falls to respond to the hormone.  
       [0267] Hormones can be classified biochemically as polypeptides, steroids, eicosanoids, or amines. Polypeptides, which include diverse hormones such as insulin and growth hormone, vary in size and function and are often synthesized as inactive precursors that are processed intracellularly into mature, active forms. Amines, which include epinephrine and dopamine, are ammo acid derivatives that function in neuroendocrine signaling. Steroids, which include the cholesterol-derived hormones estrogen and testosterone, function in sexual development and reproduction. Eicosanoids, which include prostaglandins and prostacyclins, are fatty acid derivatives that function in a variety of processes. Most polypeptides and some amines are soluble in the circulation where they are highly susceptible to proteolytic degradation within seconds after their secretion. Steroids and lipids are insoluble and must be transported in the circulation by carrier proteins. The following discussion will focus primarily on polypeptide hormones.  
       [0268] Hormones secreted by the hypothalamus and pituitary gland play a critical role in endocrine function by coordinately regulating hormonal secretions from other endocrine glands in response to neural signals. Hypothalamic hormones include thyrotropin-releasing hormone, gonadotropin-releasing hormone, somatostatin, growth-hormone releasing factor, corticotropin-releasing hormone, substance P, dopamine, and prolactin-releasing hormone. These hormones directly regulate the secretion of hormones from the anterior lobe of the pituitary. Hormones secreted by the anterior pituitary include adrenocorticotropic hormone (ACTH), melanocyte-stimulating hormone, somatotropic hormones such as growth hormone and prolactin, glycoprotein hormones such as thyroid-stimulating hormone, luteinizing hormone (LH), and follicle-stimulating hormone (FSH), β-lipotropin, and β-endorphins. These hormones regulate hormonal secretions from the thyroid, pancreas, and adrenal glands, and act directly on the reproductive organs to slate ovulation and spermatogenesis. The posterior pituitary synthesizes and secretes antidiuretic hormone (ADH, vasopressin) and oxytocin.  
       [0269] Disorders of the hypothalamus and pituitary often result from lesions such as primary brain tumors, adenomas, infarction associated with pregnancy, hypophysectomy, aneurysms, vascular malformations, thrombosis, infections, immunological disorders, and complications due to head trauma. Such disorders have profound effects on the function of other endocrine glands. Disorders associated with hypopituitarism include hypogonadism, Sheehan syndrome, diabetes insipidus, Kallian&#39;s disease, Hand-Schuller-Christian disease, Letterer-Siwe disease, sarcoidosis, empty sella syndrome, and dwarfism Disorders associated with hyperpituitarism include acromegaly, giantism, and syndrome of inappropriate ADH secretion (SIADH), often caused by benign adenomas.  
       [0270] Hormones secreted by the thyroid and parathyroid primarily control metabolic rates and the regulation of serum calcium levels, respectively. Thyroid hormones include calcitonin, somatostatin, and thyroid hormone. The parathyroid secretes parathyroid hormone. Disorders associated with hypothyroidism include goiter, myxedema, acute thyroiditis associated with bacterial infection, subacute thyroiditis associated with viral infection, autoimmune thyroiditis (Hashimoto&#39;s disease), and cretinism Disorders associated with hyperthyroidism include thyrotoxicosis and its various forms, Grave&#39;s disease, pretibial myxedema, toxic multinodular goiter, thyroid carcinoma, and Plummer&#39;s disease. Disorders associated with hyperparathyroidism include Conn disease (chronic hypercalemia) leading to bone resorption and parathyroid hyperplasia.  
       [0271] Hormones secreted by the pancreas regulate blood glucose levels by modulating the rates of carbohydrate, fat, and protein metabolism. Pancreatic hormones include insulin, glucagon, amylin, γ-aminobutyric acid, gastrin, somatostatin, and pancreatic polypeptide. The principal disorder associated with pancreatic dysfunction is diabetes mellitus caused by insufficient insulin activity. Diabetes mellitus is generally classified as either Type I (insulin-dependent, juvenile diabetes) or Type II (non-insulin-dependent, adult diabetes). The treatment of both forms by insulin replacement therapy is well known. Diabetes mellitus often leads to acute complications such as hypoglycemia (insulin shock), coma, diabetic ketoacidosis, lactic acidosis, and chronic complications leading to disorders of the eye, kidney, skin, bone, joint, cardiovascular system, nervous system, and to decreased resistance to infection.  
       [0272] The anatomy, physiology, and diseases related to hormonal function are reviewed in McCance, K. L. and S. E. Huether (1994)  Pathophysiology: The Biological Basis for Disease in Adults and Children , Mosby-Year Book, Inc., St Louis Mo.; Greenspan, F. S. and J. D. Baxter (1994)  Basic and Clinical Endocrinology , Appleton and Lange, East Norwalk Conn.  
       [0273] Growth Factors  
       [0274] Growth factors are secreted proteins that mediate intercellular communication. Unlike hormones, which travel great distances via the circulatory system, most growth factors are primarily local mediators that act on neighboring cells. Most growth factors contain a hydrophobic N-terminal signal peptide sequence which directs the growth factor into the secretory pathway. Most growth factors also undergo post-translational modifications within the secretory pathway. These modifications can include proteolysis, glycosylation, phosphorylation, and intramolecular disulfide bond formation. Once secreted, growth factors bind to specific receptors on the surfaces of neighboring target cells, and the bound receptors trigger intracellular signal transduction pathways. These signal transduction pathways elicit specific cellular responses in the target cells. These responses can include the modulation of gene expression and the stimulation or inhibition of cell division, cell differentiation, and cell motility.  
       [0275] Growth factors fan into at least two broad and overlapping classes. The broadest class includes the large polypeptide growth factors, which are wide-ranging in their effects. These factors include epidermal growth factor (EGF), fibroblast growth factor (FGF), transforming growth factors (TGF-β), insulin-like growth factor (IGF), nerve growth factor (NGF), and platelet-derived growth factor (PDGF), each defining a family of numerous related factors. The large polypeptide growth factors, with the exception of NGF, act as mitogens on diverse cell types to stimulate wound healing, bone synthesis and remodeling, extracellular matrix synthesis, and proliferation of epithelial, epidermal, and connective tissues. Members of the TGF-β, EGF, and FGF families also function as inductive signals in the differentiation of embryonic tissue. NGF functions specifically as a neurotrophic factor, promoting neuronal growth and differentiation.  
       [0276] Another class of growth factors includes the hematopoietic growth factors, which are narrow in their target specificity. These factors stimulate the proliferation and differentiation of blood cells such as B-lymphocytes, T-lymphocytes, erythrocytes, platelets, eosinophils, basophils, neutrophils, macrophages, and their stem cell precursors. These factors include the colony-stimulating factors (G-CSF, M-CSF, GM-CSF, and CSF1-3), erythropoietin, and the cytokines. The cytokines are specialized hematopoietic factors secreted by cells of the immune system and are discussed in detail below.  
       [0277] Growth factors play critical roles in neoplastic transformation of cells in vitro and in tumor progression in vivo. Overexpression of the large polypeptide growth factors promotes the proliferation and transformation of cells in culture. Inappropriate expression of these growth factors by tumor cells in vivo may contribute to tumor vascularization and metastasis. Inappropriate activity of hematopoietic growth factors can result in anemias, leukemias, and lymphomas. Moreover, growth factors are both structurally and functionally related to oncoproteins, the potentially cancer-causing products of proto-oncogenes. Certain FGF and PDGF family members are themselves homologous to oncoproteins, whereas receptors for some members of the EGF, NGF, and FGF families are encoded by proto-oncogenes. Growth factors also affect the transcriptional regulation of both proto-oncogenes and oncosuppressor genes (Pimentel, E. (1994)  Handbook of Growth Factors , CRC Press, Ann Arbor Mich.; McKay, I. and I. Leigh, eds. (1993)  Growth Factors: A Practical Approach , Oxford University Press, New York N.Y.; Habenicht, A, ed. (1990)  Growth Factors. Differentiation Factor, and Cytokines , Springer-Verlag, New York N.Y.).  
       [0278] In addition, some of the large polypeptide growth factors play crucial roles in the induction of the primordial germ layers in the developing embryo. This induction ultimately results in the formation of the embryonic mesoderm, ectoderm, and endoderm which in turn provide the framework for the entire adult body plan. Disruption of this inductive process would be catastrophic to embryonic development.  
       [0279] Small Peptide Factors—Neuropeptides and Vasomediators  
       [0280] Neuropeptides and vasomediators (NP/VM comprise a family of small peptide factors, typically of 20 amino acids or less. These factors generally function in neuronal excitation and inhibition of vasoconstriction/vasodilation, muscle contraction, and hormonal secretions from the brain and other endocrine tissues. Included in this family are neuropeptides and neuropeptide hormones such as bombesin, neuropeptide Y, neurotensin, neuromedin N, melanocortins, opioids, galanin, somatostatin, tachykinns, urotensin II and related peptides involved in smooth muscle stimulation, vasopressin, vasoactive intestinal peptide, and circulatory system-borne signaling molecules such as angiotensin, complement, calcitonin, endothelins, formyl-methionyl peptides, glucagon, cholecystokinin, gastrin, and many of the peptide hormones discussed above. NP/VMs can transduce signals directly, modulate the activity or release of other neurotransmitters and hormones, and act as catalytic enzymes in signaling cascades. The effects of NP/VMs range from extremely brief to long-lasting. (Reviewed in Martin, C. R. et al. (1985) Endocrine Physiology, Oxford University Press, New York N.Y., pp. 57-62.)  
       [0281] Cytokines  
       [0282] Cytokines comprise a family of signaling molecules that modulate the immune system and the inflammatory response. Cytokines are usually secreted by leukocytes, or white blood cells, in response to injury or infection. Cytokines function as growth and differentiation factors that act primarily on cells of the immune system such as B- and T-lymphocytes, monocytes, macrophages, and granulocytes. Like other signaling molecules, cytokines bind to specific plasma membrane receptors and trigger intracellular signal transduction pathways which alter gene expression patterns. There is considerable potential for the use of cytokines in the treatment of inflammation and immune system disorders.  
       [0283] Cytokine structure and function have been extensively characterized in vitro. Most cytokines are small polypeptides of about 30 kilodaltons or less. Over 50 cytokines have been identified from human and rodent sources. Examples of cytokine subfamilies include the interferons (IFN-α, -β, and -γ), the interleukins (IL1-IL13), the tumor necrosis factors (TNF-αand -β), and the chemokines. Many cytokines have been produced using recombinant DNA techniques, and the activities of individual cytokines have been determined in vitro. These activities include regulation of leukocyte proliferation, differentiation, and motility.  
       [0284] The activity of an individual cytokine in vitro may not reflect the full scope of that cytokine&#39;s activity in vivo. Cytokines are not expressed individually in vivo but are instead expressed in combination with a multitude of other cytokines when the organism is challenged with a stimulus. Together, these cytokines collectively modulate the immune response in a manner appropriate for that particular stimulus. Therefore, the physiological activity of a cytokine is determined by the stimulus itself and by complex interactive networks among co expressed cytokines which may demonstrate both synergistic and antagonistic relationships.  
       [0285] Chemokines comprise a cytokine subfamily with over 30 members. (Reviewed in Wells, T. N. C. and M. C. Peitsch (1997) J. Leukoc. Biol. 61:545-550.) Chemokines were initially identified as chemotactic proteins that recruit monocytes and macrophages to sites of inflammation Recent evidence indicates that chemokines may also play key roles in hematopoiesis and HIV-1 infection. Chemokines are small proteins which range from about 6-15 kilodaltons in molecular weight. Chemokines are further classified as C, CC, CXC, or CX 3 C based on the number and position of critical cysteine residues. The CC chemokines, for example, each contain a conserved motif consisting of two consecutive cysteines followed by two additional cysteines which occur downstream at 24- and 16-residue intervals, respectively (ExPASy PROSITE database, documents PS00472 and PDOC00434). The presence and spacing of these four cysteine residues are highly conserved, whereas the intervening residues diverge significantly. However, a conserved tyrosine located about 15 residues downstream of the cysteine doublet seems to be important for chemotactic activity. Most of the human genes encoding CC chemokines are clustered on chromosome 17, although there are a few examples of CC chemokine genes that map elsewhere. Other chemokines include lymphotactin (C chemokine); macrophage chemotactic and activating factor (MCAF/MCP-1; CC chemokine); platelet factor 4 and IL-8 (CXC chemokines); and fractalkine and neurotractin (CX 3 C chemokines). (Reviewed in Luster, A. D. (1998) N. Engl. J. Med. 338:436445.)  
       [0286] Receptor Molecules  
       [0287] The term receptor describes proteins that specifically recognize other molecules. The category is broad and includes proteins with a variety of functions. The bulk of receptors are cell surface proteins which bind extracellular ligands and produce cellular responses in the areas of growth, differentiation, endocytosis, and immune response. Other receptors facilitate the selective transport of proteins out of the endoplasmic reticulum and localize enzymes to particular locations in the cell. The term may also be applied to proteins which act as receptors for ligands with known or unknown chemical composition and which interact with other cellular components. For example, the steroid hormone receptors bind to and regulate transcription of DNA  
       [0288] Regulation of cell proliferation, differentiation, and migration is important for the formation and function of tissues. Regulatory proteins such as growth factors coordinately control these cellular processes and act as mediators in cell-cell signaling pathways. Growth factors are secreted proteins that bind to specific cell-surface receptors on target cells. The bound receptors trigger intracellular signal transduction pathways which activate various downstream effectors that regulate gene expression, cell division, cell differentiation, cell motility, and other cellular processes.  
       [0289] Cell surface receptors are typically integral plasma membrane proteins. These receptors recognize hormones such as catecholamines; peptide hormones; growth and differentiation factors; small peptide factors such as thyrotropin-releasing hormone; galanin, somatostatin, and tachykinins; and circulatory system-borne signaling molecules. Cell surface receptors on immune system cells recognize antigens, antibodies, and major histocompatibility complex (MHC)-bound peptides. Other cell surface receptors bind ligands to be internalized by the cell. This receptor-mediated edocytosis functions in the uptake of low density lipoproteins (LDL), transferrin, glucose- or mannose-terminal glycoproteins, galactose-terminal glycoproteins, immunoglobulins, phosphovitellogenins, fibrin, proteinase-inhibitor complexes, plasminogen activators, and thrombospondin (Lodish, H. et al. (1995)  Molecular Cell Biology , Scientific American Books, New York N.Y., p. 723; Mikhailenko, I. et al. (1997) J. Biol. Chem. 272:67846791).  
       [0290] Receptor Protein Kinases  
       [0291] Many growth factor receptors, including receptors for epidermal growth factor, platelet-derived growth factor, fibroblast growth factor, as well as the growth modulator α-thrombin, contain intrinsic protein kinase activities. When growth factor binds to the receptor, it triggers the autophosphorylation of a serine, threonine, or tyrosine residue on the receptor. These phosphorylated sites are recognition sites for the binding of other cytoplasmic signaling proteins. These proteins participate in signaling pathways that eventually link the initial receptor activation at the cell surface to the activation of a specific intracellular target molecule. In the case of tyrosine residue autophosphorylation, these signaling proteins contain a common domain referred to as a Src homology (SH) domain. SH2 domains and SH3 domains are found in phospholipase C-γ, PI-3-K p85 regulatory subunit, Ras-GTPase activating protein, and pp60 c-src  (Lowenstein, E. J. et al. (1992) Cell  70:431-442 ). The cytokine family of receptors share a different common binding domain and include transmembrane receptors for growth hormone (GM), interleukins, erythropoietin, and prolactin.  
       [0292] Other receptors and second messenger-binding proteins have intrinsic serine/threonine protein kinase activity. These include activin/TGF-β/BMP-superfamily receptors, calcium- and diacylglycerol-activated/phospolipid-dependent protein kinase (PK-C), and RNA ant protein kinase (PK-R). In addition, other serine/threonine protein kinases, including nematode Twitchin, have fibronectin-like, immunoglobulin C2-like domains.  
       [0293] G-Protein Coupled Receptors  
       [0294] G-protein coupled receptors (GPCRs) are integral membrane proteins characterized by the presence of seven hydrophobic transmembrane domains which span the plasma membrane and form a bundle of antiparallel alpha (a) helices. These proteins range in size from under 400 to over 1000 amino acids (Strosberg, A. D. (1991) Eur. J. Biochem. 196:1-10; Coughlin, S. R. (1994) Curr. Opin. Cell Biol. 6:191-197). The amino-terminus of the GPCR is extracellular, of variable length and often glycosylated; the carboxy-terminus is cytoplasmic and generally phosphorylated. Extracellular loops of the GPCR alternate with intracellular loops and link the transmembrane domains. The most conserved domains of GPCRs are the transmembrane domains and the first two cytoplasmic loops. The transmembrane domains account for structural and functional features of the receptor. In most cases, the bundle of a helices forms a binding pocket. In addition, the extracellular N-terminal segment or one or more of the three extracellular loops may also participate in ligand binding. Ligand binding activates the receptor by inducing a conformational change in intracellular portions of the receptor. The activated receptor, in turn, interacts with an intracellular heterotrimeric guanine nucleotide binding (G) protein complex which mediates further intracellular signing activities, generally the production of second messengers such as cyclic AMP (cAMP), phospholipase C, inositol triphosphate, or interactions with ion channel proteins (Baldwin, J. M. (1994) Curr. Opin. Cell Biol. 6:180-190).  
       [0295] GPCRs include those for acetylcholine, adenosine, epinephrine and norepinephrine, bombesin, bradykinin, chemokines, dopamine, endothelin, γ-aminobutyric acid (GABA), follicle-stimulating hormone (FSH), glutamate, gonadotropin-releasing hormone (GnRH), hepatocyte growth factor, histamine, leukotrienes, melanocortins, neuropeptide Y, opioid peptides, opsins, prostanoids, serotonin, somatostatin, tachykinins, thrombin, thyrotropin-releasing hormone (TRH), vasoactive intestinal polypeptide family, vasopressin and oxytocin, and orphan receptors.  
       [0296] GPCR mutations, which may cause loss of function or constitutive activation, have been associated with numerous human diseases (Coughlin, supra). For instance, retinitis pigmentosa may arise from mutations in the rhodopsin gene. Rhodopsin is the retinal photoreceptor which is located within the discs of the eye rod cell. Parma, J. et al. (1993, Nature 365:649-651) report that somatic activating mutations in the thyrotropin receptor cause hyperfunctioning thyroid adenomas and suggest that certain GPCRs susceptible to constitutive activation may behave as protooncogenes.  
       [0297] Nuclear Receptors  
       [0298] Nuclear receptors bind small molecules such as hormones or second messengers, leading to increased receptor-binding affinity to specific chromosomal DNA elements. In addition the affinity for other nuclear proteins may also be altered. Such binding and protein-protein interactions may regulate and modulate gene expression. Examples of such receptors include the steroid hormone receptors family, the retinoic acid receptors family, and the thyroid hormone receptors family.  
       [0299] Ligand-Gated Receptor Ion Channels  
       [0300] Ligand-gated receptor ion channels fall into two categories. The first category, exacellular ligand-gated receptor ion channels (ELGs), rapidly transduce neurotransmitter-binding events into electrical signals, such as fast synaptic neurotransmission. ELG function is regulated by post-translational modification. The second category, intracellular ligand-gated receptor ion channels (ILGs), are activated by many intracellular second messengers and do not require post-translational modification(s) to effect a channel-opening response.  
       [0301] ELGs depolarize excitable cells to the threshold of action potential generation. In non-excitable cells, ELGs permit a limited calcium ion-influx during the presence of agonist. ELGs include channels directly gated by neurotransmitters such as acetylcholine, L-glutamate, glycine, ATP, serotonin, GABA, and histamine. ELG genes encode proteins having strong structural and functional similarities. ILGs are encoded by distinct and unrelated gene families and include receptors for cAMP, cGMP, calcium ions, ATP, and metabolites of arachidonic acid  
       [0302] Macrophage Scavenger Receptors  
       [0303] Macrophage scavenger receptors with broad ligand specificity may participate in the binding of low density lipoproteins (LDL) and foreign antigens. Scavenger receptors types I and II are trimeric membrane proteins with each subunit containing a small N-terminal intracellular domain, a transmembrane domain, a large extracellular domain, and a C-terminal cysteine-rich domain. The extracellular domain contains a short spacer domain, an α-helical coiled-coil domain, and a triple helical collagenous domain. These receptors have been shown to bind a spectrum of ligands, including chemically modified lipoproteins and albumin, polyribonucleotides, polysaccharides, phospholipids, and asbestos (Matsumoto, A. et al. (1990) Proc. Natl. Acad. Sci. USA 87:9133-9137; Elomaa, 0. et al. (1995) Cell 80:603-609). The scavenger receptors are thought to play a key role in atherogenesis by mediating uptake of modified LDL in arterial walls, and in host defense by binding bacterial endotoxins, bacteria, and protozoa.  
       [0304] T-Cell Receptors  
       [0305] T cells play a dual role in the immune system as effectors and regulators, coupling antigen recognition with the transmission of signals that induce cell death in infected cells and stimulate proliferation of other immune cells. Although a population of T cells can recognize a wide range of different antigens, an individual T cell can only recognize a single antigen and only when it is presented to the T cell receptor (TCR) as a peptide complexed with a major histocompatibility molecule (MHC) on the surface of an antigen presenting cell. The TCR on most T cells consists of immunoglobulin-like integral membrane glycoproteins containing two polypeptide subunits, a and A, of similar molecular weight. Both TCR subunits have an extracellular domain containing both variable and constant regions, a transmembrane domain that traverses the membrane once, and a short intracellular domain (Saito, H. et al. (1984) Nature 309:757-762). The genes for the TCR subunits are constructed through somatic rearrangement of different gene segments. Interaction of antigen in the proper MHC context with the TCR initiates signaling cascades that induce the proliferation, maturation, and function of cellular components of the immune system (Weiss, A. (1991) Annu. Rev. Gene. 25:487-510). Rearrangements in TCR genes and alterations in TCR expression have been noted in lymphomas, leukemias, autoimmune disorders, and immunodeficiency disorders (Aisenberg, A. C. et al. (1985) N. Engl. J. Med. 313:529-533; Weiss, supra).  
       [0306] Intracellular Signaling Molecules  
       [0307] Intracellular signaling is the general process by which cells respond to extracellular signals (hormones, neurotransmitters, growth and differentiation factors, etc.) through a cascade of biochemical reactions that begins with the binding of a signaling molecule to a cell membrane receptor and ends with the activation of an intracellular target molecule. Intermediate steps in the process involve the activation of various cytoplasmic proteins by phosphorylation via protein kinases, and their deactivation by protein phosphatases, and the eventual translocation of some of these activated proteins to the cell nucleus where the transcription of specific genes is triggered. The intracellular signaling process regulates an types of cell functions including cell proliferation, cell differentiation, and gene transcription, and involves a diversity of molecules including protein kinases and phosphatases, and second messenger molecules, such as cyclic nucleotides, calcium-calmodulin, inositol, and various mitogens, that regulate protein phosphorylation.  
       [0308] Protein Phosphorylation  
       [0309] Protein kinases and phosphatases play a key role in the intracellular signaling process by controlling the phosphorylation and activation of various signaling proteins. The high energy phosphate for this reaction is generally transferred from the adenosine triphosphate molecule (ATP) to a particular protein by a protein kinase and removed from that protein by a protein phosphatase. Protein kinases are roughly divided into two groups: those that phosphorylate tyrosine residues (protein tyrosine kinases, PTK) and those that phosphorylate serine or threonine residues (serine/threonine kinases, STK). A few protein kinases have dual specificity for serine/threonine and tyrosine residues. Almost all kinases contain a conserved 250-300 amino acid catalytic domain containing specific residues and sequence motifs characteristic of the kinase family (Hardie, G. and S. Hanks (1995)  The Protein Kinase Facts Books , Vol 1:7-20, Academic Press, San Diego Calif.).  
       [0310] STKs include the second messenger dependent protein kinases such as the cyclic-AMP dependent protein kinases (PKA), involved in mediating hormone-induced cellular responses; calcium-calmodulin (CaM) dependent protein kinases, involved in regulation of smooth muscle contraction, glycogen breakdown, and neurotransmission; and the mitogen-activated protein kinases (MAP) which mediate signal transduction from the cell surface to the nucleus via phosphorylation cascades. Altered PKA expression is implicated in a variety of disorders and diseases including cancer, thyroid disorders, diabetes, atherosclerosis, and cardiovascular disease (Isselbacher, K. J. et al. (1994)  Harrison&#39;s Principles of Internal Medicine , McGraw-Hill, New York N.Y., pp.  416-431, 1887 ).  
       [0311] PTKs are divided into transmembrane, receptor PTKs and nontransmembrane, non-receptor PTKs. Transmembrane PTKs are receptors for most growth factors. Non-receptor PTKs lack transmembrane regions and, instead, form complexes with the intracellular regions of cell surface receptors. Receptors that function through non-receptor PTKs include those for cytokines and hormones (growth hormone and prolactin) and antigen-specific receptors on T and B lymphocytes. Many of these PTKs were first identified as the products of mutant oncogenes in cancer cells in which their activation was no longer subject to normal cellular controls. In fact, about one third of the known oncogenes encode PTKs, and it is well known that cellular transformation (oncogenesis) is often accompanied by increased tyrosine phosphorylation activity (Charbonneau, H. and N. K. Tonks (1992) Annu. Rev. Cell Biol. 8:463493).  
       [0312] An additional family of protein kinases previously thought to exist only in procaryotes is the histidine protein kinase family (HPK). BPKs bear little homology with mammalian STKs or PTKs but have distinctive sequence motifs of their own (Davie, J. R. et al. (1995) J. Biol. Chem. 270:19861-19867). A histidine residue in the N-terminal half of the molecule (region I) is an autophosphorylation site. Three additional motifs located in the C-terminal half of the molecule include an invariant asparagine residue in region II and two glycine-rich loops characteristic of nucleotide binding domains in regions III and IV. Recently a branched chain alpha-ketoacid dehydrogenase kinase has been found with characteristics of HPK in rat (Davie, supra).  
       [0313] Protein phosphatases regulate the effects of protein kinases by removing phosphate groups from molecules previously activated by kinases. The two principal categories of protein phosphatases are the protein (serine/threonine) phosphatases (PPs) and the protein tyrosine phosphatases (PTPs). PPs dephosphorylate phosphoserine/threonine residues and are important regulators of many cAMP-mediated hormone responses (Cohen, P. (1989) Annu. Rev. Biochem. 58:453-508). PTPs reverse the effects of protein tyrosine kinases and play a significant role in cell cycle and cell signaling processes (Charbonneau, supra). As previously noted, many PTKs are encoded by oncogenes, and oncogenesis is often accompanied by increased tyrosine phosphorylation activity. It is therefore possible that PTPs may prevent or reverse cell transformation and the growth of various cancers by controlling the levels of tyrosine phosphorylation in cells. This hypothesis is supported by studies showing that overexpression of PTPs can suppress transformation in cells, and that specific inhibition of PTPs can enhance cell transformation (Charbonneau, supra).  
       [0314] Phospholipid and Inositol-Phosohate Signaling  
       [0315] Inositol phospholipids (phosphoinositides) are involved in an intracellular signaling pathway that begins with binding of a signaling molecule to a G-protein linked receptor in the plasma membrane. This leads to the phosphorylation of phosphatidylinositol (PI) residues on the inner side of the plasma membrane to the biphosphate state (PIP 2 ) by inositol kinases. Simultaneously, the G-protein linked receptor binding stimulates a trimeric G-protein which in turn activates a phosphoinositide-specific phospholipase C-β. Phospholipase C-β then cleaves PIP 2  into two products, inositol triphosphate (IP 3 ) and diacylglycerol. These two products act as mediators for separate signaling events. IP 3  diffuses through the plasma membrane to induce calcium release from the endoplasmic reticulum (ER), while diacylglycerol remains in the membrane and helps activate protein kinase C, an STK that phosphorylates selected proteins in the target cell. The calcium response initiated by IP 3  is terminated by the dephosphorylation of IP 3  by specific inositol phosphatases. Cellular responses that are mediated by this pathway are glycogen breakdown in the liver in response to vasopressin, smooth muscle contraction in response to acetylcholine, and thrombin-induced platelet aggregation.  
       [0316] Cyclic Nucleotide Signaling  
       [0317] Cyclic nucleotides (cAMP and cGMP) function as intracellular second messengers to transduce a variety of extracellular signals including hormones, light, and neurotransmitters. In particular, cyclic-AMP dependent protein kinases (PKA) are thought to account for all of the effects of cAMP in most mammalian cells, including various hormone-induced cellular responses. Visual excitation and the phototransmission of light signals in the eye is controlled by cyclic-GMP regulated, Ca 2+ -specific channels. Because of the importance of cellular levels of cyclic nucleotides in mediating these various responses, regulating the synthesis and breakdown of cyclic nucleotides is an important matter. Thus adenylyl cyclase, which synthesizes cAMP from AMP, is activated to increase cAMP levels in muscle by binding of adrenaline to β-andrenergic receptors, while activation of guanylate cyclase and increased cGMP levels in photoreceptors leads to reopening of the Ca 2+ -specific channels and recovery of the dark state in the eye. In contrast, hydrolysis of cyclic nucleotides by cAMP and cGMP-specific phosphodiesterases (PDEs) produces the opposite of these and other effects mediated by increased cyclic nucleotide levels. PDEs appear to be particularly important in the regulation of cyclic nucleotides, considering the diversity found in this family of proteins. At least seven families of mammalian PDEs (PDE1-7) have been identified based on substrate specificity and affinity, sensitivity to cofactors, and sensitivity to inhibitory drugs (Beavo, J. A. (1995) Physiological Reviews 75:725-748). PDE inhibitors have been found to be particularly useful in treating various clinical disorders. Rolipram, a specific inhibitor of PDE4, has been used in the treatment of depression, and similar inhibitors are undergoing evaluation as anti-inflammatory agents. Theophylline is a nonspecific PDE inhibitor used in the treatment of bronchial asthma and other respiratory diseases (Banner, K. H. and C. P. Page (1995) Eur. Respir. J. 8:996-1000).  
       [0318] G-Protein Signaling  
       [0319] Guanine nucleotide binding proteins (G-proteins) are critical mediators of signal transduction between a particular class of extracelluar receptors, the G-protein coupled receptors (GPCR), and intracellular second messengers such as cAMP and Ca 2+ . G-proteins are linked to the cytosolic side of a GPCR such that activation of the GPCR by ligand binding stimulates binding of the G-protein to GTP, inducing an “active” state in the G-protein. In the active state, the G-protein acts as a signal to trigger other events in the cell such as the increase of cAMP levels or the release of Ca 2+  into the cytosol from the ER, which, in turn, regulate phosphorylation and activation of other intracellular proteins. Recycling of the G-protein to the inactive state involves hydrolysis of the bound GTP to GDP by a GTPase activity in the G-protein. (See Alberts, B. et al. (1994)  Molecular Biology of the Cell , Garland Publishing, Inc., New York N.Y., pp.734-759.) Two structurally distinct classes of G-proteins are recognized: heterotrimeric G-proteins, consisting of three different subunits, and monomeric, low molecular weight (LMW), G-proteins consisting of a single polypeptide chain.  
       [0320] The three polypeptide subunits of heterotrimeric G-proteins are the α, β, and γ subunits. The α subunit binds and hydrolyzes GTP. The β and γ subunits form a tight complex that anchors the protein to the inner side of the plasma membrane. The β subunits, also known as G-β proteins or β transducins, contain seven tandem repeats of the WD-repeat sequence motif, a motif found in many proteins with regulatory functions. Mutations and variant expression of β transducin proteins are linked with various disorders (Neer, E. J. et al. (1994) Nature 371:297-300; Margottin, F. et al. (1998) Mol. Cell 1:565-574).  
       [0321] LMW GTP-proteins are GTPases which regulate cell growth, cell cycle control, protein secretion, and intracellular vesicle interaction. They consist of single polypeptides which, like the a subunit of the heterotrimeric G-proteins, are able to bind and hydrolyze GTP, thus cycling between an inactive and an active state. At least sixty members of the LMW G-protein superfamily have been identified and are currently grouped into the six subfamilies of ras, rho, arf, sar1, ran, and rab. Activated ras genes were initially found in human cancers, and subsequent studies conformed that ras function is critical in determining whether cells continue to grow or become differentiated. Other members of the LMW G-protein superfamily have roles in signal transduction that vary with the function of the activated genes and the locations of the G-proteins.  
       [0322] Guanine nucleotide exchange factors regulate the activities of LMW G-proteins by determining whether GTP or GDP is bound. GTPase-activating protein (GAP) binds to GTP-ras and induces it to hydrolyze GTP to GDP. In contrast, guanine nucleotide releasing protein (GNRP) binds to GDP-ras and induces the release of GDP and the binding of GTP.  
       [0323] Other regulators of G-protein signaling (RGS) also exist that act primarily by negatively regulating the G-protein pathway by an unknown mechanism (Druey, KM. et al. (1996) Nature 379:742-746). Some 15 members of the RGS family have been identified. RGS family members are related structurally through similarities in an approximately 120 amino acid region termed the RGS domain and functionally by their ability to inhibit the interleukin (cytokine) induction of MAP kinase in cultured mammalian 293T cells (Druey, supra).  
       [0324] Calcium Signaling Molecules  
       [0325] Ca +2  is another second messenger molecule that is even more widely used as an intracellular mediator than cAMP. Two pathways exist by which Ca +2  can enter the cytosol in response to extracellular signals: One pathway acts primarily in nerve signal transduction where Ca +2  enters a nerve terminal through a voltage-gated Ca +2  channel. The second is a more ubiquitous pathway in which Ca +2  is released from the ER into the cytosol in response to binding of an extracellular signaling molecule to a receptor. Ca 2+  directly activates regulatory enzymes, such as protein kinase C, which trigger signal transduction pathways. Ca 2+  also binds to specific Ca 2+ -binding proteins (CBPs) such as calmodulin (CaM) which then activate multiple target proteins in the cell including enzymes, membrane transport pumps, and ion channels. CaM interactions are involved in a multitude of cellular processes including, but not limited to, gene regulation, DNA synthesis, cell cycle progression, mitosis, cytokinesis, cytoskeletal organization, muscle contraction, signal transduction, ion homeostasis, exocytosis, and metabolic regulation (Celio, M. R. et al. (1996)  Guidebook to Calcium-binding Proteins , Oxford University Press, Oxford, UK, pp. 15-20). Some CBPs can serve as a storage depot for Ca 2+  in an inactive state. Calsequestrin is one such CBP that is expressed in isoforms specific to cardiac muscle and skeletal muscle. It is suggested that calsequestrin binds Ca 2+  in a rapidly exchangeable state that is released during Ca 2+ -signaling conditions (Celio, M. R. et al. (1996)  Guidebook to Calcium-binding Proteins , Oxford University Press, New York N.Y., pp. 222-224).  
       [0326] Cyclins  
       [0327] Cell division is the fundamental process by which all living things grow and reproduce. In most organisms, the cell cycle consists of three principle steps; interphase, mitosis, and cytokinesis. Interphase, involves preparations for cell division, replication of the DNA and production of essential proteins. In mitosis, the nuclear material is divided and separates to opposite sides of the cell. Cytokinesis is the final division and fission of the cell cytoplasm to produce the daughter cells.  
       [0328] The entry and exit of a cell from mitosis is regulated by the synthesis and destruction of a family of activating proteins called cyclins. Cyclins act by binding to and activating a group of cyclin-dependent protein kinases (Cdks) which then phosphorylate and activate selected proteins involved in the mitotic process. Several types of cyclins exist (Ciechanover, A (1994) Cell 79:13-21.) Two principle types are mitotic cyclin, or cyclin B, which controls entry of the cell into mitosis, and G1 cyclin, which controls events that drive the cell out of mitosis.  
       [0329] Signal Complex Scaffolding Proteins  
       [0330] Ceretain proteins in intracellular signaling pathways serve to link or cluster other proteins involved in the signaling cascade. A conserved protein domain called the PDZ domain has been identified in various membrane-associated signaling proteins. This domain has been implicated in receptor and ion channel clustering and in the targeting of multiprotein signaling complexes to specialized functional regions of the cytosolic face of the plasma membrane. (For a review of PDZ domain-containing proteins, see Ponting, C. P. et al. (1997) Bioessays  19:469-479 .) A large proportion of PDZ domains are found in the eukaryotic MAGUK (membrane-associated guanylate kinase) protein family, members of which bind to the intracellular domains of receptors and channels. However, PDZ domains are also found in diverse membrane-localized proteins such as protein tyrosine phosphatases, serine/threonine kinases, G-protein cofactors, and synapse-associated proteins such as syntrophins and neuronal nitric oxide synthase (nNOS). Generally, about one to three PDZ domains are found in a given protein, although up to nine PDZ domains have been identified in a single protein.  
       [0331] Membrane Transport Molecules  
       [0332] The plasma membrane acts as a barrier to most 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. A variety of human inherited diseases are caused by a mutation in a transport protein. For example, 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.  
       [0333] Transport proteins are multi-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 carrier 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.  
       [0334] 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 epithelium contains 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 (SGLT1), 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. NIS plays a crucial role in the evaluation, diagnosis, and treatment of various thyroid pathologies because it is the molecular basis for radioiodide thyroid-imaging techniques and for specific targeting of radioisotopes to the thyroid gland (Levy, O. et al. (1997) Proc. Natl. Acad. Sci. USA 94:5568-5573). SMVT is expressed in the intestinal mucosa, kidney, and placenta, and is implicated in the transport of the water-soluble vitamins, e.g., biotin and pantothenate (Prasad, P. D. et al. (1998) J. Biol. Chem. 273:7501-7506).  
       [0335] Transporters play a major role in the regulation of pH, excretion of drugs, and the cellular K + /Na +  balance. Monocarboxylate anion transporters are proton-coupled symporters with a broad substrate specificity that includes L-lactate, pyruvate, and the ketone bodies acetate, acetoacetate, and beta-hydroxybutyrate. At least seven isoforms have been identified to date. The isoforms are predicted to have twelve transmembrane (TM) helical domains with a large intracellular loop between TM6 and TM7, and play a critical role in maintaining intracellular pH by removing the protons that are produced stoichiometrically with lactate during glycolysis. The best characterized H(+)-monocarboxylate transporter is that of the erythrocyte membrane, which transports L-lactate and a wide range of other aliphatic monocarboxylates. Other cells possess H(+)-linked monocarboxylate transporters with differing substrate and inhibitor selectivities. In particular, cardiac muscle and tumor cells have transporters that differ in their K m  values for certain substrates, including stereoselectivity for L-over D-lactate, and in their sensitivity to inhibitors. There are Na(+)-monocarboxylate cotransporters on the luminal surface of intestinal and kidney epithelia, which allow the uptake of lactate, pyruvate, and ketone bodies in these tissues. In addition, there are specific and selective transporters for organic cations and organic anions in organs including the kidney, intestine and liver. Organic anion transporters are selective for hydrophobic, charged molecules with electron-attracting side groups. Organic cation transporters, such as the ammonium transporter, mediate the secretion of a variety of drugs and endogenous metabolites, and contribute to the maintenance of intercellular pH. (Poole, R. C. and A. P. Halestrap (1993) Am J. Physiol. 264:C761-C782; Price, N. T. et al. (1998) Biochnol. J. 329:321-328; and Martinelle, Ky. and I. Haggstrom (1993) J. Biotechnol. 30: 339-350.)  
       [0336] The largest and most diverse family of transport proteins known is 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-molecales” form homo- and heterodimers, such as Tap1 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 (Taglight, D. and S. Michaelis (1998) Meth. Enzymol. 292:131-163).  
       [0337] Transport of fatty acids across the plasma membrane can occur by diffusion, a high capacity, low affinity process. However, under normal physiological conditions a significant fraction of fatty acid transport appears to occur via a high affinity, low capacity protein-mediated transport process. Fatty acid transport protein (FATP), an integral membrane protein with four transmembrane segments, is expressed in tissues exhibiting high levels of plasma membrane fatty acid flux, such as muscle, heart, and adipose. Expression of FATP is upregulated in 3T3-L1 cells during adipose conversion, and expression in COS7 fibroblasts elevates uptake of long-chain fatty acids (Hui, T. Y. et al. (1998) J. Biol. Chem. 273:27420-27429).  
       [0338] Ion Channels  
       [0339] 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. 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&#39;s concentration gradient Gated ion channels allow passive flow of an ion down the ion&#39;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.  
       [0340] Ion transporters generate and maintain the resting electrical potential of a cell. Utilizing the energy derived from ATP hydrolysis, they transport ions against the ion&#39;s concentration gradient. These transmembrane ATPases are divided into three families. The phosphorylated (P) class ion transporters, including Na + -K +  ATPase, Ca 2+ -ATPase, and H + -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 Ca +  are low and cytosolic concentration of K +  is high. The vacuolar (V) class of ion transporters includes 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 i ).  
       [0341] The resting potential of the cell is utilized in many processes involving carrier proteins and gated ion channels. Carrier proteins utilize the resting potential to transport molecules into and out of the cell. Amino acid and glucose transport into many cells is linked to sodium ion co-transport (symport) so that the movement of Na +  down an electrochemical gradient drives transport of the other molecule up a concentration gradient. Similarly, cardiac muscle links transfer of Ca 2+  out of the cell with transport of Nat into the cell (antiport).  
       [0342] 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 (S1, S2, S3, S4, S5, and S6). The center of the barrel forms a pore lined by α-helices or β-strands. The side chains of the amino acid residues comprising the α-helices or β-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.  
       [0343] 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.  
       [0344] Voltage-gated Na +  and K +  channels are necessary for the function of electrically excitable cells, such as nerve and muscle cells. Action potentials, which lead to neurotransmittter release and muscle contraction, arise from large, transient changes in the permeability of the membrane to Na +  and K +  ions. Depolarization of the membrane beyond the threshold level opens voltage-gated Na +  channels. Sodium ions flow into the cell, further depolarizing the membrane and opening more voltage-gated Na +  channels, which propagates the depolarization down the length of the cell. Depolarization also opens voltage-gated potassium channels. Consequently, potassium ions flow outward, which leads to repolarization of the membrane. Voltage-gated channels utilize charged residues in the fourth transmembrane segment (S4) to sense voltage change. The open state lasts only about 1 millisecond, at which tire the channel spontaneously converts into an inactive state that cannot be opened irrespective of the membrane potential. Inactivation is mediated by the channel&#39;s N-terminus, which acts as a plug that closes the pore. The transition from an inactive to a closed state requires a return to resting potential.  
       [0345] Voltage-gated Na +  channels are heterotrimeric complexes composed of a 260 kDa pore forming α subunit that associates with two smaller auxiliary subunits, β1 and β2. The β2 subunit is an integral membrane glycoprotein that contains an extracellular Ig domain, and its association with a and β1 subunits correlates with increased functional expression of the channel, a change in its gating properties, and an increase in whole cell capacitance due to an increase in membrane surface area. (Isom, L. L. et al. (1995) Cell 83:433442.)  
       [0346] Voltage-gated Ca 2+  channels are involved in presynaptic neurotransmitter release, and heart and skeletal muscle contraction. The voltage-gated Ca 2+  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 α 2 δ and β subunits modulate the voltage-dependence, gating properties, and the current amplitude of the channel. These subunits are encoded by at least six α 1 , one α 2 δ, and four β genes. A fourth subunit, γ, 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.)  
       [0347] Chloride channels are necessary in endocrine secretion and in regulation of cytosolic and organelle pH. In secretory epithelial cells, Cl −  enters the cell across a basolateral membrane through an Na + , K + /Cl −  cotransporter, accumulating in the cell above its electrochemical equilibrium concentration. Secretion of Cl −  from the apical surface, in response to hormonal stimulation, leads to flow of Na +  and water into the secretory lumen. The cystic fibrosis transmembrane conductance regulator (CFTR) is a chloride channel encoded by the gene for cystic fibrosis, a common fatal genetic disorder in humans. Loss of CFTR function decreases transepithelial water secretion and, as a result, the layers of mucus that coat the respiratory tree, pancreatic ducts, and intestine are dehydrated and difficult to clear. The resulting blockage of these sites leads to pancreatic insufficiency, “meconium ileus”, and devastating “chronic obstructive pulmonary disease” (AI-Awqati, Q. et al. (1992) J. Exp. Biol. 172:245-266).  
       [0348] Many intracellular organelles contain H + -ATPase pumps that generate transmembrane pH and electrochemical differences by moving protons from the cytosol to the organelle lump. If the membrane of the organelle is permeable to other ions, then the electrochemical gradient can be abrogated without affecting the pH differential. In fact, removal of the electrochemical barrier allows more H +  to be pumped across the membrane, increasing the pH differential. Cl −  is the sole counterion of H +  translocation in a number of organelles, including chromaffin granules, Golgi vesicles, lysosomes, and endosomes. Functions that require a low vacuolar pH include uptake of small molecules such as biogenic amines in chromaffin granules, processing of vacuolar constituents such as pro-hormones by proteolytic enzymes, and protein degradation in lysosomes (Al-Awqati, supra).  
       [0349] 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 γ-aminobutyric acid (GABA) and glycine, leading to hyperpolarization of the membrane and the subsequent generation of an action potential.  
       [0350] Ligand-gated channels can be regulated by intracellular second messengers. Calcium-activated K +  channels are gated by internal calcium ions. In nerve cells, an influx of calcium during depolarization opens K +  channels to modulate the magnitude of the action potential ([shi, T. M. et al. (1997) Proc. Natl. Acad. Sci. USA 94:11651-11656). Cyclic nucleotide-gated (CNG) channels are gated by cytosolic cyclic nucleotides. The best examples of these are the cAMP-gated Na +  channels involved in olfaction and the cGMP-gated cation channels involved in vision. Both systems involve ligand-mediated activation of a G-protein coupled receptor which then alters the level of cyclic nucleotide within the cell.  
       [0351] 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, supra). 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).  
       [0352] Disease Correlation  
       [0353] 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&#39;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).  
       [0354] Protein Modification and Maintenance Molecules  
       [0355] The cellular processes regulating modification and maintenance of protein molecules coordinate their conformation, stabilization, and degradation. Each of these processes is mediated by key enzymes or proteins such as proteases, protease inhibitors, transferases, isomerases, and molecular chaperones.  
       [0356] Proteases  
       [0357] Proteases cleave proteins and peptides at the peptide bond that forms the backbone of the peptide and protein chain Proteolytic processing is essential to cell growth, differentiation, remodeling, and homeostasis as well as inflammation and immune response. Typical protein half-lives range from hours to a few days, so that within all living cells, precursor proteins are being cleaved to their active form, signal sequences proteolytically removed from targeted proteins, and aged or defective proteins degraded by proteolysis. Proteases function in bacterial, parasitic, and viral invasion and replication within a host. Four principal categories of mammalian proteases have been identified based on active site structure, mechanism of action, and overall three-dimensional structure. (Beynon, R. J. and J. S. Bond (1994)  Proteolytic Enzymes: A Practical Approach , Oxford University Press, New York N.Y., pp. 1-5).  
       [0358] The serine proteases (SPs) have a serine residue, usually within a conserved sequence, in an active site composed of the serine, an aspartate, and a histidine residue. SPs include the digestive enzymes trypsin and chymotrypsin, components of the complement cascade and the blood-clotting cascade, and enzymes that control extracellular protein degradation. The main SP sub-families are trypases, which cleave after arginine or lysine; aspartases, which cleave after aspartate; chymases, which cleave after phenylalanine or leucine; metases, which cleavage after methionine; and serases which cleave after serine. Enterokinase, the initiator of intestinal digestion, is a serine protease found in the intestinal brush border, where it cleaves the acidic propeptide from trypsinogen to yield active trypsin (Kitamoto, Y. et al. (1994) Proc. Natl. Acad. Sci. USA 91:7588-7592). Prolylcarboxypeptidase, a lysosomal serine peptidase that cleaves peptides such as angiotensin II and III and [des-Arg9] bradykinin, shares sequence homology with members of both the serine carboxypeptidase and prolylendopeptidase families (Tan, F. et al. (1993) J. Biol. Chem. 268:16631-16638).  
       [0359] Cysteine proteases (CPs) have a cysteine as the major catalytic residue at an active site where catalysis proceeds via an intermediate thiol ester and is facilitated by adjacent histidine and aspartic acid residues. CPs are involved in diverse cellular processes ranging from the processing of precursor proteins to intracellular degradation. Mammalian CPs include lysosomal cathepsins and cytosolic calcium activated proteases, calpains. CPs are produced by monocytes, macrophages and other cells of the immune system which migrate to sites of inflammation and secrete molecules involved in tissue repair. Overabundance of these repair molecules plays a role in certain disorders. In autoimmune diseases such as rheumatoid arthritis, secretion of the cysteine peptidase cathepsin C degrades collagen, laminin, elastin and other structural proteins found in the extracellular matrix of bones.  
       [0360] Aspartic proteases are members of the cathepsin family of lysosomal proteases and include pepsin A, gastricsin, chymosin, renin, and cathepsins D and E. Aspartic proteases have a pair of aspartic acid residues in the active site, and are most active in the pH 2-3 range, in which one of the aspartate residues is ionized, the other un-ionized. Aspartic proteases include bacterial penicillopepsin, mammalian pepsin, renin, chymosin, and certain fungal proteases. Abnormal regulation and expression of cathepsins is evident in various inflammatory disease states. In cells isolated from inflamed synovia, the mRNA for stromelysin, cytokines, TEMP-1, cathepsin, gelatinase, and other molecules is preferentially expressed. Expression of cathepsins L and D is elevated in synovial tissues from patients with rheumatoid arthritis and osteoarthritis. Cathepsin L expression may also contribute to the influx of mononuclear cells which exacerbates the, destruction of the rheumatoid synovium (Keyszer, G. M. (1995) Arthritis Rheum. 38:976-984.) The increased expression and differential regulation of the, cathepsins are linked to the metastatic potential of a variety of cancers and as such are of therapeutic and prognostic interest (Chambers, A. F. et al. (1993) Crit. Rev. Oncog. 4:95-114).  
       [0361] Metalloproteaes have active sites that include two glutamic acid residues and one histidine residue that serve as binding sites for Zinc. Carboxypeptidases A and B are the principal mammalian metalloproteases. Both are exoproteases of similar structure and active sites. Carboxypeptidase A, like chymotrypsin prefers C-terminal aromatic and aliphatic side chains of hydrophobic nature, whereas carboxypeptidase B is directed toward basic arginine and lysine residues. Glycoprotease (GCP), or O-sialoglycoprotein endopeptidase, is a metallopeptidase which specifically cleaves O-sialoglycoproteins such as glycophorin A. Another metallopeptidase, placental leucine aminopeptidase (P-LAP) degrades several peptide hormones such as oxytocin and vasopressin, suggesting a role in maintaining homeostasis during pregnancy, and is expressed in several tissues (Rogi, T. et al. (1996) J. Biol. Chem. 271:56-61).  
       [0362] Ubiquitin proteases are associated with the ubiquitin conjugation system (UCS), a major pathway for the degradation of cellular proteins in eukaryotic cells and some bacteria. The UCS mediates the elimination of abnormal proteins and regulates the half-lives of important regulatory proteins that control cellular processes such as gene transcription and cell cycle progression. In the UCS pathway, proteins targeted for degradation are conjugated to a ubiquitin, a small heat stable protein. The ubiquitinated protein is then recognized and degraded by proteasome, a large, multisubunit proteolytic enzyme complex, and ubiquitin is released for reutilization by ubiquitin protease. The UCS is implicated in the degradation of mitotic cyclic kinases, oncoproteins, tumor suppressor genes such as p53, viral proteins, cell surface receptors associate with signal transduction, transcriptional regulators, and mutated or damaged proteins (Ciechanover, k (1994) Cell 79:13-21). A murine proto-oncogene, Unp, encodes a nuclear ubiquitin protease whose overexpression leads to oncogenic transformation of NIH3T3 cells, and the human homolog of this gene is consistently elevated in small cell tumors and adenocarcinomas of the lung (Gray, D. A. (1995) Oncogene 10:2179-2183).  
       [0363] Signal Peptidases  
       [0364] The mechanism for the translocation process into the endoplasmic reticulum (ER) involves the recognition of an N-terminal signal peptide on the elongating protein. The signal peptide direct the protein and attached ribosome to a receptor on the ER membrane. The polypeptide chain passes through a pore in the ER membrane into the lumen while the N-terminal signal peptide remains attached at the membrane surface. The process is completed when signal peptidase located inside the ER cleaves the signal peptide from the protein and releases the protein into the lumen.  
       [0365] Protease Inhibitors  
       [0366] Protease inhibitors and other regulators of protease activity control the activity and effects of proteases. Protease inhibitors have been shown to control pathogenesis in animal models of proteolytic disorders (Murphy, G. (1991) Agents Actions Suppl. 35:69-76). Low levels of the cystatins, low molecular weight inhibitors of the cysteine proteases, correlate with malignant progression of tumors. (Calkins, C. et al (1995) Biol. Biochem. Hoppe Seyler 376:71-80). Serpins are inhibitors of mammalian plasma serine proteases. Many serpins serve to regulate the blood clotting cascade and/or the complement cascade in mammals. Sp32 is a positive regulator of the mammalian acrosomal protease, acrosin, that binds the proenzyme, proacrosin, and thereby aides in packaging the enzyme into the acrosomnal matrix (Baba, T. et al. (1994) 1. Biol. Chem. 269:10133-10140). The Kunitz family of serine protease inhibitors are characterized by one or more “Kunitz domains” containing a series of cysteine residues that are regularly spaced over approximately 50 amino acid residues and form three intrachain disulfide bonds. Members of this family include aprotinin, tissue factor pathway inhibitor (TFPI-1 and TFPI-2), inter-α-trypsin inhibitor, and bikunin. (Marlor, C. W. et al. (1997) J. Biol. Chem. 272:12202-12208.) Members of this family are potent inhibitors (in the nanomolar range) against serine proteases such as kallikrein and plasmin. Aprotinin has clinical utility in reduction of perioperative blood loss.  
       [0367] A major portion of all proteins synthesized in eukaryotic cells are synthesized on the cytosolic surface of the endoplasmic reticulum (ER). Before these immature proteins are distributed to other organelles in the cell or are secreted, they must be transported into the interior lumen of the ER where post-translational modifications are performed. These modifications include protein folding and the formation of disulfide bonds, and N-linked glycosylations.  
       [0368] Protein Isomerases  
       [0369] Protein folding in the ER is aided by two principal types of protein isomerases, protein disulfide isomerase (PDI), and peptidyl-prolyl isomerase (PPI). PDI catalyzes the oxidation of free sulfhydryl groups in cysteine residues to form intramolecular disulfide bonds in proteins. PPI, an enzyme that catalyzes the isomerization of certain proline imidic bonds in oligopeptides and proteins, is considered to govern one of the rate limiting steps in the folding of many proteins to their final functional conformation. The cyclophilins represent a major class of PPI that was originally identified as the major receptor for the immunosuppressive drug cyclosporin A (Handschumacher, R. E. et al. (1984) Science  226: 544-547 ).  
       [0370] Protein Glycosylation  
       [0371] The glycosylation of most soluble secreted and membrane-bound proteins by oligosaccharides linked to asparagine residues in proteins is also performed in the ER. This reaction is catalyzed by a membrane-bound enzyme, oligosaccharyl transferase. Although the exact purpose of this “N-linked” glycosylation is unknown, the presence of oligosaccharides tends to make a glycoprotein resistant to protease digestion. In addition, oligosaccharides attached to cell-surface proteins called selectins are known to function in cell-cell adhesion processes (Alberts, B. et al. (1994)  Molecular Biology of the Cell , Garland Publishing Co., New York N.Y., p.608). “O-linked” glycosylation of proteins also occurs in the ER by the addition of N-acetylgalactosamine to the hydroxyl group of a serine or threonine residue followed by the sequential addition of other sugar residues to the first. This process is catalysed by a series of glycosyltransferases each specific for a particular donor sugar nucleotide and acceptor molecule (Lodish, H. et al. (1995)  Molecular Cell Biology , W. H. Freeman and Co., New York N.Y., pp.700-708). In many cases, both N- and O-linked oligosaccharides appear to be required for the secretion of proteins or the movement of plasma membrane glycoproteins to the cell surface.  
       [0372] An additional glycosylation mechanism operates in the ER specifically to target lysosomal enzymes to lysosomes and prevent their secretion. Lysosomal enzymes in the ER receive an N-linked oligosaccharide, like plasma membrane and secreted proteins, but are then phosphorylated on one or two mannose residues. The phosphorylation of mannose residues occurs in two steps, the first step being the addition of an N-acetylglucosamine phosphate residue by N-acetylglucosamine phosphotransferase, and the second the removal of the N-acetylglucosamine group by phosphodiesterase. The phosphorylated mannose residue then targets the lysosomal enzyme to a mannose 6-phosphate receptor which transports it to a lysosome vesicle (Lodish, supra, pp.708-711).  
       [0373] Chaperones  
       [0374] Molecular chaperones are proteins that aid in the proper folding of immature proteins and refolding of improperly folded ones, the assembly of protein subunits, and in the transport of unfolded proteins across membranes. Chaperones are also called heat-shock proteins (hsp) because of their tendency to be expressed in dramatically increased amounts following brief exposure of cells to elevated temperatures. This latter property most likely reflects their need in the refolding of proteins that have become denatured by the high temperatures. Chaperones may be divided into several classes according to their location, function, and molecular weight, and include hsp60, TCP1, hsp70, hsp40 (also called DnaJ), and hsp90. For example, hsp90 binds to steroid hormone receptors, represses transcription in the absence of the ligand, and provides proper folding of the ligand-binding domain of the receptor in the presence of the hormone (Burston, S. G. and A. R. Clarke (1995) Essays Biochem. 29:125-136). Hsp60 and hsp70 chaperones aid in the transport and folding of newly synthesized proteins. Hsp70 acts early in protein folding, binding a newly synthesized protein before it leaves the ribosome and transporting the protein to the mitochondria or ER before releasing the folded protein. Hsp60, along with hsp10, binds misfolded proteins and gives them the opportunity to refold correctly. All chaperones share an affinity for hydrophobic patches on incompletely folded proteins and the ability to hydrolyze ATP. The energy of ATP hydrolysis is used to release the hsp-bound protein in its properly folded state (Alberts, supra, pp 214, 571-572).  
       [0375] Nucleic Acid Synthesis and Modification Molecules  
       [0376] Polymerases  
       [0377] DNA and RNA replication are critical processes for cell replication and function. DNA and RNA replication are mediated by the enzymes DNA and RNA polymerase, respectively, by a “templating” process in which the nucleotide sequence of a DNA or RNA strand is copied by complementary base-pairing into a complementary nucleic acid sequence of either DNA or RNA. However, there are fundamental differences between the two processes.  
       [0378] DNA polymerase catalyzes the stepwise addition of a deoxyribonucleotide to the 3′-OH end of a polynucleotide strand (the primer strand) that is paired to a second (template) strand. The new DNA strand therefore grows in the 5′ to 3′ direction (Alberts, B. et al. (1994) The Molecular Biology of the Cell , Garland Publishing Inc., New York N.Y., pp. 251-254). The substrates for the polymerization reaction are the corresponding deoxynucleotide triphosphates which must base-pair with the correct nucleotide on the template strand in order to be recognized by the polymerase. Because DNA exists as a double-stranded helix, each of the two strands may serve as a template for the formation of a new complementary strand. Each of the two daughter cells of the dividing cell therefore inherits a new DNA double helix containing one old and one new strand. Thus, DNA is said to be replicated “semiconservatively” by DNA polymerase. In addition to the synthesis of new DNA, DNA polymerase is also involved in the repair of damaged DNA as discussed below under “Ligases.” 
       [0379] In contrast to DNA polymerase, RNA polymerase uses a DNA template strand to “transcribe” DNA into RNA using ribonucleotide triphosphates as substrates. Like DNA polymerization, RNA polymerization proceeds in a 5′ to 3′ direction by addition of a ribonucleoside monophosphate to the 3′-OH end of a growing RNA chain DNA transcription generates messenger RNAs (mRNA) that carry information for protein synthesis, as well as the transfer, ribosomal, and other RNAs that have structural or catalytic functions. In eukaryotes, three discrete RNA polymerases synthesize the three different types of RNA (Alberts, supra, pp. 367-368). RNA polymerase I makes the large ribosomal RNAs, RNA polymerase II makes the mRNAs that will be translated into proteins, and RNA polymerase III makes a variety of small, stable RNAs, including 5S ribosomal RNA and the transfer RNAs (tRNA). In all cases, RNA synthesis is initiated by binding of the RNA polymerase to a promoter region on the DNA and synthesis begins at a start site within the promoter. Synthesis is completed at a broad, general stop or termination region in the DNA where both the polymerase and the completed RNA chain are released.  
       [0380] Ligases  
       [0381] DNA repair is the process by which accidental base changes, such as those produced by oxidative damage, hydrolytic attack, or uncontrolled methylation of DNA are corrected before replication or transcription of the DNA can occur. Because of the efficiency of the DNA repair process, fewer than one in one thousand accidental base changes causes a mutation (Alberts, sutra, pp. 245-249). The three steps common to most types of DNA repair are (1) excision of the damaged or altered base or nucleotide by DNA nucleases, leaving a gap; (2) insertion of the correct nucleotide in this gap by DNA polymerase using the complementary strand as the template; and (3) sealing the break left between the inserted nucleotide(s) and the existing DNA strand by DNA ligase. In the last reaction, DNA ligase uses the energy from ATP hydrolysis to activate the 5′ end of the broken phosphodiester bond before forming the new bond with the 3′-OH of the DNA strand. In Bloom&#39;s syndrome, an inherited human disease, individuals are partially deficient in DNA ligation and consequently have an increased incidence of cancer (Alberts, supra, p. 247).  
       [0382] Nucleases  
       [0383] Nucleases comprise both enzymes that hydrolyze DNA (DNase) and RNA (RNase). They serve different purposes in nucleic acid metabolism. Nucleases hydrolyze the phosphodiester bonds between adjacent nucleotides either at internal positions (endonucleases) or at the terminal 3′ or, 5′ nucleotide positions (exonucleases). A DNA exonuclease activity in DNA polymerase, for example, serves to remove improperly paired nucleotides attached to the 3′-OH end of the growing DNA strand by the polymerase and thereby serves a “proofreading” function. As mentioned above, DNA endonuclease activity is involved in the excision step of the DNA repair process.  
       [0384] RNases also serve a variety of functions. For example, RNase P is a ribonucleoprotein enzyme which cleaves the 5′ end of pre-tRNAs as part of their maturation process. RNase H digests the RNA strand of an RNA/DNA hybrid Such hybrids occur in cells invaded by retroviruses, and RNase H is an important enzyme in the retroviral replication cycle. Pancreatic RNase secreted by the pancreas into the intestine hydrolyzes RNA present in ingested foods. RNase activity in serum and cell extracts is elevated in a variety of cancers and infectious diseases (Schein, C. H. (1997) Nat Biotechnol. 15:529-536). Regulation of RNase activity is being investigated as a means to control tumor angiogenesis, allergic reactions, viral infection and replication, and fungal infections.  
       [0385] Methylases  
       [0386] Methylation of specific nucleotides occurs in both DNA and RNA, and serves different functions in the two macromolecules. Methylation of cytosine residues to form 5-methyl cytosine in DNA occurs specifically at CG sequences which are base-paired with one another in the DNA double-helix. This pattern of methylation is passed from generation to generation during DNA replication by an enzyme called “maintenance methylase” that acts preferentially on those CG sequences that are base-paired with a CG sequence that is already methylated. Such methylation appears to distinguish active from inactive genes by preventing the binding of regulatory proteins that “turn on”, the gene, but permit the binding of proteins that inactivate the gene (Alberts, supra, pp.  448-451 ). In RNA metabolism, “tRNA methylase” produces one of several nucleotide modifications in tRNA that affect the conformation and base-pairing of the molecule and facilitate the recognition of the appropriate mRNA codons by specific tRNAs. The primary methylation pattern is the dimethylation of guanine residues to form N,N-dimethyl guanine.  
       [0387] Helicases and Single-Stranded Binding Proteins  
       [0388] Helicases are enzymes that destabilize and unwind double helix structures in both DNA and RNA. Since DNA replication occurs more or less simultaneously on both strands, the two strands must first separate to generate a replication “fork” for DNA polymerase to act on. Two types of replication proteins contribute to this process, DNA helicases and single-stranded binding proteins. DNA helicases hydrolyze ATP and use the energy of hydrolysis to separate the DNA strands. Single-stranded binding proteins (SSBs) then bind to the exposed DNA strands without covering the bases, thereby temporarily stabilizing them for templating by the DNA polymerase (Alberts, supra, pp. 255-256).  
       [0389] RNA helicases also alter and regulate RNA conformation and secondary structure. Like the DNA helicases, RNA helicases utilize energy derived from ATP hydrolysis to destabilize and unwind RNA duplexes. The most well characterized and ubiquitous family of RNA helicases is the DEAD-box family, so named for the conserved B-type ATP-binding motif which is diagnostic of proteins in this family. Over 40 DEAD-box helicases have been identified in organisms as diverse as bacteria, insects, yeast, amphibians, mammals, and plants. DEAD-box helicases function in diverse processes such as translation initiation, splicing, ribosome assembly, and RNA editing, transport, and stability. Some DEAD-box helicases play tissue- and stage-specific roles in spermatogenesis and embryogenesis. Overexpression of the DEAD-box 1 protein (DDX1) may play a role in the progression of neuroblastoma (Nb) and retinoblastoma (Rb) tumors (Godbout, R. et al. (1998) J. Biol. Chem. 273:21161-21168). These observations suggest that DDX1 may promote or enhance tumor progression by altering the normal secondary structure and expression levels of RNA in cancer cells. Other DEAD-box helicases have been implicated either directly or indirectly in tumorigenesis (Discussed in Godbout, supra). For example, murine p68 is mutated in ultraviolet light-induced tumors, and human DDX6 is located at a chromosomal breakpoint associated with B-cell lymphoma Similarly, a chimeric protein comprised of DDX10 and NUP98, a nucleoporin protein, may be involved in the pathogenesis of certain myeloid malignancies.  
       [0390] Topoisomerases  
       [0391] Besides the need to separate DNA strands prior to replication, the two strands must be “unwound” from one another prior to their separation by DNA helicases. This function is performed by proteins known as DNA topoisomerases. DNA topoisomerase effectively acts as a reversible nuclease that hydrolyzes a phosphodiesterase bond in a DNA strand, permitting the two strands to rotate freely about one another to remove the strain of the helix, and then rejoins the original phosphodiester bond between the two strands. Two types of DNA topoisomerase exist, types I and II. DNA Topoisomerase I causes a single-strand break in a DNA helix to allow the rotation of the two strands of the helix about the remaining phosphodiester bond in the opposite strand DNA topoisomerase II causes a transient break in both strands of a DNA helix where two double helices cross over one another. This type of topoisomerase can efficiently separate two interlocked DNA circles (Alberts, supra, pp.260-262). Type II topoisomerases are largely confined to proliferating cells in eukaryotes, such as cancer cells. For this reason they are targets for anticancer drugs. Topoisomerase II has been implicated in multi-drug resistance (MDR) as it appears to aid in the repair of DNA damage inflicted by DNA binding agents such as doxorubicin and vincristine.  
       [0392] Recombinases  
       [0393] Genetic recombination is the process of rearranging DNA sequences within an organism&#39;s genome to provide genetic variation for the organism in response to changes in the environment DNA recombination allows variation in the particular combination of genes present in an individual&#39;s genome, as well as the timing and level of expression of these genes (see Alberts, supra, pp. 263-273). Two broad classes of genetic recombination are commonly recognized, general recombination and site-specific recombination. General recombination involves genetic exchange between any homologous pair of DNA sequences usually located on two copies of the same chromosome. The process is aided by enzymes called recombinases that “nick” one strand of a DNA duplex more or less randomly and permit exchange with the complementary strand of another duplex. The process does not normally change the arrangement of genes on a chromosome. In site-specific recombination, the recombinase recognizes specific nucleotide sequences present in one or both of the recombining molecules. Base-pairing is not involved in this form of recombination and therefore does not require DNA homology between the recombining molecules. Unlike general recombination, this form of recombination can alter the relative positions of nucleotide sequences in chromosomes.  
       [0394] Splicing Factors  
       [0395] Various proteins are necessary for processing of transcribed RNAs in the nucleus. Pre-mRNA processing steps include capping at the 5′ end with methylguanosine, polyadenylating the 3′ end, and splicing to remove introns. The primary RNA transcript from DNA is a faithful copy of the gene containing both exon and intron sequences, and the latter sequences must be cut out of the RNA transcript to produce an mRNA that codes for a protein. This “splicing” of the mRNA sequence takes place in the nucleus with the aid of a large, multicomponent ribonucleoprotein complex known as a spliceosome. The spliceosomal complex is composed of five small nuclear ribonucleoprotein particles (snRNPs) designated U1, U2, U4, U5, and U6, and a number of additional proteins. Each snRNP contains a single species of snRNA and about ten proteins. The RNA components of some snRNPs recognize and base pair with intron consensus sequences. The protein components mediate spliceosome assembly and the splicing reaction. Autoantibodies to snRNP proteins are found in the blood of patients with systemic lupus erythematosus (Stryer, L. (1995)  Biochemistry , W. H. Freeman and Company, New York N.Y., p. 863).  
       [0396] Adhesion Molecules  
       [0397] The surface of a cell is rich in transmembrane proteoglycans, glycoproteins, glycolipids, and receptors. These macromolecules mediate adhesion with other cells and with components of the extracellular matrix (ECM). The interaction of the cell with its surroundings profoundly influences cell shape, strength, flexibility, motility, and adhesion. These dynamic properties are intimately associated with signal transduction pathways controlling cell proliferation and differentiation, tissue construction, and embryonic development.  
       [0398] Cadherins  
       [0399] Cadherins comprise a family of calcium-dependent glycoproteins that function in mediating cell-cell adhesion in virtually all solid tissues of multicellular organisms. These proteins share multiple repeats of a cadherin-specific motif, and the repeats form the folding units of the cadherin extracellular domain. Cadherin molecules cooperate to form focal contacts, or adhesion plaques, between adjacent epithelial cells. The cadherin family includes the classical cadherins and protocadherins. Classical cadherins include the E-cadherin, N-cadherin, and P-cadherin subfamilies. E-cadherin is present on many types of epithelial cells and is especially important for embryonic development. N-cadherin is present on nerve, muscle, and lens cells and is also critical for embryonic development P-cadherin is present on cells of the placenta and epidermis. Recent studies report that protocadherins are involved in a variety of cell-cell interactions (Suzuki, S. T. (1996) J. Cell Sci. 109:2609-2611). The intracellular anchorage of cadherins is regulated by their dynamic association with catenins, a family of cytoplasmic signal transduction proteins associated with the actin cytoskeleton. The anchorage of cadherins to the actin cytoskeleton appears to be regulated by protein tyrosine phosphorylation, and the cadherins are the target of phosphorylation-induced junctional disassembly (Aberle, H. et al. (1996) J. Cell. Biochem. 61:514-523).  
       [0400] Integrins  
       [0401] Integrins are ubiquitous transmembrane adhesion molecules that link the ECM to the internal cytoskeleton. Integrins are composed of two noncovalently associated transmembrane glycoprotein subunits called α and β. Integrins function as receptors that play a role in signal transduction. For example, binding of integrin to its extracellular ligand may stimulate changes in intracellular calcium levels or protein kinase activity (Sjaastad, M. D. and W. J. Nelson (1997) BioEssays 19:47-55). At least ten cell surface receptors of the integrin family recognize the ECM component fibronectin, which is involved in many different biological processes including cell migration and embryogenesis (Johansson, S. et al. (1997) Front. Biosci. 2:D126-D146).  
       [0402] Lectins  
       [0403] Lectins comprise a ubiquitous family of extracellular glycoproteins which bind cell surface carbohydrates specifically and reversibly, resulting in the agglutination of cells (reviewed in Drickamer, K. and M. E. Taylor (1993) Annu. Rev. Cell Biol. 9:237-264). This function is particularly important for activation of the immune response. Lectins mediate the agglutination and mitogenic stimulation of lymphocytes at sites of inflammation (Lasky, L. A. (1991) J. Cell. Biochem. 45:139-146; Paietta, E. et al. (1989) J. Immunol. 143:2850-2857).  
       [0404] Lectins are further classified into subfamilies based on carbohydrate-binding specificity and other criteria. The galectin subfamily, in particular, includes lectins that bind β-galactoside carbohydrate moieties in a thio]-dependent manner (reviewed in Hadari, Y. R. et al. (1998) J. Biol. Chem. 270:3447-3453). Galectins are widely expressed and developmentally regulated. Because all galectins lack an N-terminal signal peptide, it is suggested that galectins are externalized through an a typical secretory mechanism. Two classes of galectins have been defined based on molecular weight and oligomerization properties. Small galectins form homodimers and are about 14 to 16 kilodaltons in mass, while large galectins are monomeric and about 29-37 kilodaltons.  
       [0405] Galectins contain a characteristic carbohydrate recognition domain (CRD). The CRD is about 140 amino acids and contains several stretches of about 1-10 amino acids which are highly conserved among all galectins. A particular 6-amino acid motif within the CRD contains conserved tryptophan and arginine residues which are critical for carbohydrate binding. The CRD of some galectins also contains cysteine residues which may be important for disulfide bond formation. Secondary structure predictions indicate that the CRD forms several β-sheets.  
       [0406] Galectins play a number of roles in diseases and conditions associated with cell-cell and cell-matrix interactions. For example, certain galectins associate with sites of inflammation and bind to cell surface immunoglobulin E molecules. In addition, galectins may play an important role in cancer metastasis. Galectin overexpression is correlated with the metastatic potential of cancers in humans and mice. Moreover, anti-galectin antibodies inhibit processes associated with cell transformation, such as cell aggregation and anchorage-independent growth (See, for example, Su, Z.-Z. et al. (1996) Proc. Natl. Acad. Sci. USA 93:7252-7257).  
       [0407] Selectins  
       [0408] Selectins, or LEC-CAMs, comprise a specialized lectin subfamily involved primarily in inflammation and leukocyte adhesion (Reviewed in Lasky, supra). Selectins mediate the recruitment of leukocytes from the circulation to sites of acute inflammation and are expressed on the surface of vascular endothelial cells in response to cytokine signaling. Selectins bind to specific ligands on the leukocyte cell membrane and enable the leukocyte to adhere to and migrate along the endothelial surface. Binding of selectin to its ligand leads to polarized rearrangement of the actin cytoskeleton and stimulates signal transduction within the leukocyte (Brenner, B. et al. (1997) Biochem. Biophys. Res. Commun 231:802-807;, Hidari, K. I. et al. (1997) J. Biol. Chem. 272:28750-28756). Members of the selectin family possess three characteristic motifs: a lectin or carbohydrate recognition domain; an epidermal growth factor-like domain; and a variable number of short consensus repeats (scr or “sushi” repeats) which are also present in complement regulatory proteins. The selectins include lymphocyte adhesion molecule-1 (Lam-1 or L-selectin), endothelial leukocyte adhesion molecule-1 (ELAM-1 or E-selectin), and granule membrane protein-140 (GMP-140 or P-selectin) (Johnston, G. I. et al. (1989) Cell 56:1033-1044).  
       [0409] Antigen Recognition Molecules  
       [0410] All vertebrates have developed sophisticated and complex immune systems that provide protection from viral, bacterial, fungal, and parasitic infections. A key feature of the immune system is its ability to distinguish foreign molecules, or antigens, from “self” molecules. This ability is mediated primarily by secreted and transmembrane proteins expressed by leukocytes (white blood cells) such as lymphocytes, granulocytes, and monocytes. Most of these proteins belong to the immunoglobulin (1 g) superfamily, members of which contain one or more repeats of a conserved structural domain. This Ig domain is comprised of antiparallel β sheets joined by a disulfide bond in an arrangement called the Ig fold. Members of the Ig superfamily include T-cell receptors, major histocompatibility (MHC) proteins, antibodies, and immune cell-specific surface markers such as CD4, CD8, and CD28.  
       [0411] MHC proteins are cell surface markers that bind to and present foreign antigens to T cells. MHC molecules are classified as either class I or class II. Class I MHC molecules (MHC I) are expressed on the surface of almost all cells and are involved in the presentation of antigen to cytotoxic T cells. For example, a cell infected with virus will degrade intracellular viral proteins and express the protein fragments bound to MHC I molecules on the cell surface. The MHC 1/antigen complex is recognized by cytotoxic T-cells which destroy the infected cell and the virus within Class II MHC molecules are expressed primarily on specialized antigen-presenting cells of the immune system, such as B-cells and macrophages. These cells ingest foreign proteins from the extracellular fluid and express MHC II/antigen complex on the cell surface. This complex activates helper T-cells, which then secrete cytokines and other factors that stimulate the immune response. MHC molecules also play an important role in organ rejection following transplantation. Rejection occurs when the recipient&#39;s T-cells respond to foreign MHC molecules on the transplanted organ in the same way as to self MHC molecules bound to foreign antigen. (Reviewed in Alberts, B. et al. (1994)  Molecular Biology of the Cell , Garland Publishing, New York N.Y., pp. 1229-1246.)  
       [0412] Antibodies, or immunoglobulins, are either expressed on the surface of B-cells or secreted by B-cells into the circulation. Antibodies bind and neutralize foreign antigens in the blood and other extracellular fluids. The prototypical antibody is a tetramer consisting of two identical heavy polypeptide chains (H-chains) and two identical light polypeptide chains (L-chains) interlinked by disulfide bonds. This arrangement confers the characteristic Y-shape to antibody molecules. Antibodies are classified based on their H-chain composition. The five antibody classes, IgA, IgD, IgE, IgG and IgM, are defined by the α, δ, ε, γ, and μ H-chain types. There are two types of L-chains, κ and λ, either of which may associate as a pair with any H-chain pair. IgG, the most common class of antibody found in the circulation, is tetrameric, while the other classes of antibodies are generally variants or multimers of this basic structure.  
       [0413] H-chains and L-chains each contain an N-terminal variable region and a C-terminal constant region. The constant region consists of about 110 amino acids in L-chains and about 330 or 440 amino acids in H-chains. The amino acid sequence of the constant region is nearly identical among H- or L-chains of a particular class. The variable region consists of about 110 amino acids in both H- and L-chains. However, the amino acid sequence of the variable region differs among H- or L-chains of a particular class. Within each H- or L-chain variable region are three hypervariable regions of extensive sequence diversity, each consisting of about 5 to 10 amino acids. In the antibody molecule, the H- and L-chain hypervariable regions come together to form the antigen recognition site. (Reviewed in Alberts, supra, pp. 1206-1213 and 1216-1217.)  
       [0414] Both H-chains and L-chains contain repeated Ig domains. For example, a typical H-chain contains four Ig domains, three of which occur within the constant region and one of which occurs within the variable region and contributes to the formation of the antigen recognition site. Likewise, a typical L-chain contains two Ig domains, one of which occurs within the constant region and one of which occurs within the variable region.  
       [0415] The immune system is capable of recognizing and responding to any foreign molecule that enters the body. Therefore, the immune system must be armed with a full repertoire of antibodies against all potential antigens. Such antibody diversity is generated by somatic rearrangement of gene segments encoding variable and constant regions. These gene segments are joined together by site-specific recombination which occurs between highly conserved DNA sequences that flank each gene segment. Because there are hundreds of different gene segments, millions of unique genes can be generated combinatorially. In addition, imprecise joining of these segments and an unusually high rate of somatic mutation within these segments further contribute to the generation of a diverse antibody population.  
       [0416] T-cell receptors are both structurally and functionally related to antibodies. (Reviewed in Alberts, supra, pp. 1228-1229.) T-cell receptors are cell surface proteins that bind foreign antigens and mediate diverse aspects of the immune response. A typical T-cell receptor is a heterodimer comprised of two disulfide-linked polypeptide chains called α and β. Each chain is about 280 amino acids in length and contains one variable region and one constant region. Each variable or constant region folds into an Ig domain. The variable regions from the α and β chains come together in the heterodimer to form the antigen recognition site. Tell receptor diversity is generated by somatic rearrangement of gene segments encoding the α and β chains. TV receptors recognize small peptide antigens that are expressed on the surface of antigen-presenting cells and pathogen-infected cells. These peptide antigens are presented on the cell surface in association with major histocompatibility proteins which provide the proper context for antigen recognition.  
       [0417] Secreted and Extracellular Matrix Molecules  
       [0418] Protein secretion is essential for cellular function. Protein secretion is mediated by a signal peptide located at the amino terminus of the protein to be secreted. The signal peptide is comprised of about ten to twenty hydrophobic amino acids which target the nascent protein from the ribosome to the endoplasmic reticulum (ER). Proteins targeted to the ER may either proceed through the secretory pathway or remain in any of the secretory organelles such as the ER, Golgi apparatus, or lysosomes.  
       [0419] Proteins that transit through the secretory pathway are either secreted into the extracellular space or retained in the plasma membrane. Secreted proteins are often synthesized as inactive precursors that are activated by post-translational processing events during transit through the secretory pathway. Such events include glycosylation, proteolysis, and removal of the signal peptide by a signal peptidase. Other events that may occur during protein transport include chaperone-dependent unfolding and folding of the nascent protein and interaction of the protein with a receptor or pore complex. Examples of secreted proteins with amino terminal signal peptides include receptors, extracellular matrix molecules, cytokines, hormones, growth and differentiation factors, neuropeptides, vasomediators, ion channels, transporters/pumps, and proteases. (Reviewed in Alberts, B. et al. (1994)  Molecular Biology of The Cell , Garland Publishing, New York N.Y., pp. 557-560, 582-592.)  
       [0420] The extracellular matrix (EC) is a complex network of glycoproteins, polysaccharides, proteoglycans, and other macromolecules that are secreted from the cell into the extracellular space. The ECM remains in close association with the cell surface and provides a supportive meshwork that profoundly influences cell shape, motility, strength, flexibility, and adhesion. In fact, adhesion of a cell to its surrounding matrix is required for cell survival except in the case of metastatic tumor cells, which have overcome the need for cell-ECM anchorage. This phenomenon suggests that the ECM plays a critical role in the molecular mechanisms of growth control and metastasis. (Reviewed in Ruoslahti, E. (1996) Sci. Am 275:72-77.) Furthermore, the ECM determines the structure and physical properties of connective tissue and is particularly important for morphogenesis and other processes associated with embryonic development and pattern formation.  
       [0421] The collagens comprise a family of ECM proteins that provide structure to bone, teeth, skin, ligaments, tendons, cartilage, blood vessels, and basement membranes. Multiple collagen proteins have been identified. Three collagen molecules fold together in a triple helix stabilized by interchain disulfide bonds. Bundles of these triple helices then associate to form fibrils. Collagen primary structure consists of hundreds of (Gly-X-Y) repeats where about a third of the X and Y residues are Pro. Glycines are crucial to helix formation as the bulkier amino acid sidechains cannot fold into the triple helical conformation. Because of these strict sequence requirements, mutations in collagen genes have severe consequences. Osteogenesis imperfecta patients have brittle bones that fracture easily; in severe cases patients die in utero or at birth Ehlers-Danlos syndrome patients have hyperelastic skin, hypermobile joints, and susceptibility to aortic and intestinal rupture. Chondrodysplasia patients have short stature and ocular disorders. Alport syndrome patients have hematuria, sensorineural deafness, and eye lens deformation. (Isselbacher, K. J. et al. (1994)  Harrison&#39;s Principles of Internal Medicine , McGraw-Hill, Inc., New York N.Y., pp. 2105-2117; and Creighton, T. E. (1984)  Proteins. Structures and Molecular Principles , W. H. Freeman and Company, New York N.Y., pp. 191-197.)  
       [0422] Elastin and related proteins confer elasticity to tissues such as skin, blood vessels, and lungs. Elastin is a highly hydrophobic protein of about 750 amino acids that is rich in proline and glycine residues. Elastin molecules are highly cross-linked, forming an extensive extracellular network of fibers and sheets. Elastin fibers are surrounded by a sheath of microfibrils which are composed of a number of glycoproteins, including fibrillin. Mutations in the gene encoding fibrillin are responsible for Marfan&#39;s syndrome, a genetic disorder characterized by defects in connective tissue. In severe cases, the aortas of afflicted individuals are prone to rupture. (Reviewed in Alberts, supra, pp.  984-986 .)  
       [0423] Fibronectin is a large ECM glycoprotein found in all vertebrates. Fibronectin exists as a dimer of two subunits, each containing about 2,500 amino acids. Each subunit folds into a rod-like structure containing multiple domains. The domains each contain multiple repeated modules, the most common of which is the type III fibronectin repeat. The type III fibronectin repeat is about 90 amino acids in length and is also found in other ECM proteins and in some plasma membrane and cytoplasmic proteins. Furthermore, some type III fibronectin repeats contain a characteristic tripeptide consisting of Arginine-Glycine-Aspartic acid (RGD). The RGD sequence is recognized by the integrin family of cell surface receptors and is also found in other ECM proteins. Disruption of both copies of the gene encoding fibronectin causes early embryonic lethality in mice. The mutant embryos display extensive morphological defects, including defects in the formation of the notochord, somites, heart, blood vessels, neural tube, and extraembryonic structures. (Reviewed in Alberts, supra, pp. 986-987.)  
       [0424] Laminin is a major glycoprotein component of the basal lamina which underlies and supports epithelial cell sheets. Laminin is one of the first ECM proteins synthesized in the developing embryo. Laminin is an 850 kilodalton protein composed of three polypeptide chains joined in the shape of a cross by disulfide bonds. Laminin is especially important for angiogenesis and in particular, for guiding the formation of capillaries. (Reviewed in Alberts, supra, pp. 990-991.)  
       [0425] There are many other types of proteinaceous ECM components, most of which can be classified as proteoglycans. Proteoglycans are composed of unbranched polysaccharide chains (glycosaminoglycans) attached to protein cores. Common proteoglycans include aggrecan, betaglycan, decorin, perlecan, serglycin, and syndecan-1. Some of these molecules not only provide mechanical support, but also bind to extracellular signaling molecules, such as fibroblast growth factor and transforming growth factor β, suggesting a role for proteoglycans in cell-cell communication and cell growth (Reviewed in Alberts, supra, pp. 973-978.) Likewise, the glycoproteins tenascin-C and tenascin-R are expressed in developing and lesioned neural tissue and provide stimulatory and anti-adhesive (inhibitory) properties, respectively, for axonal growth (Faissner, A (1997) Cell Tissue Res. 290:331-341.)  
       [0426] Cytoskeletal Molecules  
       [0427] The cytoskeleton is a cytoplasmic network of protein fibers that mediate cell shape, structure, and movement. The cytoskeleton supports the cell membrane and forms tracks along which organelles and other elements move in the cytosol. The cytoskeleton is a dynamic structure that allows cells to adopt various shapes and to carry out directed movements. Major cytoskeletal fibers include the microtubules, the microfilaments, and the intermediate filaments. Motor proteins, including myosin, dynein, and kinesin, drive movement of or along the fibers. The motor protein dynamin drives the formation of membrane vesicles. Accessory or associated proteins modify the structure or activity of the fibers while cytoskeletal membrane anchors connect the fibers to the cell membrane.  
       [0428] Tubulins  
       [0429] Microtubules, cytoskeletal fibers with a diameter of about 24 nm, have multiple roles in the cell. Bundles of microtubules form cilia and flagella, which are whip-like extensions of the cell membrane that are necessary for sweeping materials across an epithelium and for swimming of sperm, respectively. Marginal bands of microtubules in red blood cells and platelets are important for these cells&#39; pliability. Organelles, membrane vesicles, and proteins are transported in the cell along tracks of microtubules. For example, microtubules run through nerve cell axons, allowing bi-directional transport of materials and membrane vesicles between the cell body and the nerve terminal. Failure to supply the nerve terminal with these vesicles blocks the transmission of neural signals. Microtubules are also critical to chromosomal movement during cell division. Both stable and short-lived populations of microtubules exist in the cell.  
       [0430] Microtubules are polymers of GTP-binding tubulin protein subunits. Each subunit is a heterodimer of α- and β-tubulin, multiple isoforms of which exist The hydrolysis of GTP is linked to the addition of tubulin subunits at the end of a microtubule. The subunits interact head to tail to form protofilaments; the protofilaments interact side to side to form a microtubule. A microtubule is polarized, one end ringed with α-tubulin and the other with β-tubulin, and the two ends differ in their rates of assembly. Generally, each microtubule is composed of 13 protofilaments although 11 or 15 protofilament-microtubules are sometimes found. Cilia and flagella contain doublet microtubules. Microtubules grow from specialized structures known as centrosomes or microtubule-organizing centers (MTOCs). MTOCs may contain one or two centrioles, which are pinwheel arrays of triplet microtubules. The basal body, the organizing center located at the base of a cilium or flagellum, contains one centriole. Gamma tubulin present in the MTOC is important for nucleating the polymerization of α- and β-tubulin heterodimers but does not polymerize into microtubules.  
       [0431] Microtubule-Associated Proteins  
       [0432] Microtubule-associated proteins (MAPs) have roles in the assembly and stabilization of microtubules. One major family of MAPs, assembly MAPs, can be identified in neurons as well as non-neuronal cells. Assembly MAPs are responsible for cross-linking microtubules in the cytosol These MAPs are organized into two domains: a basic microtubule-binding domain and an acidic projection domain. The projection domain is the binding site for membranes, intermediate filaments, or other microtubules. Based on sequence analysis, assembly MAPs can be further grouped into two types: Type I and Type II. Type I MAPs, which include MAP1A and MAP1B, are large, filamentous molecules that co-purify with microtubules and are abundantly expressed in brain and testes. Type I MAPs contain several repeats of a positively-charged amino acid sequence motif that binds and neutralizes negatively charged tubulin, leading to stabilization of microtubules. MAP1A and MAP1B are each derived from a single precursor polypeptide that is subsequently proteolytically processed to generate one heavy chain and one light chain  
       [0433] Another light chain, LC3, is a 16.4 kDa molecule that binds MAP1A, MAP1B, and microtubules. It is suggested that LC3 is synthesized from a source other than the MAP1A or MAP1B transcripts, and that the expression of LC3 may be important in regulating the microtubule binding activity of MAP1A and MAP1B during cell proliferation (Mann, S. S. et al. (1994) J. Biol. Chem. 269:11492-11497).  
       [0434] Type II MAPs, which include MAP2a, MAP2b, MAP2c, MAP4, and Tau, are characterized by three to four copies of an 18-residue sequence in the microtubule-binding domain MAP2a, MAP2b, and MAP2c are found only in dendrites, MAP4 is found in non-neuronal cells, and Tau is found in axons and dendrites of nerve cells. Alternative splicing of the Tau mRNA leads to the existence of multiple forms of Tau protein. Tau phosphorylation is altered in neurodegenerative disorders such as Alzheimer&#39;s disease, Pick&#39;s disease, progressive supranuclear palsy, corticobasal degeneration, and familial frontotemporal dementia and Parkinsonism linked to chromosome 17. The altered Tau phosphorylation leads to a collapse of the microtubule network and the formation of intraneuronal Tau aggregates (Spillantini, M. G. and M. Goedert (1998) Trends Neurosci. 21:428-433).  
       [0435] The protein pericentrin is found in the MTOC and has a role in microtubule assembly.  
       [0436] Actins  
       [0437] Microfilaments, cytoskeletal filaments with a diameter of about 7-9 nm, are vital to cell locomotion, cell shape, cell adhesion, cell division, and muscle contraction. Assembly and disassembly of the microfilaments allow cells to change their morphology. Microfilaments are the polymerized form of actin, the most abundant intracellular protein in the eukaryotic cell. Human cells contain six isoforms of actin. The three α-actins are found in different kinds of muscle, nonmuscle, β-actin and nonmuscle γ-actin are found in nonmuscle cells, and another y-actin is found in intestinal smooth muscle cells. G-actin, the monomeric form of actin, polymerizes into polarized, helical F-actin filaments, accompanied by the hydrolysis of ATP to ADP. Actin filaments associate to form bundles and networks, providing a framework to support the plasma membrane and determine cell shape. These bundles and networks are connected to the cell membrane. In muscle cells, thin filaments containing actin slide past thick filaments containing the motor protein myosin during contraction. A family of actin-related proteins exist that are not part of the actin cytoskeleton, but rather associate with microtubules and dynein.  
       [0438] Actin-Associated Proteins  
       [0439] Actin-associated proteins have roles in cross-linking, severing, and stabilization of actin filaments and in sequestering actin monomers. Several of the actin-associated proteins have multiple functions. Bundles and networks of actin filaments are held together by actin cross-lining proteins. These proteins have two actin-binding sites, one for each filament. Short cross-lining proteins promote bundle formation while longer, more flexible cross-linking proteins promote network formation. Calmodulin-like calcium-binding domains in actin cross-linking proteins allow calcium regulation of cross-linking. Group I cross-linking proteins have unique actin-binding domains and include the 30 kD protein, EF-1a, fascin, and scruin. Group II cross-lining proteins have a 7,000-MW actin-binding domain and include villin and dematin. Group III cross-lining proteins have pairs of a 26,000-MW actin-binding domain and include fimbrin, spectrin, dystrophin, ABP 120, and filamin  
       [0440] Severing proteins regulate the length of actin filaments by breaking them into short pieces or by blocking their ends. Severing proteins include gCAP39, severin (fragmin), gelsolin, and villin. Capping proteins can cap the ends of actin filaments, but cannot break filaments. Capping proteins include CapZ and tropomodulin. The proteins thymosin and profilin sequester actin monomers in the cytosol, allowing a pool of unpolymerized actin to exist. The actin-associated proteins tropomyosin, troponin, and caldesmon regulate muscle contraction in response to calcium.  
       [0441] Intermediate Filaments and Associated Proteins  
       [0442] Intermediate filaments (IFs) are cytoskeletal fibers with a diameter of about 10 nm, intermediate between that of microfilaments and microtubules. IFs serve structural roles in the cell, reinforcing cells and organizing cells into tissues. IFs are particularly abundant in epidermal cells and in neurons. IFs are extremely stable, and, in contrast to microfilaments and microtubules, do not function in cell motility.  
       [0443] Five types of IF proteins are known in mammals. Type I and Type II proteins are the acidic and basic keratins, respectively. Heterodimers of the acidic and basic keratins are the building blocks of keratin IFs. Keratins are abundant in soft epithelia such as skin and cornea, hard epithelia such as nails and hair, and in epithelia that line internal body cavities. Mutations in keratin genes lead to epithelial diseases including epidermolysis bullosa simplex, bullous congenital ichthyosiform erytroderma (epidermolytic hyperkeratosis), non-epidermolytic and epidermolytic palmoplantar keratodenna, ichthyosis bullosa of Siemens, pachyonychia congenita, and white sponge nevus. Some of these diseases result in severe skin blistering. (See, e.g., Wawersik, M. et al. (1997) J. Biol. Chem. 272:32557-32565; and Corden L. D. and W. H. McLean (1996) Exp. Dermatol. 5:297-307.)  
       [0444] Type III IF proteins include desmin, glial fibrillary acidic protein, vimentin, and peipherin. Desmin filaments in muscle cells link myofibrils into bundles and stabilize sarcomeres in contracting muscle. Glial fibrillary acidic protein filaments are found in the glial cells that surround neurons and astrocytes. Vimentin filaments are found in blood vessel endothelial cells, some epithelial cells, and mesenchymal cells such as fibroblasts, and are commonly associated with microtubules. Vimentin filaments may have roles in keeping the nucleus and other organelles in place in the cell. Type IV IFs include the neurofilaments and nestin. Neurofilaments, composed of three polypeptides NF-L, NF-M, and NF-H, are frequently associated with microtubules in axons. Neurofilaments are responsible for the radial growth and diameter of an axon, and ultimately for the speed of nerve impulse transmission. Changes in phosphorylation and metabolism of neurofilaments are observed in neurodegenerative diseases including amyotrophic lateral sclerosis, Parkinson&#39;s disease, and Alzheimer&#39;s disease (Julien, J. P. and W. E. Mushynski (1998) Prog. Nucleic Acid Res. Mol. Biol. 61:1-23). Type V IFs, the lamins, are found in the nucleus where they support the nuclear membrane.  
       [0445] IFs have a central α-helical rod region interrupted by short nonhelical linker segments. The rod region is bracketed, in most cases, by non-helical head and tail domains. The rod regions of intermediate filament proteins associate to form a coiled-coil dimer. A highly ordered assembly process leads from the dimers to the IFs. Neither ATP nor GTP is needed for IF assembly, unlike that of microfilaments and microtubules.  
       [0446] IF-associated proteins (IFAPs) mediate the interactions of IFs with one another and with other cell structures. IFAPs cross-link IFs into a bundle, into a network, or to the plasma membrane, and may cross-link IFs to the microfilament and microtubule cytoskeleton. Microtubules and IFs are in particular closely associated. IFAPs include BPAG1, plakoglobin, desmoplakin I, desmoplakin II, plectin, ankyrin, filaggrin, and lamin B receptor.  
       [0447] Cytoskeletal-Membrane Anchors  
       [0448] Cytoskeletal fibers are attached to the plasma membrane by specific proteins. These attachments are important for maintaining cell shape and for muscle contraction. In erythrocytes, the spectrin-actin cytoskeleton is attached to cell membrane by three proteins, band 4.1, ankyrin, and adducin. Defects in this attachment result in abnormally shaped cells which are more rapidly degraded by the spleen, leading to anemia. In platelets, the spectrin-actin cytoskeleton is also linked to the membrane by ankyrin; a second actin network is anchored to the membrane by filamin In muscle cells the protein dystrophin links actin filaments to the plasma membrane; mutations in the dystrophin gene lead to Duchenne muscular dystrophy. In adherens junctions and adhesion plaques the peripheral membrane proteins α-actin and vinculin attach actin filaments to the cell membrane.  
       [0449] IFs are also attached to membranes by cytoskeletal-membrane anchors. The nuclear lamina is attached to the inner surface of the nuclear membrane by the lamin B receptor. Vimentin IFs are attached to the plasma membrane by ankyrin and plectin. Desmosome and hemidesmosome membrane junctions hold together epithelial cells of organs and skin. These membrane junctions allow shear forces to be distributed across the entire epithelial cell layer, thus providing strength and rigidity to the epithelium. IFs in epithelial cells are attached to the desmosome by plakoglobin and desmoplakins. The proteins that link IFs to hemidesmosomes are not known. Desmin IFs surround the sarcomere in muscle and are linked to the plasma membrane by paranemin, synemin, and ankyrin.  
       [0450] Myosin-related Motor Proteins  
       [0451] Myosins are actin-activated ATPases, found in eukaryotic cells, that couple hydrolysis of ATP with motion. Myosin provides the motor function for muscle contraction and intracellular movements such as phagocytosis and rearrangement of cell contents during mitotic cell division (cytokinesis). The contractile unit of skeletal muscle, termed the sarcomere, consists of highly ordered arrays of thin actin-containing filaments and thick myosin-containing filaments. Crossbridges form between the thick and thin filaments, and the ATP-dependent movement of myosin heads within the thick filaments pulls the thin filaments, shortening the sarcomere and thus the muscle fiber.  
       [0452] Myosins are composed of one or two heavy chains and associated light chains. Myosin heavy chains contain an amino-terminal motor or head domain, a neck that is the site of light-chain binding, and a carboxy-terminal tail domain. The tail domains may associate to form an α-helical coiled coil. Conventional myosins, such as those found in muscle tissue, are composed of two myosin heavy-chain subunits, each associated with two light-chain subunits that bind at the neck region and play a regulatory role. Unconventional myosins, believed to function in intracellular motion, may contain either one or two heavy chains and associated light chains. There is evidence for about 25 myosin heavy chain genes in vertebrates, more than half of them unconventional.  
       [0453] Dynein-related Motor Proteins  
       [0454] Dyneins are (−) end-directed motor proteins which act on microtubules. Two classes of dyneins, cytosolic and axonemal, have been identified. Cytosolic dyneins are responsible for translocation of materials along cytoplasmic microtubules, for example, transport from the nerve terminal to the cell body and transport of endocytic vesicles to lysosomes. Cytoplasmic dyneins are also reported to play a role in mitosis. Axonemal dyneins are responsible for the beating of flagella and cilia. Dynein on one microtubule doublet walks along the adjacent microtubule doublet This sliding force produces bending forces that cause the flagellum or cilium to beat Dyneins have a native mass between 1000 and 2000 kDa and contain either two or three force-producing heads driven by the hydrolysis of ATP. The heads are linked via stalks to a basal domain which is composed of a highly variable number of accessory intermediate and light chains.  
       [0455] Kinesin-related Motor Proteins  
       [0456] Kinesins are (+) end-directed motor proteins which act on microtubules. The prototypical kinesin molecule is involved in the transport of membrane-bound vesicles and organelles. This function is particularly important for axonal transport in neurons. Kinesin is also important in all cell types for the transport of vesicles from the Golgi complex to the endoplasmic reticulum. This role is critical for maintaining the identity and functionality of these secretory organelles.  
       [0457] Kinesins define a ubiquitous, conserved family of over 50 proteins that can be classified into at least 8 subfamilies based on primary amino acid sequence, domain structure, velocity of movement, and cellular function. (Reviewed in Moore, J. D. and S. A. Endow (1996) Bioessays 18:207-219; and Hoyt, A. M. (1994) Curr. Opin. Cell Biol. 6:63-68.) The prototypical kinesin molecule is a heterotetramer comprised of two heavy polypeptide chains (KHCs) and two light polypeptide chains (KLCs). The KHC subunits are typically referred to as “kinesin.” KHC is about 1000 amino acids in length, and KLC is about 550 amino acids in length Two KHCs dimerize to form a rod-shaped molecule with three distinct regions of secondary structure. At one end of the molecule is a globular motor domain that functions in ATP hydrolysis and microtubule binding. Kinesin motor domains are highly conserved and share over 70% identity. Beyond the motor domain is an α-helical coiled-coil region which mediates dimerization. At the other end of the molecule is a fan-shaped tail that associates with molecular cargo. The tail is formed by the interaction of the KHC C-termini with the two KLCs.  
       [0458] Members of the more divergent subfamilies of kinesins are called kinesin-related proteins (KRPs), many of which function during mitosis in eukaryotes (Hoyt, supra). Some KRPs are required for assembly of the mitotic spindle. In vivo and in vitro analyses suggest that these KRPs exert force on microtubules that comprise the mitotic spindle, resulting in the separation of spindle poles. Phosphorylation of KRP is required for this activity. Failure to assemble the mitotic spindle results in abortive mitosis and chromosomal aneuploidy, the latter condition being characteristic of cancer cells. In addition, a unique KRP, centromere protein E, localizes to the kinetochore of human mitotic chromosomes and may play a role in their segregation to opposite spindle poles.  
       [0459] Dynamin-related Motor Proteins  
       [0460] Dynamin is a large GTPase motor protein that functions as a “molecular pinchase,” generating a mechanochemical force used to sever membranes. This activity is important in forming clathrin-coated vesicles from coated pits in endocytosis and in the biogenesis of synaptic vesicles in neurons. Binding of dynamin to a membrane leads to dynamin&#39;s self-assembly into spirals that may act to constrict a flat membrane surface into a tubule. GTP hydrolysis induces a change in conformation of the dynamin polymer that pinches the membrane tubule, leading to severing of the membrane tubule and formation of a membrane vesicle. Release of GDP and inorganic phosphate leads to dynamin disassembly. Following disassembly the dynamin may either dissociate from the membrane or remain associated to the vesicle and be transported to another region of the cell. Three homologous dynamin genes have been discovered, in addition to several dynamin-related proteins. Conserved dynamin regions are the N-terminal GTP-binding domain, a central pleckstrin homology domain that binds membranes, a central coiled-coil region that may activate dynamin&#39;s GTPase activity, and a C-terminal proline-rich domain that contains several motifs that bind SH3 domains on other proteins. Some dynamin-related proteins do not contain the pleckstrin homology domain or the proline-rich domain. (See McNiven, M. A. (1998) Cell 94:151-154; Scaife, R. N. and R. L. Margolis (1997) Cell. Signal. 9:395401.)  
       [0461] The cytoskeleton is reviewed in Lodish, H. et al. (1995)  Molecular Cell Biology , Scientific American Books, New York N.Y.  
       [0462] Ribosomal Molecules  
       [0463] Ribosomal RNAs (rRNAs) are assembled, along with ribosomal proteins, into ribosomes, which are cytoplasmic particles that translate messenger RNA into polypeptides. The eukaryotic ribosome is composed of a 60S (large) subunit and a 40S (small) subunit, which together form the 80S ribosome. In addition to the 18S, 28S, 5S, and 5.8S rRNAs, the ribosome also contains more than fifty proteins. The ribosomal proteins have a prefix which denotes the subunit to which they belong, either L (large) or S (small). Ribosomal protein activities include binding rRNA and organizing the conformation of the junctions between rRNA helices (Woodson, S. A. and N. B. Leontis (1998) Curr. Opin. Stuct. Biol.  8:294-300 ; Ramakrishnan, V. and S. W. White (1998) Trends Biochem. Sci. 23:208-212.) Three important sites are identified on the ribosome. The aminoacyl-tRNA site (A site) is where charged tRNAs (with the exception of the initiator-tRNA) bind on arrival at the ribosome. The peptidyl-tRNA site (P site) is where new peptide bonds are formed, as well as where the initiator tRNA binds. The exit site E site) is where deacylated tRNAs bind prior to their release from the ribosome. (The ribosome is reviewed in Stryer, L. (1995)  Biochemistry  W. H. Freeman and Company, New York N. Y., pp. 888-908; and Lodish, H. et al. (1995)  Molecular Cell Biology  Scientific American Books, New York N.Y. pp. 119-138.)  
       [0464] Chromatin Molecules  
       [0465] The nuclear DNA of eukaryotes is organized into chromatin. Two types of chromatin are observed: euchromatin, some of which may be transcribed, and heterochromatin so densely packed that much of it is inaccessible to transcription. Chromatin packing thus serves to regulate protein expression in eukaryotes. Bacteria lack chromatin and the chromatin-packing level of gene regulation. The fundamental unit of chromatin is the nucleosome of 200 DNA base pairs associated with two copies each of histones H2A, H2B, H3, and H4. Adjascent nucleosomes are linked by another class of histones, H1. Low molecular weight non-histone proteins called the high mobility group (ERG), associated with chromatin, may function in the unwinding of DNA and stabilization of single-stranded DNA. Chromodomain proteins function in compaction of chromatin into its transcriptionally silent heterochromatin form.  
       [0466] During mitosis, all DNA is compacted into heterochromatin and transcription ceases. Transcription in interphase begins with the activation of a region of chromatin Active chromatin is decondensed. Decondensation appears to be accompanied by changes in binding coefficient, phosphorylation and acetylation states of chromatin histones. HMG proteins HMG13 and HMG17 selectively bind activated chromatin. Topoisomerases remove superheilcal tension on DNA The activated region decondenses, allowing gene regulatory proteins and transcription factors to assemble on the DNA.  
       [0467] Patterns of chromatin structure can be stably inherited, producing heritable patterns of gene expression. In mammals, one of the two X chromosomes in each female cell is inactivated by condensation to heterochromatin during zygote development. The inactive state of this chromosome is inherited, so that adult females are mosaics of clusters of paternal-X and maternal-X clonal cell groups. The condensed X chromosome is reactivated in meiosis.  
       [0468] Chromatin is associated with disorders of protein expression such as thalassemia, a genetic anemia resulting from the removal of the locus control region (LCR) required for decondensation of the globin gene locus.  
       [0469] For a review of chromatin structure and function see Alberts, B. et al. (1994)  Molecular Cell Biology , third edition, Garland Publishing, Inc., New York N.Y., pp. 351- 354, 433-439.    
       [0470] Electron Transfer Associated Molecules  
       [0471] Electron carriers such as cytochromes accept electrons from NADH or FADH 2  and donate them to other electron carriers. Most electron-transferring proteins, except ubiquinone, are prosthetic groups such as flavins, heme, FeS clusters, and copper, bound to inner membrane proteins. Adrenodoxin, for example, is an FeS protein that forms a complex with NADPH:adrenodoxin reductase and cytochrome p450. Cytochromes contain a heme prosthetic group, a porphyrin ring containing a tightly bound iron atom. Electron transfer reactions play a crucial role in cellular energy production.  
       [0472] Energy is produced by the oxidation of glucose and fatty acids. Glucose is initially converted to pyruvate in the cytoplasm. Fatty acids and pyruvate are transported to the mitochondria for complete oxidation to CO 2  coupled by enzymes to the transport of electrons from NADH and FADH 2  to oxygen and to the synthesis of ATP (oxidative phosphorylation) from ADP and P i .  
       [0473] Pyruvate is transported into the mitochondria and converted to acetyl-CoA for oxidation via the citric acid cycle, involving pyruvate dehydrogenase components, dihydrolipoyl transacetylase, and dihydrolipoyl dehydrogenase. Enzymes involved in the citric acid cycle include: citrate synthetase, aconitases, isocitrate dehydrogenase, alpha-ketoglutarate dehydrogenase complex including transsuccinylases, succinyl CoA synthetase, succinate dehydrogenase, fumarases, and malate dehydrogenase. Acetyl CoA is oxidized to CO 2  with concomitant formation of NADH, FADH 2 , and GTP. In oxidative phosphorylation, the transfer of electrons from NADH and FADH 2  to oxygen by dehydrogenases is coupled to the synthesis of ATP from ADP and P i  by the F 0 F 1  ATPase complex in is the mitochondrial inner membrane. Enzyme complexes responsible for electron transport and ATP synthesis include the F 0 F 1  ATPase complex, ubiquinone(CoQ)-cytocbrome c reductase, ubiquinone reductase, cytochrome b, cytocbrome c 1 , FeS protein, and cytochrome c oxidase.  
       [0474] ATP synthesis requires membrane transport enzymes including the phosphate transporter and the ATP-ADP antiport protein. The ATP-binding casette (ABC) superfamily has also been suggested as belonging to the mitochondrial transport group (Hogue, D. L. et al. (1999) J. Mol. Biol. 285:379-389). Brown fat uncoupling protein dissipates oxidative energy as heat, and may be involved the fever response to infection and trauma (Cannon, B. et al. (1998) Ann. NY Acad. Sci. 856:171-187).  
       [0475] Mitochondria are oval-shaped organelles comprising an outer membrane, a tightly folded inner membrane, an intermembrane space between the outer and inner membranes, and a matrix inside the inner membrane. The outer membrane contains many porin molecules that allow ions and charged molecules to enter the intermembrane space, while the inner membrane contains a variety of transport proteins that transfer only selected molecules. Mitochondria are the primary sites of energy production in cells.  
       [0476] Mitochondria contain a small amount of DNA. Human mitochondrial DNA encodes 13 proteins, 22 tRNAs, and 2 rRNAs. Mitochondrial-DNA encoded proteins include NADH-Q reductase, a cytochrome reductase subunit, cytochrome oxidase subunits, and ATP synthase subunits.  
       [0477] Electron-transfer reactions also occur outside the mitochondria in locations such as the endoplasmic reticulum, which plays a crucial role in lipid and protein biosynthesis. Cytochrome b5 is a central electron donor for various reductive reactions occurring on the cytoplasmic surface of liver endoplasmic reticulum. Cytochrome b5 has been found in Golgi, plasma, endoplasmic reticulum (ER), and microbody membranes.  
       [0478] For a review of mitochondrial metabolism and regulation, see Lodish, H. et al. (1995)  Molecular Cell Biology , Scientific American Books, New York N.Y., pp. 745-797 and Stryer (1995)  Biochemistry , W. H. Freeman and Co., San Francisco Calif., pp 529-558, 988-989.  
       [0479] The majority of mitochondrial proteins are encoded by nuclear genes, are synthesized on cytosolic ribosomes, and are imported into the mitochondria. Nuclear-encoded proteins which are destined for the mitochondrial matrix typically contain positively-charged amino terminal signal sequences. Import of these preproteins from the cytoplasm requires a multisubunit protein complex in the outer membrane known as the translocase of outer mitochondrial membrane (TOM; previously designated MOM; Pfanner, N. et al. (1996) Trends Biochem. Sci. 21:51-52) and at least three inner membrane proteins which comprise the translocase of inner mitochondrial membrane (TIM; previously designated MIM; Pfanner, supra). An inside-negative membrane potential across the inner mitochondrial membrane is also required for preprotein import. Preproteins are recognized by surface receptor components of the TOM complex and are translocated through a proteinaceous pore formed by other TOM components. Proteins targeted to the matrix are then recognized by the import machinery of the TIM complex. The import systems of the outer and inner membranes can function independently (Segui-Real, B. et al. (1993) EMBO J. 12:2211-2218).  
       [0480] Once precursor proteins are in the mitochondria, the leader peptide is cleaved by a signal peptidase to generate the mature protein. Most leader peptides are removed in a one step process by a protease termed mitochondrial processing peptidase (MPP) (Paces, V. et al. (1993) Proc. Natl. Acad. Sci. USA 90:5355-5358). In some cases a two-step process occurs in which MPP generates an intermediate precursor form which is cleaved by a second enzyme, mitochondrial intermediate peptidase, to generate the mature protein.  
       [0481] Mitochondrial dysfunction leads to impaired calcium buffering, generation of free radicals that may participate in deleterious intracellular and extracellular processes, changes in mitochondrial permeability and oxidative damage which is observed in several neurodegenerative diseases. Neurodegenerative diseases linked to mitochondrial dysfunction include some forms of Alzheimer&#39;s disease, Friedreich&#39;s ataxia, familial amyotrophic lateral sclerosis, and Huntington&#39;s disease (Beal, M. F. (1998) Biochim. Biophys. Acta 1366:211-213). The myocardium is heavily dependent on oxidative metabolism, so mitochondrial dysfunction often leads to heart disease (DiMauro, S. and M. Hirano (1998) Curr. Opin Cardiol 13:190-197). Mitochondria are implicated in disorders of cell proliferation, since they play an important role in a cell&#39;s decision to proliferate or self-destruct through apoptosis. The oncoprotein Bcl-2, for example, promotes cell proliferation by stabilizing mitochondrial membranes so that apoptosis signals are not released (Susin, S. A. (1998) Biochim. Biophys. Acta 1366:151-165).  
       [0482] Transcription Factor Molecules  
       [0483] Multicellular organisms are comprised of diverse cell types that differ dramatically both in structure and function. The identity of a cell is determined by its characteristic pattern of gene expression, and different cell types express overlapping but distinctive sets of genes throughout development. Spatial and temporal regulation of gene expression is critical for the control of cell proliferation, cell differentiation, apoptosis, and other processes that contribute to organismal development. Futhermore, gene expression is regulated in response to extracellular signals that mediate cell-cell communication and coordinate the activities of different cell types. Appropriate gene regulation also ensures that cells function efficiently by expressing only those genes whose functions are required at a given time.  
       [0484] Transcriptional regulatory proteins are essential for the control of gene expression. Some of these proteins function as transcription factors that initiate, activate, repress, or terminate gene transcription. Transcription factors generally bind to the promoter, enhancer, and upstream regulatory regions of a gene in a sequence-specific manner, although some factors bind regulatory elements with or downstream of a gene&#39;s coding region. Transcription factors may bind to a specific region of DNA singly or as a complex with other accessory factors. (Reviewed in Lewin, B. (1990)  Genes IV , Oxford University Press, New York N.Y., and Cell Press, Cambridge Mass., pp. 554-570.)  
       [0485] The double helix structure and repeated sequences of DNA create topological and chemical features which can be recognized by transcription factors. These features are hydrogen bond donor and acceptor groups, hydrophobic patches, major and minor grooves, and regular, repeated stretches of sequence which induce distinct bends in the helix. Typically, transcription factors recognize specific DNA sequence motifs of about 20 nucleotides in length. Multiple, adjacent transcription factor-binding motifs may be required for gene regulation.  
       [0486] Many transcription factors incorporate DNA-binding structural motifs which comprise either α helices or β sheets that bind to the major groove of DNA. Four well-characterized structural motifs are helix-turn-helix, zinc finger, leucine zipper, and helix-loop-helix. Proteins containing these motifs may act alone as monomers, or they may form homo- or heterodimers that interact with DNA.  
       [0487] The helix-turn-helix motif consists of two a helices connected at a fixed angle by a short chain of amino acids. One of the helices binds to the major groove. Helix-turn-helix motifs are exemplified by the homeobox motif which is present in homeodomain proteins. These proteins are critical for specifying the anterior-posterior body axis during development and are conserved throughout the animal kingdom. The Antennapedia and Ultrabithorax proteins of  Drosophila melanogaster  are prototypical homeodomain proteins (Pabo, C. O. and R. T. Sauer (1992) Annu. Rev. Biochem. 61:1053-1095).  
       [0488] The zinc finger motif, which binds zinc ions, generally contains tandem repeats of about 30 amino acids consisting of periodically spaced cysteine and histidine residues. Examples of this sequence pattern, designated C2H2 and C3HC4 (“RING” finger), have been described Lewin, supra). Zinc finger proteins each contain an α helix and an antiparallel β sheet whose proximity and conformation are maintained by the zinc ion. Contact with DNA is made by the arginine prece ding the α helix and by the second, third, and sixth residues of the α helix. Variants of the zinc finger motif include poorly defined cysteine-rich motifs which bind zinc or other metal ions. These motifs may not contain histidine residues and are generally nonrepetitive.  
       [0489] The leucine zipper motif comprises a stretch of amino acids rich in leucine which can form an amphipathic α helix. This structure provides the basis for dimerization of two leucine zipper proteins. The region adjacent to the leucine zipper is usually basic, and upon protein dimerization, is optimally positioned for binding to the major groove. Proteins containing such motifs are generally referred to as bZIP transcription factors.  
       [0490] The helix-loop-helix motif (HLH) consists of a short α helix connected by a loop to a longer α helix. The loop is flexible and allows the two helices to fold back against each other and to bind to DNA. The transcription factor Myc contains a prototypical HLH motif.  
       [0491] Most transcription factors contain characteristic DNA binding motifs, and variations on the above motifs and new motifs have been and are currently being characterized (Faisst, S. and S. Meyer (1992) Nucleic Acids Res. 20:3-26).  
       [0492] Many neoplastic disorders in humans can be attributed to inappropriate gene expression. Malignant cell growth may result from either excessive expression of tumor promoting genes or insufficient expression of tumor suppressor genes (Cleary, M. L. (1992) Cancer Surv. 15:89-104). Chromosomal translocations may also produce chimeric loci which fuse the coding sequence of one gene with the regulatory regions of a second unrelated gene. Such an arrangement likely results in inappropriate gene transcription, potentially contributing to malignancy.  
       [0493] In addition, the immune system responds to infection or trauma by activating a cascade of events that coordinate the progressive selection, amplification, and mobilization of cellular defense mechanisms. A complex and balanced program of gene activation and repression is involved in this process. However, hyperactivity of the immune system as a result of improper or insufficient regulation of gene expression may result in considerable tissue or organ damage. This damage is well documented in immunological responses associated with arthritis, allergens, heart attack, stroke, and infections (Isselbacher, K. J. et al. (1996)  Harrison&#39;s Principles of Internal Medicine , 13/e, McGraw Hill, Inc. and Teton Data Systems Software).  
       [0494] Furthermore, the generation of multicellular organisms is based upon the induction and coordination of cell differentiation at the appropriate stages of development Central to this process is differential gene expression, which confers the distinct identities of cells and tissues throughout the body. Failure to regulate gene expression during development can result in developmental disorders. Human developmental disorders caused by mutations in zinc finger-type transcriptional regulators include: urogenenital developmental abnormalities associated with WT1; Greig cephalopolysyndactyly, Pallister-Hall syndrome, and postaxial polydactyly type A (GLI3); and Townes-Brocks syndrome, characterized by anal, renal, limb, and ear abnormalities (SALL1) (Engelkamp, D. and V. van Heyningen (1996) Curr. Opin. Genet Dev. 6:334342; Kohlhase, J. et al. (1999) Am. J. Hum. Genet 64:435445).  
       [0495] Cell Membrane Molecules  
       [0496] Eukaryotic cells are surrounded by plasma membranes which enclose the cell and maintain an environment inside the cell that is distinct from its surroundings. In addition, eukaryotic organisms are distinct from prokaryotes in possessing many intracellular organelle and vesicle structures. Many of the metabolic reactions which distinguish eukaryotic biochemistry from prokaryotic biochemistry take place within these structures. The plasma membrane and the membranes surrounding organelles and vesicles are composed of phosphoglycerides, fatty acids, cholesterol, phospholipids, glycolipids, proteoglycans, and proteins. These components confer identity and functionality to the membranes with which they associate.  
       [0497] Integral Membrane Proteins  
       [0498] 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 α-helical conformation. TM proteins are classified as bitopic (Types I and I) and polytopic (Types III and IV) (Singer, S. J. (1990) Annu. Rev. Cell Biol. 6:247-296). Bitopic proteins span the membrane once while polytopic proteins contain multiple membrane-spanning segments. TM proteins function as cell-surface receptors, receptor-interacting proteins, transporters of ions or metabolites, ion channels, cell anchoring proteins, and cell type-specific surface antigens.  
       [0499] 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, ROD, NOR, 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 targeted cancer treatment of 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), that mediate interactions with extracellular or intracellular molecules.  
       [0500] G-Protein Coupled Receptors  
       [0501] G-protein coupled receptors (GPCR) are 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 cytoplasmic C-terminus. Three extracellular loops alternate with three intracellular loops to link the seven transmembrane regions. Cysteine disulfide bridges connect the second and third extracellular loops. The most conserved regions of GPCRs are the transmembrane regions and the first two cytoplasmic 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 Receptor Facts Book , Academic Press, San Diego Calif., pp. 2-6). Mutations and changes in transcriptional activation of GPCR-encoding genes have been associated with neurological disorders such as schizophrenia, Parkinson&#39;s disease, Alzheimer&#39;s disease, drug addiction, and feeding disorders.  
       [0502] Scavenger Receptors  
       [0503] Macrophage scavenger receptors with broad ligand specificity may participate in the binding of low density lipoproteins (LDL) and foreign antigens. Scavenger receptors types I and II are trimeric membrane proteins with each subunit containing a small N-terminal intracellular domain, a transmembrane domain, a large extracellular domain, and a C-terminal cysteine-rich domain. The extracellular domain contains a short spacer region, an α-helical coiled-coil region, and a triple helical collagen-like region. These receptors have been shown to bind a spectrum of ligands, including chemically modified lipoproteins and albumin, polyribonucleotides, polysaccharides, phospholipids, and asbestos (Matsumoto, A. et al. (1990) Proc. Natl. Acad. Sci. USA 87:9133-9137; and Elomaa, 0. et al. (1995) Cell 80:603-609). The scavenger receptors are thought to play a key role in atherogenesis by mediating uptake of modified LDL in arterial walls, and in host defense by binding bacterial endotoxins, bacteria, and protozoa  
       [0504] Tetraspan Family Proteins  
       [0505] The transmembrane 4 superfamily (TM4SF) or tetraspan family is a multigene family encoding type III integral membrane proteins (Wright, AD. and M. G. Tomlinson (1994) Immunol.  
       [0506] Today 15:588-594). The TM4SF is comprised of membrane proteins which traverse the cell membrane four times. Members of the TM4SF include platelet and endothelial cell membrane proteins, melanoma-associated antigens, leukocyte surface glycoproteins, colonal carcinoma antigens, tumor-associated antigens, and surface proteins of the schistosome parasites (Jankowski, S. A. (1994) Oncogene 9:1205-1121). Members of the TM4SF share about 25-30% amino acid sequence identity with one another.  
       [0507] A number of TM4SF members have been implicated in signal transduction, control of cell adhesion, regulation of cell growth and proliferation, including development and oncogenesis, and cell motility, including tumor cell metastasis. Expression of TM4SF proteins is associated with a variety of tumors and the level of expression may be altered when cells are growing or activated.  
       [0508] Tumor Antigens  
       [0509] Tumor antigens are cell surface molecules that are differentially expressed in tumor cells relative to normal cells. Tumor antigens distinguish tumor cells immunologically from normal cells and provide diagnostic and therapeutic targets for human cancers (Takagi, S. et al. (1995) Int. J. Cancer 61:706-715; Liu, E. et al. (1992) Oncogene 7:1027-1032).  
       [0510] Leukocyte Antigens  
       [0511] Other types of cell surface antigens include those identified on leukocytic cells of the immune system. These antigens have been identified using systematic, monoclonal antibody (mAb)-based “shot gun” techniques. These techniques have resulted in the production of hundreds of mAbs directed against unknown cell surface leukocytic antigens. These antigens have been grouped into “clusters of differentiation” based on common immunocytochemical localization patterns in various differentiated and undifferentiated leukocytic cell types. Antigens in a given cluster are presumed to identify a single cell surface protein and are assigned a “cluster of differentiation” or “CD” designation. Some of the genes encoding proteins identified by CD antigens have been cloned and verified by standard molecular biology techniques. CD antigens have been characterized as both transmembrane proteins and cell surface proteins anchored to the plasma membrane via covalent attachment to fatty acid-containing glycolipids such as glycosylphosphatidylinositol (GPI). (Reviewed in Barclay, A. N. et al. (1995)  The Leucocyte Antigen Facts Book , Academic Press, San Diego Calif., pp. 17-20.)  
       [0512] Ion Channels  
       [0513] Ion channels are found in the plasma membranes of virtually every cell in the body. For example, chloride channels mediate a variety of cellular functions including regulation of membrane potentials and absorption and secretion of ions across epithelial membranes. Chloride channels also regulate the pH of organelles such as the Golgi apparatus and endosomes (see, e.g., Greger, R. (1988)  
       [0514] Annu. Rev. Physiol. 50:111-122). Electrophysiological and pharmacological properties of chloride channels, including ion conductance, current-voltage relationships, and sensitivity to modulators, suggest that different chloride channels exist in muscles, neurons, fibroblasts, epithelial cells, and lymphocytes.  
       [0515] Many ion channels have sites for phosphorylation by one or more protein kinases including protein kinase A, protein kinase C, tyrosine kinase, and casein kinase II, all of which regulate ion channel activity in cells. Inappropriate phosphorylation of proteins in cells has been linked to changes in cell cycle progression and cell differentiation. Changes in the cell cycle have been linked to induction of apoptosis or cancer. Changes in cell differentiation have been linked to diseases and disorders of the reproductive system, immune system, skeletal muscle, and other organ systems.  
       [0516] Proton Pumps  
       [0517] Proton ATPases comprise a large class of membrane proteins that use the energy of ATP hydrolysis to generate an electrochemical proton gradient across a membrane. The resultant gradient may be used to transport other ions across the membrane (Na + , K + , or Cl − ) or to maintain organelle pH. Proton ATPases are further subdivided into the mitochondrial F-ATPases, the plasma membrane ATPases, and the vacuolar ATPases. The vacuolar ATPases establish and maintain an acidic pH within various organelles involved in the processes of endocytosis and exocytosis (Mellman, I. et al. (1986) Annu. Rev. Biochem 55:663-700).  
       [0518] Proton-coupled, 12 membrane-spanning domain transporters such as PEPT 1 and PEPT 2 are responsible for gastrointestinal absorption and for renal reabsorption of peptides using an electrochemical H +  gradient as the driving force. Another type of peptide transporter, the TAP transporter, is a heterodimer consisting of TAP 1 and TAP 2 and is associated with antigen processing. Peptide antigens are transported across the membrane of the endoplasmic reticulum by TAP so they can be expressed on the cell surface in association with MHC molecules. Each TAP protein consists of multiple hydrophobic membrane spanning segments and a highly conserved ATP-binding cassette (Boll, M. et al. (1996) Proc. Natl. Acad. Sci. USA  93:284-289 ). Pathogenic microorganisms, such as herpes simplex virus, may encode inhibitors of TAP-mediated peptide transport in order to evade immune surveillance (Marusina, K and J. J. Manaco (1996) Curr. Opin.  
       [0519] Hematol. 3:19-26).  
       [0520] ABC Transporters  
       [0521] The ATP-binding cassette (ABC) transporters, also called the “traffic ATPases”, comprise a superfamily of membrane proteins that mediate transport and channel functions in prokaryotes and eukaryotes (Higgins, C. F. (1992) Annu. Rev. Cell Biol. 8:67-113). ABC proteins share a similar overall structure and significant sequence homology. All ABC proteins contain a conserved domain of approximately two hundred amino acid residues which includes one or more nucleotide binding domains. Mutations in ABC transporter genes are associated with various disorders, such as hyperbilirubinemia II/Dubin-Johnson syndrome, recessive Stargardt&#39;s disease, X-linked adrenoleukodystrophy, multidrug resistance, celiac disease, and cystic fibrosis.  
       [0522] Peripheral and Anchored Membrane Proteins  
       [0523] Some membrane proteins are not membrane-spanning but are attached to the plasma membrane via membrane anchors or interactions with integral membrane proteins. Membrane anchors are covalently joined to a protein post-translationally and include such moieties as prenyl, myristyl, and glycosylphosphatidyl inositol groups. Membrane localization of peripheral and anchored proteins is important for their function in processes such as receptor-mediated signal transduction. For example, prenylation of Ras is required for its localization to the plasma membrane and for its normal and oncogenic functions in signal transduction.  
       [0524] Vesicle Coat Proteins  
       [0525] Intercellular communication is essential for the development and survival of multicellular organisms. Cells communicate with one another through the secretion and uptake of protein signaling molecules. The uptake of proteins into the cell is achieved by the endocytic pathway, in which the interaction of extracellular signaling molecules with plasma membrane receptors results in the formation of plasma membrane-derived vesicles that enclose and transport the molecules into the cytosol. These transport vesicles fuse with and mature into endosomal and lysosomal (digestive) compartments. The secretion of proteins from the cell is achieved by exocytosis, in which molecules inside of the cell proceed through the secretory pathway. In this pathway, molecules transit from the ER to the Golgi apparatus and finally to the plasma membrane, where they are secreted from the cell.  
       [0526] Several steps in the transit of material along the secretory and endocytic pathways require the formation of transport vesicles. Specifically, vesicles form at the transitional endoplasmic reticulum (tER), the rim of Golgi cisternae, the face of the Trans-Golgi Network (TGN), the plasma membrane (M), and tubular extensions of the endosomes. Vesicle formation occurs when a region of membrane buds off from the donor organelle. The membrane-bound vesicle contains proteins to be transported and is surrounded by a proteinaceous coat, the components of which are recruited from the cytosol. Two different classes of coat protein have been identified. Clathrin coats form on vesicles derived from the TGN and PM, whereas coatomer (COP) coats form on vesicles derived from the ER and Golgi. COP coats can be further classified as COPI, involved in retrograde traffic through the Golgi and from the Golgi to the ER, and COPII, involved in anterograde traffic from the ER to the Golgi (Mellman supra).  
       [0527] In clathrin-based vesicle formation, adapter proteins bring vesicle cargo and coat proteins together at the surface of the budding membrane. Adapter protein-1 and -2 select cargo from the TGN and plasma membrane, respectively, based on molecular information encoded on the cytoplasmic tail of integral membrane cargo proteins. Adapter proteins also recruit clathrin to the bud site. Clathrin is a protein complex consisting of three large and three small polypeptide chains arranged in a three-legged structure called a triskelion. Multiple triskelions and other coat proteins appear to self-assemble on the membrane to form a coated pit. This assembly process may serve to deform the membrane into a budding vesicle. GTP-bound ADP-ribosylation factor (Arf) is also incorporated into the coated assembly. Another small G-protein, dynamin, forms a ring complex around the neck of the forming vesicle and may provide the mechanochemical force to seal the bud, thereby releasing the vesicle. The coated vesicle complex is then transported through the cytosol. During the transport process, Arf-bound GTP is hydrolyzed to GDP, and the coat dissociates from the transport vesicle (West, M. A. et al. (1997) J. Cell Biol. 138:1239-1254).  
       [0528] Vesicles which bud from the ER and the Golgi are covered with a protein coat similar to the clathrin coat of endocytic and TGN vesicles. The coat protein (COP) is assembled from cytosolic precursor molecules at specific budding regions on the organelle. The COP coat consists of two major components, a G-protein (Arf or Sar) and coat protomer (coatomer). Coatomer is an equimolar complex of seven proteins, termed alpha-, beta-, beta′-, gamma-, delta-, epsilon- and zeta-COP. The coatomer complex binds to dilysine motifs contained on the cytoplasmic tails of integral membrane proteins. These include the KKXX retrieval motif of membrane proteins of the ER and dibasic/diphenylamine motifs of members of the p24 family. The p24 family of type I membrane proteins represent the major membrane proteins of COPI vesicles (Harter, C. and F. T. Wieland (1998) Proc. Natl. Acad. Sci. USA 95:11649-11654).  
       [0529] Organelle Associated Molecules  
       [0530] Eukaryotic cells are organized into various cellular organelles which has the effect of separating specific molecules and their functions from one another and from the cytosol. Within the cell, various membrane structures surround and define these organelles while allowing them to interact with one another and the cell environment through both active and passive transport processes. Important cell organelles include the nucleus, the Golgi apparatus, the endoplasmic reticulum, mitochondria, peroxisomes, lysosomes, endosomes, and secretory vesicles.  
       [0531] Nucleus  
       [0532] The cell nucleus contains all of the genetic information of the cell in the form of DNA, and the components and machinery necessary for replication of DNA and for transcription of DNA into RNA. (See Alberts, B. et al. (1994)  Molecular Biology of the Cell , Garland Publishing Inc., New York N.Y., pp. 335-399.) DNA is organized into compact structures in the nucleus by interactions with various DNA-binding proteins such as histones and non-histone chromosomal proteins. DNA-specific nucleases, DNAses, partially degrade these compacted structures prior to DNA replication or transcription. DNA replication takes place with the aid of DNA helicases which unwind the double-stranded DNA helix, and DNA polymerases that duplicate the separated DNA strands.  
       [0533] Transcriptional regulatory proteins are essential for the control of gene expression. Some of these proteins function as transcription factors that initiate, activate, repress, or terminate gene transcription. Transcription factors generally bind to the promoter, enhancer, and upstream regulatory regions of a gene in a sequence-specific manner, although some factors bind regulatory elements within or downstream of a gene&#39;s coding region. Transcription factors may bind to a specific region of DNA singly or as a complex with other accessory factors. (Reviewed in Lewin, B. (1990)  Genes IV , Oxford University Press, New York N.Y., and Cell Press, Cambridge Mass., pp. 554570.) Many transcription factors incorporate DNA-binding structural motifs which comprise either α helices or β sheets that bind to the major groove of DNA. Four well-characterized structural motifs are helix-turn-helix, zinc finger, leucine zipper, and helix-loop-helix. Proteins containing these motifs may act alone as monomers, or they may form homo- or heterodimers that interact with DNA.  
       [0534] Many neoplastic disorders in humans can be attributed to inappropriate gene expression. Malignant cell growth may result from either excessive expression of tumor promoting genes or insufficient expression of tumor suppressor genes (Cleary, M. L. (1992) Cancer Surv. 15:89-104). Chromosomal translocations may also produce chimeric loci which fuse the coding sequence of one gene with the regulatory regions of a second unrelated gene. Such an arrangement likely results in inappropriate gene transcription, potentially contributing to malignancy.  
       [0535] In addition, the immune system responds to infection or trauma by activating a cascade of events that coordinate the progressive selection, amplification, and mobilization of cellular defense mechanisms. A complex and balanced program of gene activation and repression is involved in this process. However, hyperactivity of the immune system as a result of improper or insufficient regulation of gene expression may result in considerable tissue or organ damage. This damage is well documented in immunological responses associated with arthritis, allergens, heart attack, stroke, and infections (Isselbacher, K. J. et al. (1996)  Harrison&#39;s Principles of Internal Medicine,  13/e, McGraw Hill, Inc. and Teton Data Systems Software).  
       [0536] Transcription of DNA into RNA also takes place in the nucleus catalyzed by RNA polymerases. Three types of RNA polymerase exist RNA polymerase I makes large ribosomal RNAs, while RNA polymerase m makes a variety of small, stable RNAs including 5S ribosomal RNA and the transfer RNAs (tRNA). RNA polymerase II transcribes genes that will be translated into proteins. The primary transcript of RNA polymerase II is called heterogenous nuclear RNA (hnRNA), and must be further processed by splicing to remove non-coding sequences called introns. RNA splicing is mediated by small nuclear ribonucleoprotein complexes, or snRNPs, producing mature messenger RNA (mRNA) which is then transported out of the nucleus for translation into proteins.  
       [0537] Nucleolus  
       [0538] The nucleolus is a highly organized subcompartment in the nucleus that contains high concentrations of RNA and proteins and functions mainly in ribosomal RNA synthesis and assembly (Alberts, et al. supra, pp. 379-382). Ribosomal RNA (rRNA) is a structural RNA that is complexed with proteins to form ribonucleoprotein structures called ribosomes. Ribosomes provide the platform on which protein synthesis takes place.  
       [0539] Ribosomes are assembled in the nucleolus initially from a large, 45S rRNA combined with a variety of proteins imported from the cytoplasm, as well as smaller, 5S rRNAs. Later processing of the immature ribosome results in formation of smaller ribosomal subunits which are transported from the nucleolus to the cytoplasm where they are assembled into functional ribosomes.  
       [0540] Endoplasmic Reticulum  
       [0541] In eukaryotes, proteins are synthesized within the endoplasmic reticulum (ER), delivered from the ER to the Golgi apparatus for post-translational processing and sorting, and transported from the Golgi to specific intracellular and extracellular destinations. Synthesis of integral membrane proteins, secreted proteins, and proteins destined for the lumen of a particular organelle occurs on the rough endoplasmic reticulum (ER). The rough ER is so named because of the rough appearance in electron micrographs imparted by the attached ribosomes on which protein synthesis proceeds. Synthesis of proteins destined for the ER actually begins in the cytosol with the synthesis of a specific signal peptide which directs the growing polypeptide and its attached ribosome to the ER membrane where the signal peptide is removed and protein synthesis is completed. Soluble proteins destined for the ER lumen, for secretion, or for transport to the lumen of other organelles pass completely into the ER lumen. Transmembrane proteins destined for the ER or for other cell membranes are translocated across the ER membrane but remain anchored in the lipid bilayer of the membrane by one or more membrane-spanning α-helical regions.  
       [0542] Translocated polypeptide chains destined for other organelles or for secretion also fold and assemble in the ER lumen with the aid of certain “resident” ER proteins. Protein folding in the ER is aided by two principal types of protein isomerases, protein disulfide isomerase (PDI), and peptidyl-prolyl isomerase (PPI). PDI catalyzes the oxidation of free sulfhdryl groups in cysteine residues to form intramolecular disulfide bonds in proteins. PPI, an enzyme that catalyzes the isomerization of certain proline imide bonds in oligopeptides and proteins, is considered to govern one of the rate limiting steps in the folding of many proteins to their final functional conformation. The cyclophilins represent a major class of PPI that was originally identified as the major receptor for the immunosuppressive drug cyclosporin A (Handschumacher, R. E. et al. (1984) Science 226:544-547). Molecular “chaperones” such as BiP (binding protein) in the ER recognize incorrectly folded proteins as well as proteins not yet folded into their final form and bind to them, both to prevent improper aggregation between them, and to promote proper folding.  
       [0543] The “N-linked” glycosylation of most soluble secreted and membrane-bound proteins by oligosacchrides linked to asparagine residues in proteins is also performed in the ER. This reaction is catalyzed by a membrane-bound enzyme, oligosaccharyl transferase.  
       [0544] Golgi Apparatus  
       [0545] The Golgi apparatus is a complex structure that lies adjacent to the ER in eukaryotic cells and serves primarily as a sorting and dispatching station for products of the ER (Alberts, et al. supra, pp. 600-610). Additional posttranslational processing, principally additional glycosylation, also occurs in the Golgi. Indeed, the Golgi is a major site of carbohydrate synthesis, including most of the glycosaminoglycans of the extracellular matrix. N-linked oligosaccharides, added to proteins in the ER, are also further modified in the Golgi by the addition of more sugar residues to form complex N-linked oligosaccharides. “O-linked” glycosylation of proteins also occurs in the Golgi by the addition of N-acetylgalactosamine to the hydroxyl group of a serine or threonine residue followed by the sequential addition of other sugar residues to the first. This process is catalyzed by a series of glycosyltransferases each specific for a particular donor sugar nucleotide and acceptor molecule (Lodish, H. et al. (1995)  Molecular Cell Biology , W. H. Freeman and Co., New York N.Y., pp.700-708). In many cases, both N- and O-linked oligosaccharides appear to be required for the secretion of proteins or the movement of plasma membrane glycoproteins to the cell surface.  
       [0546] The terminal compartment of the Golgi is the Trans-Golgi Network (TGN), where both membrane and lumenal proteins are sorted for their final destination. Transport (or secretory) vesicles destined for intracellular compartments, such as lysosomes, bud off of the TGN. Other transport vesicles bud off containing proteins destined for the plasma membrane, such as receptors, adhesion molecules, and ion channels, and secretory proteins, such as hormones, neurotransmitters, and digestive enzymes.  
       [0547] Vacuoles  
       [0548] The vacuole system is a collection of membrane bound compartments in eukaryotic cells that functions in the processes of endocytosis and exocytosis. They include phagosomes, lysosomes, endosomes, and secretory vesicles. Endocytosis is the process in cells of internalizing nutrients, solutes or small particles (pinocytosis) or large particles such as internalized receptors, viruses, bacteria, or bacterial toxins (phagocytosis). Exocytosis is the process of transporting molecules to the cell surface. It facilitates placement or localization of membrane-bound receptors or other membrane proteins and secretion of hormones, neurotransmitters, digestive enzymes, wastes, etc.  
       [0549] A common property of all of these vacuoles is an acidic pH environment ranging from approximately pH 4.5-5.0. This acidity is maintained by the presence of a proton ATPase that uses the energy of ATP hydrolysis to generate an electrochemical proton gradient across a membrane (Mellman, I. et al. (1986) Annu. Rev. Biochem. 55:663-700). Eukaryotic vacuolar proton ATPase (vp-ATPase) is a multimeric enzyme composed of 3-10 different subunits. One of these subunits is a highly hydrophobic polypeptide of approximately 16 kDa that is similar to the proteolipid component of vp-ATPases from eubacteria, fungi, and plant vacuoles (Mandel, M. et al. (1988) Proc. Natl. Acad. Sci. USA 85:5521-5524). The 16 kDa proteolipid component is the major subunit of the membrane portion of vp-ATPase and functions in the transport of protons across the membrane.  
       [0550] Lysosomes  
       [0551] Lysosomes are membranous vesicles containing various hydrolytic enzymes used for the controlled intracellular digestion of macromolecules. Lysosomes contain some 40 types of enzymes including proteases, nucleases, glycosidases, lipases, phospholipases, phosphatases, and sulfatases, all of which are acid hydrolases that function at a pH of about 5. Lysosomes are surrounded by a unique membrane containing transport proteins that allow the final products of macromolecule degradation, such as sugars, amino acids, and nucleotides, to be transported to the cytosol where they may be either excreted or reutilized by the cell. A vp-ATPase, such as that described above, maintains the acidic environment necessary for hydrolytic activity (Alberts, supra, pp.610-611).  
       [0552] Endosomes  
       [0553] Endosomes are another type of acidic vacuole that is used to transport substances from the cell surface to the interior of the cell in the process of endocytosis. Like lysosomes, endosomes have an acidic environment provided by a vp-ATPase (Alberts et al. supra pp. 610-618). Two types of endosomes are apparent based on tracer uptake studies that distinguish their time of formation in the cell and their cellular location. Early endosomes are found near the plasma membrane and appear to function primarily in the recycling of internalized receptors back to the cell surface. Late endosomes appear later in the endocytic process close to the Golgi apparatus and the nucleus, and appear to be associated with delivery of endocytosed material to lysosomes or to the TGN where they may be recycled. Specific proteins are associated with particular transport vesicles and their target compartments that may provide selectivity in targeting vesicles to their proper compartments. A cytosolic prenylated GTP-binding protein, Rab, is one such protein. Rabs 4, 5, and 11 are associated with the early endosome, whereas Rabs 7 and 9 associate with the late endosome.  
       [0554] Mitochondria  
       [0555] Mitochondria are oval-shaped organelles comprising an outer membrane, a tightly folded inner membrane, an intermembrane space between the outer and inner membranes, and a matrix inside the inner membrane. The outer membrane contains many porin molecules that allow ions and charged molecules to enter the intermembrane space, while the inner membrane contains a variety of transport proteins that transfer only selected molecules. Mitochondria are the primary sites of energy production in cells.  
       [0556] Energy is produced by the oxidation of glucose and fatty acids. Glucose is initially converted to pyruvate in the cytoplasm. Fatty acids and pyruvate are transported to the mitochondria for complete oxidation to CO 2  coupled by enzymes to the transport of electrons from NADH and FADH 2  to oxygen and to the synthesis of ATP (oxidative phosphorylation) from ADP and P i , Pyruvate is transported into the mitochondria and converted to acetyl-CoA for oxidation via the citric acid cycle, involving pyruvate dehydrogenase components, dihydrolipoyl transacetylase, and dihydrolipoyl dehydrogenase. Enzymes involved in the citric acid cycle include: citrate synthetase, aconitases, isocitrate dehydrogenase, alpha-ketoglutarate dehydrogenase complex including transsuccinylases, succinyl CoA synthetase, succinate dehydrogenase, fumarases, and malate dehydrogenase. Acetyl CoA is oxidized to CO 2  with concomitant formation of NADH, FADH 2 , and GTP. In oxidative phosphorylation, the transfer of electrons from NADH and FADH 2  to oxygen by dehydrogenases is coupled to the synthesis of ATP from ADP and P i  by the F 0 F 1  ATPase complex in the mitochondrial inner membrane. Enzyme complexes responsible for electron transport and ATP synthesis include the F 0 F 1  ATPase complex, ubiquinone(CoQ)-cytochrome c reductase, ubiquinone reductase, cytochrome b, cytochrome c 1 , FeS protein, and cytochrome c oxidase.  
       [0557] Peroxisomes  
       [0558] Peroxisomes, like mitochondria, are a major site of oxygen utilization. They contain one or more enzymes, such as catalase and urate oxidase, that use molecular oxygen to remove hydrogen atoms from specific organic substrates in an oxidative reaction that produces hydrogen peroxide (Alberts, supra, pp. 574577). Catalase oxidizes a variety of substrates including phenols, formic acid, formaldehyde, and alcohol and is important in peroxisomes of liver and kidney cells for detoxifying various toxic molecules that enter the bloodstream. Another major function of oxidative reactions in peroxisomes is the breakdown of fatty acids in a process called β oxidation. β oxidation results in shortening of the alkyl chain of fatty acids by blocks of two carbon atoms that are converted to acetyl CoA and exported to the cytosol for reuse in biosynthetic reactions.  
       [0559] Also like mitochondria, peroxisomes import their proteins from the cytosol using a specific signal sequence located near the C-terminus of the protein. The importance of this import process is evident in the inherited human disease Zellweger syndrome, in which a defect in importing proteins into perixosomes leads to a perixosomal deficiency resulting in severe abnormalities in the brain liver, and kidneys, and death soon after birth. One form of this disease has been shown to be due to a mutation in the gene encoding a perixosomal integral membrane protein called peroxisome assembly factor-1.  
       [0560] The discovery of new human molecules satisfies a need in the art by providing new compositions which are useful in the diagnosis, study, prevention, and treatment of diseases associated with, as well as effects of exogenous compounds on, the expression of human molecules.  
       SUMMARY OF THE INVENTION  
       [0561] The present invention relates to nucleic acid sequences comprising human diagnostic and therapeutic polynucleotides (dithp) as presented in the Sequence Listing. The dithp uniquely identify genes encoding human structural, functional, and regulatory molecules.  
       [0562] The invention 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 NO:1-211; b) a naturally occurring polynucleotide sequence having at least 90% sequence identity to a polynucleotide sequence selected from the group consisting of SEQ ID NO:1-211; c) a polynucleotide sequence complementary to a); d) a polynucleotide sequence complementary to b); and e) an RNA equivalent of a) through d). In one alternative, the polynucleotide comprises a polynucleotide sequence selected from the group consisting of SEQ ID NO:1-211. In another alternative, the polynucleotide comprises at least 60 contiguous nucleotides of a polynucleotide sequence selected from the group consisting of a) a polynucleotide sequence selected from the group consisting of SEQ ID NO: 1-211; b) a naturally occurring polynucleotide sequence having at least 90% sequence identity to a polynucleotide sequence selected from the group consisting of SEQ ID NO:1-211; c) a polynucleotide sequence complementary to a); d) a polynucleotide sequence complementary to b); and e) an RNA equivalent of a) through d). The invention further provides a composition for the detection of expression of human diagnostic and therapeutic polynucleotides comprising at least one 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: 1-211; b) a naturally occurring polynucleotide sequence having at least 90% sequence identity to a polynucleotide sequence selected from the group consisting of SEQ ID NO:1-211; c) a polynucleotide sequence complementary to a); d) a polynucleotide sequence complementary to b); and e) an RNA equivalent of a) through d); and a detectable label.  
       [0563] The invention also provides a method for detecting a target polynucleotide in a sample, said target polynucleotide comprising a polynucleotide sequence selected from the group consisting of a) a polynucleotide sequence selected from the group consisting of SEQ ID NO:1-211; b) a naturally occurring polynucleotide sequence having at least 90% sequence identity to a polynucleotide sequence selected from the group consisting of SEQ ID NO:1-211; c) a polynucleotide sequence complementary to a); d) a polynucleotide sequence complementary to b); and e) an RNA equivalent of a) through d). The method comprises a) amplifying said target polynucleotide or a 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.  
       [0564] The invention also provides a method for detecting a target polynucleotide in a sample, said target polynucleotide comprising a polynucleotide sequence selected from the group consisting of a) a polynucleotide sequence selected from the group consisting of SEQ ID NO:1-211; b) a naturally occurring polynucleotide sequence having at least 90% sequence identity to a polynucleotide sequence selected from the group consisting of SEQ ID NO:1-211; c) a polynucleotide sequence complementary to a); d) a polynucleotide sequence complementary to b); and e) an RNA equivalent of a) through 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, 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 30 contiguous nucleotides. In another alternative, the probe comprises at least 60 contiguous nucleotides.  
       [0565] The invention further provides a recombinant polynucleotide comprising a promoter sequence operably linked to 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:1-211; b) a naturally occurring polynucleotide sequence having at least 90% sequence identity to a polynucleotide sequence selected from the group consisting of SEQ ID NO:1-211; c) a polynucleotide sequence complementary to a); d) a polynucleotide sequence complementary to b); and e) an RNA equivalent of a) through d). 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. In a further alternative, the invention provides a method for producing a human diagnostic and therapeutic polypeptide, the method comprising a) culturing a cell under conditions suitable for expression of the human diagnostic and therapeutic polypeptide, wherein said cell is transformed with the recombinant polynucleotide, and b) recovering the human diagnostic and therapeutic polypeptide so expressed.  
       [0566] The invention also provides a purified human diagnostic and therapeutic polypeptide (DITHP) encoded by at least one polynucleotide comprising a polynucleotide sequence selected from the group consisting of SEQ ID NO:1-211. Additionally, the invention provides an isolated antibody which specifically binds to the human diagnostic and therapeutic polypeptide. The invention further provides a method of identifying a test compound which specifically binds to the human diagnostic and therapeutic polypeptide, the method comprising the steps of a) providing a test compound; b) combining the human diagnostic and therapeutic polypeptide with the test compound for a sufficient time and under suitable conditions for binding; and c) detecting binding of the human diagnostic and therapeutic polypeptide to the test compound, thereby identifying the test compound which specifically binds the human diagnostic and therapeutic polypeptide.  
       [0567] The invention further provides a microarray wherein at least one element of the microarray is an isolated polynucleotide comprising at least 60 contiguous nucleotides 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 NO:1-211; b) a naturally occurring polynucleotide sequence having at least 90% sequence identity to a polynucleotide sequence selected from the group consisting of SEQ ID NO:1-211; c) a polynucleotide sequence complementary to a); d) a polynucleotide sequence complementary to b); and e) an RNA equivalent of a) through d). The invention also provides a method for generating a transcript image of a sample which contains polynucleotides. The method comprises a) labeling the polynucleotides of the sample, b) contacting the elements of the microarray with the labeled polynucleotides of the sample under conditions suitable for the formation of a hybridization complex, and c) quantifying the expression of the polynucleotides in the sample.  
       [0568] Additionally, the invention provides a method for screening a compound for effectiveness in altering expression of a target polynucleotide, wherein said target polynucleotide comprises a polynucleotide sequence selected from the group consisting of a) a polynucleotide sequence selected from the group consisting of SEQ ID NO:1-211; b) a naturally occurring polynucleotide sequence having at least 90% sequence identity to a polynucleotide sequence selected from the group consisting of SEQ ID NO:1-211; c) a polynucleotide sequence complementary to a); d) a polynucleotide sequence complementary to b); and e) an RNA equivalent of a) through d). The method comprises a) exposing a sample comprising the target polynucleotide to a compound, and b) detecting altered expression of the target polynucleotide, and c) comparing the expression of the target polynucleotide in the presence of varying amounts of the compound and in the absence of the compound  
       [0569] The invention further provides a method for assessing toxicity of a test compound, said method comprising a) treating a biological sample containing nucleic acids with the test compound; b) hybridizing the nucleic acids of the treated biological sample with a probe comprising at least 20 contiguous nucleotides of a polynucleotide comprising a polynucleotide sequence selected from the group consisting of i) a polynucleotide sequence selected from the group consisting of SEQ ID NO:1-211; ii) a naturally occurring polynucleotide sequence having at least 90% sequence identity to a polynucleotide sequence selected from the group consisting of SEQ ID NO:1-211; iii) a polynucleotide sequence complementary to i), iv) a polynucleotide sequence complementary to ii), and v) an RNA equivalent of i)-iv). Hybridization occurs under conditions whereby a specific hybridization complex is formed between said probe and a target polynucleotide in the biological sample, said target polynucleotide comprising a polynucleotide sequence selected from the group consisting of i) a polynucleotide sequence selected from the group consisting of SEQ ID NO:1-211; ii) a naturally occurring polynucleotide sequence having at least 90% sequence identity to a polynucleotide sequence selected from the group consisting of SEQ ID NO:1-211; iii) a polynucleotide sequence complementary to i), iv) a polynucleotide sequence complementary to ii), and v) an RNA equivalent of i)-iv), and alternatively, the target polynucleotide comprises a fragment of a polynucleotide sequence selected from the group consisting of i-v above; c) quantifying the amount of hybridization complex; and d) comparing the amount of hybridization complex in the treated biological sample with the amount. of hybridization complex in an untreated biological sample, wherein a difference in the amount of hybridization complex in the treated biological sample is indicative of toxicity of the test compound. The invention further 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: 212-422 , 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: 212-422 , c) a biologically active fragment of an amino acid sequence selected from the group consisting of SEQ ID NO: 212-422 , and d) an inmmunogenic fragment of an amino acid sequence selected from the group consisting of SEQ ID NO: 212-422 . In one alternative, the invention provides an isolated polypeptide comprising the amino acid sequence of SEQ ID NO: 212-422.    
       DESCRIPTION OF THE TABLES  
       [0570] Table 1 shows the sequence identification numbers (SEQ ID NO:s) and template identification numbers (template IDs) corresponding to the polynucleotides of the present invention, along with their GenBank hits (GI Numbers), probability scores, and functional annotations corresponding to the GenBank hits.  
       [0571] Table 2 shows the sequence identification numbers (SEQ ID NO:s) and template identification numbers (template IDs) corresponding to the polynucleotides of the present invention, along with polynucleotide segments of each template sequence as defined by the indicated “start” and “stop” nucleotide positions. The reading frames of the polynucleotide segments and the Pfam bits, Pfam descriptions, and E-values corresponding to the polypeptide domains encoded by the polynucleotide segments are indicated.  
       [0572] Table 3 shows the sequence identification numbers (SEQ ID NO:s) and template identification numbers (template IDs) corresponding to the polynucleotides of the present invention, along with polynucleotide segments of each template sequence as defined by the indicated “start and “stop” nucleotide positions. The reading frames of the polynucleotide segments are shown, and the polypeptides encoded by the polynucleotide segments constitute either signal peptide (SP) or transmembrane (TM) domains, as indicated. The membrane topology of the encoded polypeptide sequence is indicated, the N-terminus (N) listed as being oriented to either the cytosolic (in) or non-cytosolic (out) side of the cell membrane or organelle.  
       [0573] Table 4 shows the sequence identification numbers (SEQ ID NO:s) corresponding to the polynucleotides of the present invention, along with component sequence identification numbers (component IDs) corresponding to each template. The component sequences, which were used to assemble the template sequences, are defined by the indicated “start” and “stop” nucleotide positions along each template.  
       [0574] Table 5 shows the tissue distribution profiles for the templates of the invention Table 6 shows the sequence identification numbers (SEQ ID NO:s) corresponding to the polypeptides of the present invention, along with the reading frames used to obtain the polypeptide segments, the lengths of the polypeptide segments, the “start” and “stop” nucleotide positions of the polynucleotide sequences used to define the encoded polypeptide segments, the GenBank hits (GI Numbers), probability scores, and functional annotations corresponding to the GenBank hits.  
       [0575] Table 7 summarizes the bioinformatics tools which are useful for analysis of the polynucleotides of the present invention. The first column of Table 7 lists analytical tools, programs, and algorithms, 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).  
       DETAILED DESCRIPTION OF THE INVENTION  
       [0576] Before the nucleic acid sequences and methods are presented, it is to be understood that this invention is not limited to the particular machines, methods, and materials described. Although particular embodiments are described, machines, methods, and materials similar or equivalent to these embodiments may be used to practice the invention. The preferred machines, methods, and materials set forth are not intended to limit the scope of the invention which is limited only by the appended claims.  
       [0577] The singular forms “a”, “an”, and “the” include plural reference unless the context clearly dictates otherwise. AR technical and scientific terms have the meanings commonly understood by one of ordinary skill in the art. All publications are incorporated by reference for the purpose of describing and disclosing the cell lines, vectors, and methodologies which are presented and which might be used in connection with the invention. Nothing in the specification is to be construed as an admission that the invention is not entitled to antedate such disclosure by virtue of prior invention.  
       [0578] Definitions  
       [0579] As used herein, the lower case “ditmp” refers to a nucleic acid sequence, while the upper case “DrnHp” refers to an amino acid sequence encoded by dithp. A “full-length” dithp refers to a nucleic acid sequence containing the entire coding region of a gene endogenously expressed in human tissue.  
       [0580] “Adjuvants” are materials such as Freund&#39;s adjuvant, mineral gels (aluminum hydroxide), and surface active substances (lysolecithin, pluronic polyols, polyanions, peptides, oil emulsions, keyhole limpet hemocyanin, and dinitrophenol) which may be administered to increase a host&#39;s immunological response.  
       [0581] “Allele” refers to an alternative form of a nucleic acid sequence. Alleles result from a “mutation,” a change or an alternative reading of the genetic code. Any given gene may have none, one, or many allelic forms. Mutations which give rise to alleles include deletions, additions, or substitutions of nucleotides. Each of these changes may occur alone, or in combination with the others, one or more times in a given nucleic acid sequence. The present invention encompasses allelic dithp.  
       [0582] “Amino acid sequence” refers to a peptide, a polypeptide, or a protein of either natural or synthetic origin. The amino acid sequence is not limited to the complete, endogenous amino acid sequence and may be a fragment, epitope, variant, or derivative of a protein expressed by a nucleic acid sequence.  
       [0583] “Amplification” refers to the production of additional copies of a sequence and is carried out using polymerase chain reaction (PCR) technologies well known in the art.  
       [0584] “Antibody” refers to intact molecules as well as to fragments thereof, such as Fab, F(ab′) 2 , and Fv fragments, which are capable of binding the epitopic determine. Antibodies that bind DITHP polypeptides can be prepared using intact polypeptides or using fragments containing small peptides of interest as the immunizing antigen. The polypeptide or peptide 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.  
       [0585] “Antisense sequence” refers to a sequence capable of specifically hybridizing to a target sequence. The antisense sequence may include DNA, RNA, or any nucleic acid mimic or analog such as 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.  
       [0586] “Antisense sequence” refers to a sequence capable of specifically hybridizing to a target sequence. The antisense sequence can be DNA, RNA, or any nucleic acid mimic or analog.  
       [0587] “Antisense technology” refers to any technology which relies on the specific hybridization of an antisense sequence to a target sequence.  
       [0588] A “bin” is a portion of computer memory space used by a computer program for storage of data, and bounded in such a manner that data stored in a bin may be retrieved by the program.  
       [0589] “Biologically active” refers to an amino acid sequence having a structural, regulatory, or biochemical function of a naturally occurring amino acid sequence.  
       [0590] “Clone joining” is a process for combining gene bins based upon the bins&#39; containing sequence information from the same clone. The sequences may assemble into a primary gene transcript as well as one or more splice variants.  
       [0591] “Complementary” describes the relationship between two single-stranded nucleic acid sequences that anneal by base-pairing (5′-A-G-T-3′ pairs with its complement 3′-T-C-A-5).  
       [0592] A “component sequence” is a nucleic acid sequence selected by a computer program such as PHRED and used to assemble a consensus or template sequence from one or more component sequences.  
       [0593] A “consensus sequence” or “template sequence” is a nucleic acid sequence which has been assembled from overlapping sequences, using a computer program for fragment assembly such as the GEL VIEW fragment assembly system (Genetics Computer Group (GCG), Madison Wis.) or using a relational database management system (RDMS).  
       [0594] “Conservative amino acid substitutions” are those substitutions that, when made, 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 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                      
 
       [0595] Conservative 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 target site, or (c) the bulk of the side chain  
       [0596] “Deletion” refers to a change in either a nucleic or amino acid sequence in which at least one nucleotide or amino acid residue, respectively, is absent  
       [0597] “Derivative” refers to the chemical modification of a nucleic acid sequence, such as by replacement of hydrogen by an alkyl, acyl, amino, hydroxyl, or other group.  
       [0598] The terms “element” and “array element” refer to a polynucleotide, polypeptide, or other chemical compound having a unique and defined position on a microarray.  
       [0599] “E-value” refers to the statistical probability that a match between two sequences occurred by chance.  
       [0600] A “fragment” is a unique portion of dithp or DITHP 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 10 to 1000 contiguous amino acid residues or nucleotides. 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 amino acid residues or nucleotides 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 and the figures, may be encompassed by the present embodiments.  
       [0601] A fragment of dithp comprises a region of unique polynucleotide sequence that specifically identifies dithp, for example, as distinct from any other sequence in the same genome. A fragment of dithp is useful, for example, in hybridization and amplification technologies and in analogous methods that distinguish dithp from related polynucleotide sequences. The precise length of a fragment of dithp and the region of dithp to which the fragment corresponds are routinely determinable by one of ordinary skill in the art based on the intended purpose for the fragment  
       [0602] A fragment of DITHP is encoded by a fragment of dithp. A fragment of DITHP comprises a region of unique amino acid sequence that specifically identifies DITHP. For example, a fragment of DITHP is useful as an immunogenic peptide for the development of antibodies that specifically recognize DITHP. The precise length of a fragment of DITHP and the region of DITHP to which the fragment corresponds are routinely determinable by one of ordinary skill in the art based on the intended purpose for the fragment.  
       [0603] A “full length” nucleotide sequence is one containing at least a start site for translation to a protein sequence, followed by an open reading frame and a stop site, and encoding a “full length” polypeptide.  
       [0604] “Hit” refers to a sequence whose annotation will be used to describe a given template. Criteria for selecting the top hit are as follows: if the template has one or more exact nucleic acid matches, the top hit is the exact match with highest percent identity. If the template has no exact matches but has significant protein hits, the top hit is the protein hit with the lowest E-value. If the template has no significant protein hits, but does have significant non-exact nucleotide hits, the top hit is the nucleotide hit with the lowest E-value.  
       [0605] “Homology” refers to sequence similarity either between a reference nucleic acid sequence and at least a fragment of a dithp or between a reference amino acid sequence and a fragment of a DITHP.  
       [0606] “Hybridization” refers to the process by which a strand of nucleotides anneals with a complementary strand through base pairing. Specific hybridization is an indication that two nucleic acid sequences share a high degree of identity. Specific hybridization complexes form under defined annealing conditions, and remain hybridized after the “washing” step. The defined hybridization conditions include the annealing conditions and the washing step(s), the latter of which 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 probes that are not perfectly matched. Permissive conditions for annealing of nucleic acid sequences are routinely determinable and may be consistent among hybridization experiments, whereas wash conditions may be varied among experiments to achieve the desired stringency.  
       [0607] Generally, stringency of hybridization is expressed with reference to the temperature under which the wash step is carried out. Generally, such wash temperatures are selected to be about 5° C. to 20° C. lower than the thermal melting point (T m ) for the specific sequence at a defined ionic strength and pH. The T m  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 T m  and conditions for nucleic acid hybridization is well known and can be found in Sambrook et al., 1989,  Molecular Cloning: A Laboratory Manual,  2 nd  ed., vol. 1-3, Cold Spring Harbor Press, Plainview N.Y.; specifically see volume 2, chapter 9.  
       [0608] High stringency conditions for hybridization between polynucleotides of the present invention include wash conditions of 68° C. in the presence of about 0.2×SSC and about 0.1% SDS, for 1 hour. Alternatively, temperatures of about 65° C., 60° C., or 55° C. may be used. SSC concentration may be varied from about 0.2 to 2×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, denatured salmon sperm DNA at about 100-200 μg/ml. Useful variations on these conditions will be readily apparent to those skilled 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 resultant proteins.  
       [0609] Other parameters, such as temperature, salt concentration; and detergent concentration may be varied to achieve the desired stringency. Denaturants, such as formamide at a concentration of about 35-50% v/v, may also be used under particular circumstanes, such as RNA:DNA hybridizations. Appropriate hybridization conditions are routinely determinable by one of ordinary skill in the art.  
       [0610] “Immunogenic” describes the potential for a natural, recombinant, or synthetic peptide, epitope, polypeptide, or protein to induce antibody production in appropriate animals, cells, or cell lines.  
       [0611] “Insertion” or “addition” refers to a change in either a nucleic or amino acid sequence in which at least one nucleotide or residue, respectively, is added to the sequence.  
       [0612] “Labeling” refers to the covalent or noncovalent joining of a polynucleotide, polypeptide, or antibody with a reporter molecule capable of producing a detectable or measurable signal.  
       [0613] Microarray” is any arrangement of nucleic acids, amino acids, antibodies, etc., on a substrate. The substrate may be a solid support such as beads, glass, paper, nitrocellulose, nylon, or an appropriate membrane.  
       [0614] “Linkers” are short stretches of nucleotide sequence which may be added to a vector or a dithp to create restriction endonuclease sites to facilitate cloning. “Polylinkers” are engineered to incorporate multiple restriction enzyme sites and to provide for the use of enzymes which leave 5′ or 3′ overhangs (e.g., BamHI, EcoRI, and HindIII) and those which provide blunt ends (e.g., EcoRV, SnaBI, and StuI).  
       [0615] “Naturally occurring” refers to an endogenous polynucleotide or polypeptide that may be isolated from viruses or prokaryotic or eukaryotic cells.  
       [0616] “Nucleic acid sequence” refers to the specific order of nucleotides joined by phosphodiester bonds in a linear, polymeric arrangement. Depending on the number of nucleotides, the nucleic acid sequence can be considered an oligomer, oligonucleotide, or polynucleotide. The nucleic acid can be DNA, RNA, or any nucleic acid analog, such as PNA, may be of genomic or synthetic origin, may be either double-stranded or single-stranded, and can represent either the sense or antisense (complementary) strand.  
       [0617] “Oligomer” refers to a nucleic acid sequence of at least about 6 nucleotides and as many as about 60 nucleotides, preferably about 15 to 40 nucleotides, and most preferably between about 20 and 30 nucleotides, that may be used in hybridization or amplification technologies. Oligomers may be used as, e.g., primers for PCR, and are usually chemically synthesized.  
       [0618] “Operably “inked” refers to the situation in which a first nucleic acid sequence is placed in a functional relationship with the second nucleic acid sequence. For instance, a promoter is operably lined to a coding sequence if the promoter affects the transcription or expression of the coding sequence. Generally, 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.  
       [0619] “Peptide nucleic acid” (PNA) refers to a DNA mimic in which nucleotide bases are attached to a pseudopeptide backbone to increase stability. PNAs, also designated antigene agents, can prevent gene expression by targeting complementary messenger RNA  
       [0620] The phrases “percent identify” 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.  
       [0621] Percent identity between polynucleotide sequences ma” 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 Wis.). CLUSTAL V is described in Higgins, D. G. and Sharp, P. M. (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 sequence pairs.  
       [0622] 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 “blasta,” that is used to determine alignment between a known polynucleotide sequence and other sequences on 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/. 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 2.0.9 (May 7, 1999) set at default parameters. Such default parameters may be, for example:  
       [0623] Matrx: BLOSUM62  
       [0624] Reward for match: 1  
       [0625] Penalty for mismatch: −2  
       [0626] Open Gap: 5 and Extension Gap: 2 penalties  
       [0627] Gap x drop-off: 50  
       [0628] Expect: 10  
       [0629] Word Size: 11  
       [0630] Filter: on  
       [0631] 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 figures or Sequence Listings, may be used to describe a length over which percentage identity may be measured  
       [0632] Nucleic acid sequences that do not show a high degree of identity may nevertheless encode similar amino acid sequences due to the degeneracy of the genetic code. It is understood that changes in nucleic acid sequence can be made using this degeneracy to produce multiple nucleic acid sequences that all encode substantially the same protein.  
       [0633] 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 hydrophobicity and acidity of the substituted residue, thus preserving the structure (and therefore function) of the folded polypeptide.  
       [0634] 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 (descried 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.  
       [0635] 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 2.0.9 (May 7, 1999) with blastp set at default parameters. Such default parameters may be, for example.  
       [0636] Matrix: BLOSUM62  
       [0637] Open Gap: 11 and Extension Gap: 1 penalty  
       [0638] Gap x drop-off 50  
       [0639] Expect: 10  
       [0640] Word Size: 3  
       [0641] Filter: on  
       [0642] 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 figures or Sequence Listings, may be used to describe a length over which percentage identity may be measured.  
       [0643] “Post-translational modification” of a DITHP 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 win vary by cell type depending on the enzymatic milieu and the DITHP.  
       [0644] “Probe” refers to dithp 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 polymerase enzyme. Primer pairs can be used for amplification (and identification) of a nucleic acid sequence, e.g., by the polymerase chain reaction (PCR).  
       [0645] 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, 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 figures and Sequence Listing, may be used.  
       [0646] Methods for preparing and using probes and primers are described in the references, for example Sambrook et al., 1989,  Molecular Cloning: A Laboratory Manual,  2 nd  ed., vol. 1-3, Cold Spring Harbor Press, Plainview N.Y.; Ausubel et al., 1987,  Current Protocols in Molecular Biology , Greene Publ. Assoc. &amp; Wiley-Intersciences, New York N.Y.; Innis et al., 1990,  PCR Protocols. A Guide to Methods and Applications , Academic Press, San Diego Calif. 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 Mass.).  
       [0647] 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 T.x.) 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 Mass.) 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&#39;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.  
       [0648] “Purified” refers to molecules, either polynucleotides or polypeptides that are isolated or separated from their natural environment and are at least 60% free, preferably at least 75% free, and most preferably at least 90% free from other compounds with which they are naturally associated.  
       [0649] 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.  
       [0650] 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.  
       [0651] “Regulatory element” refers to a nucleic acid sequence from nontranslated regions of a gene, and includes enhancers, promoters, introns, and 3′ untranslated regions, which interact with host proteins to carry out or regulate transcription or translation  
       [0652] “Reporter” molecules are chemical or biochemical moieties used for labeling a nucleic acid, an amino acid, or an antibody. They include radionuclides; enzymes; fluorescent, chemiluminescent, or chromogenic agents; substrates; cofactors; inhibitors; magnetic particles; and other moieties known in the art.  
       [0653] 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.  
       [0654] “Sample” is used in its broadest sense. Samples may contain nucleic or amino acids, antibodies, or other materials, and may be derived from any source (e.g., bodily fluids including, but not limited to, saliva, blood, and urine; chromosome(s), organelles, or membranes isolated from a cell; genomic DNA, RNA, or cDNA in solution or bound to a substrate; and cleared cells or tissues or blots or imprints from such cells or tissues).  
       [0655] “Specific binding” or “specifically binding” refers to the interaction between a protein or peptide and its agonist, antibody, antagonist, or other binding partner. 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 containing 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.  
       [0656] “Substitution” refers to the replacement of at least one nucleotide or amino acid by a different nucleotide or amino acid.  
       [0657] “Substrate” refers to any suitable rigid or semi-rigid support including, e.g., membranes, filters, chips, slides, wafers, fibers, magnetic or nonmagnetic beads, gels, tubing, plates, polymers, microparticles or 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.  
       [0658] A “transcript image” refers to the collective pattern of gene expression by a particular tissue or cell type under given conditions at a given time.  
       [0659] “Transformation” refers to a process by which exogenous DNA enters a recipient cell. Transformation may occur under natural or artificial conditions using various methods well known in the art. Transformation may rely on any known method for the insertion of foreign nucleic acid sequences into a prokaryotic or eukaryotic host cell. The method is selected based on the host cell being transformed.  
       [0660] “Transformants” include 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 cells which transiently express inserted DNA or RNA.  
       [0661] 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, and 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.  
       [0662] A “variant” of a particular nucleic acid sequence is defined as a nucleic acid sequence having at least 25% sequence identity to the particular nucleic acid sequence over a certain length of one of the is nucleic acid sequences using blastn with the “BLAST 2 Sequences” tool Version 2.0.9 (May 7, 1999) set at default parameters. Such a pair of nucleic acids may show, for example, at least 30%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95% or even at least 98% or greater sequence identity over a certain defined length. The variant may result in “conservative” amino acid changes which do not affect structural and/or chemical properties. A variant may be described as, for example, an “allelic” (as defined above), “splice,” “species,” or “polymorphic” variant. A splice variant may have significant identity to a reference molecule, but will generally have a greater or lesser number of polynucleotides due to alternate splicing of exons daring 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 base. The presence of SNPs may be indicative of, for example, a certain population, a disease state, or a propensity for a disease state.  
       [0663] In an alternative, variants of the polynucleotides of the present invention may be generated through recombinant methods. One possible method is a DNA shuffling technique such as MOLECULARBREEDING (Maxygen Inc., Santa Clara Calif.; described in U.S. Pat. No. 5,837,458; Chang, C.-C. et al. (1999) Nat Biotechnol. 17:793-797; Christians, F. C. et al. (1999) Nat Biotechnol. 17:259-264; and Crameri, A et al. (1996) Nat Biotechnol. 14:315-319) to alter or improve the biological properties of DITHP, 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.  
       [0664] A “variant” t  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 2.0.9 (May 7, 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  
       [0665] In a particular embodiment, cDNA sequences derived from human tissues and cell lines were aligned based on nucleotide sequence identity and assembled into “consensus” or “template” sequences which are designated by the template identification numbers (template IDs) in column 2 of Table 1.  
       [0666] The sequence identification numbers (SEQ ID NO:s) corresponding to the template IDs are shown in column 1. The template sequences have similarity to GenBank sequences, or “hits,” as designated by the GI Numbers in column 3. The statistical probability of each GenBank hit is indicated by a probability score in column 4, and the functional annotation corresponding to each GenBank hit is listed in column 5.  
       [0667] The invention incorporates the nucleic acid sequences of these templates as disclosed in the Sequence Listing and the use of these sequences in the diagnosis and treatment of disease states characterized by defects in human molecules. The invention further utilizes these sequences in hybridization and amplification technologies, and in particular, in technologies which assess gene expression patterns correlated with specific cells or tissues and their responses in vivo or in vitro to pharmaceutical agents, toxins, and other treatments. In this manner, the sequences of the present invention are used to develop a transcript image for a particular cell or tissue.  
       [0668] Derivation of Nucleic Acid Sequences  
       [0669] cDNA was isolated from libraries constructed using RNA derived from normal and diseased human tissues and cell lines. The human tissues and cell lines used for cDNA library construction were selected from a broad range of sources to provide a diverse population of cDNAs representative of gene transcription throughout the human body. Descriptions of the human tissues and cell lines used for cDNA library construction are provided in the LIFESEQ database (Incyte Genomics, Inc. (Incyte), Palo Alto Calif.). Human tissues were broadly selected from, for example, cardiovascular, dermatologic, endocrine, gastrointestinal, hematopoietic/immune system, musculoskeletal, neural, reproductive, and urologic sources.  
       [0670] Cell lines used for cDNA library construction were derived from, for example, leukemic cells, teratocarcinomas, neuroepitheliomas, cervical carcinoma, lung fibroblasts, and endothelial cells. Such cell lines include, for example, THP-1, Jurkat, HUVEC, hNT2, WI38, HeLa, and other cell lines commonly used and available from public depositories (American Type Culture Collection, Manassas Va.). Prior to mRNA isolation, cell lines were untreated, treated with a pharmaceutical agent such as 5′-aza-2′-deoxycytidine, treated with an activating agent such as lipopolysaccharide in the case of leukocytic cell lines, or, in the case of endothelial cell lines, subjected to shear stress.  
       [0671] Sequencing of the cDNAs  
       [0672] Methods for DNA sequencing are well known in the art. Conventional enzymatic methods employ the Klenow fragment of DNA polymerase I, SEQUENASE DNA polymerase (U.S. Biochemical Corporation, Cleveland Ohio), Taq polymerase (Applied Biosystems, Foster City Calif.), thermostable 17 polymerase (Amersham Pharmacia Biotech, Inc. (Amersham Pharmacia Biotech), Piscataway N.J.), or combinations of polymerases and proofreading exonucleases such as those found in the ELONGASE amplification system (Life Technologies Inc. (Life Technologies), Gaithersburg Md.), to extend the nucleic acid sequence from an oligonucleotide primer annealed to the DNA template of interest Methods have been developed for the use of both single-stranded and double-stranded templates. Chain termination reaction products may be electrophoresed on urea-polyacrylamide gels and detected either by autoradiography (for radioisotope-labeled nucleotides) or by fluorescence (for fluorophore-labeled nucleotides). Automated methods for mechanized reaction preparation, sequencing, and analysis using fluorescence detection methods have been developed Machines used to prepare cDNAs for sequencing can include the MICROLAB 2200 liquid transfer system (Hamilton Company (Hamilton), Reno Nev.), Peltier thermal cycler (PTC200; MJ Research, Inc. (MJ Research), Watertown Mass.), and ABI CATALYST 800 thermal cycler (Applied Biosystems). Sequencing can be carried out using, for example, the ABI 373 or 377 (Applied Biosystems) or MEGABACE 1000 (Molecular Dynamics, Inc. (Molecular Dynamics), Sunnyvale Calif.) DNA sequencing systems, or other automated and manual sequencing systems well known in the art.  
       [0673] The nucleotide sequences of the Sequence Listing have been prepared by current, state-of-the-art, automated methods and, as such, may contain occasional sequencing errors or unidentified nucleotides. Such unidentified nucleotides are designated by an N. These infrequent unidentified bases do not represent a hindrance to practicing the invention for those skilled in the art. Several methods employing standard recombinant techniques may be used to correct errors and complete the missing sequence information. (See, e.g., those described in Ausubel, F. M. et al. (1997)  Short Protocols in Molecular Bioloy , John Wiley &amp; Sons, New York N.Y.; and Sambrook, J. et al. (1989)  Molecular Cloning A Laboratory Manual , Cold Spring Harbor Press, Plainview N.Y.)  
       [0674] Assembly of cDNA Sequences  
       [0675] Human polynucleotide sequences may be assembled using programs or algorithms well known in the art. Sequences to be assembled are related, wholly or in part, and may be derived-from a single or many different transcripts. Assembly of the sequences can be performed using such programs as PHRAP (Phils Revised Assembly Program) and the GELVIEW fragment assembly system (GCG), or other methods known in the art.  
       [0676] Alternatively, cDNA sequences are used as “component” sequences that are assembled into “template” or “consensus” sequences as follows. Sequence chromatograms are processed, verified, and quality scores are obtained using PHRED. Raw sequences are edited using an editing pathway known as Block 1 (See, e.g., the LIFESEQ Assembled User Guide, Incyte Genomics, Palo Alto, Calif.). A series of BLAST comparisons is performed and low-information segments and repetitive elements (e.g., dinucleotide repeats, Alu repeats, etc.) are replaced by “n&#39;s”, or masked, to prevent spurious matches. Mitochondrial and ribosomal RNA sequences are also removed. The processed sequences are then loaded into a relational database management system (RDMS) which assigns edited sequences to existing templates, if available. When additional sequences are added into the RDMS, a process is initiated which modifies existing templates or creates new templates from works in progress (i.e., nonfinal assembled sequences) containing queued sequences or the sequences themselves. After the new sequences have been assigned to templates, the templates can be merged into bins. If multiple templates exist in one bin, the bin can be split and the templates reannotated.  
       [0677] Once gene bins have been generated based upon sequence alignments, bins are “clone joined” based upon clone information. Clone joining occurs when the 5′ sequence of one clone is present in one bin and the 3′ sequence from the same clone is present in a different bin, indicating that the two bins should be merged into a single bin. Only bins which share at least two different clones are merged.  
       [0678] A resultant template sequence may contain either a partial or a full length open reading frame, or all or part of a genetic regulatory element. This variation is due in part to the fact that the full length cDNAs of many genes are several hundred, and sometimes several thousand, bases in length. With current technology, cDNAs comprising the coding regions of large genes cannot be cloned because of vector limitations, incomplete reverse transcription of the mRNA, or incomplete “second strand” synthesis. Template sequences may be extended to include additional contiguous sequences derived from the parent RNA transcript using a variety of methods known to those of skill in the art. Extension may thus be used to achieve the full length coding sequence of a gene.  
       [0679] Analysis of the cDNA Sequences  
       [0680] The cDNA sequences are analyzed using a variety of programs and algorithms which are well known in the art. (See, e.g., Ausubel, 1997, sutra, Chapter 7.7; Meyers, R. A. (Ed.) (1995)  Molecular Biology and Biotechnology , Wiley VCH, New York N.Y., pp. 856-853; and Table 7.) These analyses comprise both reading frame determinations, e.g., based on triplet codon periodicity for particular organisms (Fickett, J. W. (1982) Nucleic Acids Res. 10:5303-5318); analyses of potential start and stop codons; and homology searches.  
       [0681] Computer programs known to those of skill in the art for performing computer-assisted searches for amino acid and nucleic acid sequence similarity, include, for example, Basic Local Alignment Search Tool (BLAST; Altschul, S. F. (1993) J. Mol. Evol. 36:290-300; Altschul, S. F. et al. (1990) J. Mol. Biol. 215:403410). BLAST is especially useful in determining exact matches and comparing two sequence fragments of arbitrary but equal lengths, whose alignment is locally maximal and for which the alignment score meets or exceeds a threshold or cutoff score set by the user (Karlin, S. et al. (1988) Proc. Natl. Acad. Sci. USA 85:841-845). Using an appropriate search tool (e.g., BLAST or HMM), GenBank, SwissProt, BLOCKS, PFAM and other databases may be searched for sequences containing regions of homology to a query dithp or DITHP of the present invention.  
       [0682] Other approaches to the identification, assembly, storage, and display of nucleotide and polypeptide sequences are provided in “Relational Database for Storing Biomolecule Infomation,” U.S. Ser. No. 08/947,845, filed Oct. 9, 1997; “Project-Based Full-Length Biomolecular Sequence Database,” U.S. Ser. No. 08/811,758, filed Mar. 6, 1997; and “Relational Database and System for Storing Information Relating to Biomolecular Sequences,” U.S. Ser. No. 09/034,807, filed Mar. 4, 1998, all of which are incorporated by reference herein in their entirety.  
       [0683] Protein hierarchies can be assigned to the putative encoded polypeptide based on, e.g., motif, BLAST, or biological analysis. Methods for assigning these hierarchies are described, for example, in “Database System Employing Protein Function Hierarchies for Viewing Biomolecular Sequence Data,” U.S. Ser. No. 08/812,290, filed Mar. 6, 1997, incorporated herein by reference.  
       [0684] Identification of Human Diagnostic and Therapeutic Molecules Encoded by dithp  
       [0685] The identities of the DITHP encoded by the dithp of the present invention were obtained by analysis of the assembled cDNA sequences. SEQ ID NO:212, SEQ ID NO:213, SEQ ID NO:214, SEQ ID NO:215, SEQ ID NO:216, SEQ ID NO:217, SEQ ID NO:218, SEQ ID NO:219, SEQ ID NO:220, SEQ ID NO:221, SEQ ID NO:222, and SEQ ID NO:223, encoded by SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, and SEQ ID NO:12, respectively, are, for example, human enzyme molecules.  
       [0686] SEQ ID NO:224, SEQ ID NO:225, SEQ ID NO:226, SEQ ID NO:227, and SEQ ID NO:228, encoded by SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, and SEQ ID NO:17, respectively, are, for example, receptor molecules.  
       [0687] SEQ ID NO:229, SEQ ID NO:230, SEQ ID NO:231, SEQ ID NO:232, SEQ ID NO:233, SEQ ID NO:234, SEQ ID NO:235, SEQ ID NO:236, SEQ ID NO:237, SEQ ID NO:238, SEQ ID NO:239, SEQ ID NO:240, and SEQ ID NO:241, encoded by 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, and SEQ ID NO:30, respectively, are, for example, intracellular signaling molecules.  
                          SEQ ID NO:242, SEQ ID NO:243, SEQ ID NO:244, SEQ ID NO:245, SEQ ID NO:246,           SEQ ID NO:247, SEQ ID NO:248, SEQ ID NO:249, SEQ ID NO:250, SEQ ID NO:251, SEQ ID       NO:252, SEQ ID NO:253, SEQ ID NO:254, SEQ ID NO:255, SEQ ID NO:256, SEQ ID NO:257,       SEQ ID NO:258, SEQ ID NO:259, SEQ ID NO:260, SEQ ID NO:261, SEQ ID NO:262, SEQ ID       NO:263, SEQ ID NO:264, SEQ ID NO:265, SEQ ID NO:266, SEQ ID NO:267, SEQ ID NO:268,       SEQ ID NO:269, SEQ ID NO:270, SEQ ID NO:271, SEQ ID NO:272, SEQ ID NO:273, SEQ ID       NO:274, SEQ ID NO:275, SEQ ID NO:276, SEQ ID NO:277, SEQ ID NO:278, SEQ ID NO:279,       SEQ ID NO:280, SEQ ID NO:281, SEQ ID NO:282, SEQ ID NO:283, SEQ ID NO:284, SEQ ID       NO:285, SEQ ID NO:286, SEQ ID NO:287, SEQ ID NO:288, SEQ ID NO:289, SEQ ID NO:290,       SEQ ID NO:291, SEQ ID NO:292, SEQ ID NO:293, SEQ ID NO:294, SEQ ID NO:295, SEQ ID       NO:296, SEQ ID NO:297, SEQ ID NO:298, SEQ ID NO:299, SEQ ID NO:300, SEQ ID NO:301,       SEQ ID NO:302, SEQ ID NO:303, SEQ ID NO:304, SEQ ID NO:305, SEQ ID NO:306, SEQ ID       NO:307, SEQ ID NO:308, and SEQ ID NO:309, encoded by 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:37, SEQ ID NO:38, SEQ       ID NO:39, SEQ ID NO 40, SEQ ID NO:41, SEQ ID NO:42, SEQ ID NO:43, SEQ ID NO:44, SEQ       ID NO:45, SEQ ID NO:46, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:49, SEQ ID NO:50, SEQ       ID NO:51, SEQ ID NO:52, SEQ ID NO:53, SEQ ID NO:54, 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:S0, SEQ       ID NO:81, SEQ ID NO:82, SEQ ID NO:83, SEQ ID NO:84, SEQ ID NO:85, SEQ ID NO:86, SEQ       ID NO:87, SEQ ID NO:88, SEQ ID NO:89, SEQ ID NO:90, SEQ ID NO:91, SEQ ID NO:92, SEQ       ID NO:93, SEQ ID NO:94, SEQ ID NO:95, SEQ ID NO:96, SEQ ID NO:97, and SEQ ID NO:98,          
 
       [0688] respectively, are, for example, transcription factor molecules.  
       [0689] SEQ ID NO:310, SEQ ID NO:311, SEQ ID NO:312, SEQ ID NO:313, SEQ ID NO:314, SEQ ID NO:315, SEQ ID NO:316, and SEQ ID NO:317, encoded by SEQ ID NO:99, SEQ ID NO:100, SEQ ID NO:101, SEQ ID NO:102, SEQ ID NO:103, SEQ ID NO:104, SEQ ID NO:105, and SEQ ID NO:106, respectively, are, for example, membrane transport molecules.  
       [0690] SEQ ID NO:318, SEQ ID NO:319, SEQ ID NO:320, SEQ ID NO:321, SEQ ID NO:322,  
       [0691] SEQ ID NO:323, SEQ ID NO:324, SEQ ID NO:325, SEQ ID NO:326, and SEQ ID NO:327, encoded by SEQ ID NO:107, SEQ ID NO:108, SEQ ID NO:109, SEQ ID NO:110, SEQ ID NO:111, SEQ ID NO:112, SEQ ID NO:113, SEQ ID NO:114, SEQ ID NO:115, and SEQ ID NO:116, respectively, are, for example, protein modification and maintenance molecules.  
       [0692] SEQ ID NO:328, SEQ ID NO:329, SEQ ID NO:330, SEQ ID NO:331, SEQ ID NO:332, SEQ ID NO:333, SEQ ID NO:334, SEQ ID NO:335, SEQ ID NO:336, SEQ ID NO:337, SEQ ID NO:338, SEQ ID NO:339, SEQ ID NO:340, and SEQ ID NO:341, encoded by SEQ ID NO:117, SEQ ID NO:118, SEQ ID NO:119, SEQ ID NO:120, SEQ ID NO:121, SEQ ID NO:122, SEQ ID NO:123, SEQ ID NO:124, SEQ ID NO:125, SEQ ID NO:126, SEQ ID NO:127, SEQ ID NO:128, SEQ ID NO:129, and SEQ ID NO:130, respectively, are, for example, nucleic acid synthesis and modification molecules.  
       [0693] SEQ ID NO:342, encoded by SEQ ID NO:131 is, for example, an adhesion molecule. SEQ ID NO:343, SEQ ID NO:344, SEQ ID NO:345, SEQ ID NO:346, SEQ ID NO:347,  
       [0694] SEQ ID NO:348, and SEQ ID NO:349, encoded by SEQ ID NO:132, SEQ ID NO:133, SEQ ID NO:134, SEQ ID NO:135, SEQ ID NO:136, SEQ ID NO:137, and SEQ ID NO:138, respectively, are, for example, antigen recognition molecules.  
       [0695] SEQ ID NO:350, SEQ ID NO:351, SEQ ID NO:352, and SEQ ID NO:353, encoded by SEQ ID NO:139, SEQ ID NO:140, SEQ ID NO:141, and SEQ ID NO:142, respectively, are, for example, electron transfer associated molecules.  
       [0696] SEQ ID NO:354, SEQ ID NO:355, SEQ ID NO:356, SEQ ID NO:357, SEQ ID NO:358, and SEQ ID NO:359, encoded by SEQ ID NO:143, SEQ ID NO:144, SEQ ID NO:145, SEQ ID NO:146, SEQ ID NO:147, and SEQ ID NO:148, respectively, are, for example, secreted/extracellular matrix molecules.  
       [0697] SEQ ID NO:360, SEQ ID NO:361, SEQ ID NO:362, SEQ ID NO:363, SEQ ID NO:364, SEQ ID NO:365, SEQ ID NO:366, SEQ ID NO:367, SEQ ID NO:368, and SEQ ID NO:369, encoded by SEQ ID NO:149, SEQ ID NO:150, SEQ ID NO:151, SEQ ID NO:152, SEQ ID NO:153, SEQ ID NO:154, SEQ ID NO:155, SEQ ID NO:156, SEQ ID NO:157, and SEQ ID NO:158, respectively, are, for example, cytoskeletal molecules.  
       [0698] SEQ ID NO:370, SEQ ID NO:371, SEQ ID NO:372, and SEQ ID NO:373, encoded by SEQ ID NO:159, SEQ ID NO:160, SEQ ID NO:161, and SEQ ID NO:162, respectively, are, for example, cell membrane molecules.  
       [0699] SEQ ID NO:374, SEQ ID NO:375, SEQ ID NO:376, SEQ ID NO:377, SEQ ID NO:378, SEQ ID NO:379, SEQ ID NO:380, SEQ ID NO:381, SEQ ID NO:382, SEQ ID NO:383, SEQ ID NO:384, SEQ ID NO:385, SEQ ID NO:386, SEQ ID NO:387, SEQ ID NO:388, SEQ ID NO:389, SEQ ID NO:390, SEQ ID NO:391, and SEQ ID NO:392, encoded by SEQ ID NO:163, SEQ ID NO:164, SEQ ID NO:165, SEQ ID NO:166, SEQ ID NO:167, SEQ ID NO:168, SEQ ID NO:169, SEQ ID NO:170, SEQ ID NO:171, SEQ ID NO:172, SEQ ID NO:173, SEQ ID NO:174, SEQ ID NO:175, SEQ ID NO:176, SEQ ID NO:177, SEQ ID NO:178, SEQ ID NO:179, SEQ ID NO:180, and SEQ ID NO:181, respectively, are, for example, ribosomal molecules.  
       [0700] SEQ ID NO:393, SEQ ID NO:394, SEQ ID NO:395, SEQ ID NO:396, SEQ ID NO:397, SEQ ID NO:398, SEQ ID NO:399, SEQ ID NO:400, SEQ ID NO:401, SEQ ID NO:402, and SEQ ID NO:403, encoded by SEQ ID NO:182, SEQ ID NO:183, SEQ ID NO:184, SEQ ID NO:185, SEQ ID NO:186, SEQ ID NO:187, SEQ ID NO:188, SEQ ID NO:189, SEQ ID NO:190, SEQ ID NO:191, and SEQ ID NO:192, respectively, are, for example, organelle associated molecules.  
       [0701] SEQ ID NO:404, SEQ ID NO:405, SEQ ID NO:406, SEQ ID NO:407, SEQ ID NO:408, SEQ ID NO:409, SEQ ID NO:410, SEQ ID NO:411, SEQ ID NO-0.412, SEQ ID NO:413, and SEQ ID NO:414, encoded by SEQ ID NO:193, SEQ ID NO:194, SEQ ID NO:195, SEQ ID NO:196, SEQ ID NO:197, SEQ ID NO:198, SEQ ID NO:199, SEQ ID NO:200, SEQ ID NO:201, SEQ ID NO:202, and SEQ ID NO:203, respectively, are; for example, biochemical pathway molecules.  
       [0702] SEQ ID NO:415, SEQ ID NO:416, SEQ ID NO:417, SEQ ID NO:418, SEQ ID NO:419,  
       [0703] SEQ ID NO:420, SEQ ID NO:421, and SEQ ID NO:422, encoded by SEQ ID NO:204, SEQ ID NO:205, SEQ ID NO:206, SEQ ID NO:207, SEQ ID NO:208, SEQ ID NO:209, SEQ ID NO:210, and SEQ ID NO:211, respectively, are, for example, molecules associated with growth and development  
       [0704] Sequences of Hunan Diagnostic and Therapeutic Molecules  
       [0705] The dithp of the present invention may be used for a variety of diagnostic and therapeutic purposes. For example, a dithp may be used to diagnose a particular condition, disease, or disorder associated with human molecules. Such conditions, diseases, and disorders include, but are not limited to, a cell proliferative disorder, such as actinic keratosis, arteriosclerosis, atherosclerosis, bursitis, cirrhosis, hepatitis, mixed connective tissue disease (MCTD), myelofibrosis, paroxysmal nocturnal hemoglobinuria, polycythemia vera, psoriasis, primary thrombocythemia, and cancers including adenocarcinoma, leukemia, lymphoma, melanoma, myeloma, sarcoma, teratocarcinoma, and, in particular, a cancer of the adrenal gland, bladder, bone, bone marrow, brain, breast cervix, gall bladder, ganglia, gastrointestinal tract, heart, kidney, liver, lung, muscle, ovary, pancreas, parathyroid, penis, prostate, salivary glands, skin, spleen, testis, thymus, thyroid, and uterus; an autoimmune/inflammatory disorder, such as inflammation, actinic keratosis, acquired immunodeficiency syndrome (AIDS), Addison&#39;s disease, adult respiratory distress syndrome, allergies, ankylosing spondylitis, amyloidosis, anemia, arteriosclerosis, asthma, atherosclerosis, autoimmune hemolytic anemia, autoimmune thyroiditis, bronchitis, bursitis, cholecystitis, cirrhosis, contact dermatitis, Crohn&#39;s disease, atopic dermatitis, dermatomyositis, diabetes mellitus, emphysema, erythroblastosis fetalis, erythema nodosum, atrophic gastritis, glomerulonephritis, Goodpasture&#39;s syndrome, gout, Graves&#39; disease, Hashimoto&#39;s thyroiditis, paroxysmal nocturnal hemoglobinuria, hepatitis, hypereosinophilia, irritable bowel syndrome, episodic lymphopenia with lymphocytotoxins, mixed connective tissue disease (MCTD), multiple sclerosis, myasthenia gravis, myocardial or pericardial inflammation, myelofibrosis, osteoarthritis, osteoporosis, pancreatitis, polycythemia vera, polymyositis, psoriasis, Reiter&#39;s syndrome, rheumatoid arthritis, scleroderma, Sjögren&#39;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, trauma, and hematopoietic cancer including lymphoma, leukemia, and myeloma; an infection caused by a viral agent classified as adenovinis, arenavirus, bunyavirus, calicivirus, coronavirus, filovirus, hepadnavirus, herpesvirus, flavivirus, orthomyxovirus, parvovirus, papovavirus, paramyxovirus, picornavirus, poxvirus, reovirus, retrovirus, rhabdovirus, or togavirus; an infection caused by a bacterial agent classified as pneumococcus, staphylococcus, streptococcus, bacillus, corynebacterium, clostridium, meningococcus, gonococcus, listeria, moraxella, kingella, haemophilus, legionella, bordetella, gram-negative enterobacterium including shigella, salmonella, or campylobacter, pseudomonas, vibrio, brucella, francisella, yersinia, bartonella, norcardium, actinomyces, mycobacterium, spirochaetale, rickettsia, chlamydia, or mycoplasma; an infection caused by a fuingal agent classified as aspergillus, blastomyces, dermatophytes, cryptococcus, coccidioides, malasezzia, histoplasma, or other mycosis-causing fungal agent; and an infection caused by a parasite classified as plasmodium or malaria-causing, parasitic entamoeba, leisbmania, trypanosoma, toxoplasma, pneumocystis carinii, intestinal protozoa such as giardia, trichomonas, tissue nematode such as trichinella, intestinal nematode such as ascaris, lymphatic filarial nematode, trematode such as schistosoma, and cestrode such as tapeworm; a developmental disorder such as renal tubular acidosis, anemia, Cushing&#39;s syndrome, achondroplastic dwarfism, Duchenne and Becker muscular dystrophy, epilepsy, gonadal dysgenesis, WAGR syndrome (Wilms&#39; tumor, aniridia, genitourinary abnormalities, and mental retardation), Smith-Magenis syndrome, myelodysplastic syndrome, hereditary mucoepithelial dysplasia, hereditary keratodermas, hereditary neuropathies such as Charcot-Marie-Tooth disease and neurofibromatosis, hypothyroidism, hydrocephalus, seizure disorders such as Syndenham&#39;s chorea and cerebral palsy, spina bifida, anencephaly, craniorachischisis, congenital glaucoma, cataract, and sensorineural hearing loss; an endocrine disorder such as a disorder of the hypothalamus and/or pituitary resulting from lesions such as a primary brain tumor, adenoma, infarction associated with pregnancy, hypophysectomy, aneurysm, vascular malformation, thrombosis, infection, immunological disorder, and complication due to head trauma; a disorder associated with hypopituitarism including hypogonadism, Sheehan syndrome, diabetes insipidus, Kallman&#39;s disease, Hand-Schuller-Christian disease, Letterer-Siwe disease, sarcoidosis, empty sella syndrome, and dwarfism; a disorder associated with hyperpituitarism including acromegaly, giantism, and syndrome of inappropriate antidiuretic hormone (ADH) secretion (SIADH) often caused by benign adenoma; a disorder associated with hypothyroidism including goiter, myxedema, acute thyroiditis associated with bacterial infection, subacute thyroiditis associated with viral infection, autoimmune thyroiditis (Hashimoto&#39;s disease), and cretinism; a disorder associated with hyperthyroidism including thyrotoxicosis and its various forms, Grave&#39;s disease, pretibial myxedema, toxic multinodular goiter, thyroid carcinoma, and Plummer&#39;s disease; a disorder associated with hyperparathyroidism including Corn disease (chronic hypercalemia); a pancreatic disorder such as Type I or Type II diabetes mellitus and associated complications; a disorder associated with the adrenals such as hyperplasia, carcinoma, or adenoma of the adrenal cortex, hypertension associated with alkalosis, amyloidosis, hypokalemia, Gushing&#39;s disease, Liddle&#39;s syndrome, and Arnold-Healy-Gordon syndrome, pheochromocytoma tumors, and Addison&#39;s disease; a disorder associated with gonadal steroid hormones such as: in women, abnormal prolactin production, infertility, endometriosis, perturbation of the menstrual cycle, polycystic ovarian disease, hyperprolactinemia, isolated gonadotropin deficiency, amenorrhea, galactorrhea, hermaphroditism, hirsutism and virilization, breast cancer, and, in post-menopausal women, osteoporosis; and, in men, Leydig cell deficiency, male climacteric phase, and germinal cell aplasia, a hypergonadal disorder associated with Leydig cell tumors, androgen resistance associated with absence of androgen receptors, syndrome of 5 α-reductase, and gynecomastia; a metabolic disorder such as Addison&#39;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, pseudovitamin D-deficiency rickets; disorders of carbohydrate metabolism such as congenital type II dyseryhropoietic anemia, diabetes, insulin-dependent diabetes mellitus, non-insulin-dependent diabetes mellitus, fructose-1,6diphosphatase deficiency, galactosemia, glucagonoma, hereditary fructose intolerance, hypoglycemia, mannosidosis, neuraminidase deficiency, obesity, galactose epimerase deficiency, glycogen storage diseases, lysosomal storage diseases, fructosuria, pentosuria, and inherited abnormalities of pyruvate metabolism; disorders of lipid metabolism such as fatty liver, cholestasis, primary biliary cirrhosis, carnitine deficiency, carnitine palmitoyltransferase deficiency, myoadenylate deaminase deficiency, hypertriglyceridemia, lipid storage disorders such Fabry&#39;s disease, Gaucher&#39;s disease, Niemann-Pick&#39;s disease, metachromatic leukodystrophy, adrenoleukodystrophy, GM 2  gangliosidosis, and ceroid lipofuscinosis, abetalipoproteinemia, Tangier disease, hyperlipoproteinemia, diabetes mellitus, lipodystrophy, lipomatoses, acute panniculitis, disseminated fat necrosis, adiposis dolorosa, lipoid adrenal hyperplasia, minimal change disease, lipomas, atherosclerosis, hypercholesterolemia, hypercholesterolemia with hypertriglyceridemia, primary hypoalphalipoproteinemia, hypothyroidism, renal disease, liver disease, lecithin:cholesterol acyltransferase deficiency, cerebrotendinous xanthomatosis, sitosterolemia, hypocholesterolemia, Tay-Sachs disease, Sandhoff&#39;s disease, hyperlipidemia, hyperlipemia, lipid myopathies, and obesity; and disorders of copper metabolism such as Menke&#39;s disease, Wilson&#39;s disease, and Ehlers-Danlos syndrome type IX; a neurological disorder such as epilepsy, ischemic cerebrovascular disease, stroke, cerebral neoplasms, Alzheimer&#39;s disease, Pick&#39;s disease, Huntington&#39;s disease, dementia, Parkinson&#39;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 disorder of the central nervous system, cerebral palsy, a neuroskeletal disorder, an autonomic nervous system disorder, a cranial nerve disorder, a spinal cord disease, muscular dystrophy and other neuromuscular disorder, a peripheral nervous system disorder, dermatomyositis and polymyositis, inherited, metabolic, endocrine, and toxic myopathy, myasthenia gravis, periodic paralysis, a mental disorder including mood, anxiety, and schizophrenic disorders, seasonal affective disorder (SAD), akathesia, amnesia, catatonia, diabetic neuropathy, tardive dyskinesia, dystonias, paranoid psychoses, postherpetic neuralgia, and Tourette&#39;s disorder; a gastrointestinal disorder including ulcerative colitis, gastric and duodenal ulcers, cystinuria, dibasicaminoaciduria, hypercystinuria, lysinuria, hartnup disease, tryptophan malabsorption, methionine malabsorption, hissidinuria, iminoglycinuria, dicarboxylicaminoaciduria, cystinosis, renal glycosuria, hypouricemia, familial hypophophatemic rickets, congenital chloridorrhea, distal renal tubular acidosis, Menkes&#39; disease, Wilson&#39;s disease, lethal diarrhea, juvenile pernicious anemia, folate malabsorption, adrenoleukodystrophy, hereditary myoglobinuria, and Zellweger syndrome; a transport disorder such as akinesia, amyotrophic lateral sclerosis, ataxia telangiectasia, cystic fibrosis, Becker&#39;s muscular dystrophy, Bell&#39;s palsy, Charcot-Marie Tooth disease, diabetes mellitus, diabetes insipidus, diabetic neuropathy, Duchenne muscular dystrophy, hyperkalemic periodic paralysis, normokalemic periodic paralysis, Parkinson&#39;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, tachyarrymia, hypertension, Long QT syndrome, myocarditis, cardiomyopathy, nemaline myopathy, centronuclear myopathy, lipid myopathy, mitochondrial myopathy, thyrotoxic myopathy, ethanol myopathy, dermatomyositis, inclusion body myositis, infectious myositis, and polymyositis, neurological disorders associated with transport, e.g., Alzheimer&#39;s disease, amnesia, bipolar disorder, dementia, depression, epilepsy, Tourette&#39;s disorder, paranoid psychoses, and schizophrenia, and other disorders associated with transport, e.g., neurofibromatosis, postherpetic neuralgia, trigeminal neuropathy, sarcoidosis, sickle cell anemia, cataracts, infertility, pulmonary artery stenosis, sensorineural autosomal deafness, hyperglycemia, hypoglycemia, Grave&#39;s disease, goiter, glucose-galactose malabsorption syndrome, hypercholesterolemia, Cushing&#39;s disease, and Addison&#39;s disease; and a connective tissue disorder such as osteogenesis imperfecta, Ehlers-Danlos syndrome, chondrodysplasias, Marfan syndrome, Alport syndrome, familial aortic aneurysm, achondroplasia, mucopolysaccharidoses, osteoporosis, osteopetrosis, Paget&#39;s disease, rickets, osteomalacia, hyperparathyroidism, renal osteodystrophy, osteonecrosis, osteomyelitis, osteoma, osteoid osteoma, osteoblastoma, osteosarcoma, osteochondroma, chondroma, chondroblastoma, chondromyxoid fibroma, chondrosarcoma, fibrous cortical defect, nonossifying fibroma, fibrous dysplasia, fibrosarcoma, malignant fibrous histiocytoma, Ewing&#39;s sarcoma, primitive neuroectodermal tumor, giant cell tumor, osteoarthritis, rheumatoid arthritis, ankylosing spondyloarthritis, Reiter&#39;s syndrome, psoriatic arthritis, enteropathic arthritis, infectious arthritis, gout, gouty arthritis, calcium pyrophosphate crystal deposition disease, ganglion, synovial cyst, villonodular synovitis, systemic sclerosis, Dupuytren&#39;s contracture, hepatic fibrosis, lupus erythematosus, mixed connective tissue disease, epidermolysis bullosa simplex, bullous congenital ichthyosiform erythroderma (epidermolytic hyperkeratosis), non-epidermolytic and epidermolytic palmoplantar keratoderma, ichthyosis bullosa of Siemens, pachyonycnia congenita, and white sponge nevus. The dithp can be used to detect the presence of, or to quantify the amount of, a dithp-related polynucleotide in a sample. This information is then compared to information obtained from appropriate reference samples, and a diagnosis is established. Alternatively, a polynucleotide complementary to a given dithp can inhibit or inactivate a therapeutically relevant gene related to the dithp.  
       [0706] Analysis of dithp Expression Patterns  
       [0707] The expression of dithp may be routinely assessed by hybridization-based methods to determine, for example, the tissue-specificity, disease-specificity, or developmental stage-specificity of dithp expression. For example, the level of expression of dithp may be compared among different cell types or tissues, among diseased and normal cell types or tissues, among cell types or tissues at different developmental stages, or among cell types or tissues undergoing various treatments. This type of analysis is useful, for example, to assess the relative levels of dithp expression in fully or partially differentiated cells or tissues, to determine if changes in dithp expression levels are correlated with the development or progression of specific disease states, and to assess the response of a cell or tissue to a specific therapy, for example, in pharmacological or toxicological studies. Methods for the analysis of dithp expression are based on hybridization and amplification technologies and include membrane-based procedures such as northern blot analysis, high-throughput procedures that utilize, for example, microarrays, and PCR-based procedures.  
       [0708] Hybridization and Genetic Analysis  
       [0709] The dithp, their fragments, or complementary sequences, may be used to identify the presence of and/or to determine the degree of similarity between two (or more) nucleic acid sequences. The dithp may be hybridized to naturally occurring or recombinant nucleic acid sequences under appropriately selected temperatures and salt concentrations. Hybridization with a probe based on the nucleic acid sequence of at least one of the dithp allows for the detection of nucleic acid sequences, including genomic sequences, which are identical or related to the dithp of the Sequence Listing. Probes may be selected from non-conserved or unique regions of at least one of the polynucleotides of SEQ ID NO:1-211 and tested for their ability to identity or amplify the target nucleic acid sequence using standard protocols.  
       [0710] Polynucleotide sequences that are capable of hybridizing, in particular, to those shown in SEQ ID NO:1-211 and fragments thereof, can be identified using various conditions of stringency. (See, e.g., Wahl, G. M. and S. L. Berger (1987) Methods Enzymol.  152:399-407 ; Kimmel, AR. (1987) Methods Enzymol. 152:507-511.) Hybridization conditions are discussed in “Definitions.” 
       [0711] A probe for use in Southern or northern hybridization may be derived from a fragment of a dithp sequence, or its complement, that is up to several hundred nucleotides in length and is either single-stranded or double-stranded. Such probes may be hybridized in solution to biological materials such as plasmids, bacterial, yeast, or human artificial chromosomes, cleared or sectioned tissues, or to artificial substrates containing dithp. Microarrays are particularly suitable for identifying the presence of and detecting the level of expression for multiple genes of interest by examining gene expression correlated with, e.g., various stages of development, treatment with a drug or compound, or disease progression. An array analogous to a dot or slot blot may be used to arrange and link polynucleotides to the surface of a substrate using one or more of the following: mechanical (vacuum), chemical, thermal, or UV bonding procedures. Such an array may contain any number of dithp and may be produced by hand or by using available devices, materials, and machines.  
       [0712] Microarrays may be prepared, used, and analyzed using methods known in the art. (See, e.g., Brennan, T. M. et al. (1995) U.S. Pat. No. 5,474,796; Schena, M. et al. (1996) Proc. Natl. Acad. Sci. USA 93:10614-10619; Baldeschweiler et al. (1995) PCT application WO95/251116; Shalon, D. et al. (1995) PCT application WO95/35505; Heller, R. A. et al. (1997) Proc. Natl. Acad. Sci. USA 94:2150-2155; and Heller, M. J. et al. (1997) U.S. Pat. No. 5,605,662.)  
       [0713] Probes may be labeled by either PCR or enzymatic techniques using a variety of commercially available reporter molecules. For example, commercial kits are available for radioactive and chemiluminescent labeling (Amersham Pharmacia Biotech) and for alkaline phosphatase labeling (Life Technologies). Alternatively, dithp may be cloned into commercially available vectors for the production of RNA probes. Such probes may be transcribed in the presence of at least one labeled nucleotide (e.g.,  32 P-ATP, Amersham Pharmacia Biotech).  
       [0714] Additionally the polynucleotides of SEQ ID NO:1-211 or suitable fragments thereof can be used to isolate full length cDNA sequences utilizing hybridization and/or amplification procedures well known in the art, e.g., cDNA library screening, PCR amplification, etc. The molecular cloning of such full length cDNA sequences may employ the method of cDNA library screening with probes using the hybridization, stringency, washing, and probing strategies described above and in Ausubel, supra, Chapters 3, 5, and 6. These procedures may also be employed with genomic libraries to isolate genomic sequences of dithp in order to analyze, e.g., regulatory elements.  
       [0715] Genetic Mapping  
       [0716] Gene identification and mapping are important in the investigation and treatment of almost all conditions, diseases, and disorders. Cancer, cardiovascular disease, Alzheimer&#39;s disease, arthritis, diabetes, and mental illnesses are of particular interest. Each of these conditions is more complex than the single gene defects of sickle cell anemia or cystic fibrosis, with select groups of genes being predictive of predisposition for a particular condition, disease, or disorder. For example, cardiovascular disease may result from malfunctioning receptor molecules that fail to clear cholesterol from the bloodstream, and diabetes may result when a particular individual&#39;s immune system is activated by an infection and attacks the insulin-producing cells of the pancreas. In some studies, Alzieimer&#39;s disease has been linked to a gene on chromosome 21; other studies predict a different gene and location. Mapping of disease genes is a complex and reiterative process and generally proceeds from genetic linkage analysis to physical mapping.  
       [0717] As a condition is noted among members of a family, a genetic linkage map traces parts of chromosomes that are inherited in the same pattern as the condition. Statistics link the inheritance of particular conditions to particular regions of chromosomes, as defined by RFLP or other markers. (See, for example, Lander, E. S. and Botstein, D. (1986) Proc. Natl. Acad. Sci. USA 83:7353-7357.) Occasionally, genetic markers and their locations are known from previous studies. More often, however, the markers are simply stretches of DNA that differ among individuals. Examples of genetic linkage maps can be found in various scientific journals or at the Online Mendelian Inheritance in Man (OMIM) World Wide Web site.  
       [0718] In another embodiment of the invention, dithp sequences may be used to generate hybridization probes useful in chromosomal mapping of naturally occurring genomic sequences. Either coding or noncoding sequences of dithp may be used, and in some instances, noncoding sequences may be preferable over coding sequences. For example, conservation of a dithp coding sequence among members of a multi-gene family may potentially cause undesired cross hybridization during chromosomal mapping. The sequences may be mapped to a particular chromosome, to a specific region of a chromosome, or to artificial chromosome constructions, e.g., human artificial chromosomes (HACs), yeast artificial chromosomes (YACs), bacterial artificial chromosomes (BACs), bacterial P1 constructions, or single chromosome cDNA libraries. (See, e.g., Harrington, J. J. et al. (1997) Nat Genet. 15:345-355; Price, C. M. (1993) Blood Rev. 7:127-134; and Trask, B. J. (1991) Trends Genet. 7:149-154.)  
       [0719] Fluorescent in situ hybridization (FISH) may be correlated with other physical chromosome mapping techniques and genetic map data. (See, e.g., Meyers, supra, pp. 965-968.) Correlation between the location of dithp on a physical chromosomal map and a specific disorder, or a predisposition to a specific disorder, may help define the region of DNA associated with that disorder. The dithp sequences may also be used to detect polymorphisms that are genetically linked to the inheritance of a particular condition, disease, or disorder.  
       [0720] In situ hybridization of chromosomal preparations and genetic mapping techniques, such as linkage analysis using established chromosomal markers, may be used for extending existing 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 number or arm of the corresponding human chromosome is not known. These new marker sequences can be mapped to human chromosomes and may provide valuable information to investigators searching for disease genes using positional cloning or other gene discovery techniques. Once a disease or syndrome has been crudely correlated by genetic linkage with a particular genomic region, e.g., ataxia-telangiectasia to 11q22-23, any sequences mapping to that area may represent associated or regulatory genes for further investigation. (See, e.g., Gatti, R. A. et al. (1988) Nature 336:577-580.) The nucleotide sequences of the subject invention may also be used to detect differences in chromosomal architecture due to translocation, inversion, etc., among normal, carrier, or affected individuals.  
       [0721] Once a disease-associated gene is mapped to a chromosomal region, the gene must be cloned in order to identify mutations or other alterations (e.g., translocations or inversions) that may be correlated with disease. This process requires a physical map of the chromosomal region containing the disease-gene of interest along with associated markers. A physical map is necessary for determining the nucleotide sequence of and order of marker genes on a particular chromosomal region. Physical mapping techniques are well known in the art and require the generation of overlapping sets of cloned DNA fragments from a particular organelle, chromosome, or genome. These clones are analyzed to reconstruct and catalog their order. Once the position of a marker is determined, the DNA from that region is obtained by consulting the catalog and selecting clones from that region. The gene of interest is located through positional cloning techniques using hybridization or similar methods.  
       [0722] Diagnostic Uses  
       [0723] The dithp of the present invention may be used to design probes useful in diagnostic assays. Such assays, well known to those skilled in the art, may be used to detect or confirm conditions, disorders, or diseases associated with abnormal levels of dithp expression. Labeled probes developed from dithp sequences are added to a sample under hybridizing conditions of desired stringency. In some instances, dithp, or fragments or oligonucleotides derived from dithp, may be used as primers in amplification steps prior to hybridization. The amount of hybridization complex formed is quantified and compared with standards for that cell or tissue. If dithp expression varies significantly from the standard, the assay indicates the presence of the condition, disorder, or disease. Qualitative or quantitative diagnostic methods may include northern, dot blot, or other membrane or dip-stick based technologies or multiple-sample format technologies such as PCR, enzyme-linked immunosorbent assay (ELISA)-like, pin, or chip-based assays.  
       [0724] The probes described above may also be used to monitor the progress of conditions, disorders, or diseases associated with abnormal levels of dithp expression, or to evaluate the efficacy of a particular therapeutic treatment. The candidate probe may be identified from the dithp that are specific to a given human tissue and have not been observed in GenBank or other genome databases. Such a probe may be used in animal studies, preclinical tests, clinical trials, or in monitoring the treatment of an individual patient. In a typical process, standard expression is established by methods well known in the art for use as a basis of comparison, samples from patients affected by the disorder or disease are combined with the probe to evaluate any deviation from the standard profile, and a therapeutic agent is administered and effects are monitored to generate a treatment profile. Efficacy is evaluated by determining whether the expression progresses toward or returns to the standard normal pattern. Treatment profiles may be generated over a period of several days or several months. Statistical methods well known to those skilled in the art may be use to determine the significance of such therapeutic agents.  
       [0725] The polynucleotides are also useful for identifying individuals from minute biological samples, for example, by matching the RFLP pattern of a sample&#39;s DNA to that of an individual&#39;s DNA The polynucleotides of the present invention can also be used to determine the actual base-by-base DNA sequence of selected portions of an individual&#39;s genome. These sequences can be used to prepare PCR primers for amplifying and isolating such selected DNA, which can ten be sequenced. Using this technique, an individual can be identified through a unique set of DNA sequences. Once a unique ID database is established for an individual, positive identification of that individual can be made from extremely small tissue samples.  
       [0726] In a particular aspect, oligonucleotide primers derived from the dithp of the invention 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 dithp 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 (is SNP), are capable of identifying polymorphisms by comparing the sequences 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 Calif.).  
       [0727] DNA-based identification techniques are critical in forensic technology. DNA sequences taken from very small biological samples such as tissues, e.g., hair or skin, or body fluids, e.g., blood, saliva, semen, etc., can be amplified using, e.g., PCR, to identify individuals. (See, e.g., Erlich, H. (1992)  PCR Technology , Freeman and Co., New York N.Y.). Similarly, polynucleotides of the present invention can be used as polymorphic markers.  
       [0728] There is also a need for reagents capable of identifying the source of a particular tissue. Appropriate reagents can comprise, for example, DNA probes or primers prepared from the sequences of the present invention that are specific for particular tissues. Panels of such reagents can identify tissue by species and/or by organ type. In a similar fashion, these reagents can be used to screen tissue cultures for contamination.  
       [0729] The polynucleotides of the present invention can also be used as molecular weight markers on nucleic acid gels or Southern blots, as diagnostic probes for the presence of a specific mRNA in a particular cell type, in the creation of subtracted cDNA libraries which aid in the discovery of novel polynucleotides, in selection and synthesis of oligomers for attachment to an array or other support, and as an antigen to elicit an immune response.  
       [0730] Disease Model Systems Using dithp  
       [0731] The dithp of the invention 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. Pat. Nos. 5,175,383 and 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.  
       [0732] The dithp of the invention 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).  
       [0733] The dithp of the invention 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 dithp 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 dithp, resulting, e.g., in the secretion of DITHP in its milk, may also serve as a convenient source of that protein (Janne, J. et al. (1998) Biotechnol. Annu. Rev. 4:55-74).  
       [0734] Screening Assays  
       [0735] DITHP encoded by polynucleotides of the present invention may be used to screen for molecules that bind to or are bound by the encoded polypeptides. The binding of the polypeptide and the molecule may activate (agonist), increase, inhibit (antagonist), or decrease activity of the polypeptide or the bound molecule. Examples of such molecules include antibodies, oligonucleotides, proteins (e.g., receptors), or small molecules.  
       [0736] Preferably, the molecule is closely related to the natural ligand of the polypeptide, e.g., a ligand or fragment thereof, a natural substrate, or a structural or functional mimetic. (See, Coligan et al., (1991)  Current Protocols in Immunology  1(2): Chapter 5.) Similarly, the molecule can be closely related to the natural receptor to which the polypeptide binds, or to at least a fragment of the receptor, e.g., the active site. In either case, the molecule can be rationally designed using known techniques.  
       [0737] Preferably, the screening for these molecules involves producing appropriate cells which express the polypeptide, either as a secreted protein or on the cell membrane. Preferred cells include cells from mammals, yeast, Drosophila, or  E. coli . Cells expressing the polypeptide or cell membrane fractions which contain the expressed polypeptide are then contacted with a test compound and binding, stimulation, or inhibition of activity of either the polypeptide or the molecule is analyzed.  
       [0738] An assay may simply test binding of a candidate compound to the polypeptide, wherein binding is detected by a fluorophore, radioisotope, enzyme conjugate, or other detectable label. Alternatively, the assay may assess binding in the presence of a labeled competitor.  
       [0739] Additionally, the assay can be carried out using cell-free preparations, polypeptide/molecule affixed to a solid support, chemical libraries, or natural product mixtures. The assay may also simply comprise the steps of mixing a candidate compound with a solution containing a polypeptide, measuring polypeptide/molecule activity or binding, and comparing the polypeptide/molecule activity or binding to a standard.  
       [0740] Preferably, an ELISA assay using, e.g., a monoclonal or polyclonal antibody, can measure polypeptide level in a sample. The antibody can measure polypeptide level by either binding, directly or indirectly, to the polypeptide or by competing with the polypeptide for a substrate.  
       [0741] All of the above assays can be used in a diagnostic or prognostic context. The molecules discovered using these assays can be used to treat disease or to bring about a particular result in a patient (e.g., blood vessel growth) by activating or inhibiting the polypeptide/molecule. Moreover, the assays can discover agents which may inhibit or enhance the production of the polypeptide from suitably manipulated cells or tissues.  
       [0742] Transcript Imaging and Toxicological Testing  
       [0743] Another embodiment relates to the use of dithp to develop a transcript image of a tissue or cell type. A transcript image represents the global pattern of gene expression by a particular tissue or cell type. Global gene expression patterns are analyzed by quantifying the number of expressed genes and their relative abundance under given conditions and at a given time. (See Seilhamer et al., “Comparative Gene Transcript Analysis,” U.S. Pat. No. 5,840,484, expressly incorporated by reference herein.) Thus a transcript image may be generated by hybridizing the polynucleotides of the present invention or their complements to the totality of transcripts or reverse transcripts of a particular tissue or cell type. In one embodiment, the hybridization takes place in high-throughput format, wherein the polynucleotides of the present invention or their complements comprise a subset of a plurality of elements on a microarray. The resultant transcript image would provide a profile of gene activity pertaining to human molecules for diagnostics and therapeutics.  
       [0744] Transcript images which profile dithp expression may be generated using transcripts isolated from tissues, cell lines, biopsies, or other biological samples. The transcript image may thus reflect dithp expression in vivo, as in the case of a tissue or biopsy sample, or in vitro, as in the case of a cell line.  
       [0745] Transcript images which profile dithp expression may also be used in conjunction with in vitro model systems and preclinical evaluation of pharmaceuticals, as well as toxicological testing of industrial and naturally-occurring environmental compounds. All compounds induce characteristic gene expression patterns, frequently termed molecular fingerprints or toxicant signatures, which are indicative of mechanisms of action and toxicity (Nuwaysir, E. F. et al. (1999) Mol. Carcinog. 24:153-159; Steiner, S. and Anderson, N. L. (2000) Toxicol. Lett. 112-113:467-71, expressly incorporated by reference herein). If a test compound has a signature similar to that of a compound with known toxicity, it is likely to share those toxic properties. These fingerprints or signatures are most useful and refined when they contain expression information from a large number of genes and gene families. Ideally, a genome-wide measurement of expression provides the highest quality signature. Even genes whose expression is not altered by any tested compounds are important as well, as the levels of expression of these genes are used to normalized the rest of the expression data. The normalization procedure is useful for comparison of expression data after treatment with different compounds. While the assignment of gene function to elements of a toxicant signature aids in interpretation of toxicity mechanisms, knowledge of gene function is not necessary for the statistical matching of signatures which leads to prediction of toxicity. (See, for example, Press Release 00-02 from the National Institute of Environmental Health Sciences, released Feb. 29, 2000, available at http:/www.niehs.nih.gov/oc./news/toxchip.htm). Therefore, it is important and desirable in toxicological screening using toxicant signatures to include all expressed gene sequences.  
       [0746] In one embodiment, the toxicity of a test compound is assessed by treating a biological sample containing nucleic acids with the test compound. Nucleic acids that are expressed in the treated biological sample are hybridized with one or more probes specific to the polynucleotides of the present invention, so that transcript levels corresponding to the polynucleotides of the present invention may be quantified. The transcript levels in the treated biological sample are compared with levels in an untreated biological sample. Differences in the transcript levels between the two samples are indicative of a toxic response caused by the test compound in the treated sample.  
       [0747] Another particular embodiment relates to the use of DITHP encoded by polynucleotides of the present invention to analyze the proteome of a tissue or cell type. The term proteome refers to the global pattern of protein expression in a particular tissue or cell type. Each protein component of a proteome can be subjected individually to further analysis. Proteome expression patterns, or profiles, are analyzed by quantifying the number of expressed proteins and their relative abundance under given conditions and at a given time. A profile of a cell&#39;s proteome may thus be generated by separating and analyzing the polypeptides of a particular tissue or cell type. In one embodiment, the separation is achieved using two-dimensional gel electrophoresis, in which proteins from a sample are separated by isoelectric focusing in the first dimension, and then according to molecular weight by sodium dodecyl sulfate slab gel electrophoresis in the second dimension (Steiner and Anderson, supra). The proteins are visualized in the gel as discrete and uniquely positioned spots, typically by staining the gel with an agent such as Coomassie Blue or silver or fluorescent stains. The optical density of each protein spot is generally proportional to the level of the protein in the sample. The optical densities of equivalently positioned protein spots from different samples, for example, from biological samples either treated or untreated with a test compound or therapeutic agent, are compared to identify any changes in protein spot density related to the treatment. The proteins in the spots are partially sequenced using, for example, standard methods employing chemical or enzymatic cleavage followed by mass spectrometry. The identity of the protein in a spot may be determined by comparing its partial sequence, preferably of at least 5 contiguous amino acid residues, to the polypeptide sequences of the present invention. In some cases, further sequence data may be obtained for definitive protein identification.  
       [0748] A proteomic profile may also be generated using antibodies specific for DITHP to quantify the levels of DITHP expression. In one embodiment, the antibodies are used as elements on a microarray, and protein expression levels are quantified by exposing the microarray to the sample and detecting the levels of protein bound to each array element (Lueking, A. et al. (1999) Anal. Biochem. 270:103-11; Mendoze, L. G. et al. (1999) Biotechniques 27:778-88). Detection may be performed by a variety of methods known in the art, for example, by reacting the proteins in the sample with a thiol- or amino-reactive fluorescent compound and detecting the amount of fluorescence bound at each array element.  
       [0749] Toxicant signatures at the proteome level are also useful for toxicological screening, and should be analyzed in parallel with toxicant signatures at the transcript level. There is a poor correlation between transcript and protein abundances for some proteins in some tissues (Anderson, N. L. and Seilhamer, J. (1997) Electrophoresis 18:533-537), so proteome toxicant signatures may be useful in the analysis of compounds which do not significantly affect the transcript image, but which alter the proteomic profile. In addition, the analysis of transcripts in body fluids is difficult, due to rapid degradation of mRNA, so proteomic profiling may be more reliable and informative in such cases.  
       [0750] In another embodiment, the toxicity of a test compound is assessed by treating a biological sample containing proteins with the test compound. Proteins that are expressed in the treated biological sample are separated so that the amount of each protein can be quantified. The amount of each protein is compared to the amount of the corresponding protein in an untreated biological sample. A difference in the amount of protein between the two samples is indicative of a toxic response to the test compound in the treated sample. Individual proteins are identified by sequencing the amino acid residues of the individual proteins and comparing these partial sequences to the DITHP encoded by polynucleotides of the present invention.  
       [0751] In another embodiment, the toxicity of a test compound is assessed by treating a biological sample containing proteins with the test compound. Proteins from the biological sample are incubated with antibodies specific to the DITHP encoded by polynucleotides of the present invention. The amount of protein recognized by the antibodies is quantified. The amount of protein in the treated biological sample is compared with the amount in an untreated biological sample. A difference in the amount of protein between the two samples is indicative of a toxic response to the test compound in the treated sample.  
       [0752] Transcript images may be used to profile dithp expression in distinct tissue types. This process can be used to determine human molecule activity in a particular tissue type relative to this activity in a different tissue type. Transcript images may be used to generate a profile of dithp expression characteristic of diseased tissue. Transcript images of tissues before and after treatment may be used for diagnostic purposes, to monitor the progression of disease, and to monitor the efficacy of drug treatments for diseases which affect the activity of human molecules.  
       [0753] Transcript images of cell lines can be used to assess human molecule activity and/or to identity cell lines that lack or misregulate this activity. Such cell lines may then be treated with pharmaceutical agents, and a transcript image following treatment may indicate the efficacy of these agents in restoring desired levels of this activity. A similar approach may be used to assess the toxicity of pharmaceutical agents as reflected by undesirable changes in human molecule activity. Candidate pharmaceutical agents may be evaluated by comparing their associated transcript images with those of pharmaceutical agents of known effectiveness.  
       [0754] Antisense Molecules  
       [0755] The polynucleotides of the present invention are useful in antisense technology. Antisense technology or therapy relies on the modulation of expression of a target protein through the specific binding of an antisense sequence to a target sequence encoding the target protein or directing its expression. (See, e.g., Agrawal, S., ed. (1996)  Antisense Therapeutics , Humana Press Inc., Totawa N.J.; Alama, A. et al. (1997) Pharmacol. Res. 36(3):171-178; Crooke, S. T. (1997) Adv. Pharmacol. 40:149; Sharma, H. W. and R. Narayanan (1995) Bioessays 17(12):1055-1063; and Lavrosky, Y. et al. (1997) Biochem. Mol. Med. 62(1):11-22.) An antisense sequence is a polynucleotide sequence capable of specifically hybridizing to at least a portion of the target sequence. Antisense sequences bind to cellular mRNA and/or genomic DNA, affecting translation and/or transcription. Antisense sequences can be DNA, RNA, or nucleic acid mimics and analogs. (See, e.g., Rossi, J. J. et al (1991) Antisense Res. Dev. 1(3):285-288; Lee, R. et al. (1998) Biochemistry 37(3):900-1010; Pardridge, W. M. et al. (1995) Proc. Natl. Acad. Sci. USA 92(12):5592-5596; and Nielsen, P. E. and Haaima, G. (1997) Chem Soc. Rev. 96:73-78.) Typically, the binding which results in modulation of expression occurs through hybridization or binding of complementary base pairs. Antisense sequences can also bind to DNA duplexes through specific interactions in the major groove of the double helix.  
       [0756] The polynucleotides of the present invention and fragments thereof can be used as antisense sequences to modify the expression of the polypeptide encoded by dithp. The antisense sequences can be produced ex vivo, such as by using any of the ABI nucleic acid synthesizer series (Applied Biosystems) or other automated systems known in the art. Antisense sequences can also be produced biologically, such as by transforming an appropriate host cell with an expression vector containing the sequence of interest. (See, e.g., Agrawal, supra.)  
       [0757] 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 F. M. et al. (1995)  Current Protocols in Molecular Biology , John Wiley &amp; Sons, New York N.Y.; Uckert, W. and W. Walther (1994) Pharmacol. Ther. 63(3):323-347.) Other gene delivery mechanisms include liposome-derived systems, artificial viral envelopes, and other systems known in the art. (See, e.g., Rossi, J. J. (1995) Br. Med. Bull. 51(1):217-225; Boado, R. J. et al. (1998) J. Pharm. Sci 87(11):1308-1315; and Morris, M. C. et al. (1997) Nucleic Acids Res. 25(14): 2730-2736 .)  
       [0758] Expression  
       [0759] In order to express a biologically active DITHP, the nucleotide sequences encoding DITHP or fragments 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. Methods which are well known to those skilled in the art may be used to construct expression vectors containing sequences encoding DITHP 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, supra, Chapters 4, 8, 16, and 17; and Ausubel, supra, Chapters 9, 10, 13, and 16.)  
       [0760] A variety of expression vector/host systems may be utilized to contain and express sequences encoding DITHP. 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 (mammalian) cell systems. (See, e.g., Sambrook, supra; Ausubel, 1995, supra, 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) Hume Gene Ther. 7:1937-1945; Takamatsu, N. (1987) EMBO J. 6:307-311; Coruzzi, G. et al. (1984) EMBO J. 3:1671-1680; Broglie, 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 N.Y., 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. Nail. 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, I. M. and N. Somia (1997) Nature 389:239-242.) The invention is not limited by the host cell employed.  
       [0761] For long term production of recombinant proteins in mammalian systems, stable expression of DITHP in cell lines is preferred. For example, sequences encoding DITHP 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. Any number of selection systems may be used to recover transformed cell lines. (See, e.g., Wigler, M. et al. (1977) Cell 11:223-232; Lowy, I. et al. (1980) Cell 22:817-823.; 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; Hartman, S. C. and R. C. Mulligan (1988) Proc. Natl. Acad. Sci. USA 85:8047-8051; Rhodes, C. A. (1995) Methods Mol. Biol. 55:121-131.)  
       [0762] Therapeutic Uses of dithp  
       [0763] The dithp of the invention 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)-X1 disease characterized by X-linked inheritance (Cavazzana-Calvo, M. et al. (2000) Science 288:669-672), severe combined immunodeficiency syndrome associated with an inherited adenosine deaminase (ADA) deficiency (Blaese, R. M. et al. (1995) Science  270:475-480 ; Bordignon, C. et al. (1995) Science  270:470-475 ), cystic fibrosis (Zabner, J. et al. (1993) Cell 75:207-216; Crystal, R. G. et al. (1995) Hun Gene Therapy 6:643-666; Crystal, R. G. et al. (1995) Hum. Gene Therapy 6:667-703), thalassemias, familial hypercholesterolemia, and hemophilia resulting from Factor VIII or Factor IX deficiencies (Crystal, R. G. (1995) Science  270:404-410 ; Verma, I. M. 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 (IV) (Baltimore, D. (1988) Nature 335:395-396; Poeschla, E. et al. (1996) Proc. Natl. Acad. Sci. USA. 93:11395-11399), hepatitis B or C virus (HBV, HCV); fungal parasites, such as  Candida albicans  and  Paracoccidioides brasiliensis ; and protozoan parasites such as  Plasmodium falciparum  and  Trypanosoma cruzi ). In the case where a genetic deficiency in dithp expression or regulation causes disease, the expression of dithp from an appropriate population of transduced cells may alleviate the clinical manifestations caused by the genetic deficiency.  
       [0764] In a further embodiment of the invention, diseases or disorders caused by deficiencies in dithp are treated by constructing mammalian expression vectors comprising dithp and introducing these vectors by mechanical means into dithp 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 Anderson, W. F. (1993) Annu. Rev. Biochem 62:191-217; Ivics, Z. (1997) Cell 91:501-510; Boulay, J. -L. and Recipon, H. (1998) Curr. Opin. Biotechnol. 9:445-450).  
       [0765] Expression vectors that may be effective for the expression of dithp include, but are not limited to, the PCDNA 3.1, EPITAG, PRCCMV2, PREP, PVAX vectors (Invitrogen, Carlsbad Calif.), PCMV-SCRIPT, PCMV-TAG, PEGSH/PERV (Stratagene, La Jolla Calif.), and PTET-OFF, PTET-ON, PTRE2, PTRE2-LUC, PTK-HYG (Clontech, Palo Alto Calif.). The dithp of the invention may be expressed using (i) a constitutively active promoter, (e.g., from cytomegalovirus (CMV), Rous sarcoma virus (RSV), SV40 virus, thymidine kinase (TK), or β-actin genes), (ii) an inducible promoter (e.g., the tetracycline-regulated promoter (Gossen, M. and Bujard, H. (1992) Proc. Natl. Acad. Sci. U.S.A. 89:5547-5551; Gossen, M. et al., (1995) Science 268:1766-1769; Rossi, F. M. V. and Blau, H. N. (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 Blau, H. M. supra), or (iii) a tissue-specific promoter or the native promoter of the endogenous gene encoding DITHP from a normal individual.  
       [0766] 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 Eb, A. J. (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.  
       [0767] In another embodiment of the invention, diseases or disorders caused by genetic defects with respect to dithp expression are treated by constructing a retrovirus vector consisting of (i) dithp under the control of an independent promoter or the retrovirus long terminal repeat (LTR) promoter, (ii) appropriate RNA packaging signals, and (ii) 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. U.S.A. 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 Miller, A. D. (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. Pat. No. 5,910,434 to Rigg (“Method for obtaining retrovirus packaging cell lines producing high transducing efficiency retroviral supernatant”) discloses a method for obtaining retrovirus packaging cell lines and is hereby incorporated by reference. Propagation of retrovirus vectors, transduction of a population of cells (e.g., CD4 +  T-cells), and the return of transduced cells to a patient are procedures well known to persons skilled in the art of gene therapy and have been well documented (Ranga, U. et al. (1997) J. Virol. 71:7020-7029; Bauer, G. et al. (1997) Blood 89:2259-2267; Bonyhadi, M. L. (1997) J. Virol.  71:4707-4716 ; Ranga, U. et ,al. (1998) Proc. Natl. Acad. Sci. U.S.A. 95:1201-1206; Su, L. (1997) Blood 89:2283-2290).  
       [0768] In the alternative, an adenovirus-based gene therapy delivery system is used to deliver dithp to cells which have one or more genetic abnormalities with respect to the expression of dithp. 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. Pat. No. 5,707,618 to Armentano (“Adenovirus vectors for gene therapy”), hereby incorporated by reference. For adenoviral vectors, see also Antinozzi, P. A. et al. (1999) Annu. Rev. Nutr. 19:511-544 and Verma, I. M. and Somia, N. (1997) Nature 18:389:239-242, both incorporated by reference herein.  
       [0769] In another alternative, a herpes-based, gene therapy delivery system is used to deliver dithp to target cells which have one or more genetic abnormalities with respect to the expression of dithp. The use of herpes simplex virus (HSV)-based vectors may be especially valuable for introducing dithp 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. Pat. No. 5,804,413 to DeLuca (“Herpes simplex virus strains for gene transfer”), which is hereby incorporated by reference. U.S. Pat. No. 5,804,413 teaches the use of recombinant HSV d92 which consists of a genome containing at least one exogenous gene to be transferred to a cell under the control of the appropriate promoter for purposes including human gene therapy. Also taught by this patent are the construction and use of recombinant HSV strains deleted for ICP4, ICP27 and ICP22. For HSV vectors, see also Goins, W. F. et al. 1999 J. Virol. 73:519-532 and Xu, H. et al., (1994) Dev. Biol. 163:152-161, hereby incorporated by reference. The manipulation of cloned herpesvirus sequences, the generation of recombinant virus following the transfection of multiple plasmids containing different segments of the large herpesvirus genomes, the growth and propagation of herpesvirus, and the infection of cells with herpesvirus are techniques well known to those of ordinary skill in the art.  
       [0770] In another alternative, an alphavirus (positive, single-stranded RNA virus) vector is used to deliver dithp 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 Li, K. -J. (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 dithp into the alphavirus genome in place of the capsid-coding region results in the production of a large number of dithp RNAs and the synthesis of high levels of DITHP 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 228:7483). The wide host range of alphaviruses will allow the introduction of dithp 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 skin in the art.  
       [0771] Antibodies  
       [0772] Anti-DITHP antibodies may be used to analyze protein expression levels. Such antibodies include, but are not limited to, polyclonal, monoclonal, chimeric, single chain, and Fab fragments. For descriptions of and protocols of antibody technologies, see, e.g., Pound J. D. (1998)  Immunochemical Protocols , Humana Press, Totowa, N.J.  
       [0773] The amino acid sequence encoded by the dithp of the Sequence Listing may be analyzed by appropriate software (e.g., LASERGENE NAVIGATOR software, DNASTAR) to determine regions of high immunogenicity. The optimal sequences for immunization are selected from the C-terminus, the N-terminus, and those intervening, hydrophilic regions of the polypeptide which are likely to be exposed to the external environment when the polypeptide is in its natural conformation. Analysis used to select appropriate epitopes is also described by Ausubel (1997, supra, Chapter 11.7). Peptides used for antibody induction do not need to have biological activity; however, they must be antigenic. Peptides used to induce specific antibodies may have an amino acid sequence consisting of at least five amino acids, preferably at least 10 amino acids, and most preferably at least 15 amino acids. A peptide which mimics an antigenic fragment of the natural polypeptide may be fused with another protein such as keyhole limpet hemocyanin (KLH; Sigma, St. Louis Mo.) for antibody production. A peptide encompassing an antigenic region may be expressed from a dithp, synthesized as described above, or purified from human cells.  
       [0774] Procedures well known in the art may be used for the production of antibodies. Various hosts including mice, goats, and rabbits, may be immunized by injection with a peptide. Depending on the host species, various adjuvants may be used to increase immunological response.  
       [0775] In one procedure, peptides about 15 residues in length may be synthesized using an ABI 431A peptide synthesizer (Applied Biosystems) using fmoc-chemistry and coupled to KLH (Sigma) by reaction with M-maleimidobenzoyl-N-hydroxysuccinimide ester (Ausubel, 1995, supra). Rabbits are immunized with the peptide-KLH complex in complete Freund&#39;s adjuvant. The resulting antisera are tested for antipeptide activity by binding the peptide to plastic, blocking with 1% bovine serum albumin (BSA), reacting with rabbit antisera, washing, and reacting with radioiodinated goat anti-rabbit IgG. Antisera with antipeptide activity are tested for anti-DITHP activity using protocols well known in the art, including ELISA, radioimmunoassay (RIA), and immunoblotting.  
       [0776] In another procedure, isolated and purified peptide may be used to immunize mice (about 100 μg of peptide) or rabbits (about 1 mg of peptide). Subsequently, the peptide is radioiodinated and used to screen the immunized animals&#39; B-lymphocytes for production of antipeptide antibodies. Positive cells are then used to produce hybridomas using standard techniques. About 20 mg of peptide is sufficient for labeling and screening several thousand clones. Hybridomas of interest are detected by screening with radioiodinated peptide to identify those fusions producing peptide-specific monoclonal antibody. In a typical protocol, wells of a multi-well plate (FAST, Becton-Dickinson, Palo Alto, Calif.) are coated with affinty-purified, specific rabbit-anti-mouse (or suitable anti-species IgG) antibodies at 10 mg/ml. The coated wells are blocked with 1% BSA and washed and exposed to supernatants from hybridomas. After incubation, the wells are exposed to radiolabeled peptide at 1 mg/ml.  
       [0777] Clones producing antibodies bind a quantity of labeled peptide that is detectable above background. Such clones are expanded and subjected to 2 cycles of cloning. Cloned hybridomas are injected into pristane-treated mice to produce ascites, and monoclonal antibody is purified from the ascitic fluid by affinity chromatography on protein A (Amersham Pharmacia Biotech). Several procedures for the production of monoclonal antibodies, including in vitro production, are described in Pound (supra). Monoclonal antibodies with antipeptide activity are tested for anti-DITHP activity using protocols well known in the art, including ELISA, RIA, and immunoblotting.  
       [0778] Antibody fragments containing specific binding sites for an epitope may also be generated. For example, such fragments include, but are not limited to, the F(ab′) 2  fragments produced by pepsin digestion of the antibody molecule, and the Fab fragments generated by reducing the disulfide bridges of the F(ab′) 2  fragments. Alternatively, construction of Fab expression libraries in filamentous bacteriophage allows rapid and easy identification of monoclonal fragments with desired specificity (Pound, supra, Chaps. 45-47). Antibodies generated against polypeptide encoded by dithp can be used to purify and characterize full-length DITHP protein and its activity, binding partners, etc.  
       [0779] Assays Using Antibodies  
       [0780] Anti-DITHP antibodies may be used in assays to quantify the amount of DRIP found in a particular human cell. Such assays include methods utilizing the antibody and a label to detect expression level under normal or disease conditions. The peptides and antibodies of the invention may be used with or without modification or labeled by joining them, either covalently or noncovalently, with a reporter molecule.  
       [0781] Protocols for detecting and measuring protein expression using either polyclonal or monoclonal antibodies are well known in the art. Examples include ELISA, RIA, and fluorescent activated cell sorting (FACS). Such immunoassays typically involve the formation of complexes between the DITHP and its specific antibody and the measurement of such complexes. These and other assays are described in Pound (supra).  
       [0782] Without ft 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.  
       [0783] The disclosures of all patents, applications, and publications mentioned above and below, including U.S. Ser. No. 60/184,777, U.S. Ser. No. 60/184,797, U.S. Ser. No. 60/184,698, U.S. Ser. No. 60/184,770, U.S. Ser. No. 60/184,774, U.S. Ser. No. 60/184,693, U.S. Ser. No. 60/184,771,U.S. Ser. No. 60/184,813, U.S. Ser. No. 60/184,773, U.S. Ser. No. 60/184,776, U.S. Ser. No. 60/184,769, U.S. Ser. No. 60/184,768, U.S. Ser. No. 60/184,837, U.S. Ser. No. 60/184,697, U.S. Ser. No. 60/184,841, U.S. Ser. No. 60/184,772, U.S. Ser. No. 60/185,213, U.S. Ser. No. 60/185,216, U.S. Ser. No. 60/204,863, U.S. Ser. No. 60/205,221, U.S. Ser. No. 60/204,815, U.S. Ser. No. 60/203,785, U.S. Ser. No. 60/204,821, U.S. Ser. No. 60/204,908, U.S. Ser. No. 60/204,226, U.S. Ser. No. 60/204,525, U.S. Ser. No. 60/205,285, U.S. Ser. No. 60/205,232, U.S. Ser. No. 60/205,323, U.S. Ser. No. 60/205,287, U.S. Ser. No. 60/205,324, and U.S. Ser. No. 60/205,286, are hereby expressly incorporated by reference. 
     
    
    
     EXAMPLES  
     [0784] I. Construction of cDNA Libraries  
     [0785] RNA was purchased from CLONTECH Laboratories, Inc. Palo Alto Calif.) or isolated from various tissues. 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 CsCl cushions or extracted with chloroform. RNA was precipitated with either isopropanol or sodium acetate and ethanol, or by other routine methods.  
     [0786] Phenol extraction and precipitation of RNA were repeated as necessary to increase RNA purity. In most cases, RNA was treated with DNase. For most libraries, poly(A+) RNA was isolated using oligo dm-coupled paramagnetic particles (Promega Corporation (Promega), Madison Wis.), OLIGOTEX latex particles (QIAGEN, Inc. (QIAGEN), Valencia Calif.), 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, Inc., Austin IX).  
     [0787] 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 Cloning Systems, Inc. (Stratagene), La Jolla Calif.) or SUPERSCRIPT plasmid system (Life Technologies), using the recommended procedures or similar methods known in the art. (See, e.g., Ausubel, 1997, supra, Chapters 5.1 through 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 SEPHAROSE S1000, SEPHAROSE CL2B, or SEPHAROSE C14B 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), PSPORT1 plasmid (Life Technologies), PCDNA2.1 plasmid nitrogen, Carlsbad Calif.), PBK-CMV plasmid (Stratagene), or pINCY (Incyte Genomics, Palo Alto Calif.), or derivatives thereof. Recombinant plasmids were transformed into competent  E. coli  cells including XL1-Blue, XL1-BlueMRF, or SOLR from Stratagene or DH5α, DH10B, or ElectroMAX DH10B from Life Technologies.  
     [0788] II. Isolation of cDNA Clones  
     [0789] Plasmids 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: the Magic or WIZARD Minipreps DNA purification system (Promega); the AGTC Miniprep purification kit (Edge BioSystems, Gaithersburg Md.); and the QIAWELL 8, QIAWELL 8 Plus, and QIAWELL 8 Ultra plasmid purification systems or the R.E.A.L. PREP 96 plasmid purification kit (QIAGEN). Following precipitation, plasmids were resuspended in 0.1 ml of distilled water and stored, with or without lyophilization, at 4° C.  
     [0790] 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, Inc. (Molecular Probes), Eugene Oreg.) and a FLUOROSKAN II fluorescence scanner (Labsystems Oy, Helsinki, Finland).  
     [0791] III. Sequencing and Analysis  
     [0792] cDNA sequencing reactions were processed using standard methods or high-throughput instrumentation such as the ABI CATALYST 800 thermal cycler (Applied Biosystems) or the PTC-200 thermal cycler (MJ Research) in conjunction with the HYDRA microdispenser (Robbins Scientific Corp., Sunnyvale Calif.) or the MICROLAB 2200 liquid transfer system (Hamilton). 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 (Applied 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 (Applied 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, Chapter 7.7). Some of the cDNA sequences were selected for extension using the techniques disclosed in Example VIII.  
     [0793] IV. Assembly and Analysis of Sequences  
     [0794] Component sequences from chromatograms were subject to PHRED analysis and assigned a quality score. The sequences having at least a required quality score were subject to various pre-processing editing pathways to eliminate, e.g., low quality 3′ ends, vector and linker sequences, polyA tails, Alu repeats, mitochondrial and ribosomal sequences, bacterial contamination sequences, and sequences smaller than 50 base pairs. In particular, low-information sequences and repetitive elements (e.g., dinucleotide repeats, Alu repeats, etc.) were replaced by “n&#39;s”, or masked, to prevent spurious matches.  
     [0795] Processed sequences were then subject to assembly procures in which the sequences were assigned to gene bins (ins). Each sequence could only belong to one bin. Sequences in each gene bin were assembled to produce consensus sequences (templates). Subsequent new sequences were added to existing bins using BLASTn (v. 1.4 WashU) and CROSSMATCH. Candidate pairs were identified as all BLAST hits having a quality score greater than or equal to 150. Alignments of at least 82% local identity were accepted into the bin. The component sequences from each bin were assembled using a version of PHRAP. Bins with several overlapping component sequences were assembled using DEEP PHRAP. The orientation (sense or antisense) of each assembled template was determined based on the number and orientation of its component sequences. Template sequences as disclosed in the sequence listing correspond to sense strand sequences (the “forward” reading frames), to the best determination. The complementary (antisense) strands are inherently disclosed herein The component sequences which were used to assemble each template consensus sequence are listed in Table 4, along with their positions along the template nucleotide sequences.  
     [0796] Bins were compared against each other and those having local similarity of at least 82% were combined and reassembled. Reassembled bins having templates of insufficient overlap (less than 95% local identity) were re-split. Assembled templates were also subject to analysis by STITCHER/EXON MAPPER algorithms which analyze the probabilities of the presence of splice variants, alternatively spliced exons, splice junctions, differential expression of alternative spliced genes across tissue types or disease states, etc. These resulting bins were subject to several rounds of the above assembly procedures.  
     [0797] Once gene bins were generated based upon sequence alignments, bins were clone joined based upon clone information. If the 5′ sequence of one clone was present in one bin and the 3′ sequence from the same clone was present in a different bin, it was likely that the two bins actually belonged together in a single bin. The resulting combined bins underwent assembly procedures to regenerate the consensus sequences.  
     [0798] The final assembled templates were subsequently annotated using the following procedure. Template sequences were analyzed using BLASTn (v2.0, NCBI) versus gbpri (GenBank version 120). “Hits” were defined as an exact match having from 95% local identity over 200 base pairs through 100% local identity over 100 base pairs, or a homolog match having an E-value, i.e. a probability score, of ≦1×10 −8 . The hits were subject to frameshift FASTx versus GENPEPT (GenBank version 120). (See Table 7). In this analysis, a homolog match was defined as having an E-value of ≦1×10 −8 . The assembly method used above was described in “System and Methods for Analyzing Biomolecular Sequences,” U.S. Ser. No. 09/276,534, filed Mar. 25, 1999, and the LIFESEQ Gold user manual (Incyte) both incorporated by reference herein.  
     [0799] Following assembly, template sequences were subjected to motif, BLAST, and functional analyses, and categorized in protein hierarchies using methods described in, e.g., “Database System Employing Protein Function Hierarchies for Viewing Biomolecular Sequence Data,” U.S. Ser. No. 08/812,290, filed Mar. 6, 1997; “Relational Database for Storing Biomolecule Information,” U.S. Ser. No. 08/947,845, filed Oct. 9, 1997; “Project-Based Full-Length Biomolecular Sequence Database,” U.S. Ser. No. 08/811,758, filed Mar. 6, 1997; and “Relational Database and System for Storing Information Relating to Biomolecular Sequences,” U.S. Ser. No. 09/034,807, filed Mar. 4, 1998, all of which are incorporated by reference herein.  
     [0800] The template sequences were further analyzed by translating each template in all three forward reading frames and searching each translation against the Pfam database of hidden Markov model-based protein families and domains using the HOMER software package (available to the public from Washington University School of Medicine, St. Louis Mo.). Regions of templates which, when translated, contain similarity to Pfam consensus sequences are reported in Table 2, along with descriptions of Pfam protein domains and families. Only those Pfam hits with an E-value of ≦1×10 −3  are reds (See also World Wide Web site http:/pfam.wustl.edu/for detailed descriptions of Pfam protein domains and families.)  
     [0801] Additionally, the template sequences were translated in all three forward reading frames, and each translation was searched against hidden Markov models for signal peptides using the HMMER software package. Construction of hidden Markov models and their usage in sequence analysis has been described. (See, for example, Eddy, S. R. (1996) Curr. Opin. Str. Biol. 6:361-365.) Oly those signal peptide hits with a cutoff score of 11 bits or greater are reported. A cutoff score of 11 bits or greater corresponds to at least about 91-94% true-positives in signal peptide prediction. Template sequences were also translated in all three forward reading frames, and each translation was searched against TMAP, a program that uses weight matrices to delineate transmembrane segments on protein sequences and determine orientation, with respect to the cell cytosol (Persson, B. and P. Argos (1994) J. Mol. Biol. 237:182-192; Persson, B. and P. Argos (1996) Protein Sci. 5:363-371). Regions of templates which, when translated, contain similarity to signal peptide or transmembrane consensus sequences are reported in Table 3.  
     [0802] The results of HMMER analysis as reported in Tables 2 and 3 may support the results of BLAST analysis as reported in Table 1 or may suggest alternative or additional properties of template-encoded polypeptides not previously uncovered by BLAST or other analyses.  
     [0803] Template sequences are further analyzed using the bioinformatics tools listed in Table 7, or using sequence analysis software known in the art such as MACDNASIS PRO software (Hitachi Software Engineering, South San Francisco Calif.) and LASERGENE software (DNASTAR). Template sequences may be further queried against public databases such as the GenBank rodent, mammalian, vertebrate, prokaryote, and eukaryote databases.  
     [0804] The template sequences were translated to derive the corresponding longest open reading frame as presented by the polypeptide sequences. Alternatively, a polypeptide of the invention may begin at any of the methionine residues within the full length translated polypeptide. Polypeptide sequences were subsequently analyzed by querying against the GenBank protein database (GENPEPT, (GenBank version 121)). Full length polynucleotide sequences are also analyzed using MACDNASIS PRO software (Hitachi Software Engineering, South San Francisco Calif.) and LASERGENE software (DNASTAR). Polynucleotide and polypeptide sequence alignments are generated using 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.  
     [0805] Table 6 shows sequences with homology to the polypeptides of the invention as identified by BLAST analysis against the GenBank protein (GENPEPT) database. Column 1 shows the polypeptide sequence identification number (SEQ ID NO:) for the polypeptide segments of the invention. Column 2 shows the reading frame used in the translation of the polynucleotide sequences encoding the polypeptide segments. Column 3 shows the length of the translated polypeptide segments. Columns 4 and 5 show the start and stop nucleotide positions of the polynucleotide sequences encoding the polypeptide segments. Column 6 shows the GenBank identification number (GI Number) of the nearest GenBank homolog. Column 7 shows the probability score for the match between each polypeptide and its GenBank homolog. Column 8 shows the annotation of the GenBank homolog.  
     [0806] V. Analysis of Polynucleotide Expression  
     [0807] 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.)  
     [0808] 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   ×   Percent                 Identity       5   ×   minimum                   {       length                   (     Seq   .              1     )       ,     length                   (     Seq   .              2     )         }                     
 
     [0809] 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 normalize 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:  
     [0810] VI. Tissue Distribution Profiling  
     [0811] A tissue distribution profile is determined for each template by compiling the cDNA library tissue classifications of its component cDNA sequences. Each. component sequence, is derived from a cDNA library constructed from a human tissue. Each human tissue is classified into one of the following categories: cardiovascular system; connective tissue; digestive system; embryonic structures; endocrine system; exocrine glands; genitalia, female; genitalia, male; germ cells; hemic and immune system; liver; musculoskeletal system; nervous system; pancreas; respiratory system; sense organs; skin; stomatognathic system; unclassified/mixed; or urinary tract. Template sequences, component sequences, and cDNA library/tissue information are found in the LIFESEQ GOLD database (Incyte Genomics, Palo Alto Calif.).  
     [0812] Table 5 shows the tissue distribution profile for the templates of the invention. For each template, the three most frequently observed tissue categories are shown in column 3, along with the percentage of component sequences belonging to each category. Only tissue categories with percentage values of ≧10% are shown. A tissue distribution of “widely distributed” in column 3 indicates percentage values of &lt;10% in all tissue categories.  
     [0813] VII. Transcript Image Analysis  
     [0814] Transcript images are generated as described in Seilhamer et al., “Comparative Gene Transcript Analysis,” U.S. Pat. No. 5,840,484, incorporated herein by reference.  
     [0815] VIII. Extension of Polynucleotide Sequences and Isolation of a Full-length cDNA  
     [0816] Oligonucleotide primers designed using a dithp of the Sequence Listing are used to extend the nucleic acid sequence. One primer is synthesized to initiate 5′ extension of the template, and the other primer, to initiate 3′ extension of the template. The initial primers may be designed using OLIGO 4.06 software (National Biosciences, Inc. (National Biosciences), Plymouth Minn.), 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 dimerzations are avoided. Selected human cDNA libraries are used to extend the sequence. If more than one extension is necessary or desired, additional or nested sets of primers are designed.  
     [0817] High fidelity amplification is obtained by PCR using methods well known in the art PCR is performed in 96-well plates using the PTC-200 thermal cycler (MJ Research). The reaction mix contains DNA template, 200 nmol of each primer, reaction buffer containing Mg 2+ , (NH 4 ) 2 SO 4 , and β-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+ are 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.  
     [0818] The concentration of DNA in each well is determined by dispensing 100 μl PICOGREEN quantitation reagent (0.25% (v/v); Molecular Probes) dissolved in 1× Tris-EDTA CE) and 0.5 μl of undiluted PCR product into each well of an opaque fluorimeter plate (Corning Incorporated (Corning), Corning N.Y.), allowing the DNA to bind to the reagent. The plate is scanned in a FLUOROSKAN II Labsystems Oy) to measure the fluorescence of the sample and to quantify the concentration of DNA A 5 μl to 10 μl aliquot of the reaction mixture is analyzed by electrophoresis on a 1% agarose mini-gel to determine which reactions are successful in extending the sequence.  
     [0819] The extended nucleotides are desalted and concentrated, transferred to 384-well plates, digested with CviJI cholera virus endonuclease (Molecular Biology Research, Madison Wis.), and sonicated or sheared prior to religation into pUC 18 vector (Amersham Pharmacia Biotech). For shotgun sequencing, the digested nucleotides are separated on low concentration (0.6 to 0.8%) agarose gels, fragments are excised, and agar digested with AGAR ACE (Promega). Extended clones are reilgated using T4 ligase (New England Biolabs, Inc., Beverly Mass.) 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 are selected on antibiotic-containing media, individual colonies are picked and cultured overnight at 37° C. in 384-well plates in LB/2x carbenicillin liquid media.  
     [0820] The cells are lysed, and DNA is 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., 2min; Step 5: steps 2, 3, and 4 repeated 29 times; Step 6: 72° C., 5 min; Step 7: storage at 4° C. DNA is quantified by PICOGREEN reagent (Molecular Probes) as described above. Samples with low DNA recoveries are reamplified using the same conditions as described above. Samples are 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 (Applied Biosystems).  
     [0821] In like manner, the dithp is used to obtain regulatory sequences (promoters, introns, and enhancers) using the procedure above, oligonucleotides designed for such extension, and an appropriate genomic library.  
     [0822] IX. Labeling of Probes and Southern Hybridization Analyses  
     [0823] Hybridization probes derived from the dithp of the Sequence Listing are employed for screening cDNAs, mRNAs, or genomic DNA. The labeling of probe nucleotides between 100 and 1000 nucleotides in length is specifically described, but essentially the same procedure may be used with larger cDNA fragments. Probe sequences are labeled at room temperature for 30 minutes using a T4 polynucleotide kinase, γ 32 P-ATP, and 0.5× One-Phor-All Plus (Amersham Pharmacia Biotech) buffer and purified using a ProbeQuant G-50 Microcolumn (Amersham Pharmacia Biotech). The probe mixture is diluted to 10 7  dpm/μg/ml hybridization buffer and used in a typical membrane-based hybridization analysis.  
     [0824] The DNA is digested with a restriction endonuclease such as Eco RV and is electrophoresed through a 0.7% agarose gel. The DNA fragments are transferred from the agarose to nylon membrane (NYTRAN Plus, Schleicher &amp; Schuell, Inc., Keene N. H.) using procedures specified by the manufacturer of the membrane. Prehybridization is carried out for three or more hours at 68° C., and hybridization is carried out overnight at 68° C. To remove non-specific signals, blots are sequentially washed at room temperature under increasingly stringent conditions, up to 0.1× saline sodium citrate (SSC) and 0.5% sodium dodecyl sulfate. After the blots are placed in a PHOSPHORIMAGER cassette (Molecular Dynamics) or are exposed to autoradiography film, hybridization patterns of standard and experimental lanes are compared. Essentially the same procedure is employed when screening RNA.  
     [0825] X. Chromosome Mapping of dithp  
     [0826] The cDNA sequences which were used to assemble SEQ ID NO:1-211 are 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 match SEQ ID NO:1-211 are assembled into clusters of contiguous and overlapping sequences using assembly algorithms such as PHRAP (Table 7). Radiation hybrid and genetic mapping data available from public resources such as the Stanford Human Genome Center (SHGC), Whitehead Institute for Genome Research (WIGR), and Généthon are used to determine if any of the clustered sequences have been previously mapped. Inclusion of a mapped sequence in a cluster will result 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 NO:1-211 are described as ranges, or intervals, of human chromosomes. The map position of an interval, in centiMorgans, is measured relative to the terminus of the chromosome&#39;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 Généthon which provide boundaries for radiation hybrid markers whose sequences were included in each of the clusters.  
     [0827] XI. Microarray Analysis  
     [0828] Probe Preparation from Tissue or Cell Samples  
     [0829] Total RNA is isolated from tissue samples using the guanidinium thiocyanate method and polyA +  RNA is purified using the oligo (dT) cellulose method Each polyA +  RNA sample is reverse transcribed using MMLV reverse-transcriptase, 0.05 pg/μl oligo-dT primer (21mer), 1×first strand buffer, 0.03 units/μl RNase inhibitor, 500 μM dATP, 500 μM dGTP, 500 μM d=TP, 40 μM dCTP, 40 μM dCTP-Cy3 (BDS) or dCTP-Cy5 (Amersham Pharmacia Biotech). The reverse transcription reaction is performed in a 25 ml volume containing 200 ng polyA +  RNA with GEMBRIGHT kits (Incyte). Specific control polyA +  RNAs are synthesized by in vitro transcription from non-coding yeast genomic DNA (W. Lei, unpublished). As quantitative controls, the control mRNAs at 0.002 ng, 0.02 ng, 0.2 ng, and 2 ng are diluted into reverse transcription reaction at ratios of 1:100,000, 1:10,000, 1:1000, 1:100 (w/w) to sample mRNA respectively. The control mRNAs are diluted into reverse transcription reaction at ratios of 1:3, 3:1, 1:10, 10:1, 1:25, 25:1 (w/w) to sample mRNA differential expression patterns. After incubation at 370 C for 2 br, each reaction sample (one with Cy3 and another with Cy5 labeling) is treated with 2.5 ml of 0.5M sodium hydroxide and incubated for 20 minutes at 85° C. to the stop the reaction and degrade the RNA Probes are purified using two successive CHROMA SPIN 30 gel filtration spin columns (CLONTECH Laboratories, Inc. (CLONTECH), Palo Alto Calif.) 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 probe is then dried to completion using a SpeedVAC (Savant Instruments Inc., HoIbrook N.Y.) and resuspended in 14 μl 5×SSC/0.2%  
     [0830] SDS.  
     [0831] Microarray Preparation  
     [0832] 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 μg. Amplified array elements are then purified using SEPHACRYL-400 (Amersham Pharmacia Biotech).  
     [0833] 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.  
     [0834] Array elements are applied to the coated glass substrate using a procedure described in U.S. Pat. No. 5,807,522, incorporated herein by reference. 1 μl of the array element DNA, at an average concentration of 100 ng/μl, is loaded into the open capillary printing element by a high-speed robotic apparatus. The apparatus then deposits about 5 nl of array element sample per slide.  
     [0835] 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, Mass.) for 30 minutes at 60° C. followed by washes in 0.2% SDS and distilled water as before.  
     [0836] Hybridization  
     [0837] Hybridization reactions contain 9 μl of probe mixture consisting of 0.2 μg each of Cy3 and Cy5 labeled cDNA synthesis products in 5×SSC, 0.2% SDS hybridization buffer. The probe mixture is heated to 65° C. for 5 minutes and is aliquoted onto the microarray surface and covered with an 1.8 cm 2  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 μl of 5×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 (1×SSC, 0.1% SDS), three times for 10 minutes each at 45° C. in a second wash buffer (0.1×SSC), and dried  
     [0838] Detection  
     [0839] Reporter-labeled hybridization complexes are detected with a microscope equipped with an Innova 70 mixed gas 10 W laser (Coherent, Inc., Santa Clara Calif.) capable of generating spectral lines at 488 nm for excitation of Cy3 and at 6321n for excitation of Cy5. The excitation laser light is focused on the array using a 20×microscope objective (Nikon, Inc., Melville N.Y.). 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×1.8 cm array used in the present example is scanned with a resolution of 20 micrometers.  
     [0840] 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 R1477, Hamamatsu Photonics Systems, Bridgewater N.J.) 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 Cy5. 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.  
     [0841] The sensitivity of the scans is typically calibrated using the signal intensity generated by a cDNA control species added to the probe mix 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 probes 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.  
     [0842] 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, Mass.) 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&#39;s emission spectrum.  
     [0843] 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 win 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).  
     [0844] XII. Complementary Nucleic Acids  
     [0845] Sequences complementary to the dithp are used to detect, decrease, or inhibit expression of the naturally occurring nucleotide. The use of oligonucleotides comprising from about 15 to 30 base pairs is typical in the art. However, smaller or larger sequence fragments can also be used. Appropriate oligonucleotides are designed from the dithp using OLIGO 4.06 software (National Biosciences) or other appropriate programs and are synthesized using methods standard in the art or ordered from a commercial supplier. To inhibit transcription, a complementary oligonucleotide is designed from the most unique 5′ sequence and used to prevent transcription factor binding to the promoter sequence. To inhibit translation, a complementary oligonucleotide is designed to prevent ribosomal binding and processing of the transcript  
     [0846] XIII. Expression of DITHP  
     [0847] Expression and purification of DITHP is accomplished using bacterial or virus-based expression systems. For expression of DITHP 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 T5 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 DITHP upon induction with isopropyl beta-D-thiogalactopyranoside (IPTG). Expression of DITHP in eukaryotic cells is achieved by infecting insect or mammalian cell lines with recombinant  Autosraphica californica  nuclear polyhedrosis virus (AcMNPV), commonly known as baculovirus. The nonessential polyhedrin gene of baculovirus is replaced with cDNA encoding DITHP 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 fruiperda  (Sf9) insect cells in most cases, or human hepatocytes, in some cases. Infection of the latter requires additional genetic modifications to baculovirus. (See e.g., Engelhard, supra; and Sandig, supra.)  
     [0848] In most expression systems, DITHP 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 japonicum , 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 DITHP at specifically engineered sites. FLAG, an 8-amino acid peptide, enables immunoaffnity purification using commercially available monoclonal and polyclonal anti-FLAG antibodies (Eastman Kodak Company, Rochester N.Y.). 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, Chapters 10 and 16). Purified DITh=obtained by these methods can be used directly in the following activity assay.  
     [0849] XIV. Demonstration of DITHP Activity  
     [0850] DITHP activity is demonstrated through a variety of specific assays, some of which are outlined below.  
     [0851] Oxidoreductase activity of DITHP is measured by the increase in extinction coefficient of NAD(P)H coenzyme at 340 nm for the measurement of oxidation activity, or the decrease in extinction coefficient of NAD(P)H coenzyme at 340 nm for the measurement of reduction activity (Dalziel, K. (1963) J. Biol. Chem. 238:2850-2858). One of three substrates may be used: Asn-βGal, biocytidine, or ubiquinone-10. The respective subunits of the enzyme reaction, for example, cytochtome c 1 -b oxidoreductase and cytochrome c, are reconstituted. The reaction mixture contains a)1-2 mg/ml DITHP; and b) 15 mM substrate, 2.4 mM NAD(P) + in 0.1 M phosphate buffer, pH 7.1 (oxidation reaction), or 2.0 mM NAD(P)H, in 0.1 M Na 2 HPO 4  buffer, pH 7.4 (reduction reaction); in a total volume of 0.1 ml. Changes in absorbance at 340 nm (A340) are measured at 23.5° C. using a recording spectrophotometer (Shimadzu Scientific Instruments, Inc., Pleasanton Calif.). The amount of NAD(P)H is stoichiometrically equivalent to the amount of substrate initially present, and the change in A 340  is a direct measure of the amount of NAD(P)H produced; ΔA 340 =6620[NADH]. Oxidoreductase activity of DITHP activity is proportional to the amount of NAD(P)H present in the assay.  
     [0852] Transferase activity of DITHP is measured through assays such as a methyl transferase assay in which the transfer of radiolabeled methyl groups between a donor substrate and an acceptor substrate is measured (Bokar, J. A. et al. (1994) J. Biol. Chem. 269:17697-17704). Reaction mixtures (50 μl final volume) contain 15 mM HEPES, pH 7.9, 1.5 mM MgCl 2 , 10 mM dithiothreitol, 3% polyvinylalcohol, 1.5 μCi [methyl- 3 H]AdoMet (0.375 μM AdoMet) (DuPont-NEN), 0.6 μg DITHP, and acceptor substrate (0.4 μg [ 35 S]RNA or 6-mercaptopurine (6-MP) to 1 mM final concentration). Reaction mixtures are incubated at 30° C. for 30 minutes, then 65° C. for 5 minutes. The products are separated by chromatography or electrophoresis and the level of methyl transferase activity is determined by quantification of methyl-3H recovery.  
     [0853] DITHP hydrolase activity is measured by the hydrolysis of appropriate synthetic peptide substrates conjugated with various chromogenic molecules in which the degree of hydrolysis is quantified by spectrophotometric (or fluorometric) absorption of the released chromophore. (Beynon, R. J. and J. S. Bond (1994)  Proteolytic Enzymes: A Practical Approach , Oxford University Press, New York N.Y., pp. 25-55) Peptide substrates are designed according to the category of protease activity as endopeptidase (serine, cysteine, aspartic proteases), animopeptidase (leucine aminopeptidase), or carboxypeptidase (Carboxypeptidase A and B, procollagen C-proteinase).  
     [0854] DITHP isomerase activity such as peptidyl prolyl cis/trans isomerase activity can be assayed by an enzyme assay described by Rahfeld, J. U., et al. (1994) (FEBS Let 352:180-184). The assay is performed at 10° C. in 35 mM HEPES buffer, pH 7.8, containing chymotrypsin (0.5 mg/ml) and DITHP at a variety of concentrations. Under these assay conditions, the substrate, Suc-Ala-Xaa-Pro-Phe-4-NA, is in equilibrium with respect to the prolyl bond, with 80-95% in trans and 5-20% in cis conformation. An aliquot (2 ul) of the substrate dissolved in dimethyl sulfoxide (10 mg/ml) is added to the reaction mixture described above. Only the cis isomer of the substrate is a substrate for cleavage by chymotrypsin. Thus, as the substrate is isomerized by DITHP, the product is cleaved by chymotrypsin to produce 4-nitroanilide, which is detected by it&#39;s absorbance at 390 nm. 4 Nitroanilide appears in a time-dependent and a DITHP concentration-dependent manner.  
     [0855] An assay for DITHP activity associated with growth and development measures cell proliferation as the amount of newly initiated DNA synthesis in Swiss mouse 3T3 cells. A plasmid containing polynucleotides encoding DITHP is transfected into quiescent 3T3 cultured cells using methods well known in the art. The transiently transfected cells are then incubated in the presence of [ 3 H]thymidine, a radioactive DNA precursor. Where applicable, varying amounts of DITHP ligand are added to the transfected cells. Incorporation of [ 3 H]thymidine into acid-precipitable DNA is measured over an appropriate time interval, and the amount incorporated is directly proportional to the amount of newly synthesized DNA.  
     [0856] Growth factor activity of DITHP is measured by the stimulation of DNA synthesis in Swiss mouse 3T3 cells (McKay, I. and L. Leigh, eds. (1993)  Growth Factors: A Practical Approach , Oxford University Press, New York N.Y.). Initiation of DNA synthesis indicates the cells&#39; entry into the mitotic cycle and their commitment to undergo later division 3T3 cells are competent to respond to most growth factors, not only those that are mitogenic, but also those that are involved in embryonic induction. This competence is possible because the in vivo specificity demonstrated by some growth factors is not necessarily inherent but is determined by the responding tissue. In this assay, varying amounts of DITHP are added to quiescent 3T3 cultured cells in the presence of [ 3 H]thymidine, a radioactive DNA precursor. DITHP for this assay can be obtained by recombinant means or from biochemical preparations. Incorporation of [ 3 H]thymidine into acid-precipitable DNA is measured over an appropriate time interval, and the amount incorporated is directly proportional to the amount of newly synthesized DNA A linear dose-response curve over at least a hundred-fold DITHP concentration range is indicative of growth factor activity. One unit of activity per milliliter is defined as the concentration of DITHP producing a 50% response level, where 100% represents maximal incorporation of [ 3 H]thymidine into acid-precipitable DNA.  
     [0857] Alternatively, an assay for cytokine activity of DITHP measures the proliferation of leukocytes. In this assay, the amount of tritiated thymidine incorporated into newly synthesized DNA is used to estimate proliferative activity. Varying amounts of DITHP are added to cultured leukocytes, such as granulocytes, monocytes, or lymphocytes, in the presence of [ 3 H]thymidine, a radioactive DNA precursor. DITHP for this assay can be obtained by recombinant means or from biochemical preparations. Incorporation of [ 3 H]thymidine into acid-precipitable DNA is measured over an appropriate time interval, and the amount incorporated is directly proportional to the amount of newly synthesized DNA. A linear dose-response curve over at least a hundred-fold DITHP concentration range is indicative of DITHP activity. One unit of activity per milliliter is conventionally defined as the concentration of DITHP producing a 50% response level, where 100% represents maximal incorporation of [ 3 H]thymidine into acid-precipitable DNA.  
     [0858] An alternative assay for DITHP cytokine activity utilizes a Boyden micro chamber (Neuroprobe, Cabin John M.D.) to measure leukocyte chemotaxis (Vicari, supra). In this assay, about 10 5  migratory cells such as macrophages or monocytes are placed in cell culture media in the upper compartment of the chamber. Varying dilutions of DITHP are placed in the lower compartment. The two compartments are separated by a 5 or 8 micron pore polycarbonate filter (Nucleopore, Pleasanton Calif.). After incubation at 37° C. for 80 to 120 minutes, the filters are fixed in methanol and stained with appropriate labeling agents. Cells which migrate to the other side of the filter are counted using standard microscopy. The chemotactic index is calculated by dividing the number of migratory cells counted when DITHP is present in the lower compartment by the number of migratory cells counted when only media is present in the lower compartment. The chemotactic index is proportional to the activity of DITHP.  
     [0859] Alternatively, cell lines or tissues transformed with a vector containing dithp can be assayed for DITHP activity by immunoblotting. Cells are denatured in SDS in the presence of β-mercaptoethanol, nucleic acids removed by ethanol precipitation, and proteins purified by acetone precipitation. Pellets are resuspended in 20 mM tris buffer at pH 7.5 and incubated with Protein G-Sepharose pre-coated with an antibody specific for DITHP. After washing, the Sepharose beads are boiled in electrophoresis sample buffer, and the eluted proteins subjected to SDS-PAGE. The SDS-PAGE is transferred to a nitrocellulose membrane for immunoblotting, and the DITHP activity is assessed by visualizing and quantifying bands on the blot using the antibody specific for DITHP as the primary antibody and  125 I-labeled IgG specific for the primary antibody as the secondary antibody.  
     [0860] DITHP kinase activity is measured by phosphorylation of a protein substrate using γ-labeled [ 32 p]-ATP and quantitation of the incorporated radioactivity using a radioisotope counter. DITHP is incubated with the protein substrate, [ 32 P]-ATP, and an appropriate kinase buffer. The [ 32 P] incorporated into the product is separated from free [ 32 ]-ATP by electrophoresis and the incorporated [ 32 P] is counted. The amount of [ 32 P] recovered is proportional to the kinase activity of DITHP in the assay. A determination of the specific amino acid residue phosphorylated is made by phosphoamino acid analysis of the hydrolyzed protein.  
     [0861] In the alternative, DITHP activity is measured by the increase in cell proliferation resulting from transformation of a mammalian cell line such as COS7, HeLa or CHO with an eukaryotic expression vector encoding DITHP. Eukaryotic expression vectors are commercially available, and the techniques to introduce them into cells are well known to those skilled in the art. The cells are incubated for 48-72 hours after transformation under conditions appropriate for the cell line to allow expression of DITHP. Phase microscopy is then used to compare the mitotic index of transformed versus control cells. An increase in the mitotic index indicates DITHP activity.  
     [0862] In a further alternative, an assay for DITHP signaling activity is based upon the ability of GPCR family proteins to modulate G protein-activated second messenger signal transduction pathways (e.g., cAMP; Gaudin, P. et al. (1998) J. Biol. Chem. 273:4990-4996). A plasmid encoding full length DITHP is transfected into a mammalian cell line (e.g., Chinese hamster ovary (CHO) or human embryonic kidney (HEK-293) cell lines) using methods well-known in the art. Transfected cells are grown in 12-well trays in culture medium for 48 hours, then the culture medium is discarded, and the attached cells are gently washed with PBS. The cells are then incubated in culture medium with or without ligand for 30 minutes, then the medium is removed and cells lysed by treatment with 1 M perchloric acid. The cAMP levels in the lysate are measured by radioimmunoassay using methods well-known in the art. Changes in the levels of cAMP in the lysate from cells exposed to ligand compared to those without ligand are proportional to the amount of DITHP present in the transfected cells.  
     [0863] Alternatively, an assay for DITHP protein phosphatase activity measures the hydrolysis of P-nitrophenyl phosphate (PNPP). DITHP is incubated together with PNPP in HEPES buffer pH 7.5, in the presence of 0.1% β-mercaptoethanol at 37° C. for 60 min. The reaction is stopped by the addition of 6 ml of 10 N NaOH, and the increase in light absorbance of the reaction mixture at 410 nm resulting from the hydrolysis of PNPP is measured using a spectrophotometer. The increase in light absorbance is proportional to the phosphatase activity of DITHP in the assay (Diamond, R. H. et al (1994) Mol Cell Biol 14:3752-3762).  
     [0864] An alternative assay measures DITHP-mediated G-protein signaling activity by monitoring the mobilization of Ca ++  as an indicator of the signal transduction pathway stimulation. (See, e.g., Grynkievicz, G. et al. (1985) J. Biol. Chem. 260:3440; McColl, S. et al. (1993) J. Immunol.  150:4550-4555 ; and Aussel, C. et al. (1988) J. Immunol. 140:215-220). The assay requires preloading neutrophils or T cells with a fluorescent dye such as FURA-2 or BCECF (Universal Imaging Corp, Westchester Pa.) whose emission characteristics are altered by Ca ++  binding. When the cells are exposed to one or more activating stimuli artificially (e.g., anti-CD3 antibody ligation of the T cell receptor) or physiologically (e.g., by allogeneic stimulation), Ca ++  flux takes place. This flux can be observed and quantified by assaying the cells in a fluorometer or fluorescent activated cell sorter. Measurements of Ca ++  flux are compared between cells in their normal state and those transfected with DITHP. Increased Ca ++  mobilization attributable to increased DITHP concentration is proportional to DITHP activity.  
     [0865] DITHP transport activity is assayed by measuring uptake of labeled substrates into  Xenopus laevis  oocytes. Oocytes at stages V and VI are injected with DITHP mRNA (10 ng per oocyte) and is incubated for 3 days at 18° C. in OR2 medium (82.5 mM NaCl, 2.5 mM KCl, 1 mM CaCl 2 , 1 mM MgCl 2 , 1 mM Na 2 HPO 4 , 5 mM Hepes, 3.8 mM NaOH, 50 μg/ml gentamycin, pH 7.8) to allow expression of DITHP protein. Oocytes are then transferred to standard uptake medium (100 mM NaCl, 2 mM KCl, 1 mM CaCl 2 , 1 mM MgCl 2 , 10 mM Hepes/Tris 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  3 H, 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. DITHP transport activity is proportional to the level of internalized labeled substrate.  
     [0866] DITHP transferase activity is demonstrated by a test for galactosyltransferase activity. This can be determined by measuring the transfer of radiolabeled galactose from UDP-galactose to a GlcNAc-terminated oligosaccharide chain (Kolbinger, F. et al. (1998) J. Biol. Cheri 273:58-65). The sample is incubated with 14 μl of assay stock solution (180 mM sodium cacodylate, pH 6.5, 1 mg/ml bovine serum albumin, 0.26 mM UDP-galactose, 2 μl of UDP-[ 3 H]galactose), 1 μl of MnCl 2  (500 mM), and 2.5 μl of GlcNAcβO—(CH 2 ) 8 —CO 2 Me (37 mg/ml in dimethyl sulfoxide) for 60 minutes at 37° C. The reaction is quenched by the addition of 1 ml of water and loaded on a C18 Sep-Pak cartridge (Waters), and the column is washed twice with 5 ml of water to remove unreacted UDP-[ 3 H]galactose. The [ 3 H]galactosylated GlcNAcβO—(CH 2 ) 8 —CO 2 Me remains bound to the column during the water washes and is eluted with 5 ml of methanol. Radioactivity in the eluted material is measured by liquid scintillation counting and is proportional to galactosyltransferase activity in the starting sample.  
     [0867] In the alternative, DITHP induction by heat or toxins may be demonstrated using primary cultures of human fibroblasts or human cell lines such as CCL-13, HEK293, or HEP G2 (ATCC). To heat induce DITHP expression, aliquots of cells are incubated at 42° C. for 15, 30, or 60 minutes. Control aliquots are incubated at 37° C. for the same time periods. To induce DITHP expression by toxins, aliquots of cells are treated with 100 μM arsenite or 20 mM azetidine-2-carboxylic acid for 0, 3, 6, or 12 hours. After exposure to heat, arsenite, or the amino acid analogue, samples of the treated cells are harvested and cell lysates prepared for analysis by western blot. Cells are lysed in lysis buffer containing 1% Nonidet P40, 0.15 M NaCl, 50 mM Tris-HCl, 5 mM EDTA, 2 mM N-ethylmaleimide, 2 mM phenylmethylsulfonyl fluoride, 1 mg/ml leupeptin, and 1 mg/ml pepstatin. Twenty micrograms of the cell lysate is separated on an 8% SDS-PAGE gel and transferred to a membrane. After blocking with 5% nonfat dry milk/phosphate-buffered saline for 1 h, the membrane is incubated overnight at 4° C. or at room temperature for 24 hours with a 1:1000 dilution of anti-DITHP serum in 2% nonfat dry milk/phosphate-buffered saline. The membrane is then washed and incubated with a 1:1000 dilution of horseradish peroxidase-conjugated goat anti-rabbit IgG in 2% dry milk/phosphate-buffered saline. After washing with 0.1% Tween 20 in phosphate-buffered saline, the DITHP protein is detected and compared to controls using chemiluminescence.  
     [0868] Alternatively, DITHP protease activity is measured by the hydrolysis of appropriate synthetic peptide substrates conjugated with various chromogenic molecules in which the degree of hydrolysis is quantified by spectrophotometric (or fluorometric) absorption of the released chromophore (Beynon, R. J. and J. S. Bond (1994)  Proteolytic Enzymes: A Practical Approach , Oxford University Press, New York, N.Y., pp.25-55). Peptide substrates are designed according to the category of protease activity as endopeptidase (serine, cysteine, aspartic proteases, or metalloproteases), aminopeptidase (leucine aminopeptidase), or carboxypeptidase (carboxypeptidases A and B, procollagen C-proteinase). Commonly used chromogens are 2-naphthylamine, 4-nitroaniline, and furylacrylic acid. Assays are performed at ambient temperature and contain an aliquot of the enzyme and the appropriate substrate in a suitable buffer. Reactions are carried out in an optical cuvette, and the increase/decrease in absorbance of the chromogen released during hydrolysis of the peptide substrate is measured. the change in absorbance is proportional to the DITHP protease activity in the assay.  
     [0869] In the alternative, an assay for DITHP protease activity takes advantage of fluorescence resonance energy transfer (FRET) that occurs when one donor and one acceptor fluorophore with an appropriate spectral overlap are in close proximity. A flexible peptide linker containing a cleavage site specific for PRTS is fused between a red-shifted variant (RSGFP4) and a blue variant (BFP5) of Green Fluorescent Protein. This fusion protein has spectral properties that suggest energy transfer is occurring from BFP5 to RSGFP4. When the fusion protein is incubated with DITHP, the substrate is cleaved, and the two fluorescent proteins dissociate. This is accompanied by a marked decrease in energy transfer which is quantified by comparing the emission spectra before and after the addition of DITHP (Mitra, R. D. et al (1996) Gene 173:13-17). This assay can also be performed in living cells. In this case the fluorescent substrate protein is expressed constitutively in cells and DITHP is introduced on an inducible vector so that FRET can be monitored in the presence and absence of DITHP (Sagot, I. et al (1999) FEBS Lett. 447:53-57).  
     [0870] A method to determine the nucleic acid binding activity of DITHP involves a polyacrylamide gel mobility-shift assay. In preparation for this assay, DITHP is expressed by transforming a mammalian cell line such as COS7, HeLa or CHO with a eukaryotic expression vector containing DITHP cDNA. The cells are incubated for 48-72 hours after transformation under conditions appropriate for the cell line to allow expression and accumulation of DITHP. Extracts containing solubilized proteins can be prepared from cells expressing DITHP by methods well known in the art Portions of the extract containing DITHP are added to [ 32 P]-labeled RNA or DNA. Radioactive nucleic acid can be synthesized in vitro by techniques well known in the art The mixtures are, incubated at 25° C. in the presence of RNase- and DNase-inhibitors under buffered conditions for 5-10 minutes. After incubation, the samples are analyzed by polyacrylamide gel electrophoresis followed by autoradiography. The presence of a band on the autoradiogram indicates the formation of a complex between DITHP and the radioactive transcript. A band of similar mobility will not be present in samples prepared using control extracts prepared from untransformed cells.  
     [0871] In the alternative, a method to determine the methylase activity of a DITHP measures transfer of radiolabeled methyl groups between a donor substrate and an acceptor substrate. Reaction mixtures (50 μl final volume) contain 15 mM HEPES, pH 7.9, 1.5 mM MgCl 2 , 10 mM dithiothreitol, 3% polyvinylalcohol, 1.5 μCi [methyl- 3 H]AdoMet (0.375 μM AdoMet) (DuPont-NEN), 0.6 μg DITHP, and acceptor substrate (e.g., 0.4 μg [ 35 S]RNA, or 6-mercaptopurine (6-MP) to 1 mM final concentration). Reaction mixtures are incubated at 30° C. for 30 minutes, then 65° C. for 5 minutes. Analysis of [methyl- 3 H]RNA is as follows: 1) 50 μl of 2×loading buffer (20 mM Tris-HCl, pH 7.6, 1 M LiCl, 1 mM EDTA, 1% sodium dodecyl sulphate (SDS)) and 50 μl oligo d(T)-cellulose (10 mg/ml in 1×loading buffer) are added to the reaction mixture, and incubated at ambient temperature with shaking for 30 minutes. 2) Reaction mixtures are transferred to a 96-well filtration plate attached to a vacuum apparatus. 3) Each sample is washed sequentially with three 2.4 ml aliquots of 1×oligo d(T) loading buffer containing 0.5% SDS, 0.1% SDS, or no SDS. and 4) RNA is eluted with 300 μl of water into a 96-well collection plate, transferred to scintillation vials containing liquid scintillant, and radioactivity determined. Analysis of [methyl- 3 H]6-MP is as follows: 1) 500 μl 0.5 M borate buffer, pH 10.0, and then 2.5 ml of 20% (v/v) isoamyl alcohol in toluene are added to the reaction mixtures. 2) The samples mixed by vigorous vortexing for ten seconds. 3) After centrifugation at 700 g for 10 minutes, 1.5 ml of the organic phase is transferred to scintillation vials containing 0.5 ml absolute ethanol and liquid scintillant, and radioactivity determined. and 4) Results are corrected for the extraction of 6-MP into the organic phase (approximately 41%).  
     [0872] An assay for adhesion activity of DITHP measures the disruption of cytoskeletal filament networks upon overexpression of DITHP in cultured cell lines (Rezniczek, G. A. et al. (1998) J. Cell Biol. 141:209-225). cDNA encoding DITHP is subcloned into a mammalian expression vector that drives high levels of cDNA expression. This construct is transfected into cultured cells, such as rat kangaroo PtK2 or rat bladder carcinoma 804G cells. Actin filaments and intermediate filaments such as keratin and vimentin are visualized by immunofluorescence microscopy using antibodies and techniques well known in the art. The configuration and abundance of cytoskeletal filaments can be assessed and quantified using confocal imaging techniques. In particular, the bundling and collapse is of cytoskeletal filament networks is indicative of DITHP adhesion activity.  
     [0873] Alternatively, an assay for DITHP activity measures the expression of DITHP on the cell surface. cDNA encoding DITHP is transfected into a non-leukocytic cell line. Cell surface proteins are labeled with biotin (de la Fuente, M. A. et al. (1997) Blood 90:2398-2405). Immunoprecipitations are performed using DITHP-specific antibodies, and immunoprecipitated samples are analyzed using SDS-PAGE and immunoblotting techniques. The ratio of labeled immunoprecipitant to unlabeled immunoprecipitant is proportional to the amount of DITHP expressed on the cell surface.  
     [0874] Alternatively, an assay for DITHP activity measures the amount of cell aggregation induced by overexpression of DITHP. In this assay, cultured cells such as NIH3T3 are transfected with cDNA encoding DITHP contained within a suitable mammalian expression vector under control of a strong promoter. Cotransfection with cDNA encoding a fluorescent marker protein, such as Green Fluorescent Protein (CLONTECH), is useful for identifying stable transfectants. The amount of cell agglutination, or clumping, associated with transfected cells is compared with that associated with untransfected cells. The amount of cell agglutination is a direct measure of DITHP activity.  
     [0875] DITHP may recognize and precipitate antigen from serum This activity can be measured by the quantitative precipitin reaction (Golub, E. S. et al. (1987)  Immunology: A Synthesis , Sinauer Associates, Sunderland M A, pages 113-115). DITHP is isotopically labeled using methods known in the art. Various serum concentrations are added to constant amounts of labeled DITHP. DITHP-antigen complexes precipitate out of solution and are collected by centrifugation. The amount of precipitable DITHP-antigen complex is proportional to the amount of radioisotope detected in the precipitate. The amount of precipitable DITHP-antigen complex is plotted against the serum concentration. For various serum concentrations, a characteristic precipitation curve is obtained, in which the amount of precipitable DITHP-antigen complex initially increases proportionately with increasing serum concentration, peaks at the equivalence point, and then decreases proportionately with further increases in serum concentration. Thus, the amount of precipitable DITHP-antigen complex is a measure of DITHP activity which is characterized by sensitivity to both limiting and excess quantities of antigen.  
     [0876] A microtubule motility assay for DITHP measures motor protein activity. In this assay, recombinant DITHP is immobilized onto a glass slide or similar substrate. Taxol-stabilized bovine brain microtubules (commercially available) in a solution containing ATP and cytosolic extract are perfused onto the slide. Movement of microtubules as driven by DITHP motor activity can be visualized and quantified using video-enhanced light microscopy and image analysis techniques. DITHP motor protein activity is directly proportional to the frequency and velocity of microtubule movement.  
     [0877] Alternatively, an assay for DITHP measures the formation of protein filaments in vitro. A solution of DITHP at a concentration greater than the “critical concentration” for polymer assembly is applied to carbon-coated grids. Appropriate nucleation sites may be supplied in the solution. The grids are negative stained with 0.7% (w/v) aqueous uranyl acetate and examined by electron microscopy. The appearance of filaments of approximately 25 nm (microtubules), 8 nm (actin), or 10 nm (intermediate filaments) is a demonstration of protein activity.  
     [0878] DITHP electron transfer activity is demonstrated by oxidation or reduction of NADP. Substrates such as Asn-βGal, biocytidine, or ubiquinone-10 may be used. The reaction mixture contains 1-2 mg/ml HORP, 15 mM substrate, and 2.4 mM NAD(P) + in 0.1 M phosphate buffer, pH 7.1 (oxidation reaction), or 2.0 mM NAD(P)H, in 0.1 M Na 2 HPO 4  buffer, pH 7.4 (reduction reaction); in a total volume of 0.1 ml. FAD may be included with NAD, according to methods well known in the art. Changes in absorbance are measured using a recording spectrophotometer. The amount of NAD(P)H is stoichiometrically equivalent to the amount of substrate initially present, and the change in A 340  is a direct measure of the amount of NAD(P)H produced; ΔA 340=6620 [NADH]. DITHP activity is proportional to the amount of NAD(P)H present in the assay. The increase in extinction coefficient of NAD(P)H coenzyme at 340 nm is a measure of oxidation activity, or the decrease in extinction coefficient of NAD(P)H coenzyme at 340 nm is a measure of reduction activity (Dalziel, K (1963) J. Biol. Chen 238:2850-2858).  
     [0879] DITHP transcription factor activity is measured by its ability to stimulate transcription of a reporter gene (Liu, H. Y. et al. (1997) EMBO J. 16:5289-5298). The assay entails the use of a well characterized reporter gene construct, LexA op -LacZ, that consists of LexA DNA transcriptional control elements (LexA op ) fused to sequences encoding the  E. coli  Lac enzyme. The methods for constructing and expressing fusion genes, introducing them into cells, and measuring LacZ enzyme activity, are well known to those skilled in the art. Sequences encoding DITHP are cloned into a plasmid that directs the synthesis of a fusion protein, LexA-DITHP, consisting of DITHP and a DNA binding domain derived from the LexA transcription factor. The resulting plasmid, encoding a LexA-DITHP fusion protein, is introduced into yeast cells along with a plasmid containing the LexA op -LacZ reporter gene. The amount of LacZ enzyme activity associated with LexA-DITHP transfected cells, relative to control cells, is proportional to the amount of transcription stimulated by the DITHP.  
     [0880] Chromatin activity of DITHP is demonstrated by measuring sensitivity to DNase I (Dawson, B. A. et al. (1989) J. Biol. Chem. 264:12830-12837). Samples are treated with DNase 1, followed by insertion of a cleavable biotinylated nucleotide analog, 5-[(N-biotinamido)hexanoamido-ethyl-1,3-thiopropionyl-3-aminoallyl]-2′-deoxyuridine 5′-triphosphate using nick-repair techniques well known to those skilled in the art Following purification and digestion with EcoRI restriction endonuclease, biotinylated sequences are affinity isolated by sequential binding to streptavidin and biotincellulose.  
     [0881] Another specific assay demonstrates the ion conductance capacity of DITHP using an electrophysiological assay. DITHP is expressed by transforming a mammalian cell line such as COS7. HeLa or CHO with a eukaryotic expression vector encoding DITH. Eukaryotic expression vectors are commercially available, and the techniques to introduce them into cells are well known to those skilled in the art. A small amount of a second plasmid, which expresses any one of a number of marker genes such as β-galactosidase, is co-transformed into the cells in order to allow rapid identification of those cells which have taken up and expressed the foreign DNA The cells are incubated for 48-72 hours after transformation under conditions appropriate for the cell line to allow expression and accumulation of DITHP and β-galactosidase. Transformed cells expressing β-galactosidase are stained blue when a suitable colorimetric substrate is added to the culture media under conditions that are well known in the art. Stained cells are tested for differences in membrane conductance due to various ions by electrophysiological techniques that are well known in the art. Untransformed cells, and/or cells transformed with either vector sequences alone or β-galactosidase sequences alone, are used as controls and tested in parallel. The contribution of DITHP to cation or anion conductance can be shown by incubating the cells using antibodies specific for either DITHP. The respective antibodies will bind to the extracellular side of DITHP, thereby blocking the pore in the ion channel, and the associated conductance.  
     [0882] XV. Functional Assays  
     [0883] DITHP function is assessed by expressing dithp 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 (Life Technologies) and pCR3.1 (Invitrogen Corporation, Carlsbad Calif.), both of which contain the cytomegalovirus promoter. 5-10 μg of recombinant vector are transiently transfected into a human cell line, preferably of endothelial or hematopoietic origin, using either liposome formulations or electroporation. 1-2 μg of an additional plasmid containing sequences encoding a marker protein are co-transfected.  
     [0884] 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 CD64GFP fusion protein. Flow cytometry (FCM), an automated laser optics-based technique, is used to identity transfected cells expressing GFP or CD64-GFP and to evaluate the apoptotic state of the cells and other cellular properties.  
     [0885] 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 Cytometry , Oxford, New York N.Y.  
     [0886] The influence of DITHP on gene expression can be assessed using highly purified populations of cells transfected with sequences encoding DITHP and either CD64 or CD64GFP. CD64 and CD64GFP 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, Inc., Lake Success N.Y.). mRNA can be purified from the cells using methods well known by those of skill in the art Expression of mRNA encoding DITHP and other genes of interest can be analyzed by northern analysis or microarray techniques.  
     [0887] XVI. Production of Antibodies  
     [0888] DITHP substantially purified using polyacrylamide gel electophoresis (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.  
     [0889] Alternatively, the DITHP amino acid sequence is analyzed using LASERGENE software (DNASTAR) to determine regions of high immunogenicity, and a corresponding peptide 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, Chapter 11.) Typically, peptides 15 residues in length are synthesized using an ABI 431A peptide synthesizer (Applied Biosystems) using fmoc-chemistry and coupled to KLH (Sigma) by reaction with N-maleimidobenzoyl-N-hydroxysuccinimide ester (MBS) to increase immunogenicity. (See, e.g., Ausubel, supra.) Rabbits are immunized with the peptide-KLH complex in complete Freund&#39;s adjuvant Resulting antisera are tested for antipeptide activity by, for example, binding the peptide to plastic, blocking with 1% BSA, reacting with rabbit antisera, washing, and reacting with radio-iodinated goat anti-rabbit IgG. Antisera with antipeptide activity are tested for anti-DITHP activity using protocols well known in the art, including ELISA, RIA, and immunoblotting.  
     [0890] XVII. Purification of Naturally Occurring DITHP Using Specific Antibodies  
     [0891] Naturally occurring or recombinant DITHP is substantially purified by immunoaffinity chromatography using antibodies specific for DITHP. An immunoaffinity column is constructed by covalently coupling anti-DITHP 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&#39;s instructions.  
     [0892] Media containing DITHP are passed over the immunoaffinity column, and the column is washed under conditions that allow the preferential absorbance of DITHP (e.g., high ionic strength buffers in the presence of detergent). The column is eluted under conditions that disrupt antibody/DITHP 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 DITHP is collected.  
     [0893] XVIII. Identification of Molecules Which Interact with DITHP  
     [0894] DITHP, or biologically active fragments thereof, are labeled with  125 I Bolton-Hunter reagent (See, e.g., Bolton, A. E. and W. M. Hunter (1973) Biochem. J. 133:529-539.) Candidate molecules previously arrayed in the wells of a multi-well plate are incubated with the labeled DITHP, washed, and any wells with labeled DITHP complex are assayed Data obtained using different concentrations of DITHP are used to calculate values for the number, affinity, and association of DITHP with the candidate molecules.  
     [0895] Alternatively, molecules interacting with DITHP 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).  
     [0896] DITHP may also be used in the PATHCALLING process (CuraGen Corp., New Haven Conn.) 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. Pat. No. 6,057,101).  
     [0897] All publications and patents mentioned in the above specification are herein incorporated by reference. Various modifications and variations of the described method and system 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 specific preferred embodiments, it should be understood that the invention as claimed should not be unduly limited to such specific embodiments. Indeed, various modifications of the above-described modes for carrying out the invention which are obvious to those skilled in the field of molecular biology or related fields are intended to be within the scope of the following claims.  
                               TABLE 1                          SEQ       GI               ID NO:   Template ID   Number   Probability Score   Annotation                                         1   LG:1040582.1:2000FEB18   g178480   1.00E−92   Human aldehyde reductase mRNA, complete cds.       2   LG:453570.1:2000FEB18   g2909424   2.40E−65   Glyoxalase I       3   LG:408751.3:2000FEB18   g3608122   4.40E−74   dihydropyrimidinase       4   LI:090574.1:2000FEB01   g29600   5.00E−85   carbonic anhydrase I (AA 1-261)       5   LI:229932.2:2000FEB01   g1835116   2.00E−63   acetyl-CoA synthetase       6   LI:332176.1:2000FEB01   g2104689   0   alpha glucosidase II, alpha subunit       7   LI:403248.2:2000FEB01   g63713   4.00E−23   ornithine decarboxylase       8   LG:220992.1:2000MAY19   g10435462   0   unnamed protein product ( Homo sapiens )       9   LG:1094571.1:2000MAY19   g7023634   4.00E−92   unnamed protein product ( Homo sapiens )       10   LI:350754.4:2000MAY01   g307504   0   transglutaminase E3 ( Homo sapiens )       11   LI:255828.29:2000MAY01   g189998   9.00E−65   M2-type pyruvate kinase ( Homo sapiens )       12   LI:1190263.1:2000MAY01   g2576305   1.00E−172   arylsulphatase ( Homo sapiens )       13   LG:270916.2:2000FEB18   g2088668   1.20E−11   similar to Achlya amblsexualis antheridiol steroid receptor (NID:g166306)       14   LG:999414.3:2000FEB18   g3861482   0   Human chromosome 3, olfactory receptor pseudogene cluster 1,                       complete sequence, and myosin light chain kinase (MLCK)                       pseudogene, partial sequence.       15   LG:429446.1:2000FEB18   g2358042   0   Human T-cell receptor alpha delta locus from bases 501613 to 752736                       (section 3 of 5) of the Complete Nucleotide Sequence.       16   LI:057229.1:2000FEB01   g10439739   2.00E−19   unnamed protein product ( Homo sapiens )       17   LI:351965.1:2000FEB01   g2358042   0   Human T-cell receptor alpha delta locus from bases 501613 to 752736                       (section 3 of 5) of the Complete Nucleotlde Sequence.       18   LG:068682.1:2000FEB18   g404634   1.10E−31   serine/threonine kinase       19   LG:242665.1:2000FEB18   g2117166   1.00E−160   Ras like GTPase ( Homo sapiens )       20   LG:241743.1:2000FEB18   g5763838   9.70E−49   dJ593C16.1 (ras GTPase activating protein)       21   LI:034212.1:2000FEB01   g1469876   0   The KIAA0147 gene product is related to adenylyl cyclase.       22   LG:344886.1:2000MAY19   g7008402   2.00E−89   kappa B-ras 1 ( Homo sapiens )       23   LG:228930.1:2000MAY19   g206218   4.50E−87   phospholipase C-1       24   LG:338927.1:2000MAY19   g3599940   3.00E−57   faclogenital dysplasia protein 2 ( Mus musculus )       25   LG:898771.1:2000MAY19   g508528   5.00E−58   myocyte nuclear factor ( Mus musculus )       26   LI:257664.67:2000MAY01   g183399   1.00E−142   Human guanine nucleotide-binding protein alpha-subunit gene (G-s-                       alpha), exon 3.       27   LI:001496.2:2000MAY01   g3005085   1.00E−177   hook1 protein ( Homo sapiens )       28   LI:1085273.2:2000MAY01   g1781037   0   neuronal tyrosine threonine phosphatase 1 ( Mus musculus )       29   LI:333138.2:2000MAY01   g2077934   1.00E−164   Protein Kinase ( Rattus norvegicus )       30   LI:338927.1:2000MAY01   g3599940   6.00E−45   faciogenital dysplasia protein 2 ( Mus musculus )       31   LG:335558.1:2000FEB18   g1181619   5.00E−97   a variant of TSC-22 ( Gallus gallus )       32   LG:998283.7:2000FEB18   g6683492   1.00E−105   bromodomain PHD finger transcription factor ( Homo sapiens )       33   LI:402739.1:2000FEB01   g4164151   6.00E−34   AhR repressor       34   LI:175223.1:2000FEB01   g2745892   2.00E−11   Y box transcription factor                       supported by Genscan and several ESTs: C83049 (NID:g3062006),                       AA823760 (NID:g2893628), AA215791 (NID:g1815572), AI095488       35   LG:981076.2:2000MAY19   g3924670   1.00E−59   (NID:a3434464), and AA969095 (NID:a3144275) ( Homo sapiens )       36   LI:1008973.1:2000MAY01   g6939732   2.00E−52   transcription factor Elongin A2 ( Homo sapiens )       37   LI:1190250.1:2000MAY01   g3757892   4.00E−66   enhancer of polycomb ( Mus musculus )       38   LG:021371.3:2000FEB18   g984814   1.40E−60   zinc finger protein       39   LG:475404.1:2000FEB18   g487784   5.00E−36   Human zinc finger protein ZNF136.       40   LG:979406.2:2000FEB18   g4325310   3.60E−1 1   zinc-finger protein 7       41   LG:410726.1:2000FEB1B   g6002480   2.60E−39   BWSCR2 associated zinc-finger protein BAZ2       42   LG:200005.1:2000FEB18   g1504006   3.20E−25   similarto human ZFY protein.       43   LG:1076828.1:2000FEB18   g498720   2.00E−33   Human HZF10 mRNA for zinc finger protein.       44   LG:1076931.1:2000FEB18   g498151   2.00E−52   Human mRNA for KIAA0065 gene, partial cds.       45   LG:1078121.1:2000FEB18   g186773   2.00E−47   Human Kruppel related zinc finger protein (HTF10) mRNA, complete cds.       46   LG:1079203.1:2000FEB18   g1017721   0   Human repressor transcriptional factor (ZNF85) mRNA, complete cds.       47   LG:1082586.1:2000FEB18   g7959207   1.00E−19   KIAA1473 protein ( Homo sapiens )       48   LG:1082774.1:2000FEB18   g184451   0   Human Krueppel-related DNA-binding protein (TF9 PF4) mRNA, 5′ cds.       49   LG:1082775.1:2000FEB18   g506502   3.50E−36   NK10       50   LG:1083120.1:2000FEB18   g7023216   1.00E−14   unnamed protein product ( Homo sapiens )       51   LG:1087707.1:2000FEB18   g347905   2.00E−40   Human zinc finger protein (ZNF141) mRNA, complete cds.       52   LG:1090915.1:2000FEB18   g347905   2.00E−24   Human zinc finger protein (ZNF141) mRNA, complete cds.       53   LG:1094230.1:2000FEB18   g454818   1.00E−98   Human Krueppel-related DNA-binding protein (PF4) mRNA, 5′ end.       54   LG:474848.3:2000FEB18   g498152   4.00E−16   ha0946 protein is Kruppel-related.       55   LI:251656.1:2000FEB01   g55471   0   Zfp-29       56   LI:021371.1:2000FEB01   g984814   2.00E−96   zinc finger protein       57   LI:133095.1:2000FEB01   g453376   8.00E−42   zinc finger protein PZF       58   LI:236654.2:2000FEB01   g498721   2.00E−22   zinc finger protein       59   LI:200009.1:2000FEB01   g498719   4.00E−24   zinc finger protein       60   LI:758502.1:2000FEB01   g200407   0   pMLZ-4       61   LI:344772.1:2000FEB01   g4062983   3.00E−67   Eos protein       62   LI:789445.1:2000FEB01   g1049301   2.00E−26   KRAB zinc finger protein; Method:conceptual translation supplied by       63   LI:789657.1:2000FEB01   g1020145   1.00E−53   DNA binding protein       64   LI:789808.1:2000FEB01   g288424   0   Human ZNF37A mRNA for zinc finger protein.       65   LI:792919.1:2000FEB01   g2232012   0   Human zinc finger protein (FDZF2) mRNA, complete cds.       66   LI:793949.1:2000FEB01   g1017721   3.00E−53   Human repressor transcriptional factor (ZNF85) mRNA, complete cds.       67   LI:794389.1:2000FEB01   g5640017   4.00E−45   zinc finger protein ZFP113       68   LI:796010.1:2000FEB01   g288424   0   Human ZNF37A mRNA for zinc finger protein.       69   LI:796324.1:2000FEB01   g288424   0   Human ZNF37A mRNA for zinc finger protein.       70   LI:796373.1:2000FEB01   g1020145   9.00E−36   DNA binding protein       71   LI:796415.1:2000FEB01   g498151   4.00E−28   Human mRNA for KIAA0065gene, partial cds.       72   LI:798636.1:2000FEB01   g2970037   0   Human HKL1 mRNA, complete cds.       73   LI:800045.1:2000FEB01   g538413   2.00E−55   zinc finger protein       74   LI:800680.1:2000FEB01   g7023216   7.00E−18   unnamed protein product ( Homo sapiens )       75   LI:800894.1:2000FEB01   g3342001   0   Human hematopoietic cell derived zinc finger protein mRNA, complete       76   LI:801015.1:2000FEB01   g487785   4.00E−16   zinc finger protein ZNF136 ( Homo sapiens )       77   LI:801236.1:2000FEB01   g488555   4.00E−48   zinc finger protein ZNF135       78   LI:803335.1:2000FEB01   g498152   1.00E−20   ha0946 protein is Kruppel-related.       79   U:803998.1:2000FEB01   g1017722   1.00E−53   repressor transcriptional factor       80   LI:478757.1:2000FEB01   g498151   9.00E−27   Human mRNA for KIAA0065 gene, partial cds.       81   LI:808532.1:2000FEB01   g2232012   0   Human zinc finger protein (FDZF2) mRNA, complete cds.       82   LI:443073.1:2000FEB01   g4567179   3.00E−33   BC37295_1 ( Homo sapiens )       83   LI:479671.1:2000FEB01   g487784   3.00E−38   Human zinc finger protein ZNF136.       84   LI:810078.1:2000FEB01   g498718   0   Human HZF1 mRNA for zinc finger protein.       85   LI:810224.1:2000FEB01   g288424   0   Human ZNF37A mRNA for zinc finger protein.       86   LI:817052.2:2000FEB01   g1020145   1.00E−51   DNA binding protein       87   LG:892274.1:2000MAY19   g6650686   2.00E−95   Human Y-linked zinc finger protein (ZFY) gene, complete cds.       88   LG:1080959.1:2000MAY19   g5262560   2.00E−40   hypothetical protein ( Homo sapiens )       89   LG:1054900.1:2000MAY19   g5262560   3.00E−35   hypothetical protein ( Homo sapiens )       90   LG:1077357.1:2000MAY19   g10047297   2.00E−23   KIAA1611 protein ( Homo sapiens )       91   LG:1084051.1:2000MAY19   g5931821   8.00E−79   dJ228H13.3 (zinc finger protein) ( Homo sapiens )       92   LG:1076853.1:2000MAY19   g506502   1.00E−141   NK10 ( Mus musculus )       93   LG:481631.10:2000MAY19   g7023216   1.00E−142   unnamed protein product ( Homo sapiens )       94   LG:1088431.2:2000MAY19   g7023216   7.00E−18   unnamed protein product ( Homo sapiens )       95   LI:401619.10:2000MAY01   g7959865   1.00E−18   PRO2032 ( Homo sapiens )       96   LI:1144007.1:2000MAY01   g5360097   0   putative kruppel-related zinc finger protein NY-REN-23 antigen ( Homo                           sapiens )       97   LI:331074.1:2000MAY01   g2149792   0   Roaz ( Rattus norvegicus )       98   LI:1170349.1:2000MAY01   g487787   1.00E−45   zinc finger protein ZNF140 ( Homo sapiens )       99   LG:335097.1:2000FEB18   g7020440   6.00E−25   unnamed protein product ( Homo sapiens )       100   LG:1076451.1:2000FEB18   g2088550   0   Human hereditary haemochromatosis region, histone 2A-like protein                       gene, hereditary haemochromatosis (HLA-H) gene, RoRet gene, and                       sodium phosphate transporter (NPT3) gene, complete cds.       101   LI:805478.1:2000FEB01   g2088550   0   Human hereditary haemochromatosis region, histone 2A-like protein                       gene, hereditary haemochromatosis (HLA-H) gene, RoRet gene, and                       sodium phosphate transporter (NPT3) gene. complete cds.       102   LG:101269.1:2000MAY19   g3953533   2.10E−56   inwardly rectifying potassium channel Klr5.1       103   LI:331087.1:2000MAY01   g4186073   3.00E−41   calcium channel alpha-2-delta-C subunit ( Mus musculus )       104   LI:410188.1:2000MAY01   g4836145   0   tetrodotoxin-resistant voltage-gated sodium channel ( Homo sapiens )       105   LI:1188288.1:2000MAY01   g204220   0   beta-alanine-sensitive neuronal GABA transporter ( Rattus norvegicus )       106   LI:427997.4:2000MAY01   g6996442   1.00E−48   CTL1 protein ( Homo sapiens )       107   LG:451682.1:2000FEB18   g5091520   3.00E−75   ESTs AU058081 (E30812), AU058365(E50679), AU030138(E50679)                                           oleracea mRNA for proteasome 37 kD subunit.(X96974)       108   LG:1077283.1:2000FEB18   g2565302   0   Rhesus monkey cyclophilin A mRNA, complete cds.       109   LG: 481436.5:2000FEB18   g3873707   2.60E−34   Similarity to  B. subtilis  DNAJ protein (SW: DNAJ_BACSU); cDNA EST                       yk437a1.5 comes from this gene       110   LI:793701.1:2000FEB01   g1049231   4.00E−33   Method: conceptual translation supplied by author; putative hybrid                       protein similar to HERV-H protease and HERV-E integrase (Human                       endogenous retrovirus)       111   LI:373637.1:2000FEB01   g2286123   3.00E−50   testis specific DNAj-homolog       112   LG:239368.2:2000MAY19   g4981382   1.00E−11   dnaJ protein ( Thermotoga maritima )       113   LI:053826.1:2000MAY01   g2943716   4.00E−67   25 kDa trypsin inhibitor ( Homo sapiens )       114   LI:449393.1:2000MAY01   g6957716   1.00E−128   putative chaperonin ( Arabidopsis thaliana )       115   LI:1071427.96:2000MAY01   g9956070   1.00E−144   similar to  Homo sapiens  mRNA for KIAA0723 protein with GenBank                       Accession Number AB018266.10       116   LI:336338.8:2000MAY01   g9296929   2.00E−16   protease PC6 isoform A ( Homo sapiens )       117   LG:345527.1:2000FEB18   g805296   3.40E−176   lymphocyte specific helicase       118   LG:1089383.1:2000FEB18   g2104910   3.00E−23   ORF derived from D1 leader region and integrase coding region ( Homo                           sapiens )       119   LG:1092522.1:2000FEB18   g1263080   0   Human mariner1 transposase gene, complete consensus sequence.       120   LG:1093216.1:2000FEB18   g2104910   1.00E−23   ORF derived from D1 leader region and integrase coding region ( Homo                           sapiens )       121   LI:270318.3:2000FEB01   g3880433   3.00E−12   similar to mitochondrial RNA splicing MSR4 like protein; cDNA EST                       EMBL: C09217 comes from this gene       122   LI:335671.2:2000FEB01   g805296   1.00E−83   lymphocyte specific helicase       123   LI:793758.1:2000FEB01   g2104910   4.00E−26   ORF derived from D1 leader region and integrase coding region ( Homo                           sapiens )       124   LI:803718.1:2000FEB01   g2104910   3.00E−23   ORF derived from D1 leader region and integrase coding region ( Homo                           sapiens )       125   LI:412179.1:2000FEB01   g1263080   4.00E−93   Human mariner1 transposase gene, complete consensus sequence.       126   LI:815679.1:2000FEB01   g7020440   3.00E−12   unnamed protein product ( Homo sapiens )       127   LI:481361.3:2000FEB01   g3776011   5.00E−25   RNA helicase       128   LG:247388.1:2000MAY19   g6016932   3.00E−127   dJ620E11.1a (novel Helicase C-terminal domain and SNF2 N-terminal                       domains containing protein, similar to KIAA0308)       129   LG:255789.10:2000MAY19   g37542   2.00E−57   Human mRNA for U1 small nuclear RNP-specific C protein.       130   LI:787618.1:2000MAY01   g7020440   3.00E−12   unnamed protein product ( Homo sapiens )       131   LI:331610.2:2000MAY01   g2599502   0   protocadherin 68 ( Homo sapiens )       132   LG:982697.1:2000FEB18   g10436424   1.00E−25   unnamed protein product ( Homo sapiens )       133   LG:1080896.1:2000FEB18   g5926696   0   Human genomic DNA, chromosome 6p21.3, HLA Class I region, section                       8/20.       134   LI:811341.1:2000FEB01   g5926696   0   Human genomic DNA, chromosome 6p21.3, HLA Class I region, section                       8/20.       135   LI:903225.1:2000FEB01   g5926710   0   Human genomic DNA, chromosome 6p21.3, HLA Class I region, section                       20/20.       136   LI:242079.2:2000FEB01   g5926703   0   Human genomic DNA, chromosome 6p21.3, HLA Class I region, section                       15/20.       137   LG:979580.1:2000MAY19   g9280152   7.00E−23   unnamed portein product ( Macaca fascicularis )       138   LI:1169865.1:2000MAY01   g673417   1.00E−112   class II antigen ( Homo sapiens )       139   LG:337818.2:2000FEB18   g404777   4.80E−84   cytochrome P-450 2B-Bx       140   LI:337818.1:2000FEB01   g203759   4.00E−58   cytochrome P-450(1)       141   LG:241577.4:2000MAY19   g2809498   1.50E−29   cytochrome c oxidase subunit IV       142   LG:344786.4:2000MAY19   g164981   2.00E−06   cytochrome P-450p-2 ( Oryctolagus cuniculus )       143   LI:414307.1:2000FEB01   g30095   9.00E−48   collagen subunit (alpha-1 (X)) 3       144   LI:202943.2:2000FEB01   g391663   7.00E−06   hikaru genki type 1 product       145   LI:246194.2:2000FEB01   g1405821   1.00E−05   SULFATED SURFACE GLYCOPROTEIN 185       146   LI:815961.1:2000FEB01   g292045   0   Human mucin mRNA, partial cds.       147   LG:120744.1:2000MAY19   g4582324   1.00E−168   dJ708F5.1 (PUTATIVE novel Collagen alpha 1 LIKE protein) (Homo       148   LI:757520.1:2000MAY01   g7161771   0   keratin ( Homo sapiens )       149   LG:160570.1:2000FEB18   g466548   1.00E−46   NBL4       150   LI:350398.3:2000FEB01   g3724141   1.00E−06   myosin I       151   LI:221285.1:2000FEB01   g18218   2.00E−74   spoke protein       152   LI:401605.2:2000FEB01   g1755049   1.00E−15   myosin X       153   LI:329017.1:2000FEB01   g1813638   2.00E−51   PF20       154   LI:401322.1:2000FEB01   g38076   2.00E−30   Macaque mRNA for alpha-tubulin.       155   LG:403409.1:2000MAY19   g7303061   0   Khc-73 gene product ( Drosophila melanogaster )       156   LG:233933.5:2000MAY19   g7385113   2.00E−18   ankyrin 1 ( Bos taurus )       157   LI:290344.1:2000MAY01   g1353782   0   dystrophin-related protein 2 ( Homo sapiens )       158   LI:410742.1:2000MAY01   g2290200   0   desmoglein 3 ( Mus musculus )       159   LG:406568.1:2000MAY19   g28969   5.30E−44   64 Kd autoantigen       160   LI:283762.1:2000MAY01   g1469868   0   The KIAA0143 gene product is related to a putative  C. elegans  gene                       encoded on cosmid C32D5. ( Homo sapiens )       161   LI:347687.113:2000MAY01   g387514   1.00E−123   DM-20 protein ( Mus musculus )       162   LI:1146510.1:2000MAY01   g2149291   3.00E−24   defender against death 1 protein ( Homo sapiens )       163   LG:451710.1:2000FEB18   g5816996   2.40E−42   ribosomal protein L32-like protein       164   LG:455771.1:2000FEB18   g643074   1.70E−59   putative 40S ribosomal protein s12       165   LG:452089.1:2000FEB18   g463252   1.90E−62   RL5 ribosomal protein       166   LG:246415.1:2000FEB18   g296451   0   Human mRNA for ribosomal protein S26.       167   LG:414144.10:2000FEB18   g200785   1.80E−16   ribosomal protein L7       168   LG:1101445.1:2000FEB18   g1800114   0   Human ribosomal protein L7 antisense mRNA gene, partial sequence.       169   LG:452134.1:2000FEB18   g550024   0   Human ribosomal protein S10 mRNA, complete cds.       170   LI:903021.1:2000FEB01   g36139   0   Human mRNA for ribosomal protein L7.       171   LI:246422.1:2000FEB01   g409069   0   Human mRNA for HBp15/L22, complete cds.       172   LG:449404.1:2000MAY19   g4886269   5.00E−66   putative ribosomal protein S14 ( Arabidopsis thaliana )       173   LG:449413.1:2000MAY19   g643074   1.00E−70   putative 40S ribosomal protein s12 (Fragaria x ananassa)       174   LG:450105.1:2000MAY19   g643074   6.00E−76   putative 40S ribosomal protein s12 (Fragaria x ananassa)       175   LG:460809.1:2000MAY19   g36129   4.00E−54   Human mRNA for ribosomal protein L31.       176   LG:481781.1:2000MAY19   g2331301   1.00E−130   ribosomal protein S4 type I ( Zea mays )       177   LG:1101153.1:2000MAY19   g2668748   2.00E−95   ribosomal protein L17 ( Zea mays )       178   LI:257695.20:2000MAY01   g57714   1.00E−62   ribosomal protein S16 (AA 1-146) ( Rattus rattus )       179   LI:455771.1:2000MAY01   g643074   6.00E−76   putative 40S ribosomal protein s12 (Fragaria x ananassa)       180   LI:274551.1:2000MAY01   g36145   2.00E−59   Human mRNA for ribosomal protein S12.       181   LI:035973.1:2000MAY01   g57121   8.00E−29   ribosomal protein L37 ( Rattus norvegicus )       182   LG:978427.5:2000FEB18   g545998   2.50E−67   tricarboxylate carrier (rats, liver, Peptide Mitochondrial Partial, 357 aa)       183   LG:247781.2:2000FEB18   g2352427   9.40E−29   peroxisomal Ca-dependent solute carrier       184   LI:034583.1:2000FEB01   g5815141   0   nuclear body associated kinase 1b       185   LI:333307.2:2000FEB01   g295671   0.0003   selected as a weak suppressor of a mutant of the subunit AC40 of DNA                       dependant RNA polymerase I and III       186   LI:814710.2:2000FEB01   g178281   1.00E−46   AHNAK nucleoprotein       187   LG:414732.1:2000MAY19   g183233   2.00E−54   beta-glucuronidase precursor (EC 3.2.1.31)       188   LG:413910.6:2000MAY19   g7022046   1.00E−109   unnamed protein product ( Homo sapiens )       189   LI:414732.2:2000MAY01   g183232   0   Human beta-glucuronidase mRNA, complete cds.       190   LI:900264.2:2000MAY01   g414797   2.00E−81   pyruvate dehydrogenase phosphatase ( Bos taurus )       191   LI:335593.1:2000MAY01   g3851553   5.00E−34   RNA-binding protein Nova-2 ( Homo sapiens )       192   LI:1189543.1:2000MAY01   g7025507   0   ventral neuron-specific protein 1 NOVA1 ( Mus musculus )       193   LG:455450.1:2000FEB18   g4105111   2.10E−20   dehydrin 6       194   LG:1040978.1:2000FEB18   g453189   3.30E−41   acyl carrier protein       195   LG:446649.1:2000FEB18   g181960   2.00E−35   Human endozepine (putative ligand of benzodiazepine receptor)                       mRNA, complete cds.       196   LG:132147.3:2000FEB18   g6446606   0   E3 ubiquitin ligase SMURF1 ( Homo sapiens )       197   LI:036034.1:2000FEB01   g9622856   1.00E−33   sorting nexin 15A ( Homo sapiens )       198   LG:162161.1:2000MAY19   g5823961   3.00E−87   dJ20B11.1 (ortholog of rat RSEC5 (mammalian exocyst complex subunit))                       ( Homo sapiens )       199   LG:407214.10:2000MAY19   g9963839   4.00E−54   lipase ( Homo sapiens )       200   LG:204626.1:2000MAY19   g3243240   5.10E−41   syntaxin 11       201   LI:007401.1:2000MAY01   g4512103   3.00E−81   rab 11 binding protein ( Bos taurus )       202   LI:476342.1:2000MAY01   g790641   2.00E−21   gamma-thionin ( Hordeum vulgare )       203   LI:1072759.1:2000MAY01   g2367625   4.00E−21   protein synthesis elongation factor 1-alpha ( Rhodotorula mucilaginosa )       204   LG:998857.1:2000FEB18   g2731641   6.10E−13   Fas-ligand associated factor 3       205   LG:482261.1:2000FEB18   g4003386   0   Human genomic DNA of 8p21.3-p22 anti-oncogene of hepatocellular                       colorectal and non-small cell lung cancer, segment 9/11.       206   LG:480328.1:2000FEB18   g246482   0   prohibitin (Human, mRNA, 1043 nt).       207   LG:311197.1:2000MAY19   g505033   8.00E−62   mitogen inducible gene mig-2 ( Homo sapiens )       208   LG:1054883.1:2000MAY19   g325464   0   Human endogenous retrovirus type C oncovirus sequence.       209   LG:399395.1:2000MAY19   g1177607   8.00E−10   pva1 ( Plasmodium vivax )       210   LG:380497.2:2000MAY19   g10504238   3.00E−88   hepatocellular carcinoma-related putative tumor suppressor (Homo       211   LI:272913.22:2000MAY01   g4982485   3.00E−59   apoptosis related protein APR-3 ( Homo sapiens )                  
 
     [0898]                                           TABLE 2                       SEQ ID NO:   Template ID   Start   Stop   Frame   Pfam Hit   Pfam Description   E-value                                                                1   LG:1040582.1:2000FEB18   267   539   forward 3   aldo_ket_red   Aldo/keto reductase family   2.50E−51       2   LG:453570.1:2000FEB18   186   605   forward 3   Glyoxalase   Glyoxalase   3.80E−72       3   LG:408751.3:2000FEB18   194   1345   forward 2   Dihydrooratase   Dihydroorotase-like   1.40E−19       4   LI:090574.1:2000FEB01   60   776   forward 3   carb_anhydrase   Eukaryotic-type carbonic anhydrase   9.70E−144       6   LI:332176.1:2000FEB01   2   961   forward 2   Glyco_hydro_31   Glycosyl hydrolases family 31   4.10E−144       7   LI:403248.2:2000FEB01   191   367   forward 2   Orn_DAP_Arg_deC   Pyrldoxal-dependent decarboxylase   1.40E−12       8   LG:220992.1:2000MAY19   156   1556   forward 3   Amidase   Amidase   1.10E−153       9   LG:1094571.1:2000MAY19   328   720   forward 1   FAD_Synth   Riboflavin kinase/FAD synthetase   2.30E−42       10   LI:350754.4:2000MAY01   855   1121   forward 3   Transglut_core   Transglutaminase-like superfamily   2.90E−47       10   LI:350754.4:2000MAY01   1455   2132   forward 3   Transglutamin_C   Transglutaminase family   3.20E−106       10   LI:350754.4:2000MAY01   54   413   forward 3   Transglutamin_N   Transglutaminase family   2.50E−63       11   LI:255828.29:2000MAY01   2   367   forward 2   PK   Pyruvate kinase   7.00E−71       11   LI:255828.29:2000MAY01   348   512   forward 3   PK   Pyruvate kinase   5.70E−24       12   LI:1190263.1:2000MAY01   281   1750   forward 2   Sulfatase   Sulfatase   8.60E−66       14   LG:999414.3:2000FEB18   718   1038   forward 1   7tm_1   7 transmembrane receptor   3.60E−13                               (rhodopsin family)       14   LG:999414.3:2000FEB18   1115   1453   forward 2   7tm_1   7 transmembrane receptor   4.30E−07                               (rhodopsin family)       18   LG:068682.1:2000FEB18   176   883   forward 2   pkinase   Eukaryotic protein kinase domain   1.70E−65       19   LG:242665.1:2000FEB18   190   747   forward 1   ras   Ras family   2.30E−34       20   LG:241743.1:2000FEB18   199   345   forward 1   PH   PH domain   8.00E−06       22   LG:344886.1:2000MAY19   379   957   forward 1   ras   Ras family   1.70E−17       25   LG:898771.1:2000MAY19   525   662   forward 3   Fork_head   Fork head domain   1.70E−25       28   LI:1085273.2:2000MAY01   285   1070   forward 3   DSPc   Dual specificity phosphatase,   1.30E−39                               catalytic domain       29   LI:333138.2:2000MAY01   291   1016   forward 3   pkinase   Eukaryotic protein kinase domain   2.00E−90       32   LG:998283.7:2000FEB18   370   630   forward 1   bromodomain   Bromodomain   2.60E−29       32   LG:998283.7:2000FEB18   4   153   forward 1   PHD   PHD-finger   1.90E−12       34   LI:175223.1:2000FEB01   210   431   forward 3   CSD   ‘Cold-shock’ DNA-binding domair   1.40E−18       38   LG:021371.3:2000FEB18   932   1000   forward 2   zf-C2H2   Zinc finger, C2H2 type   2.10E−04       39   LG:475404.1:2000FEB18   176   328   forward 2   KRAB   KRAB box   1.10E−15       40   LG:979406.2:2000FEB18   85   273   forward 1   KRAB   KRAB box   2.40E−34       41   LG:410726.1:2000FEB18   646   834   forward 1   KRAB   KRAB box   2.10E−17       41   LG:410726.1:2000FEB18   274   558   forward 1   SCAN   SCAN domain   8.90E−55       43   LG:1076828.1:2000FEB18   448   516   forward 1   zf-C2H2   Zinc finger, C2H2 type   2.00E−07       44   LG:1076931.1:2000FEB18   173   310   forward 2   KRAB   KRAB box   3.40E−21       45   LG:1078121.1:2000FEB18   186   374   forward 3   KRAB   KRAB box   2.80E−41       46   LG:1079203.1:2000FEB18   421   489   forward 1   zf-C2H2   Zinc finger, C2H2 type   6.00E−06       46   LG:1079203.1:2000FEB18   647   715   forward 2   zf-C2H2   Zinc finger, C2H2 type   9.20E−05       47   LG:1082586.1:2000FEB18   414   536   forward 3   KRAB   KRAB box   6.80E−12       48   LG:1082774.1:2000FEB18   138   326   forward 3   KRAB   KRAB box   1.30E−40       49   LG:1082775.1:2000FEB18   45   230   forward 3   KRAB   KRAB box   7.10E−39       49   LG:1082775.1:2000FEB18   840   908   forward 3   zf-C2H2   Zinc finger, C2H2 type   4.40E−05       50   LG:1083120.1:2000FEB18   117   266   forward 3   KRAB   KRAB box   5.10E−22       51   LG:1087707.1:2000FEB18   162   350   forward 3   KRAB   KRAB box   2.80E−40       52   LG:1090915.1:2000FEB18   129   251   forward 3   KRAB   KRAB box   7.40E−22       53   LG:1094230.1:2000FEB18   120   308   forward 3   KRAB   KRAB box   3.70E−41       54   LG:474848.3:2000FEB18   253   441   forward 1   KRAB   KRAB box   2.10E−38       55   LI:251656.1:2000FEB01   242   310   forward 2   zf-C2H2   Zinc finger, C2H2 type   3.90E−08       56   LI:021371.1:2000FEB01   717   785   forward 3   zf-C2H2   Zinc finger, C2H2 type   2.10E−04       57   LI:133095.1:2000FEB01   539   607   forward 2   zf-C2H2   Zinc finger, C2H2 type   4.30E−06       58   LI:236654.2:2000FEB01   805   873   forward 1   zf-C2H2   Zinc finger, C2H2 type   1.40E−04       59   LI:200009.1:2000FEB01   564   632   forward 3   zf-C2H2   Zinc finger, C2H2 type   1.80E−05       60   LI:758502.1:2000FEB01   633   701   forward 3   zf-C2H2   Zinc finger, C2H2 type   2.50E−07       62   LI:789445.1:2000FEB01   71   262   forward 2   KRAB   KRAB box   1.60E−27       63   LI:789657.1:2000FEB01   542   610   forward 2   zf-C2H2   Zinc finger, C2H2 type   2.60E−06       64   LI:789808.1:2000FEB01   272   340   forward 2   zf-C2H2   Zinc finger, C2H2 type   1.00E−07       64   LI:789808.1:2000FEB01   426   494   forward 3   zf-C2H2   Zinc finger, C2H2 type   3.40E−04       65   LI:792919.1:2000FEB01   31   99   forward 1   zf-C2H2   Zinc finger, C2H2 type   5.30E−06       66   LI:793949.1:2000FEB01   120   308   forward 3   KRAB   KRAB box   1.70E−41       67   LI:794389.1:2000FEB01   75   143   forward 3   zf-C2H2   Zinc finger, C2H2 type   8.70E−06       68   LI:796010.1:2000FEB01   276   344   forward 3   zf-C2H2   Zinc finger, C2H2 type   1.00E−07       68   LI:796010.1:2000FEB01   433   501   forward 1   zf-C2H2   Zinc finger, C2H2 type   3.40E−04       69   LI:796324.1:2000FEB01   290   358   forward 2   zf-C2H2   Zinc finger, C2H2 type   1.00E−07       69   LI:796324.1:2000FEB01   450   518   forward 3   zf-C2H2   Zinc finger, C2H2 type   3.40E−04       70   LI:796373.1:2000FEB01   181   249   forward 1   zf-C2H2   Zinc finger, C2H2 type   1.10E−06       71   LI:796415.1:2000FEB01   45   230   forward 3   KRAB   KRAB box   7.10E−39       72   LI:798636.1:2000FEB01   329   397   forward 2   zf-C2H2   Zinc finger, C2H2 type   2.60E−07       73   LI:800045.1:2000FEB01   364   432   forward 1   zf-C2H2   Zinc finger, C2H2 type   5.30E−07       74   LI:800680.1:2000FEB01   155   319   forward 2   KRAB   KRAB box   5.00E−21       75   LI:800894.1:2000FEB01   125   313   forward 2   KRAB   KRAB box   6.50E−40       76   LI:801015.1:2000FEB01   22   216   forward 1   KRAB   KRAB box   3.00E−24       77   LI:801236.1:2000FEB01   225   293   forward 3   zf-C2H2   Zinc finger, C2H2 type   4.40E−07       78   LI:803335.1:2000FEB01   220   408   forward 1   KRAB   KRAB box   2.10E−38       79   LI:803998.1:2000FEB01   62   130   forward 2   zf-C2H2   Zinc finger, C2H2 type   1.20E−05       80   LI:478757.1:2000FEB01   467   643   forward 2   KRAB   KRAB box   2.40E−21       81   LI:808532.1:2000FEB01   53   121   forward 2   zf-C2H2   Zinc finger, C2H2 type   5.70E−05       82   LI:443073.1:2000FEB01   176   244   forward 2   zf-C2H2   Zinc finger, C2H2 type   2.50E−05       83   LI:479671.1:2000FEB01   160   312   forward 1   KRAB   KRAB box   1.70E−19       84   LI:810078.1:2000FEB01   424   492   forward 1   zf-C2H2   Zinc finger, C2H2 type   1.80E−06       84   LI:810078.1:2000FEB01   587   655   forward 2   zf-C2H2   Zinc finger, C2H2 type   1.20E−05       85   LI:810224.1:2000FEB01   171   239   forward 3   zf-C2H2   Zinc finger, C2H2 type   1.00E−07       86   LI:817052.2:2000FEB01   901   969   forward 1   zf-C2H2   Zinc finger, C2H2 type   8.90E−08       87   LG:892274.1:2000MAY19   96   461   forward 3   dUTPase   dUTPase   9.20E−27       87   LG:892274.1:2000MAY19   489   752   forward 3   rvp   Retroviral aspartyl protease   5.30E−11       88   LG:1080959.1:2000MAY19   182   322   forward 2   KRAB   KRAB box   2.00E−16       89   LG:1054900.1:2000MAY19   78   218   forward 3   KRAB   KRAB box   2.30E−17       90   LG:1077357.1:2000MAY19   94   282   forward 1   KRAB   KRAB box   4.80E−31       91   LG:1084051.1:2000MAY19   195   263   forward 3   zf-C2H2   Zinc finger, C2H2 type   1.80E−06       92   LG:1076853.1:2000MAY19   706   774   forward 1   zf-C2H2   Zinc finger, C2H2 type   1.50E−07       93   LG:481631.10:2000MAY19   96   263   forward 3   KRAB   KRAB box   5.70E−25       93   LG:481631.10:2000MAY19   882   950   forward 3   zf-C2H2   Zinc finger, C2H2 type   1.70E−05       94   LG:1088431.2:2000MAY19   175   339   forward 1   KRAB   KRAB box   5.00E−21       96   LI:1144007.1:2000MAY01   914   1108   forward 2   KRAB   KRAB box   5.90E−05       96   LI:1144007.1:2000MAY01   323   610   forward 2   SCAN   SCAN domain   4.10E−60       97   LI:331074.1:2000MAY01   194   262   forward 2   zf-C2H2   Zinc finger, C2H2 type   1.00E−03       98   LI:1170349.1:2000MAY01   185   370   forward 2   KRAB   KRAB box   2.50E−29       98   LI:1170349.1:2000MAY01   740   808   forward 2   zf-C2H2   Zinc finger, C2H2 type   5.80E−05       102   LG:101269.1:2000MAY19   556   831   forward 1   IRK   Inward rectifier potassium channel   3.50E−65       104   LI:410188.1:2000MAY01   3760   4569   forward 1   ion_trans   Ion transport protein   3.70E−97       104   LI:410188.1:2000MAY01   4586   5314   forward 2   ion_trans   Ion transport protein   3.30E−66       105   LI:1188288.1:2000MAY01   751   1215   forward 1   SNF   Sodium:neurotransmitter   8.50E−113                               symporter family       105   LI:1188288.1:2000MAY01   423   782   forward 3   SNF   Sodium:neurotransmitter   8.60E−74                               symporter family       105   LI:1188288.1:2000MAY01   1187   1438   forward 2   SNF   Sodium:neurotransmitter   5.50E−52                               symporter family       107   LG:451682.1:2000FEB18   117   560   forward 3   proteasome   Proteasome A-type and B-type   4.40E−59       108   LG:1077283.1:2000FEB18   110   427   forward 2   pro_isomerase   Cyclophilin type peptidyl-prolyl   1.80E−37                               cis-trans isomerase       108   LG:1077283.1:2000FEB18   177   278   forward 3   pro_isomerase   Cyclophilin type peptidyl-prolyl   1.30E−18                               cis-trans isomerase       109   LG:481436.5:2000FEB18   351   539   forward 3   Dnaj   Dnaj domain   2.80E−28       111   LI:373637.1:2000FEB01   17   217   forward 2   Dnaj   Dnaj domain   6.30E−39       113   LI:053826.1:2000MAY01   834   1106   forward 3   SCP   SCP-like extracellular protein   1.10E−17       114   LI:449393.1:2000MAY01   90   788   forward 3   cpn60_TCP1   TCP-1/cpn60 chaperonin family   9.80E−66       117   LG:345527.1:2000FEB18   667   957   forward 1   helicase_C   Helicases conserved C-terminal domain   7.20E−21       117   LG:345527.1:2000FEB18   8   631   forward 2   SNF2_N   SNF2 and others N-terminal domain   7.20E−44       122   LI:335671.2:2000FEB01   188   475   forward 2   helicase_C   Heilcases conserved C-terminal domain   9.10E−13       122   LI:335671.2:2000FEB01   3   95   forward 3   SNF2_N   SNF2 and others N-terminal domain   7.10E−06       128   LG:247388.1:2000MAY19   346   600   forward 1   helicase_C   Heilcases conserved C-terminal domain   2.70E−19       128   LG:247388.1:2000MAY19   3   173   forward 3   SNF2_N   SNF2 and others N-terminal domain   1.60E−14       131   LI:331610.2:2000MAY01   1415   1699   forward 2   cadherin   Cadherin domain   6.00E−20       135   LI:903225.1:2000FEB01   603   764   forward 3   Ribosomal_L23   Ribosomal protein L23   4.80E−14       138   LI:1169865.1:2000MAY01   593   790   forward 2   ig   Immunoglobulin domain   2.30E−08       138   LI:1169865.1:2000MAY01   242   547   forward 2   MHC_II_alpha   Class II histocompatibility antigen,   1.80E−65                               alpha domain       139   LG:337818.2:2000FEB18   136   1518   forward 1   p450   Cytochrome P450   1.50E−173       140   LI:337818.1:2000FEB01   654   998   forward 3   p450   Cytochrome P450   3.50E−45       140   LI:337818.1:2000FEB01   136   384   forward 1   p450   Cytochrome P450   4.40E−27       140   LI:337818.1:2000FEB01   359   673   forward 2   p450   Cytochrome P450   5.40E−27       143   LI:414307.1:2000FEB01   590   964   forward 2   C1q   C1q domain   2.30E−38       143   LI:414307.1:2000FEB01   365   544   forward 2   Collagen   Collagen triple helix repeat (20 copies)   2.50E−10       144   LI:202943.2:2000FEB01   36   209   forward 3   sushi   Sushi domain (SCR repeat)   1.40E−09       147   LG:120744.1:2000MAY19   301   813   forward 1   vwa   von Willebrand factor type A domain   2.00E−51       148   LI:757520.1:2000MAY01   427   1362   forward 1   filament   Intermediate filament proteins   7.10E−157       149   LG:160570.1:2000FEB18   260   562   forward 2   Band_41   FERM domain (Band 4.1 family)   1.60E−22       152   LI:401605.2:2000FEB01   1   129   forward 1   myosin_head   Myosin head (motor domain)   5.90E−07       153   LI:329017.1:2000FEB01   226   336   forward 1   WD40   WD domain, G-beta repeat   5.10E−06       154   LI:401322.1:2000FEB01   156   341   forward 3   tubulin   Tubulin/FtsZ family   7.10E−20       154   LI:401322.1:2000FEB01   371   478   forward 2   tubulin   Tubulin/FtsZ family   2.50E−06       155   LG:403409.1:2000MAY19   1458   1652   forward 3   FHA   FHA domain   3.00E−04       155   LG:403409.1:2000MAY19   78   1193   forward 3   kinesin   Kinesin motor domain   6.80E−172       156   LG:233933.5:2000MAY19   258   356   forward 3   ank   Ank repeat   4.90E−06       157   LI:290344.1:2000MAY01   992   1312   forward 2   spectrin   Spectrin repeat   4.10E−07       157   LI:290344.1:2000MAY01   1361   1450   forward 2   WW   WW domain   5.40E−08       158   LI:410742.1:2000MAY01   599   889   forward 2   cadherin   Cadherin domain   1.80E−21       158   LI:410742.1:2000MAY01   1224   1520   forward 3   cadherin   Cadherin domain   9.90E−04       161   LI:347687.113:2000MAY01   214   855   forward 1   Myelin_PLP   Myelin proteolipid protein   7.10E−160                               (PLP or lipophilin)       163   LG:451710.1:2000FEB18   130   459   forward 1   Ribosomal_L32e   Ribosomal protein L32   4.80E−57       164   LG:455771.1:2000FEB18   69   473   forward 3   Ribosomal_S12   Ribosomal protein S12   6.60E−78       165   LG:452089.1:2000FEB18   107   268   forward 2   Ribosomal_L5   Ribosomal protein L5   2.40E−25       165   LG:452089.1:2000FEB18   278   577   forward 2   Ribosomal_L5_C   ribosomal L5P family C-terminus   2.70E−60       166   LG:246415.1:2000FEB18   27   365   forward 3   Ribosomal_S26e   Ribosomal protein S26e   2.40E−59       168   LG:1101445.1:2000FEB18   306   464   forward 3   Ribosomal_L30   Ribosomal protein L30p/L7e   4.20E−28       171   LI:246422.1:2000FEB01   53   397   forward 2   Ribosomal_L22e   Ribosomal L22e protein family   4.30E−28       171   LI:246422.1:2000FEB01   64   318   forward 1   Ribosomal_L22e   Ribosomal L22e protein family   7.70E−07       172   LG:449404.1:2000MAY19   175   531   forward 1   Ribosomal_S11   Ribosomal protein S11   6.90E−77       173   LG:449413.1:2000MAY19   99   368   forward 3   Ribosomal_S12   Ribosomal protein S12   6.10E−47       173   LG:449413.1:2000MAY19   367   504   forward 1   Ribosomal_S12   Ribosomal protein S12   3.20E−21       174   LG:450105.1:2000MAY19   86   490   forward 2   Ribosomal_S12   Ribosomal protein S12   6.60E−78       175   LG:460809.1:2000MAY19   3   236   forward 3   Ribosomal_L31e   Ribosomal protein L31e   6.00E−17       176   LG:481781.1:2000MAY19   243   671   forward 3   Ribosomal_S4e   Ribosomal family S4e   1.40E−97       177   LG:1101153.1:2000MAY19   89   499   forward 2   Ribosomal_L22   Ribosomal protein L22p/L17e   5.20E−76       178   LI:257695.20:2000MAY01   110   673   forward 2   Ribosomal_S9   Ribosomal protein S9/S16   1.60E−40       179   LI:455771.1:2000MAY01   69   473   forward 3   Ribosomal_S12   Ribosomal protein S12   6.60E−78       181   LI:035973.1:2000MAY01   318   479   forward 3   Ribosomal_L37e   Ribosomal protein L37e   1.60E−13       183   LG:247781.2:2000FEB18   142   426   forward 1   mito_carr   Mitochondrial carrier proteins   1.80E−24       190   LI:900264.2:2000MAY01   1151   1555   forward 2   PP2C   Protein phosphatase 2C   2.90E−11       192   LI:1189543.1:2000MAY01   1292   1447   forward 2   KH-domain   KH domain   2.50E−13       192   LI:1189543.1:2000MAY01   592   744   forward 1   KH-domain   KH domain   5.40E−13       193   LG:455450.1:2000FEB18   1   426   forward 1   dehydrin   Dehydrins   4.20E−41       194   LG:1040978.1:2000FEB18   278   481   forward 2   pp-binding   Phosphopantetheine attachment site   3.90E−14       195   LG:446649.1:2000FEB18   80   316   forward 2   ACBP   Acyl CoA binding protein   1.60E−44       196   LG:132147.3:2000FEB18   1497   2414   forward 3   HECT   HECT-domain (ubiquitin-transferase).   9.90E−138       196   LG:132147.3:2000FEB18   1065   1154   forward 3   WW   WW domain   1.50E−12       198   LG:162161.1:2000MAY19   128   385   forward 2   TIG   IPT/TIG domain   5.50E−15       200   LG:204626.1:2000MAY19   322   1212   forward 1   Syntaxin   Syntaxin   8.60E−44       202   LI:476342.1:2000MAY01   159   299   forward 3   Gamma-thionin   Gamma-thionins family   1.70E−19       205   LG:482261.1:2000FEB18   286   552   forward 1   Gag_p10   Retroviral GAG p10 protein   6.80E−31       205   LG:482261.1:2000FEB18   1044   1229   forward 3   gag_p24   gag gene protein p24 (core nucleocapsid   2.00E−15       205   LG:482261.1:2000FEB18   1375   1545   forward 1   gag_p24   gag gene protein p24 (core nucleocapsid   9.50E−15       206   LG:480328.1:2000FEB18   985   1515   forward 1   Band_7   SPFH domain/Band 7 family   2.60E−39       206   LG:480328.1:2000FEB18   49   117   forward 1   zf-C2H2   Zinc finger, C2H2 type   1.60E−06       210   LG:380497.2:2000MAY19   202   336   forward 1   G-patch   G-patch domain   7.00E−17                    
     [0899]                                       TABLE 3                                           Domain           SEQ ID NO:   Template ID   Start   Stop   Frame   Type   Topology                                                            1   LG:1040582.1:2000FEB18   31   117   forward 1   TM   N in       1   LG:1040582.1:2000FEB18   319   405   forward 1   TM   N in       1   LG:1040582.1:2000FEB18   108   155   forward 3   TM   N out       2   LG:453570.1:2000FEB18   361   447   forward 1   TM   N in       3   LG:408751.3:2000FEB18   1318   1404   forward 1   TM   N in       3   LG:408751.3:2000FEB18   1025   1099   forward 2   TM   N in       3   LG:408751.3:2000FEB18   1298   1360   forward 2   TM   N in       3   LG:408751.3:2000FEB18   1379   1441   forward 2   TM   N in       3   LG:408751.3:2000FEB18   1463   1537   forward 2   TM   N in       3   LG:408751.3:2000FEB18   1047   1133   forward 3   TM   N in       3   LG:408751.3:2000FEB18   1266   1352   forward 3   TM   N in       3   LG:408751.3:2000FEB18   1419   1469   forward 3   TM   N in       4   LI:090574.1:2000FEB01   79   144   forward 1   TM   N in       4   LI:090574.1:2000FEB01   607   678   forward 1   TM   N in       4   LI:090574.1:2000FEB01   1009   1080   forward 1   TM   N in       4   LI:090574.1:2000FEB01   497   583   forward 2   TM   N out       4   LI:090574.1:2000FEB01   743   829   forward 2   TM   N out       4   LI:090574.1:2000FEB01   1026   1085   forward 3   TM   N out       5   LI:229932.2:2000FEB01   76   162   forward 1   TM   N out       5   LI:229932.2:2000FEB01   190   276   forward 1   TM   N out       5   LI:229932.2:2000FEB01   1237   1323   forward 1   TM   N out       5   LI:229932.2:2000FEB01   68   142   forward 2   TM   N in       5   LI:229932.2:2000FEB01   335   412   forward 2   TM   N in       5   LI:229932.2:2000FEB01   758   844   forward 2   TM   N in       5   LI:229932.2:2000FEB01   1229   1288   forward 2   TM   N in       5   LI:229932.2:2000FEB01   60   146   forward 3   TM   N in       5   LI:229932.2:2000FEB01   216   302   forward 3   TM   N in       5   LI:229932.2:2000FEB01   690   752   forward 3   TM   N in       5   LI:229932.2:2000FEB01   765   827   forward 3   TM   N in       5   LI:229932.2:2000FEB01   1209   1289   forward 3   TM   N in       6   LI:332176.1:2000FEB01   343   399   forward 1   TM   N in       6   LI:332176.1:2000FEB01   1078   1131   forward 1   TM   N in       6   LI:332176.1:2000FEB01   1606   1692   forward 1   TM   N in       6   LI:332176.1:2000FEB01   2218   2274   forward 1   TM   N in       6   LI:332176.1:2000FEB01   2383   2433   forward 1   TM   N in       6   LI:332176.1:2000FEB01   110   196   forward 2   TM   N in       6   LI:332176.1:2000FEB01   1307   1378   forward 2   TM   N in       6   LI:332176.1:2000FEB01   1640   1726   forward 2   TM   N in       6   LI:332176.1:2000FEB01   1946   2005   forward 2   TM   N in       6   LI:332176.1:2000FEB01   135   200   forward 3   TM   N in       6   LI:332176.1:2000FEB01   693   752   forward 3   TM   N in       6   LI:332176.1:2000FEB01   777   839   forward 3   TM   N in       6   LI:332176.1:2000FEB01   867   929   forward 3   TM   N in       6   LI:332176.1:2000FEB01   1035   1118   forward 3   TM   N in       6   LI:332176.1:2000FEB01   1173   1253   forward 3   TM   N in       6   LI:332176.1:2000FEB01   1572   1658   forward 3   TM   N in       6   LI:332176.1:2000FEB01   2121   2180   forward 3   TM   N in       6   LI:332176.1:2000FEB01   2277   2363   forward 3   TM   N in       6   LI:332176.1:2000FEB01   2400   2456   forward 3   TM   N in       8   LG:220992.1:2000MAY19   343   393   forward 1   TM       8   LG:220992.1:2000MAY19   646   732   forward 1   TM       8   LG:220992.1:2000MAY19   1639   1725   forward 1   TM       8   LG:220992.1:2000MAY19   1879   1965   forward 1   TM       8   LG:220992.1:2000MAY19   2005   2088   forward 1   TM       8   LG:220992.1:2000MAY19   17   76   forward 2   TM   N in       8   LG:220992.1:2000MAY19   1646   1732   forward 2   TM   N in       8   LG:220992.1:2000MAY19   1850   1933   forward 2   TM   N in       8   LG:220992.1:2000MAY19   1434   1484   forward 3   TM   N out       8   LG:220992.1:2000MAY19   1734   1820   forward 3   TM   N out       8   LG:220992.1:2000MAY19   1974   2036   forward 3   TM   N out       8   LG:220992.1:2000MAY19   2067   2129   forward 3   TM   N out       8   LG:220992.1:2000MAY19   2151   2237   forward 3   TM   N out       9   LG:1094571.1:2000MAY19   781   867   forward 1   TM   N in       9   LG:1094571.1:2000MAY19   419   505   forward 2   TM   N in       9   LG:1094571.1:2000MAY19   767   853   forward 2   TM   N in       9   LG:1094571.1:2000MAY19   756   842   forward 3   TM   N in       10   LI:350754.4:2000MAY01   277   348   forward 1   TM   N in       10   LI:350754.4:2000MAY01   583   651   forward 1   TM   N in       10   LI:350754.4:2000MAY01   670   747   forward 1   TM   N in       10   LI:350754.4:2000MAY01   381   467   forward 3   TM   N in       10   LI:350754.4:2000MAY01   2469   2555   forward 3   TM   N in       12   LI:1190263.1:2000MAY01   664   735   forward 1   TM   N in       12   LI:1190263.1:2000MAY01   787   861   forward 1   TM   N in       12   LI:1190263.1:2000MAY01   901   954   forward 1   TM   N in       12   LI:1190263.1:2000MAY01   188   274   forward 2   TM   N in       12   LI:1190263.1:2000MAY01   455   508   forward 2   TM   N in       12   LI:1190263.1:2000MAY01   809   895   forward 2   TM   N in       12   LI:1190263.1:2000MAY01   1616   1663   forward 2   TM   N in       12   LI:1190263.1:2000MAY01   183   251   forward 3   TM   N in       12   LI:1190263.1:2000MAY01   648   704   forward 3   TM   N in       12   LI:1190263.1:2000MAY01   1149   1235   forward 3   TM   N in       13   LG:270916.2:2000FEB18   173   259   forward 2   TM   N out       14   LG:999414.3:2000FEB18   109   195   forward 1   TM   N out       14   LG:999414.3:2000FEB18   358   438   forward 1   TM   N out       14   LG:999414.3:2000FEB18   520   591   forward 1   TM   N out       14   LG:999414.3:2000FEB18   661   744   forward 1   TM   N out       14   LG:999414.3:2000FEB18   883   969   forward 1   TM   N out       14   LG:999414.3:2000FEB18   976   1062   forward 1   TM   N out       14   LG:999414.3:2000FEB18   302   388   forward 2   TM   N in       14   LG:999414.3:2000FEB18   533   613   forward 2   TM   N in       14   LG:999414.3:2000FEB18   992   1048   forward 2   TM   N in       14   LG:999414.3:2000FEB18   1169   1246   forward 2   TM   N in       14   LG:999414.3:2000FEB18   1307   1366   forward 2   TM   N in       14   LG:999414.3:2000FEB18   207   284   forward 3   TM   N out       14   LG:999414.3:2000FEB18   324   404   forward 3   TM   N out       14   LG:999414.3:2000FEB18   540   599   forward 3   TM   N out       14   LG:999414.3:2000FEB18   1029   1115   forward 3   TM   N out       14   LG:999414.3:2000FEB18   1167   1253   forward 3   TM   N out       14   LG:999414.3:2000FEB18   1314   1373   forward 3   TM   N out       15   LG:429446.1:2000FEB18   628   699   forward 1   TM   N out       15   LG:429446.1:2000FEB18   629   682   forward 2   TM   N in       15   LG:429446.1:2000FEB18   627   713   forward 3   TM   N in       16   LI:057229.1:2000FEB01   10   69   forward 1   TM       16   LI:057229.1:2000FEB01   118   198   forward 1   TM       16   LI:057229.1:2000FEB01   292   360   forward 1   TM       16   LI:057229.1:2000FEB01   11   67   forward 2   TM       16   LI:057229.1:2000FEB01   146   226   forward 2   TM       16   LI:057229.1:2000FEB01   290   355   forward 2   TM       16   LI:057229.1:2000FEB01   12   71   forward 3   TM   N out       16   LI:057229.1:2000FEB01   114   176   forward 3   TM   N out       17   LI:351965.1:2000FEB01   487   573   forward 1   TM       17   LI:351965.1:2000FEB01   1036   1098   forward 1   TM       17   LI:351965.1:2000FEB01   492   578   forward 3   TM   N in       17   LI:351965.1:2000FEB01   969   1055   forward 3   TM   N in       17   LI:351965.1:2000FEB01   1098   1184   forward 3   TM   N in       18   LG:068682.1:2000FEB18   707   793   forward 2   TM   N out       19   LG:242665.1:2000FEB18   10   63   forward 1   TM   N out       19   LG:242665.1:2000FEB18   12   62   forward 3   TM   N out       19   LG:242665.1:2000FEB18   333   398   forward 3   TM   N out       20   LG:241743.1:2000FEB18   43   99   forward 1   TM   N out       21   LI:034212.1:2000FEB01   1300   1365   forward 1   TM   N in       21   LI:034212.1:2000FEB01   1570   1647   forward 1   TM   N in       21   LI:034212.1:2000FEB01   2386   2472   forward 1   TM   N in       21   LI:034212.1:2000FEB01   2533   2598   forward 1   TM   N in       21   LI:034212.1:2000FEB01   2620   2706   forward 1   TM   N in       21   LI:034212.1:2000FEB01   2740   2826   forward 1   TM   N in       21   LI:034212.1:2000FEB01   719   805   forward 2   TM       21   LI:034212.1:2000FEB01   1205   1291   forward 2   TM       21   LI:034212.1:2000FEB01   1460   1546   forward 2   TM       21   LI:034212.1:2000FEB01   1685   1768   forward 2   TM       21   LI:034212.1:2000FEB01   1814   1882   forward 2   TM       21   LI:034212.1:2000FEB01   2066   2128   forward 2   TM       21   LI:034212.1:2000FEB01   2156   2218   forward 2   TM       21   LI:034212.1:2000FEB01   2540   2626   forward 2   TM       21   LI:034212.1:2000FEB01   2657   2734   forward 2   TM       21   LI:034212.1:2000FEB01   12   62   forward 3   TM   N out       21   LI:034212.1:2000FEB01   1236   1301   forward 3   TM   N out       21   LI:034212.1:2000FEB01   1590   1646   forward 3   TM   N out       21   LI:034212.1:2000FEB01   1668   1721   forward 3   TM   N out       21   LI:034212.1:2000FEB01   2130   2216   forward 3   TM   N out       21   LI:034212.1:2000FEB01   2295   2381   forward 3   TM   N out       21   LI:034212.1:2000FEB01   2436   2513   forward 3   TM   N out       21   LI:034212.1:2000FEB01   2538   2624   forward 3   TM   N out       21   LI:034212.1:2000FEB01   2667   2735   forward 3   TM   N out       22   LG:344886.1:2000MAY19   937   1002   forward 1   TM   N in       22   LG:344886.1:2000MAY19   1081   1155   forward 1   TM   N in       22   LG:344886.1:2000MAY19   1696   1782   forward 1   TM   N in       22   LG:344886.1:2000MAY19   413   463   forward 2   TM   N in       22   LG:344886.1:2000MAY19   551   637   forward 2   TM   N in       22   LG:344886.1:2000MAY19   950   1012   forward 2   TM   N in       22   LG:344886.1:2000MAY19   1031   1093   forward 2   TM   N in       22   LG:344886.1:2000MAY19   1112   1183   forward 2   TM   N in       22   LG:344886.1:2000MAY19   1271   1348   forward 2   TM   N in       22   LG:344886.1:2000MAY19   1634   1720   forward 2   TM   N in       22   LG:344886.1:2000MAY19   567   626   forward 3   TM   N in       22   LG:344886.1:2000MAY19   1011   1073   forward 3   TM   N in       22   LG:344886.1:2000MAY19   1089   1151   forward 3   TM   N in       22   LG:344886.1:2000MAY19   1707   1757   forward 3   TM   N in       23   LG:228930.1:2000MAY19   111   167   forward 3   TM   N in       24   LG:338927.1:2000MAY19   934   1020   forward 1   TM   N out       24   LG:338927.1:2000MAY19   1133   1219   forward 2   TM   N in       24   LG:338927.1:2000MAY19   1170   1250   forward 3   TM   N in       25   LG:898771.1:2000MAY19   1261   1314   forward 1   TM   N out       25   LG:898771.1:2000MAY19   1397   1450   forward 2   TM   N out       26   LI:257664.67:2000MAY01   280   366   forward 1   TM   N in       26   LI:257664.67:2000MAY01   421   498   forward 1   TM   N in       26   LI:257664.67:2000MAY01   12   71   forward 3   TM   N out       27   LI:001496.2:2000MAY01   399   473   forward 3   TM       28   LI:1085273.2:2000MAY01   2188   2274   forward 1   TM   N in       28   LI:1085273.2:2000MAY01   503   583   forward 2   TM   N out       28   LI:1085273.2:2000MAY01   2126   2194   forward 2   TM   N out       28   LI:1085273.2:2000MAY01   897   968   forward 3   TM   N in       29   LI:333138.2:2000MAY01   1930   2016   forward 1   TM   N out       29   LI:333138.2:2000MAY01   50   103   forward 2   TM       29   LI:333138.2:2000MAY01   884   940   forward 2   TM       29   LI:333138.2:2000MAY01   114   179   forward 3   TM   N out       29   LI:333138.2:2000MAY01   273   356   forward 3   TM   N out       29   LI:333138.2:2000MAY01   819   875   forward 3   TM   N out       29   LI:333138.2:2000MAY01   1581   1667   forward 3   TM   N out       30   LI:338927.1:2000MAY01   1069   1140   forward 1   TM   N in       30   LI:338927.1:2000MAY01   968   1051   forward 2   TM   N in       30   LI:338927.1:2000MAY01   1056   1118   forward 3   TM   N out       30   LI:338927.1:2000MAY01   1155   1217   forward 3   TM   N out       31   LG:335558.1:2000FEB18   518   604   forward 2   TM   N in       31   LG:335558.1:2000FEB18   614   682   forward 2   TM   N in       31   LG:335558.1:2000FEB18   761   829   forward 2   TM   N in       31   LG:335558.1:2000FEB18   798   860   forward 3   TM   N in       31   LG:335558.1:2000FEB18   882   944   forward 3   TM   N in       31   LG:335558.1:2000FEB18   966   1028   forward 3   TM   N in       32   LG:998283.7:2000FEB18   1066   1146   forward 1   TM   N in       32   LG:998283.7:2000FEB18   23   109   forward 2   TM   N in       32   LG:998283.7:2000FEB18   194   280   forward 2   TM   N in       32   LG:998283.7:2000FEB18   392   478   forward 2   TM   N in       32   LG:998283.7:2000FEB18   527   613   forward 2   TM   N in       32   LG:998283.7:2000FEB18   776   862   forward 2   TM   N in       32   LG:998283.7:2000FEB18   1064   1141   forward 2   TM   N in       32   LG:998283.7:2000FEB18   12   65   forward 3   TM   N in       32   LG:998283.7:2000FEB18   147   227   forward 3   TM   N in       32   LG:998283.7:2000FEB18   684   770   forward 3   TM   N in       32   LG:998283.7:2000FEB18   1011   1097   forward 3   TM   N in       33   LI:402739.1:2000FEB01   415   501   forward 1   TM   N in       35   LG:981076.2:2000MAY19   388   450   forward 1   TM   N in       35   LG:981076.2:2000MAY19   20   82   forward 2   TM   N out       35   LG:981076.2:2000MAY19   389   451   forward 2   TM   N out       35   LG:981076.2:2000MAY19   464   526   forward 2   TM   N out       35   LG:981076.2:2000MAY19   539   604   forward 2   TM   N out       35   LG:981076.2:2000MAY19   438   524   forward 3   TM   N in       37   LI:1190250.1:2000MAY01   530   613   forward 2   TM       37   LI:1190250.1:2000MAY01   558   635   forward 3   TM   N out       38   LG:021371.3:2000FEB18   122   208   forward 2   TM   N in       41   LG:410726.1:2000FEB18   22   108   forward 1   TM   N in       41   LG:410726.1:2000FEB18   385   471   forward 1   TM   N in       42   LG:200005.1:2000FEB18   166   222   forward 1   TM   N out       42   LG:200005.1:2000FEB18   185   232   forward 2   TM   N out       42   LG:200005.1:2000FEB18   162   248   forward 3   TM   N out       46   LG:1079203.1:2000FEB18   11   70   forward 2   TM   N in       46   LG:1079203.1:2000FEB18   125   196   forward 2   TM   N in       46   LG:1079203.1:2000FEB18   965   1051   forward 2   TM   N in       47   LG:1082586.1:2000FEB18   256   339   forward 1   TM   N in       47   LG:1082586.1:2000FEB18   248   316   forward 2   TM   N out       49   LG:1082775.1:2000FEB18   553   606   forward 1   TM   N in       50   LG:1083120.1:2000FEB18   214   291   forward 1   TM   N out       50   LG:1083120.1:2000FEB18   233   319   forward 2   TM   N out       50   LG:1083120.1:2000FEB18   252   320   forward 3   TM   N in       51   LG:1087707.1:2000FEB18   367   453   forward 1   TM   N out       51   LG:1087707.1:2000FEB18   469   531   forward 1   TM   N out       51   LG:1087707.1:2000FEB18   667   729   forward 1   TM   N out       51   LG:1087707.1:2000FEB18   742   804   forward 1   TM   N out       51   LG:1087707.1:2000FEB18   407   481   forward 2   TM   N in       51   LG:1087707.1:2000FEB18   671   739   forward 2   TM   N in       51   LG:1087707.1:2000FEB18   743   811   forward 2   TM   N in       51   LG:1087707.1:2000FEB18   570   641   forward 3   TM   N out       51   LG:1087707.1:2000FEB18   747   833   forward 3   TM   N out       52   LG:1090915.1:2000FEB18   11   61   forward 2   TM   N out       53   LG:1094230.1:2000FEB18   469   555   forward 1   TM   N out       53   LG:1094230.1:2000FEB18   449   535   forward 2   TM   N out       54   LG:474848.3:2000FEB18   445   531   forward 1   TM   N out       54   LG:474848.3:2000FEB18   456   518   forward 3   TM   N out       58   LI:236654.2:2000FEB01   221   307   forward 2   TM   N out       59   LI:200009.1:2000FEB01   1045   1131   forward 1   TM   N out       59   LI:200009.1:2000FEB01   1171   1233   forward 1   TM   N out       59   LI:200009.1:2000FEB01   1076   1162   forward 2   TM   N in       59   LI:200009.1:2000FEB01   1044   1130   forward 3   TM   N in       60   LI:758502.1:2000FEB01   286   369   forward 1   TM   N out       60   LI:758502.1:2000FEB01   755   805   forward 2   TM   N in       60   LI:758502.1:2000FEB01   780   833   forward 3   TM   N in       62   LI:789445.1:2000FEB01   9   80   forward 3   TM   N out       63   LI:789657.1:2000FEB01   854   937   forward 2   TM   N in       64   LI:789808.1:2000FEB01   347   400   forward 2   TM   N in       65   LI:792919.1:2000FEB01   176   256   forward 2   TM       65   LI:792919.1:2000FEB01   371   427   forward 2   TM       66   LI:793949.1:2000FEB01   208   282   forward 1   TM   N out       66   LI:793949.1:2000FEB01   472   558   forward 1   TM   N out       66   LI:793949.1:2000FEB01   455   541   forward 2   TM   N out       67   LI:794389.1:2000FEB01   265   333   forward 1   TM   N out       67   LI:794389.1:2000FEB01   424   477   forward 1   TM   N out       67   LI:794389.1:2000FEB01   384   455   forward 3   TM   N in       68   LI:796010.1:2000FEB01   351   404   forward 3   TM   N in       69   LI:796324.1:2000FEB01   365   418   forward 2   TM   N in       72   LI:798636.1:2000FEB01   490   543   forward 1   TM   N in       73   LI:800045.1:2000FEB01   627   701   forward 3   TM   N in       74   LI:800680.1:2000FEB01   334   411   forward 1   TM   N out       74   LI:800680.1:2000FEB01   359   421   forward 2   TM   N out       75   LI:800894.1:2000FEB01   536   592   forward 2   TM   N in       75   LI:800894.1:2000FEB01   300   374   forward 3   TM   N out       75   LI:800894.1:2000FEB01   396   482   forward 3   TM   N out       77   LI:801236.1:2000FEB01   262   318   forward 1   TM   N out       78   LI:803335.1:2000FEB01   412   498   forward 1   TM   N out       78   LI:803335.1:2000FEB01   423   485   forward 3   TM   N out       79   LI:803998.1:2000FEB01   221   307   forward 2   TM   N out       81   LI:808532.1:2000FEB01   472   558   forward 1   TM   N in       81   LI:808532.1:2000FEB01   117   203   forward 3   TM   N in       81   LI:808532.1:2000FEB01   363   443   forward 3   TM   N in       81   LI:808532.1:2000FEB01   558   623   forward 3   TM   N in       82   LI:443073.1:2000FEB01   293   379   forward 2   TM   N in       82   LI:443073.1:2000FEB01   81   152   forward 3   TM   N in       82   LI:443073.1:2000FEB01   189   260   forward 3   TM   N in       83   LI:479671.1:2000FEB01   523   579   forward 1   TM   N out       85   LI:810224.1:2000FEB01   246   299   forward 3   TM       87   LG:892274.1:2000MAY19   49   105   forward 1   TM   N out       87   LG:892274.1:2000MAY19   613   681   forward 1   TM   N out       87   LG:892274.1:2000MAY19   506   589   forward 2   TM   N in       91   LG:1084051.1:2000MAY19   301   363   forward 1   TM   N in       92   LG:1076853.1:2000MAY19   964   1050   forward 1   TM   N in       92   LG:1076853.1:2000MAY19   56   130   forward 2   TM   N out       92   LG:1076853.1:2000MAY19   741   818   forward 3   TM   N in       93   LG:481631.10:2000MAY19   298   357   forward 1   TM   N out       93   LG:481631.10:2000MAY19   598   654   forward 1   TM   N out       94   LG:1088431.2:2000MAY19   379   441   forward 1   TM   N out       94   LG:1088431.2:2000MAY19   354   431   forward 3   TM   N out       95   LI:401619.10:2000MAY01   157   219   forward 1   TM   N out       95   LI:401619.10:2000MAY01   232   294   forward 1   TM   N out       95   LI:401619.10:2000MAY01   502   576   forward 1   TM   N out       95   LI:401619.10:2000MAY01   146   232   forward 2   TM   N in       95   LI:401619.10:2000MAY01   326   412   forward 2   TM   N in       95   LI:401619.10:2000MAY01   440   490   forward 2   TM   N in       95   LI:401619.10:2000MAY01   512   580   forward 2   TM   N in       95   LI:401619.10:2000MAY01   186   257   forward 3   TM   N in       95   LI:401619.10:2000MAY01   528   599   forward 3   TM   N in       96   LI:1144007.1:2000MAY01   2833   2910   forward 1   TM   N in       96   LI:1144007.1:2000MAY01   3301   3378   forward 1   TM   N in       96   LI:1144007.1:2000MAY01   3511   3597   forward 1   TM   N in       96   LI:1144007.1:2000MAY01   3634   3696   forward 1   TM   N in       96   LI:1144007.1:2000MAY01   3736   3801   forward 1   TM   N in       96   LI:1144007.1:2000MAY01   2645   2725   forward 2   TM   N out       96   LI:1144007.1:2000MAY01   2879   2965   forward 2   TM   N out       96   LI:1144007.1:2000MAY01   3356   3433   forward 2   TM   N out       96   LI:1144007.1:2000MAY01   3476   3523   forward 2   TM   N out       96   LI:1144007.1:2000MAY01   2772   2858   forward 3   TM   N in       96   LI:1144007.1:2000MAY01   3258   3332   forward 3   TM   N in       96   LI:1144007.1:2000MAY01   4017   4097   forward 3   TM   N in       97   LI:331074.1:2000MAY01   1264   1326   forward 1   TM   N in       97   LI:331074.1:2000MAY01   1357   1419   forward 1   TM   N in       97   LI:331074.1:2000MAY01   1450   1512   forward 1   TM   N in       97   LI:331074.1:2000MAY01   1540   1626   forward 1   TM   N in       97   LI:331074.1:2000MAY01   1433   1513   forward 2   TM   N in       97   LI:331074.1:2000MAY01   1574   1660   forward 2   TM   N in       97   LI:331074.1:2000MAY01   1461   1529   forward 3   TM   N in       97   LI:331074.1:2000MAY01   1560   1646   forward 3   TM   N in       98   LI:1170349.1:2000MAY01   34   102   forward 1   TM   N in       99   LG:335097.1:2000FEB18   601   672   forward 1   TM   N out       99   LG:335097.1:2000FEB18   847   909   forward 1   TM   N out       99   LG:335097.1:2000FEB18   928   981   forward 1   TM   N out       99   LG:335097.1:2000FEB18   164   244   forward 2   TM   N out       99   LG:335097.1:2000FEB18   623   682   forward 2   TM   N out       99   LG:335097.1:2000FEB18   12   74   forward 3   TM   N in       99   LG:335097.1:2000FEB18   219   299   forward 3   TM   N in       99   LG:335097.1:2000FEB18   594   680   forward 3   TM   N in       100   LG:1076451.1:2000FEB18   94   156   forward 1   TM   N in       100   LG:1076451.1:2000FEB18   101   187   forward 2   TM   N out       100   LG:1076451.1:2000FEB18   18   98   forward 3   TM   N out       100   LG:1076451.1:2000FEB18   96   164   forward 3   TM   N out       100   LG:1076451.1:2000FEB18   216   290   forward 3   TM   N out       101   LI:805478.1:2000FEB01   83   136   forward 2   TM   N out       101   LI:805478.1:2000FEB01   212   298   forward 2   TM   N out       102   LG:101269.1:2000MAY19   655   741   forward 1   TM   N in       102   LG:101269.1:2000MAY19   650   736   forward 2   TM   N in       102   LG:101269.1:2000MAY19   96   182   forward 3   TM   N in       102   LG:101269.1:2000MAY19   249   335   forward 3   TM   N in       102   LG:101269.1:2000MAY19   663   740   forward 3   TM   N in       103   LI:331087.1:2000MAY01   251   298   forward 2   TM   N out       103   LI:331087.1:2000MAY01   237   311   forward 3   TM       104   LI:410188.1:2000MAY01   520   591   forward 1   TM   N in       104   LI:410188.1:2000MAY01   640   711   forward 1   TM   N in       104   LI:410188.1:2000MAY01   724   810   forward 1   TM   N in       104   LI:410188.1:2000MAY01   832   879   forward 1   TM   N in       104   LI:410188.1:2000MAY01   883   969   forward 1   TM   N in       104   LI:410188.1:2000MAY01   1171   1257   forward 1   TM   N in       104   LI:410188.1:2000MAY01   1303   1389   forward 1   TM   N in       104   LI:410188.1:2000MAY01   2290   2361   forward 1   TM   N in       104   LI:410188.1:2000MAY01   2389   2460   forward 1   TM   N in       104   LI:410188.1:2000MAY01   2470   2556   forward 1   TM   N in       104   LI:410188.1:2000MAY01   2635   2721   forward 1   TM   N in       104   LI:410188.1:2000MAY01   2794   2862   forward 1   TM   N in       104   LI:410188.1:2000MAY01   2878   2964   forward 1   TM   N in       104   LI:410188.1:2000MAY01   3757   3837   forward 1   TM   N in       104   LI:410188.1:2000MAY01   3871   3957   forward 1   TM   N in       104   LI:410188.1:2000MAY01   3961   4047   forward 1   TM   N in       104   LI:410188.1:2000MAY01   4111   4194   forward 1   TM   N in       104   LI:410188.1:2000MAY01   4342   4428   forward 1   TM   N in       104   LI:410188.1:2000MAY01   4492   4578   forward 1   TM   N in       104   LI:410188.1:2000MAY01   4714   4794   forward 1   TM   N in       104   LI:410188.1:2000MAY01   6439   6519   forward 1   TM   N in       104   LI:410188.1:2000MAY01   7492   7575   forward 1   TM   N in       104   LI:410188.1:2000MAY01   7783   7845   forward 1   TM   N in       104   LI:410188.1:2000MAY01   4673   4735   forward 2   TM   N in       104   LI:410188.1:2000MAY01   4766   4828   forward 2   TM   N in       104   LI:410188.1:2000MAY01   4928   5014   forward 2   TM   N in       104   LI:410188.1:2000MAY01   5231   5317   forward 2   TM   N in       104   LI:410188.1:2000MAY01   6341   6409   forward 2   TM   N in       104   LI:410188.1:2000MAY01   7655   7741   forward 2   TM   N in       104   LI:410188.1:2000MAY01   8060   8146   forward 2   TM   N in       104   LI:410188.1:2000MAY01   4776   4859   forward 3   TM   N in       104   LI:410188.1:2000MAY01   6309   6371   forward 3   TM   N in       104   LI:410188.1:2000MAY01   7704   7775   forward 3   TM   N in       105   LI:1188288.1:2000MAY01   457   519   forward 1   TM       105   LI:1188288.1:2000MAY01   841   915   forward 1   TM       105   LI:1188288.1:2000MAY01   958   1038   forward 1   TM       105   LI:1188288.1:2000MAY01   1072   1140   forward 1   TM       105   LI:1188288.1:2000MAY01   1477   1539   forward 1   TM       105   LI:1188288.1:2000MAY01   1564   1626   forward 1   TM       105   LI:1188288.1:2000MAY01   1810   1896   forward 1   TM       105   LI:1188288.1:2000MAY01   2134   2220   forward 1   TM       105   LI:1188288.1:2000MAY01   2734   2820   forward 1   TM       105   LI:1188288.1:2000MAY01   1067   1147   forward 2   TM   N out       105   LI:1188288.1:2000MAY01   1157   1243   forward 2   TM   N out       105   LI:1188288.1:2000MAY01   1313   1399   forward 2   TM   N out       105   LI:1188288.1:2000MAY01   1556   1618   forward 2   TM   N out       105   LI:1188288.1:2000MAY01   2294   2368   forward 2   TM   N out       105   LI:1188288.1:2000MAY01   435   521   forward 3   TM   N in       105   LI:1188288.1:2000MAY01   597   683   forward 3   TM   N in       105   LI:1188288.1:2000MAY01   2301   2354   forward 3   TM   N in       105   LI:1188288.1:2000MAY01   2700   2753   forward 3   TM   N in       106   LI:427997.4:2000MAY01   148   222   forward 1   TM   N in       106   LI:427997.4:2000MAY01   745   828   forward 1   TM   N in       106   LI:427997.4:2000MAY01   1192   1278   forward 1   TM   N in       106   LI:427997.4:2000MAY01   1351   1434   forward 1   TM   N in       106   LI:427997.4:2000MAY01   1450   1518   forward 1   TM   N in       106   LI:427997.4:2000MAY01   1759   1845   forward 1   TM   N in       106   LI:427997.4:2000MAY01   134   220   forward 2   TM   N in       106   LI:427997.4:2000MAY01   749   832   forward 2   TM   N in       106   LI:427997.4:2000MAY01   1031   1087   forward 2   TM   N in       106   LI:427997.4:2000MAY01   1607   1693   forward 2   TM   N in       106   LI:427997.4:2000MAY01   1730   1816   forward 2   TM   N in       106   LI:427997.4:2000MAY01   2111   2191   forward 2   TM   N in       106   LI:427997.4:2000MAY01   150   236   forward 3   TM   N in       106   LI:427997.4:2000MAY01   681   767   forward 3   TM   N in       106   LI:427997.4:2000MAY01   765   851   forward 3   TM   N in       106   LI:427997.4:2000MAY01   1068   1124   forward 3   TM   N in       106   LI:427997.4:2000MAY01   1665   1751   forward 3   TM   N in       106   LI:427997.4:2000MAY01   1782   1856   forward 3   TM   N in       107   LG:451682.1:2000FEB18   93   155   forward 3   TM       109   LG:481436.5:2000FEB18   583   669   forward 1   TM   N in       109   LG:481436.5:2000FEB18   769   834   forward 1   TM   N in       109   LG:481436.5:2000FEB18   1111   1176   forward 1   TM   N in       109   LG:481436.5:2000FEB18   575   655   forward 2   TM   N out       109   LG:481436.5:2000FEB18   764   826   forward 2   TM   N out       109   LG:481436.5:2000FEB18   1091   1153   forward 2   TM   N out       109   LG:481436.5:2000FEB18   1187   1249   forward 2   TM   N out       109   LG:481436.5:2000FEB18   84   170   forward 3   TM   N in       109   LG:481436.5:2000FEB18   753   833   forward 3   TM   N in       109   LG:481436.5:2000FEB18   1164   1241   forward 3   TM   N in       110   LI:793701.1:2000FEB01   352   405   forward 1   TM   N in       110   LI:793701.1:2000FEB01   389   475   forward 2   TM   N in       111   LI:373637.1:2000FEB01   412   498   forward 1   TM       111   LI:373637.1:2000FEB01   434   520   forward 2   TM   N out       111   LI:373637.1:2000FEB01   866   919   forward 2   TM   N out       111   LI:373637.1:2000FEB01   423   473   forward 3   TM   N in       111   LI:373637.1:2000FEB01   867   920   forward 3   TM   N in       112   LG:239368.2:2000MAY19   241   327   forward 1   TM   N out       113   LI:053825.1:2000MAY01   31   117   forward 1   TM   N out       113   LI:053826.1:2000MAY01   1102   1188   forward 1   TM   N out       113   LI:053826.1:2000MAY01   1282   1350   forward 1   TM   N out       113   LI:053826.1:2000MAY01   41   112   forward 2   TM   N out       113   LI:053826.1:2000MAY01   164   238   forward 2   TM   N out       113   LI:053826.1:2000MAY01   461   538   forward 2   TM   N out       113   LI:053826.1:2000MAY01   1130   1192   forward 2   TM   N out       113   LI:053826.1:2000MAY01   1214   1276   forward 2   TM   N out       113   LI:053826.1:2000MAY01   1307   1378   forward 2   TM   N out       113   LI:053826.1:2000MAY01   126   200   forward 3   TM   N in       113   LI:053826.1:2000MAY01   348   416   forward 3   TM   N in       113   LI:053826.1:2000MAY01   624   683   forward 3   TM   N in       113   LI:053826.1:2000MAY01   1215   1277   forward 3   TM   N in       113   LI:053826.1:2000MAY01   1290   1352   forward 3   TM   N in       115   LI:1071427.96:2000MAY01   1072   1140   forward 1   TM       115   LI:1071427.96:2000MAY01   1297   1383   forward 1   TM       115   LI:1071427.96:2000MAY01   1459   1536   forward 1   TM       115   LI:1071427.96:2000MAY01   1765   1851   forward 1   TM       115   LI:1071427.96:2000MAY01   1909   1971   forward 1   TM       115   LI:1071427.96:2000MAY01   2002   2064   forward 1   TM       115   LI:1071427.96:2000MAY01   1562   1648   forward 2   TM   N out       115   LI:1071427.96:2000MAY01   1706   1792   forward 2   TM   N out       115   LI:1071427.96:2000MAY01   1823   1885   forward 2   TM   N out       115   LI:1071427.96:2000MAY01   1913   1975   forward 2   TM   N out       115   LI:1071427.96:2000MAY01   2045   2098   forward 2   TM   N out       115   LI:1071427.96:2000MAY01   384   470   forward 3   TM   N out       115   LI:1071427.96:2000MAY01   840   926   forward 3   TM   N out       115   LI:1071427.96:2000MAY01   987   1049   forward 3   TM   N out       115   LI:1071427.96:2000MAY01   1092   1154   forward 3   TM   N out       115   LI:1071427.96:2000MAY01   1383   1454   forward 3   TM   N out       115   LI:1071427.96:2000MAY01   1599   1655   forward 3   TM   N out       115   LI:1071427.96:2000MAY01   1767   1844   forward 3   TM   N out       115   LI:1071427.96:2000MAY01   1884   1952   forward 3   TM   N out       115   LI:1071427.96:2000MAY01   2013   2099   forward 3   TM   N out       115   LI:1071427.96:2000MAY01   2127   2189   forward 3   TM   N out       116   LI:336338.8:2000MAY01   100   186   forward 1   TM   N out       116   LI:336338.8:2000MAY01   427   513   forward 1   TM   N out       116   LI:336338.8:2000MAY01   110   196   forward 2   TM       116   LI:336338.8:2000MAY01   281   367   forward 2   TM       116   LI:336338.8:2000MAY01   422   508   forward 2   TM       116   LI:336338.8:2000MAY01   354   416   forward 3   TM   N out       116   LI:336338.8:2000MAY01   432   494   forward 3   TM   N out       117   LG:345527.1:2000FEB18   46   120   forward 1   TM   N out       117   LG:345527.1:2000FEB18   917   979   forward 2   TM   N out       117   LG:345527.1:2000FEB18   1010   1072   forward 2   TM   N out       117   LG:345527.1:2000FEB18   1112   1198   forward 2   TM   N out       117   LG:345527.1:2000FEB18   96   182   forward 3   TM   N out       117   LG:345527.1:2000FEB18   474   536   forward 3   TM   N out       117   LG:345527.1:2000FEB18   552   614   forward 3   TM   N out       118   LG:1089383.1:2000FEB18   43   126   forward 1   TM   N out       118   LG:1089383.1:2000FEB18   14   100   forward 2   TM       118   LG:1089383.1:2000FEB18   140   205   forward 2   TM       118   LG:1089383.1:2000FEB18   12   59   forward 3   TM   N out       120   LG:1093216.1:2000FEB18   31   117   forward 1   TM   N out       120   LG:1093216.1:2000FEB18   151   234   forward 1   TM   N out       120   LG:1093216.1:2000FEB18   283   348   forward 1   TM   N out       120   LG:1093216.1:2000FEB18   23   109   forward 2   TM   N in       120   LG:1093216.1:2000FEB18   143   193   forward 2   TM   N in       120   LG:1093216.1:2000FEB18   48   122   forward 3   TM   N out       120   LG:1093216.1:2000FEB18   180   263   forward 3   TM   N out       122   LI:335671.2:2000FEB01   22   108   forward 1   TM   N out       122   LI:335671.2:2000FEB01   1048   1134   forward 1   TM   N out       122   LI:335671.2:2000FEB01   854   916   forward 2   TM   N in       122   LI:335671.2:2000FEB01   926   988   forward 2   TM   N in       122   LI:335671.2:2000FEB01   998   1072   forward 2   TM   N in       122   LI:335671.2:2000FEB01   399   461   forward 3   TM   N out       122   LI:335671.2:2000FEB01   480   542   forward 3   TM   N out       122   LI:335671.2:2000FEB01   576   662   forward 3   TM   N out       122   LI:335671.2:2000FEB01   1023   1085   forward 3   TM   N out       122   LI:335671.2:2000FEB01   1098   1160   forward 3   TM   N out       122   LI:335671.2:2000FEB01   1173   1235   forward 3   TM   N out       123   LI:793758.1:2000FEB01   31   117   forward 1   TM   N out       123   LI:793758.1:2000FEB01   151   234   forward 1   TM   N out       123   LI:793758.1:2000FEB01   283   348   forward 1   TM   N out       123   LI:793758.1:2000FEB01   23   109   forward 2   TM   N in       123   LI:793758.1:2000FEB01   143   193   forward 2   TM   N in       123   LI:793758.1:2000FEB01   48   122   forward 3   TM   N out       123   LI:793758.1:2000FEB01   180   263   forward 3   TM   N out       124   LI:803718.1:2000FEB01   43   126   forward 1   TM   N out       124   LI:803718.1:2000FEB01   14   100   forward 2   TM       124   LI:803718.1:2000FEB01   140   205   forward 2   TM       124   LI:803718.1:2000FEB01   12   59   forward 3   TM   N out       125   LI:412179.1:2000FEB01   328   414   forward 1   TM       125   LI:412179.1:2000FEB01   436   504   forward 1   TM       125   LI:412179.1:2000FEB01   56   115   forward 2   TM   N out       125   LI:412179.1:2000FEB01   413   475   forward 2   TM   N out       125   LI:412179.1:2000FEB01   512   574   forward 2   TM   N out       125   LI:412179.1:2000FEB01   96   176   forward 3   TM   N out       125   LI:412179.1:2000FEB01   384   446   forward 3   TM   N out       125   LI:412179.1:2000FEB01   462   524   forward 3   TM   N out       126   LI:815679.1:2000FEB01   10   84   forward 1   TM   N out       126   LI:815679.1:2000FEB01   313   399   forward 1   TM   N out       126   LI:815679.1:2000FEB01   946   1032   forward 1   TM   N out       126   LI:815679.1:2000FEB01   1171   1248   forward 1   TM   N out       126   LI:815679.1:2000FEB01   323   409   forward 2   TM   N in       126   LI:815679.1:2000FEB01   500   568   forward 2   TM   N in       126   LI:815679.1:2000FEB01   971   1021   forward 2   TM   N in       126   LI:815679.1:2000FEB01   1493   1561   forward 2   TM   N in       126   LI:815679.1:2000FEB01   15   92   forward 3   TM   N in       126   LI:815679.1:2000FEB01   285   356   forward 3   TM   N in       126   LI:815679.1:2000FEB01   690   764   forward 3   TM   N in       126   LI:815679.1:2000FEB01   993   1076   forward 3   TM   N in       126   LI:815679.1:2000FEB01   1626   1712   forward 3   TM   N in       127   LI:481361.3:2000FEB01   199   252   forward 1   TM   N out       128   LG:247388.1:2000MAY19   190   240   forward 1   TM   N out       128   LG:247388.1:2000MAY19   233   319   forward 2   TM   N out       128   LG:247388.1:2000MAY19   446   532   forward 2   TM   N out       130   LI:787618.1:2000MAY01   10   84   forward 1   TM   N in       130   LI:787618.1:2000MAY01   313   399   forward 1   TM   N in       130   LI:787618.1:2000MAY01   679   750   forward 1   TM   N in       130   LI:787618.1:2000MAY01   1018   1098   forward 1   TM   N in       130   LI:787618.1:2000MAY01   1189   1266   forward 1   TM   N in       130   LI:787618.1:2000MAY01   323   409   forward 2   TM   N out       130   LI:787618.1:2000MAY01   500   568   forward 2   TM   N out       130   LI:787618.1:2000MAY01   944   1030   forward 2   TM   N out       130   LI:787618.1:2000MAY01   1508   1582   forward 2   TM   N out       130   LI:787618.1:2000MAY01   1616   1702   forward 2   TM   N out       130   LI:787618.1:2000MAY01   15   92   forward 3   TM   N out       130   LI:787618.1:2000MAY01   285   356   forward 3   TM   N out       131   LI:331610.2:2000MAY01   91   156   forward 1   TM       131   LI:331610.2:2000MAY01   277   363   forward 1   TM       131   LI:331610.2:2000MAY01   682   744   forward 1   TM       131   LI:331610.2:2000MAY01   4126   4212   forward 1   TM       131   LI:331610.2:2000MAY01   4951   5001   forward 1   TM       131   LI:331610.2:2000MAY01   5023   5109   forward 1   TM       131   LI:331610.2:2000MAY01   5128   5190   forward 1   TM       131   LI:331610.2:2000MAY01   5407   5469   forward 1   TM       131   LI:331610.2:2000MAY01   5485   5547   forward 1   TM       131   LI:331610.2:2000MAY01   5563   5625   forward 1   TM       131   LI:331610.2:2000MAY01   5728   5805   forward 1   TM       131   LI:331610.2:2000MAY01   5896   5949   forward 1   RTM       131   LI:331610.2:2000MAY01   6268   6327   forward 1   TM       131   LI:331610.2:2000MAY01   6454   6522   forward 1   TM       131   LI:331610.2:2000MAY01   6559   6645   forward 1   TM       131   LI:331610.2:2000MAY01   7477   7539   forward 1   TM       131   LI:331610.2:2000MAY01   7552   7614   forward 1   TM       131   LI:331610.2:2000MAY01   671   724   forward 2   TM   N out       131   LI:331610.2:2000MAY01   4127   4213   forward 2   TM   N out       131   LI:331610.2:2000MAY01   4928   5011   forward 2   TM   N out       131   LI:331610.2:2000MAY01   5051   5113   forward 2   TM   N out       131   LI:331610.2:2000MAY01   5135   5197   forward 2   TM   N out       131   LI:331610.2:2000MAY01   5207   5269   forward 2   TM   N out       131   LI:331610.2:2000MAY01   5537   5611   forward 2   TM   N out       131   LI:331610.2:2000MAY01   5726   5797   forward 2   TM   N out       131   LI:331610.2:2000MAY01   5903   5989   forward 2   TM   N out       131   LI:331610.2:2000MAY01   6392   6478   forward 2   TM   N out       131   LI:331610.2:2000MAY01   6746   6814   forward 2   TM   N out       131   LI:331610.2:2000MAY01   7295   7381   forward 2   TM   N out       131   LI:331610.2:2000MAY01   7586   7633   forward 2   TM   N out       131   LI:331610.2:2000MAY01   2763   2849   forward 3   TM       131   LI:331610.2:2000MAY01   4527   4595   forward 3   TM       131   LI:331610.2:2000MAY01   5079   5165   forward 3   TM       131   LI:331610.2:2000MAY01   5445   5516   forward 3   TM       131   LI:331610.2:2000MAY01   5676   5759   forward 3   TM       131   LI:331610.2:2000MAY01   6255   6341   forward 3   TM       131   LI:331610.2:2000MAY01   6378   6464   forward 3   TM       131   LI:331610.2:2000MAY01   6624   6692   forward 3   TM       131   LI:331610.2:2000MAY01   6705   6779   forward 3   TM       131   LI:331610.2:2000MAY01   6810   6884   forward 3   TM       131   LI:331610.2:2000MAY01   7062   7133   forward 3   TM       131   LI:331610.2:2000MAY01   7677   7748   forward 3   TM       131   LI:331610.2:2000MAY01   7833   7919   forward 3   TM       132   LG:982697.1:2000FEB18   355   441   forward 1   TM   N in       132   LG:982697.1:2000FEB18   946   993   forward 1   TM   N in       132   LG:982697.1:2000FEB18   897   983   forward 3   TM   N in       132   LG:982697.1:2000FEB18   1215   1301   forward 3   TM   N in       133   LG:1080896.1:2000FEB18   367   426   forward 1   TM   N in       133   LG:1080896.1:2000FEB18   476   562   forward 2   TM   N in       133   LG:1080896.1:2000FEB18   815   901   forward 2   TM   N in       133   LG:1080896.1:2000FEB18   342   395   forward 3   TM   N in       134   LI:811341.1:2000FEB01   562   615   forward 1   TM   N out       134   LI:811341.1:2000FEB01   691   777   forward 1   TM   N out       135   LI:903225.1:2000FEB01   20   100   forward 2   TM   N out       135   LI:903225.1:2000FEB01   12   83   forward 3   TM   N out       135   LI:903225.1:2000FEB01   768   827   forward 3   TM   N out       137   LG:979580.1:2000MAY19   298   354   forward 1   TM   N in       137   LG:979580.1:2000MAY19   826   909   forward 1   TM   N in       137   LG:979580.1:2000MAY19   934   1020   forward 1   TM   N in       137   LG:979580.1:2000MAY19   233   289   forward 2   TM   N out       137   LG:979580.1:2000MAY19   338   418   forward 2   TM   N out       137   LG:979580.1:2000MAY19   201   272   forward 3   TM   N in       138   LI:1169865.1:2000MAY01   197   283   forward 2   TM   N in       138   LI:1169865.1:2000MAY01   863   949   forward 2   TM   N in       139   LG:337818.2:2000FEB18   40   117   forward 1   TM   N out       139   LG:337818.2:2000FEB18   532   618   forward 1   TM   N out       139   LG:337818.2:2000FEB18   907   993   forward 1   TM   N out       139   LG:337818.2:2000FEB18   1372   1425   forward 1   TM   N out       140   LI:337818.1:2000FEB01   40   114   forward 1   TM   N in       140   LI:337818.1:2000FEB01   401   466   forward 2   TM   N in       140   LI:337818.1:2000FEB01   852   905   forward 3   TM   N in       141   LG:241577.4:2000MAY19   496   582   forward 1   TM   N in       142   LG:344786.4:2000MAY19   19   105   forward 1   TM   N out       142   LG:344786.4:2000MAY19   14   88   forward 2   TM   N in       142   LG:344786.4:2000MAY19   173   247   forward 2   TM   N in       142   LG:344786.4:2000MAY19   21   107   forward 3   TM       143   LI:414307.1:2000FEB01   116   202   forward 2   TM   N in       144   LI:202943.2:2000FEB01   166   237   forward 1   TM   N in       144   LI:202943.2:2000FEB01   263   313   forward 2   TM   N out       144   LI:202943.2:2000FEB01   276   326   forward 3   TM   N in       146   LI:815961.1:2000FEB01   232   291   forward 1   TM   N out       146   LI:815961.1:2000FEB01   81   167   forward 3   TM   N out       146   LI:815961.1:2000FEB01   243   329   forward 3   TM   N out       146   LI:815961.1:2000FEB01   354   422   forward 3   TM   N out       146   LI:815961.1:2000FEB01   573   659   forward 3   TM   N out       146   LI:815961.1:2000FEB01   741   803   forward 3   TM   N out       147   LG:120744.1:2000MAY19   181   249   forward 1   TM   N out       147   LG:120744.1:2000MAY19   188   256   forward 2   TM       147   LG:120744.1:2000MAY19   275   328   forward 2   TM       148   LI:757520.1:2000MAY01   2140   2220   forward 1   TM   N in       148   LI:757520.1:2000MAY01   2293   2379   forward 1   TM   N in       148   LI:757520.1:2000MAY01   1988   2059   forward 2   TM   N in       148   LI:757520.1:2000MAY01   2285   2359   forward 2   TM   N in       148   LI:757520.1:2000MAY01   1677   1763   forward 3   TM       148   LI:757520.1:2000MAY01   1995   2066   forward 3   TM       149   LG:160570.1:2000FEB18   345   413   forward 3   TM   N out       149   LG:160570.1:2000FEB18   462   518   forward 3   TM   N out       151   LI:221285.1:2000FEB01   1375   1452   forward 1   TM   N out       152   LI:401605.2:2000FEB01   235   321   forward 1   TM   N in       152   LI:401605.2:2000FEB01   192   263   forward 3   TM   N in       152   LI:401605.2:2000FEB01   489   563   forward 3   TM   N in       153   LI:329017.1:2000FEB01   179   235   forward 2   TM   N in       153   LI:329017.1:2000FEB01   359   433   forward 2   TM   N in       153   LI:329017.1:2000FEB01   449   526   forward 2   TM   N in       153   LI:329017.1:2000FEB01   617   703   forward 2   TM   N in       153   LI:329017.1:2000FEB01   920   973   forward 2   TM   N in       155   LG:403409.1:2000MAY19   136   222   forward 1   TM   N out       155   LG:403409.1:2000MAY19   973   1029   forward 1   TM   N out       155   LG:403409.1:2000MAY19   1285   1371   forward 1   TM   N out       155   LG:403409.1:2000MAY19   182   268   forward 2   TM   N in       156   LG:233933.5:2000MAY19   148   234   forward 1   TM   N out       156   LG:233933.5:2000MAY19   39   125   forward 3   TM   N out       157   LI:290344.1:2000MAY01   232   312   forward 1   TM   N out       157   LI:290344.1:2000MAY01   1258   1311   forward 1   TM   N out       157   LI:290344.1:2000MAY01   3640   3714   forward 1   TM   N out       157   LI:290344.1:2000MAY01   4366   4449   forward 1   TM   N out       157   LI:290344.1:2000MAY01   4468   4548   forward 1   TM   N out       157   LI:290344.1:2000MAY01   146   226   forward 2   TM   N out       157   LI:290344.1:2000MAY01   3122   3196   forward 2   TM   N out       157   LI:290344.1:2000MAY01   3833   3919   forward 2   TM   N out       157   LI:290344.1:2000MAY01   4457   4537   forward 2   TM   N out       157   LI:290344.1:2000MAY01   4760   4846   forward 2   TM   N out       157   LI:290344.1:2000MAY01   432   503   forward 3   TM   N out       157   LI:290344.1:2000MAY01   1647   1733   forward 3   TM   N out       157   LI:290344.1:2000MAY01   3177   3248   forward 3   TM   N out       157   LI:290344.1:2000MAY01   3594   3680   forward 3   TM   N out       157   LI:290344.1:2000MAY01   3753   3815   forward 3   TM   N out       157   LI:290344.1:2000MAY01   3864   3926   forward 3   TM   N out       157   LI:290344.1:2000MAY01   4443   4526   forward 3   TM   N out       158   LI:410742.1:2000MAY01   136   210   forward 1   TM   N out       158   LI:410742.1:2000MAY01   2200   2286   forward 1   TM   N out       158   LI:410742.1:2000MAY01   2437   2514   forward 1   TM   N out       158   LI:410742.1:2000MAY01   3149   3229   forward 2   TM   N in       158   LI:410742.1:2000MAY01   3437   3505   forward 2   TM   N in       158   LI:410742.1:2000MAY01   510   578   forward 3   TM   N in       158   LI:410742.1:2000MAY01   1905   1991   forward 3   TM   N in       158   LI:410742.1:2000MAY01   2811   2897   forward 3   TM   N in       158   LI:410742.1:2000MAY01   3168   3254   forward 3   TM   N in       159   LG:406568.1:2000MAY19   490   549   forward 1   TM   N in       159   LG:406568.1:2000MAY19   1732   1818   forward 1   TM   N in       159   LG:406568.1:2000MAY19   1825   1899   forward 1   TM   N in       159   LG:406568.1:2000MAY19   1918   2004   forward 1   TM   N in       159   LG:406568.1:2000MAY19   12   59   forward 3   TM   N in       159   LG:406568.1:2000MAY19   1935   2018   forward 3   TM   N in       159   LG:406568.1:2000MAY19   2094   2174   forward 3   TM   N in       160   LI:283762.1:2000MAY01   1675   1746   forward 1   TM       160   LI:283762.1:2000MAY01   2095   2181   forward 1   TM       160   LI:283762.1:2000MAY01   2632   2718   forward 1   TM       160   LI:283762.1:2000MAY01   2830   2916   forward 1   TM       160   LI:283762.1:2000MAY01   2941   3027   forward 1   TM       160   LI:283762.1:2000MAY01   3235   3321   forward 1   TM       160   LI:283762.1:2000MAY01   3328   3414   forward 1   TM       160   LI:283762.1:2000MAY01   3592   3666   forward 1   TM       160   LI:283762.1:2000MAY01   3682   3768   forward 1   TM       160   LI:283762.1:2000MAY01   4153   4224   forward 1   TM       160   LI:283762.1:2000MAY01   4360   4434   forward 1   TM       160   LI:283762.1:2000MAY01   4594   4656   forward 1   TM       160   LI:283762.1:2000MAY01   4681   4743   forward 1   TM       160   LI:283762.1:2000MAY01   4885   4962   forward 1   TM       160   LI:283762.1:2000MAY01   5011   5061   forward 1   TM       160   LI:283762.1:2000MAY01   92   178   forward 2   TM   N in       160   LI:283762.1:2000MAY01   278   364   forward 2   TM   N in       160   LI:283762.1:2000MAY01   995   1075   forward 2   TM   N in       160   LI:283762.1:2000MAY01   1523   1597   forward 2   TM   N in       160   LI:283762.1:2000MAY01   1817   1903   forward 2   TM   N in       160   LI:283762.1:2000MAY01   2522   2599   forward 2   TM   N in       160   LI:283762.1:2000MAY01   2666   2752   forward 2   TM   N in       160   LI:283762.1:2000MAY01   2837   2887   forward 2   TM   N in       160   LI:283762.1:2000MAY01   3038   3097   forward 2   TM   N in       160   LI:283762.1:2000MAY01   3563   3625   forward 2   TM   N in       160   LI:283762.1:2000MAY01   3638   3700   forward 2   TM   N in       160   LI:283762.1:2000MAY01   4067   4144   forward 2   TM   N in       160   LI:283762.1:2000MAY01   4439   4522   forward 2   TM   N in       160   LI:283762.1:2000MAY01   4685   4765   forward 2   TM   N in       160   LI:283762.1:2000MAY01   4784   4843   forward 2   TM   N in       160   LI:283762.1:2000MAY01   4973   5050   forward 2   TM   N in       160   LI:283762.1:2000MAY01   5072   5125   forward 2   TM   N in       160   LI:283762.1:2000MAY01   693   755   forward 3   TM   N out       160   LI:283762.1:2000MAY01   765   827   forward 3   TM   N out       160   LI:283762.1:2000MAY01   840   902   forward 3   TM   N out       160   LI:283762.1:2000MAY01   1623   1694   forward 3   TM   N out       160   LI:283762.1:2000MAY01   1800   1880   forward 3   TM   N out       160   LI:283762.1:2000MAY01   2622   2708   forward 3   TM   N out       160   LI:283762.1:2000MAY01   2778   2861   forward 3   TM   N out       160   LI:283762.1:2000MAY01   3144   3230   forward 3   TM   N out       160   LI:283762.1:2000MAY01   3276   3362   forward 3   TM   N out       160   LI:283762.1:2000MAY01   3441   3527   forward 3   TM   N out       160   LI:283762.1:2000MAY01   3666   3752   forward 3   TM   N out       160   LI:283762.1:2000MAY01   4077   4163   forward 3   TM   N out       160   LI:283762.1:2000MAY01   4245   4331   forward 3   TM   N out       160   LI:283762.1:2000MAY01   4395   4481   forward 3   TM   N out       160   LI:283762.1:2000MAY01   4584   4646   forward 3   TM   N out       160   LI:283762.1:2000MAY01   4662   4724   forward 3   TM   N out       160   LI:283762.1:2000MAY01   4845   4892   forward 3   TM   N out       161   LI:347687.113:2000MAY01   319   405   forward 1   TM   N out       161   LI:347687.113:2000MAY01   463   549   forward 1   TM   N out       161   LI:347687.113:2000MAY01   733   819   forward 1   TM   N out       161   LI:347687.113:2000MAY01   1240   1293   forward 1   TM   N out       161   LI:347687.113:2000MAY01   1720   1797   forward 1   TM   N out       161   LI:347687.113:2000MAY01   1861   1908   forward 1   TM   N out       161   LI:347687.113:2000MAY01   1972   2034   forward 1   TM   N out       161   LI:347687.113:2000MAY01   2050   2112   forward 1   TM   N out       161   LI:347687.113:2000MAY01   2308   2394   forward 1   TM   N out       161   LI:347687.113:2000MAY01   977   1057   forward 2   TM   N in       161   LI:347687.113:2000MAY01   1250   1309   forward 2   TM   N in       161   LI:347687.113:2000MAY01   1730   1792   forward 2   TM   N in       161   LI:347687.113:2000MAY01   1808   1870   forward 2   TM   N in       161   LI:347687.113:2000MAY01   1886   1948   forward 2   TM   N in       161   LI:347687.113:2000MAY01   324   398   forward 3   TM   N in       161   LI:347687.113:2000MAY01   948   1034   forward 3   TM   N in       161   LI:347687.113:2000MAY01   1686   1763   forward 3   TM   N in       161   LI:347687.113:2000MAY01   1791   1874   forward 3   TM   N in       161   LI:347687.113:2000MAY01   2025   2108   forward 3   TM   N in       163   LG:451710.1:2000FEB18   502   588   forward 1   TM   N in       163   LG:451710.1:2000FEB18   453   515   forward 3   TM   N in       164   LG:455771.1:2000FEB18   199   285   forward 1   TM   N out       165   LG:452089.1:2000FEB18   695   772   forward 2   TM   N out       165   LG:452089.1:2000FEB18   708   764   forward 3   TM   N out       166   LG:246415.1:2000FEB18   196   246   forward 1   TM   N in       167   LG:414144.10:2000FEB18   589   672   forward 1   TM   N in       167   LG:414144.10:2000FEB18   615   692   forward 3   TM   N out       168   LG:1101445.1:2000FEB18   787   858   forward 1   TM   N out       168   LG:1101445.1:2000FEB18   506   592   forward 2   TM   N out       169   LG:452134.1:2000FEB18   276   326   forward 3   TM   N out       170   LI:903021.1:2000FEB01   109   162   forward 1   TM   N out       172   LG:449404.1:2000MAY19   163   219   forward 1   TM   N out       172   LG:449404.1:2000MAY19   200   280   forward 2   TM   N out       173   LG:449413.1:2000MAY19   353   439   forward 2   TM   N out       177   LG:1101153.1:2000MAY19   520   600   forward 1   TM   N in       177   LG:1101153.1:2000MAY19   585   671   forward 3   TM   N in       178   LI:257695.20:2000MAY01   433   516   forward 1   TM   N in       179   LI:455771.1:2000MAY01   199   285   forward 1   TM   N out       180   LI:274551.1:2000MAY01   81   152   forward 3   TM   N out       180   LI:274551.1:2000MAY01   216   269   forward 3   TM   N out       181   LI:035973.1:2000MAY01   622   708   forward 1   TM   N out       181   LI:035973.1:2000MAY01   596   682   forward 2   TM   N out       181   LI:035973.1:2000MAY01   588   674   forward 3   TM   N out       182   LG:978427.5:2000FEB18   221   295   forward 2   TM   N out       182   LG:978427.5:2000FEB18   365   433   forward 2   TM   N out       182   LG:978427.5:2000FEB18   198   284   forward 3   TM   N out       183   LG:247781.2:2000FEB18   22   108   forward 1   TM   N in       183   LG:247781.2:2000FEB18   1114   1200   forward 1   TM   N in       183   LG:247781.2:2000FEB18   1149   1235   forward 3   TM   N in       185   LI:333307.2:2000FEB01   24   98   forward 3   TM   N out       187   LG:414732.1:2000MAY19   40   93   forward 1   TM   N out       187   LG:414732.1:2000MAY19   156   233   forward 3   TM   N out       188   LG:413910.6:2000MAY19   385   441   forward 1   TM   N out       188   LG:413910.6:2000MAY19   886   948   forward 1   TM   N out       188   LG:413910.6.2000MAY19   104   190   forward 2   TM   N out       188   LG:413910.6:2000MAY19   387   461   forward 3   TM   N out       188   LG:413910.6:2000MAY19   921   1007   forward 3   TM   N out       189   LI:414732.2:2000MAY01   34   93   forward 1   TM   N out       189   LI:414732.2:2000MAY01   24   110   forward 3   TM   N out       189   LI:414732.2:2000MAY01   159   236   forward 3   TM   N out       190   LI:900264.2:2000MAY01   730   807   forward 1   TM   N in       190   LI:900264.2:2000MAY01   1018   1092   forward 1   TM   N in       190   LI:900264.2:2000MAY01   1294   1350   forward 1   TM   N in       190   LI:900264.2:2000MAY01   1519   1578   forward 1   TM   N in       190   LI:900264.2:2000MAY01   2311   2397   forward 1   TM   N in       190   LI:900264.2:2000MAY01   2509   2562   forward 1   TM   N in       190   LI:900264.2:2000MAY01   2752   2808   forward 1   TM   N in       190   LI:900264.2:2000MAY01   3103   3165   forward 1   TM   N in       190   LI:900264.2:2000MAY01   3178   3240   forward 1   TM   N in       190   LI:900264.2:2000MAY01   3253   3315   forward 1   TM   N in       190   LI:900264.2:2000MAY01   3424   3510   forward 1   TM   N in       190   LI:900264.2:2000MAY01   3520   3603   forward 1   TM   N in       190   LI:900264.2:2000MAY01   3883   3945   forward 1   TM   N in       190   LI:900264.2:2000MAY01   3982   4044   forward 1   TM   N in       190   LI:900264.2:2000MAY01   68   154   forward 2   TM       190   LI:900264.2:2000MAY01   188   274   forward 2   TM       190   LI:900264.2:2000MAY01   1079   1165   forward 2   TM       190   LI:900264.2:2000MAY01   2285   2359   forward 2   TM       190   LI:900264.2:2000MAY01   2732   2812   forward 2   TM       190   LI:900264.2:2000MAY01   3095   3172   forward 2   TM       190   LI:900264.2:2000MAY01   3260   3319   forward 2   TM       190   LI:900264.2:2000MAY01   3434   3505   forward 2   TM       190   LI:900264.2:2000MAY01   3515   3601   forward 2   TM       190   LI:900264.2:2000MAY01   3662   3748   forward 2   TM       190   LI:900264.2:2000MAY01   3842   3913   forward 2   TM       190   LI:900264.2:2000MAY01   3992   4063   forward 2   TM       190   LI:900264.2:2000MAY01   198   248   forward 3   TM   N in       190   LI:900264.2:2000MAY01   1080   1133   forward 3   TM   N in       190   LI:900264.2:2000MAY01   1431   1517   forward 3   TM   N in       190   LI:900264.2:2000MAY01   1518   1571   forward 3   TM   N in       190   LI:900264.2:2000MAY01   1740   1814   forward 3   TM   N in       190   LI:900264.2:2000MAY01   2409   2480   forward 3   TM   N in       190   LI:900264.2:2000MAY01   2928   2993   forward 3   TM   N in       190   LI:900264.2:2000MAY01   3096   3161   forward 3   TM   N in       190   LI:900264.2:2000MAY01   3342   3404   forward 3   TM   N in       190   LI:900264.2:2000MAY01   3447   3509   forward 3   TM   N in       190   LI:900264.2:2000MAY01   3531   3614   forward 3   TM   N in       190   LI:900264.2:2000MAY01   3987   4064   forward 3   TM   N in       191   LI:335593.1:2000MAY01   685   771   forward 1   TM   N in       191   LI:335593.1:2000MAY01   1273   1335   forward 1   TM   N in       191   LI:335593.1:2000MAY01   1366   1428   forward 1   TM   N in       191   LI:335593.1:2000MAY01   710   757   forward 2   TM   N in       191   LI:335593.1:2000MAY01   1250   1336   forward 2   TM   N in       191   LI:335593.1:2000MAY01   1358   1408   forward 2   TM   N in       191   LI:335593.1:2000MAY01   1448   1525   forward 2   TM   N in       191   LI:335593.1:2000MAY01   1604   1690   forward 2   TM   N in       191   LI:335593.1:2000MAY01   81   128   forward 3   TM   N in       191   LI:335593.1:2000MAY01   246   296   forward 3   TM   N in       191   LI:335593.1:2000MAY01   807   866   forward 3   TM   N in       191   LI:335593.1:2000MAY01   876   947   forward 3   TM   N in       191   LI:335593.1:2000MAY01   1155   1217   forward 3   TM   N in       191   LI:335593.1:2000MAY01   1233   1295   forward 3   TM   N in       191   LI:335593.1:2000MAY01   1359   1445   forward 3   TM   N in       192   LI:1189543.1:2000MAY01   1765   1842   forward 1   TM       192   LI:1189543.1:2000MAY01   1861   1935   forward 1   TM       192   LI:1189543.1:2000MAY01   2236   2307   forward 1   TM       192   LI:1189543.1:2000MAY01   2356   2442   forward 1   TM       192   LI:1189543.1:2000MAY01   2476   2544   forward 1   TM       192   LI:1189543.1:2000MAY01   2659   2712   forward 1   TM       192   LI:1189543.1:2000MAY01   3097   3174   forward 1   TM       192   LI:1189543.1:2000MAY01   3217   3288   forward 1   TM       192   LI:1189543.1:2000MAY01   3439   3492   forward 1   TM       192   LI:1189543.1:2000MAY01   860   946   forward 2   TM       192   LI:1189543.1:2000MAY01   1016   1099   forward 2   TM       192   LI:1189543.1:2000MAY01   1145   1216   forward 2   TM       192   LI:1189543.1:2000MAY01   1601   1672   forward 2   TM       192   LI:1189543.1:2000MAY01   1691   1768   forward 2   TM       192   LI:1189543.1:2000MAY01   2411   2485   forward 2   TM       192   LI:1189543.1:2000MAY01   2831   2917   forward 2   TM       192   LI:1189543.1:2000MAY01   3080   3166   forward 2   TM       192   LI:1189543.1:2000MAY01   3227   3310   forward 2   TM       192   LI:1189543.1:2000MAY01   1155   1229   forward 3   TM   N out       192   LI:1189543.1:2000MAY01   1683   1766   forward 3   TM   N out       192   LI:1189543.1:2000MAY01   1770   1838   forward 3   TM   N out       192   LI:1189543.1:2000MAY01   2019   2069   forward 3   TM   N out       192   LI:1189543.1:2000MAY01   2352   2438   forward 3   TM   N out       192   LI:1189543.1:2000MAY01   2508   2594   forward 3   TM   N out       192   LI:1189543.1:2000MAY01   3030   3101   forward 3   TM   N out       192   LI:1189543.1:2000MAY01   3183   3263   forward 3   TM   N out       192   LI:1189543.1:2000MAY01   3360   3446   forward 3   TM   N out       193   LG:455450.1:2000FEB18   422   490   forward 2   TM   N out       194   LG:1040978.1:2000FEB18   500   586   forward 2   TM   N out       194   LG:1040978.1:2000FEB18   276   332   forward 3   TM   N out       196   LG:132147.3:2000FEB18   259   345   forward 1   TM   N out       196   LG:132147.3:2000FEB18   418   504   forward 1   TM   N out       196   LG:132147.3:2000FEB18   718   780   forward 1   TM   N out       196   LG:132147.3:2000FEB18   1477   1548   forward 1   TM   N out       196   LG:132147.3:2000FEB18   1585   1647   forward 1   TM   N out       196   LG:132147.3:2000FEB18   1690   1752   forward 1   TM   N out       196   LG:132147.3:2000FEB18   2560   2637   forward 1   TM   N out       196   LG:132147.3:2000FEB18   2731   2790   forward 1   TM   N out       196   LG:132147.3:2000FEB18   2908   2976   forward 1   TM   N out       196   LG:132147.3:2000FEB18   3082   3168   forward 1   TM   N out       196   LG:132147.3:2000FEB18   3184   3243   forward 1   TM   N out       196   LG:132147.3:2000FEB18   3376   3462   forward 1   TM   N out       196   LG:132147.3:2000FEB18   1451   1531   forward 2   TM   N out       196   LG:132147.3:2000FEB18   1538   1615   forward 2   TM   N out       196   LG:132147.3:2000FEB18   2741   2827   forward 2   TM   N out       196   LG:132147.3:2000FEB18   2960   3031   forward 2   TM   N out       196   LG:132147.3:2000FEB18   3050   3112   forward 2   TM   N out       196   LG:132147.3:2000FEB18   1626   1703   forward 3   TM   N in       196   LG:132147.3:2000FEB18   2508   2594   forward 3   TM   N in       196   LG:132147.3:2000FEB18   2919   2987   forward 3   TM   N in       196   LG:132147.3:2000FEB18   3177   3263   forward 3   TM   N in       196   LG:132147.3:2000FEB18   3372   3422   forward 3   TM   N in       197   LI:036034.1:2000FEB01   157   219   forward 1   TM   N out       197   LI:036034.1:2000FEB01   395   457   forward 2   TM   N in       197   LI:036034.1:2000FEB01   479   541   forward 2   TM   N in       197   LI:036034.1:2000FEB01   563   625   forward 2   TM   N in       197   LI:036034.1:2000FEB01   647   709   forward 2   TM   N in       198   LG:162161.1:2000MAY19   372   458   forward 3   TM   N in       199   LG:407214.10:2000MAY19   34   120   forward 1   TM   N out       199   LG:407214.10:2000MAY19   44   124   forward 2   TM   N out       199   LG:407214.10:2000MAY19   203   289   forward 2   TM   N out       200   LG:204626.1:2000MAY19   19   99   forward 1   TM   N out       202   LI:476342.1:2000MAY01   39   122   forward 3   TM   N out       203   LI:1072759.1:2000MAY01   409   495   forward 1   TM   N in       203   LI:1072759.1:2000MAY01   889   951   forward 1   TM   N in       203   LI:1072759.1:2000MAY01   1387   1458   forward 1   TM   N in       203   LI:1072759.1:2000MAY01   1687   1770   forward 1   TM   N in       203   LI:1072759.1:2000MAY01   392   478   forward 2   TM   N out       203   LI:1072759.1:2000MAY01   1055   1132   forward 2   TM   N out       203   LI:1072759.1:2000MAY01   1424   1507   forward 2   TM   N out       203   LI:1072759.1:2000MAY01   1694   1768   forward 2   TM   N out       203   LI:1072759.1:2000MAY01   1191   1277   forward 3   TM   N out       203   LI:1072759.1:2000MAY01   1677   1760   forward 3   TM   N out       204   LG:998857.1:2000FEB18   1195   1281   forward 1   TM   N in       204   LG:998857.1:2000FEB18   164   226   forward 2   TM   N out       204   LG:998857.1:2000FEB18   344   400   forward 2   TM   N out       204   LG:998857.1:2000FEB18   398   460   forward 2   TM   N out       204   LG:998857.1:2000FEB18   1478   1561   forward 2   TM   N out       205   LG:482261.1:2000FEB18   19   93   forward 1   TM   N out       205   LG:482261.1:2000FEB18   890   961   forward 2   TM   N out       205   LG:482261.1:2000FEB18   1070   1123   forward 2   TM   N out       205   LG:482261.1:2000FEB18   21   89   forward 3   TM   N out       205   LG:482261.1:2000FEB18   1242   1292   forward 3   TM   N out       206   LG:480328.1:2000FEB18   436   522   forward 1   TM   N out       206   LG:480328.1:2000FEB18   568   642   forward 1   TM   N out       206   LG:480328.1:2000FEB18   769   849   forward 1   TM   N out       206   LG:480328.1:2000FEB18   967   1029   forward 1   TM   N out       206   LG:480328.1:2000FEB18   56   130   forward 2   TM   N in       206   LG:480328.1:2000FEB18   194   280   forward 2   TM   N in       206   LG:480328.1:2000FEB18   396   482   forward 3   TM   N out       206   LG:480328.1:2000FEB18   747   818   forward 3   TM   N out       207   LG:311197.1:2000MAY19   241   315   forward 1   TM   N in       207   LG:311197.1:2000MAY19   527   613   forward 2   TM   N out       208   LG:1054883.1:2000MAY19   76   129   forward 1   TM   N out       208   LG:1054883.1:2000MAY19   83   145   forward 2   TM   N out       209   LG:399395.1:2000MAY19   163   216   forward 1   TM   N out       211   LI:272913.22:2000MAY01   37   123   forward 1   TM   N in                    
     [0900]                           TABLE 4                       SEQ ID NO:   Component ID   Start   Stop                                                1   g1260446   2   316       1   6791379H1   1   397       1   g1614819   215   655       1   g1647514   244   543       2   5911492F8   1   467       2   5911492H1   1   271       2   5911492T8   303   633       3   5311056H1   591   753       3   6866213H1   784   1388       3   5659105H1   1261   1340       3   5498383R6   1291   1674       3   g5553287   1   315       3   6989857H1   1   436       3   6955370H1   22   540       3   g4534562   24   504       3   g4390046   24   500       3   g1192915   25   170       3   g2003054   31   344       3   6818987J1   33   250       3   6818431J1   33   570       3   g2003419   45   421       3   g1551472   61   213       3   6147606H1   71   625       3   6990907H1   383   885       3   6866026H1   381   973       3   7067123H1   525   1069       3   5498383F6   573   1055       3   5498383H1   573   811       4   1709025H1   198   445       4   70319971D1   397   784       4   70320592D1   412   733       4   70320769D1   462   809       4   70317687D1   464   962       4   6483492H1   184   713       4   5006925H1   88   362       4   70513533D1   496   1069       4   70317606D1   522   961       4   1296898H1   262   495       4   6338333H1   640   1159       4   g4332126   1   454       4   2659966F6   1   192       4   2659966T6   1   194       4   70513545V1   31   585       4   70514030D1   31   577       4   70514030V1   31   589       4   70513041V1   31   366       4   2659966H1   1   247       4   70512591V1   31   171       4   70512591D1   31   169       4   70318172D1   57   447       4   70513426V1   61   680       4   70320498D1   247   638       4   034207H1   1   290       4   70515532V1   24   612       4   70317785D1   295   738       4   70516168V1   24   391       4   70321122D1   370   783       4   70514482D1   24   278       4   70514482V1   24   278       4   70318245D1   25   395       4   70318040D1   29   409       4   70514959V1   1   300       5   g2958900   175   294       5   167606H1   1   173       5   60123139D2   1   307       5   60123152D1   1   368       5   60123139D1   11   156       5   6127857H1   1   475       5   g706800   1   340       5   g690740   11   385       5   4454268H1   377   645       5   60123152B1   714   1379       5   1602161H1   603   798       5   6217784H1   857   1351       5   6217935H1   859   1356       5   1473544R6   214   653       5   1473544T1   249   653       5   1473544H1   408   653       5   60123139B1   948   1362       5   60124858B2   982   1362       5   g4457589   222   601       5   1473544T6   247   569       5   g715576   191   567       5   g993694   255   560       5   2866203H1   459   557       5   g682774   196   472       5   g3213293   1   454       5   g3148379   141   356       6   4835189H1   679   932       6   g1920265   798   1061       6   4541025H1   804   1057       6   2330975H1   952   1172       6   2330975R6   952   1311       6   2430651T6   1141   1667       6   2571536T6   1144   1680       6   3878850T6   1244   1662       6   g2930491   1302   1714       6   g3844521   1455   1711       6   6623593H1   104   689       6   6623593J1   634   1236       6   4724029H1   1   133       6   5577111H1   14   200       6   g2183982   84   445       6   3878850F6   119   524       6   3878850H1   120   399       6   g2783388   239   721       6   g2785249   239   710       6   g1920471   284   439       6   g1784782   388   828       6   6551102H1   429   951       6   6550902H1   429   1042       6   6551002H1   429   957       6   g3645387   447   909       6   2430651H1   448   703       6   2430651R6   448   680       6   3712318H1   471   759       6   g981405   485   787       6   g1975879   1   219       6   g1784993   585   909       6   1467511T7   1   438       6   6326238H1   8   306       6   526662H1   25   208       6   2856544H1   34   146       7   1980062R6   2   491       7   1980059H1   1   280       7   5573392H1   30   283       8   2642346H1   1   239       8   2662640H1   2   95       8   2523793H1   11   212       8   3155877H1   12   278       8   3539049H1   13   237       8   3179390H1   13   320       8   5543678H1   15   210       8   6834974H1   15   623       8   257055R6   16   423       8   133343H1   16   157       8   484198H1   16   163       8   257055H1   17   224       8   2346879H1   18   247       8   2346879F6   18   442       8   g1985882   19   366       8   3156918H1   21   155       8   4934879H1   22   297       8   g1751206   41   388       8   4024568H1   42   189       8   258140H1   58   433       8   2346879T6   167   442       8   2313878H1   229   495       8   6081177H1   264   720       8   3773491H1   306   476       8   6413047H1   401   935       8   4212712H1   407   675       8   6918705H1   413   837       8   3014942H1   416   684       8   1807684F6   483   905       8   1803962F6   483   841       8   1803962H1   483   760       8   1807684H1   483   746       8   3876768H1   529   819       8   6024789H1   553   695       8   4062033H1   555   779       8   4634711H1   615   877       8   3014303H1   667   964       8   3014303F6   667   1096       8   1633402H1   672   885       8   5865873H1   725   997       8   6552378H1   881   1430       8   2892794H1   907   1089       8   2892794F6   907   1389       8   6406936H1   935   1476       8   4902732H1   962   1225       8   1492959H1   970   1171       8   g1686447   974   1299       8   5929258H1   974   1266       8   3509940H1   974   1230       8   4588644H1   983   1230       8   6979634H1   991   1412       8   946914H1   1010   1310       8   g5854943   1083   1460       8   5544986H1   1095   1303       8   3860122H1   1133   1400       8   g4260489   1228   1701       8   1724521H1   1385   1493       8   1724521F6   1385   1714       8   3026692H1   1429   1687       8   3881903H1   1460   1700       8   3014303T6   1504   1714       8   490059H1   1515   1763       8   1803962T6   1575   1714       8   692274H1   1583   1765       8   2313878T6   1587   1714       8   g2436491   1623   1714       8   g3277091   1654   1714       8   3319247H1   1718   1969       8   4917483H1   1916   2145       8   6826179J1   1971   2309       8   6826179H1   1971   2309       9   3088820H1   537   807       9   2012535H1   561   787       9   3232447H1   618   863       9   2536817H2   670   926       9   g2209764   832   1178       9   7260050H1   1   537       9   2457623F6   101   601       9   6987326H1   110   654       9   7032756H1   151   720       9   6457158H1   350   878       9   2890632F6   351   531       9   2893095F6   351   909       9   2893095H1   351   614       9   7166124H1   401   930       9   4760777H1   427   714       9   3160745H1   437   710       9   g2006716   479   776       9   g2209659   481   940       9   3362481H1   514   763       9   1751755H1   520   735       9   6141843H1   536   793       10   6584158H1   14   592       10   3401851H1   4   241       10   3403550H1   1   259       10   3404251H1   3   258       10   g1137215   2430   2639       10   3378612H1   2451   2638       10   6584049H1   6   562       10   3402034H1   10   229       10   g307503   16   2638       10   70779637V1   2077   2632       10   7258175H1   2121   2603       10   3961980H1   2130   2252       10   6584189T1   2152   2553       10   6584158T1   2159   2659       10   g3785069   2394   2641       10   981592T6   2170   2701       10   3385383T6   2171   2601       10   1221889T6   2202   2700       10   70779908V1   2348   2654       10   70776665V1   1884   2562       10   70775642V1   1981   2519       10   70776673V1   2022   2640       10   6584049T1   2035   2569       10   3401765H1   2050   2287       10   3401379H1   15   260       10   3404170H1   18   267       11   6905943H1   1   534       12   70562343V1   1   628       12   70560334V1   1   538       12   70559056V1   2   275       12   70560532V1   12   269       12   70559641V1   12   383       12   70559213V1   12   511       12   70561550V1   12   575       12   70561738V1   12   582       12   70559422V1   13   631       12   70559480V1   12   687       12   758040H1   12   272       12   70559415V1   13   605       12   758040R6   12   310       12   70561876V1   13   585       12   70559130V1   35   384       12   g2576304   119   2175       12   70561182V1   150   812       12   70560479V1   217   834       12   70557043V1   221   595       12   70562198V1   227   819       12   70562312V1   245   894       12   70561005V1   249   895       12   70559925V1   254   840       12   70556220V1   274   841       12   70561793V1   282   939       12   70559002V1   293   888       12   6127888H1   351   824       12   70557039V1   359   611       12   70559636V1   391   1033       12   70561656V1   409   852       12   70560742V1   507   990       12   70561641V1   594   1066       12   g873095   657   1032       12   70449484V1   674   894       12   6848717H1   732   1253       12   70558944V1   1495   2173       12   70450051V1   1550   1731       12   70561063V1   1607   2025       12   g5361507   1719   2172       12   g4735148   1778   2172       12   g4875543   1779   2195       12   758040T6   1784   2147       12   g872996   1824   2188       12   g3840560   1830   2175       12   g4333943   2114   2199       13   2734453F6   1   435       13   2734453H1   1   252       14   4860612T7   1184   1574       14   g902319   1020   1492       14   g778351   1137   1386       14   g1638619   1140   1268       14   6929341H1   724   1222       14   1675892T6   754   1203       14   1720010F6   696   1079       14   1720010H1   696   922       14   4711951H1   675   833       14   5496406H1   546   795       14   6420632H1   79   637       14   2195346F6   1   450       14   2195346H1   344   450       14   3933140F6   11   351       14   3933140H1   96   351       14   6020638H1   1   263       14   g5880310   52   255       14   g2156573   11   254       14   g2264198   30   253       14   6936789H1   11   173       14   5043956H1   11   159       15   1897573H1   1   259       15   4337619T6   137   645       15   5436383H1   598   849       16   1671029F6   1   443       16   1671029H1   72   144       16   1671029T6   388   443       16   g2238932   388   443       16   g2243675   388   443       17   g2219785   236   531       17   5099781H1   378   604       17   3254347R6   527   1132       17   5668261H1   661   889       17   4289028H1   781   1061       17   g3931955   822   1267       17   g3401348   823   1271       17   g2825326   825   1144       17   2750359R6   824   1242       17   2750359T6   847   1241       17   2351445H1   151   367       17   2750359H1   968   1241       17   2203345H1   1018   1169       17   3254347H1   979   1089       17   4822216H1   1202   1393       17   3184753F6   1   472       17   5108214T6   60   258       17   3184753T6   60   312       17   g4287070   152   617       17   g2524411   160   493       17   3184753H1   228   472       17   g2788109   236   513       18   6829315H1   314   884       18   g3109791   492   811       18   g5452473   492   650       18   g4564783   26   423       18   g5438746   1   423       18   g3805312   34   423       18   g4372490   78   423       18   g2954208   76   423       18   g2954218   142   422       18   g3307490   142   344       18   2011384H1   190   263       19   3106785H1   747   1023       19   g565430   783   1150       19   g1505888   812   1132       19   g892901   859   1159       19   g892900   1   179       19   3436153H1   1   269       19   g4737696   1   452       19   839090H1   73   301       19   6938671H1   206   685       19   6939071H1   207   731       19   g646973   427   675       19   g1277657   484   934       19   2755039H1   596   767       19   838332T6   615   1116       19   4800144H1   651   916       19   2084151H1   664   945       20   6199407H1   1   558       20   2519336F6   37   448       20   2519336H1   37   198       20   6793417H1   61   331       20   1639314H1   71   248       20   6818283H1   75   652       20   1984661R6   94   490       20   1984662H1   94   368       20   1984661T6   127   543       20   4021069H1   177   388       20   3456613H1   177   427       20   4923878H1   211   494       20   6847871H1   274   451       20   g5151844   373   585       21   g4281322   2017   2463       21   3934096H1   2040   2211       21   3934274H1   2042   2187       21   1648485T6   2045   2159       21   1648242H1   2052   2198       21   1648485H1   2052   2198       21   1648485F6   2052   2198       21   4676772H1   2056   2317       21   1004843T6   2092   2693       21   g4533314   2101   2207       21   3416826H1   2134   2365       21   5304471H1   2184   2431       21   1310089F6   2250   2718       21   1310089H1   2250   2448       21   2461918H1   2266   2461       21   g4330296   2658   2953       21   g3178479   2713   2950       21   g856243   2748   3020       21   3951063F6   1   417       21   3951063H1   1   285       21   g2114724   215   609       21   1804812F6   311   871       21   1804812H1   311   582       21   1804772H1   311   530       21   2662164H1   346   588       21   2011927H1   365   608       21   4863613H1   380   645       21   3934325H1   386   672       21   3763239H1   402   671       21   1004843R6   502   864       21   1004843H1   502   706       21   g846375   547   889       21   5810205H1   585   885       21   384851H1   652   926       21   g1861137   688   1180       21   2670050H1   722   974       21   4118973H1   762   1013       21   2744119H1   784   968       21   g1081212   787   1107       21   5576610H1   879   1134       21   6418368H1   926   1226       21   5864111H1   950   1233       21   5033538H1   997   1264       21   3511167H1   1010   1288       21   5805062H1   1021   1225       21   4873652H1   1038   1293       21   6614566H1   1064   1565       21   2645969F6   1085   1588       21   2645969H1   1085   1330       21   g2112831   1112   1336       21   3742279H1   1144   1422       21   6334985H1   1249   1754       21   5292787H2   1287   1357       21   g853299   1363   1679       21   5742830H1   1399   1713       21   777713H1   1400   1625       21   777713R6   1401   1702       21   2291319H1   1424   1651       21   3241291H1   1436   1673       21   1832630H1   1481   1750       21   1804812T6   1571   2172       21   1481768H1   1572   1793       21   3730896H1   1583   1874       21   4442950H1   1631   1773       21   2794894H1   1648   1896       21   4161422H1   1664   1913       21   4163675H1   1665   1779       21   g2787436   1672   1875       21   1898917T6   1693   2159       21   5026603H1   1695   1945       21   3726809H1   1753   2047       21   1891483T6   1803   2160       21   2293309H1   1836   2077       21   1214225H1   1850   2020       21   g856337   1855   2128       21   g3733909   1913   2198       21   g3890798   1917   2386       21   5696574H1   1925   2048       21   g1081162   1942   2201       21   2645969T6   1964   2222       21   g1860963   1970   2387       21   777713T6   1983   2420       21   g4149122   1984   2203       21   4875545H1   1994   2292       22   487652H1   1   242       22   487652R6   1   232       22   g2003010   47   328       22   6442754H1   49   581       22   6152037H1   50   234       22   7072795H1   75   624       22   3126080H1   75   234       22   491193H1   83   234       22   4751976H1   90   232       22   7100707H1   100   609       22   2292643H1   102   232       22   6140970H1   105   232       22   4066069H1   110   234       22   5007390H1   113   231       22   4773161H1   130   415       22   6993149H1   145   680       22   572575H1   150   232       22   5756677H1   180   483       22   6064882H1   346   644       22   5372689H1   367   514       22   3123257H1   367   493       22   g2022792   381   648       22   5068149H1   503   782       22   5894157H1   560   759       22   5898812H1   560   853       22   2621801H1   561   824       22   6401738H1   574   844       22   2122453H1   622   902       22   2122453F6   622   764       22   5681165H1   629   889       22   5277977H1   637   892       22   4645753H1   659   921       22   6193452H1   705   1000       22   6193441H1   705   963       22   746066H1   713   961       22   4771093H1   754   1022       22   g2054100   771   1123       22   g4762720   796   1252       22   g2321843   801   1217       22   g3017276   804   1256       22   g483457   850   1222       22   g4762705   865   1252       22   4338534H1   870   1155       22   628211H1   887   1051       22   5563275H1   922   1144       22   5286207H1   952   1210       22   6160626H1   953   1244       22   6838871H1   1013   1491       22   2400686H1   1068   1153       22   g866503   1107   1440       22   6747627H1   1122   1593       22   g920433   1143   1481       22   2294209R6   1179   1688       22   2294209H1   1179   1444       22   g1471741   1213   1600       22   g1332314   1213   1712       22   6021757H1   1214   1822       22   2189963H1   1229   1505       22   g5766946   1310   1760       22   g2524669   1309   1776       22   g2178309   1317   1448       22   g4762067   1326   1779       22   g5361693   1328   1778       22   g4074013   1332   1773       22   g5849550   1331   1762       22   6736883H1   1355   1730       22   g2183535   1356   1766       22   g4332913   1357   1772       22   g3147585   1357   1758       22   g2053942   1361   1759       22   g1332315   1371   1839       22   3918780H1   1373   1665       22   g3884754   1379   1765       22   5207656H1   1382   1616       22   2292948H1   1388   1552       22   1373715H1   1388   1603       22   4742391H1   1388   1647       22   2749392H1   1400   1627       22   g1241849   1408   1559       22   g6046888   1408   1785       22   2294209T6   1428   1945       22   g1471742   1441   1766       22   g4332659   1471   1772       22   g3842415   1473   1785       22   g2841585   1474   1756       22   g2003011   1479   1772       22   2501320H1   1513   1738       22   g3961438   1525   1988       22   g2022791   1559   1757       22   g6132921   1559   1977       22   5690247H1   1568   1843       22   g1496515   1580   1977       22   g2252260   1620   1984       22   g3739177   1650   1831       22   5338636H1   1671   1915       22   5710854H2   1686   2004       22   3223930H1   1686   1772       22   4255692H1   1701   1756       22   g1210253   1725   1980       22   g920434   1730   1946       22   g1210208   1763   1980       22   4320678H1   1841   1973       22   g884068   1924   2233       23   6991066H1   1   190       23   4753733H1   121   279       23   4753733F8   122   613       23   7065882H1   183   679       23   g5676063   251   722       23   1576986F6   270   661       23   1576986H1   270   349       23   1576986T6   275   683       23   g3843718   364   724       23   g3750491   408   722       23   3604635H1   409   584       23   g793096   600   881       24   6756043H1   101   504       24   3845553H1   368   574       24   5057288F9   382   1021       24   5057288H1   382   660       24   5065461F8   404   839       24   2359950H1   625   748       24   6112349H1   792   1081       24   5444375H1   912   1131       24   g4264503   989   1390       24   3385663T6   1011   1248       24   3845553T6   1009   1248       24   3385663F6   1   315       24   3385663H1   1   257       24   5468404H1   3   271       24   5468404F8   3   587       24   5072501H1   11   292       24   6917202H1   10   259       24   4148204T6   1176   1391       24   4148204H1   1183   1435       24   4148204F6   1183   1390       24   g4532148   1256   1361       24   213841R6   1325   1390       24   213841T6   1325   1393       24   213841H1   1325   1390       24   213833H1   1325   1390       25   7204172H1   1   536       25   6819816H1   368   566       25   6836688H1   383   960       25   6349781H2   434   616       25   7280863H1   477   880       25   g4081510   513   860       25   6862186H1   519   806       25   g5543132   541   939       25   g4196555   541   821       25   g5636120   541   982       25   g3246528   541   923       25   g5636142   541   1018       25   g2568732   577   1024       25   3747228H1   589   887       25   6600395H1   587   999       25   6978931H1   605   954       25   2240520H1   629   730       25   2240520F6   641   967       25   5626829H1   663   931       25   g2162199   663   863       25   6862047H1   699   978       25   7281807H1   809   1041       25   g2209650   815   1308       25   6216076H1   841   1322       25   7238641H1   862   978       25   6210267H1   868   970       25   7039186H1   940   1508       25   g2209760   1092   1569       26   7281748H1   1   439       26   7281705H1   1   574       27   925628R6   1574   1961       27   6466550H1   228   746       27   035057H1   483   638       27   2173285F6   571   968       27   2173285H1   571   762       27   1743077R6   577   1069       27   1743077H1   577   892       27   3344276H1   598   856       27   6579246H1   697   1025       27   5637939F8   825   1101       27   5637939H1   825   1082       27   5624530R8   836   1190       27   5637939R8   1016   1380       27   3483518H1   1163   1470       27   6817649J1   1402   1969       27   2966432H1   1421   1627       27   g3419012   1500   1961       27   g2178116   1505   1630       27   g2715860   1512   1962       27   3891858T6   1559   1901       27   3891858F6   1566   1961       27   3891858H1   1566   1796       27   925628T6   1574   1943       27   925628H1   1574   1797       27   g5113339   1766   2184       27   7286802H1   1   500       27   3490274H1   26   319       27   6817649H1   55   308       27   6779822H1   68   617       27   1343838H1   166   433       28   6827746J1   1   328       28   g4372436   2   487       28   7277307H1   1   320       28   g1109781   68   2457       28   g2987914   238   414       28   3242379H1   460   594       28   5843682H1   675   814       28   6244385H1   750   960       28   3560794H1   878   978       28   6146228H1   1389   1559       28   g5454470   1759   2219       28   g5634013   1818   2245       28   g3933775   1835   2188       28   6824121H1   1864   2239       28   6824121J1   1864   2239       28   6789580H1   1898   2361       28   g5596185   1915   2221       28   g1137662   1994   2365       28   g3917233   2020   2409       28   3222719H1   2019   2299       28   g4487317   2029   2459       28   g2878126   2045   2386       28   925319R6   2061   2458       28   925319T6   2061   2419       28   925319H1   2061   2370       28   g6463796   2103   2467       28   2175458H1   2139   2397       28   g1153903   2165   2458       28   g1114183   2165   2444       28   g3844190   2203   2458       28   6193937H1   2265   2534       28   g5364673   2276   2476       29   6450124H1   992   1374       29   70254601V1   1036   1539       29   70250522V1   1050   1299       29   70255407V1   1073   1570       29   g2004277   1088   1409       29   70247748V1   1116   1378       29   70249044V1   1117   1269       29   70256246V1   1130   1541       29   70248757V1   1132   1311       29   7331424H1   1149   1638       29   70248822V1   1157   1333       29   70656623V1   1723   2219       29   661089H1   1723   1986       29   661089R6   1723   2224       29   6444789H1   1742   2329       29   70248827V1   1769   2103       29   6551207H1   1771   2379       29   6745795H1   1775   1933       29   6977014H1   1787   2347       29   70258294V1   1791   2007       29   70254633V1   1805   2279       29   70255157V1   1838   2268       29   6746363H1   1854   2506       29   70655542V1   1865   1948       29   6561163H1   1872   2335       29   7239061H1   1879   2417       29   70247905V1   1879   2117       29   70255827V1   764   1239       29   70255250V1   842   1401       29   70248754V1   866   1233       29   70256314V1   917   1512       29   70254664V1   444   930       29   70254648V1   463   958       29   70257273V1   531   636       29   g3778732   559   1002       29   4023624H1   1   286       29   70249613V1   1   198       29   3269702H1   8   254       29   70249634V1   142   374       29   1286494H1   165   402       29   7191139H2   175   755       29   5841654H1   175   446       29   7120789H1   398   702       29   70255670V1   413   949       29   70255767V1   1672   2119       29   3272240F6   1165   1617       29   3272240H1   1166   1413       29   70257558V1   1171   1601       29   70248599V1   1202   1420       29   70250795V1   1247   1524       29   70249059V1   1304   1679       29   70250085V1   1310   1485       29   70251290V1   1314   1573       29   70255060V1   1326   1720       29   70254762V1   1330   1860       29   70257824V1   1339   1525       29   70255199V1   1341   1753       29   70250692V1   1345   1576       29   70257468V1   1347   1462       29   70251556V1   1351   1435       29   70255729V1   1361   1869       29   70255867V1   1392   1903       29   70255422V1   1366   1870       29   70254725V1   1690   2218       29   70254417V1   1671   1916       29   7033280H1   1691   2365       29   1741694R6   1410   1866       29   1741694H1   1410   1587       29   70256209V1   1414   1915       29   7079231H1   1418   1892       29   70248162V1   1486   1604       29   70254647V1   612   1111       29   7077379H1   627   1163       29   7237267H1   1907   2197       29   70660270V1   1926   2390       29   70255193V1   1976   2586       29   70656301V1   1979   2231       29   71273576V1   1979   2197       29   6620779H1   2006   2603       29   70655675V1   2023   2430       29   70255204V1   2038   2632       29   1741694T6   2052   2625       29   6121206F8   2130   2669       29   5267406H1   2070   2344       29   6120592H1   2110   2682       29   6128080H1   2110   2689       29   6120292H1   2110   2671       29   6127177H1   2110   2671       29   3269702T6   2111   2623       29   70254909V1   2152   2688       29   4001246R6   2158   2517       29   4001246H1   2158   2458       29   70659041V1   2163   2394       29   g5864796   2171   2685       29   g2913171   2184   2622       29   6752303H1   2193   2671       29   70658326V1   2194   2559       29   70256172V1   2209   2671       29   2287809R6   2232   2681       29   2287809T6   2232   2629       29   2287809H1   2232   2458       29   6834934H1   2233   2671       29   2255790H1   2260   2543       29   70657495V1   2264   2391       29   6835034H1   2288   2570       29   70254456V1   2316   2671       29   661089T6   2355   2639       29   70249168V1   2452   2682       29   6059187H1   2608   2671       29   6059196H1   2606   2682       29   70255727V1   977   1493       29   70257455V1   1692   1951       29   70257816V1   1397   1604       29   70257088V2   1659   1980       29   70255058V1   1663   2172       29   5779331H1   1663   1928       29   70248161V1   1487   1604       29   6536779H1   1490   2108       29   70254435V1   1509   2113       29   70254449V1   1538   1906       29   70257853V1   1570   1900       29   70254760V1   1571   1970       29   70255434V1   1577   2080       29   5191169H1   1609   1875       29   70257088V1   1659   1980       29   70255260V1   1694   2245       29   70255174V1   1695   2113       29   g756281   1698   2066       29   70247381V1   1704   2145       29   70659718V1   1723   2028       29   70254418V1   675   1188       29   70258031V1   699   905       29   7178470H1   651   1108       29   70256308V1   672   1122       30   70780965V1   4   464       30   2359950H1   634   758       30   70777278V1   648   1196       30   70780989V1   653   1111       30   70779952V1   697   1217       30   70777355V1   715   1195       30   70771142V1   716   1207       30   70775784V1   751   1132       30   70769785V1   764   1127       30   6112349H1   809   1108       30   70776223V1   839   1239       30   5444375H1   933   1164       30   5057288T8   997   1251       30   g4264503   1011   1388       30   3385663T6   1033   1284       30   3845553T6   1031   1284       30   70776533V1   1086   1284       30   70776877V1   1087   1284       30   4148204T6   1174   1389       30   4148204F6   1181   1388       30   4148204H1   1181   1433       30   g4532148   1254   1359       30   5072501H1   11   296       30   70770035V1   207   641       30   70769523V1   245   581       30   70779544V1   337   548       30   70776851V1   339   867       30   70779695V1   4   418       30   5468404H1   3   275       30   6756043H1   102   509       30   7190732H1   163   722       30   71087882V1   4   560       30   70777729V1   4   492       30   70781241V1   4   467       30   70780840V1   4   442       30   70779497V1   342   547       30   3845553H1   372   583       30   5057288F9   386   1043       30   5057288H1   386   669       30   70776801V1   399   933       30   5065461F8   408   858       30   70778496V1   429   737       30   70777098V1   456   752       30   70780976V1   476   1081       30   70777266V1   503   1128       30   70776726V1   513   1093       30   70772943V1   541   740       30   70777725V1   576   1166       30   70779028V1   613   766       30   213833H1   1323   1388       30   213841T6   1323   1391       30   213841R6   1323   1388       30   213841H1   1323   1388       30   6917202H1   10   263       30   3385663H1   1   261       30   5468404F8   3   596       30   3385663F6   1   319       30   70775648V1   4   416       30   70778292V1   4   577       30   70779496V1   4   568       30   70775549V1   4   538       30   70776773V1   4   576       31   g3254719   1   445       31   g5745388   1   462       31   g3959110   1   353       31   g897505   1   215       31   g5233046   1   295       31   g2929868   1   406       31   g3016800   1   358       31   6202689H1   1   548       31   g2435161   9   456       31   4707801H1   1197   1296       31   806768H1   1211   1421       31   6915008H1   1320   1761       31   3036834H1   1578   1846       31   3036834F6   1578   2016       31   5387714H1   1632   1886       31   7122625H1   443   835       31   6915116J1   622   1129       31   5092947H1   899   1177       31   5370334H1   939   1050       31   5092947F6   899   1330       31   5017752H1   959   1229       31   4707801F6   1197   1714       31   5505129H1   17   282       31   5514010H1   17   254       31   5505122H1   17   228       31   1225071H1   52   262       31   g2929769   56   178       31   6314660H1   60   613       31   2682720F6   173   606       31   6516656H1   275   736       31   6201718H1   312   599       31   2682720H1   324   606       32   3665294H1   706   823       32   g1139437   903   977       32   3663680H1   706   950       32   5040414H1   731   947       32   g1139431   904   977       32   1772915R6   911   1308       32   1772915H1   911   966       32   4849209H1   1000   1259       32   1772915T6   1024   1540       32   g4691031   1318   1588       32   6298418H1   1   314       32   3403590H1   1   232       32   6327716H1   98   415       32   g1068995   238   588       32   6921019H1   255   457       32   4307678H1   254   563       32   g1858272   267   664       32   2812311H1   503   816       32   2812294H1   505   826       32   4976432H1   536   792       32   g5037963   518   976       32   4976496H1   536   801       32   g3736768   525   978       32   g4986041   540   979       32   4124067H1   552   792       32   g4073888   527   978       32   g5526438   564   978       32   5209523H1   413   705       32   4109819H1   416   696       32   3726063H1   426   688       32   4671035H1   421   693       32   2311003H1   454   685       32   5206768H1   478   723       32   679406H1   480   737       32   829499H1   273   519       32   2655709T6   279   932       32   1494347H1   285   504       32   042027H1   310   561       32   g1291521   316   662       32   030668H1   319   524       32   g897225   340   705       32   g921054   340   660       32   g897232   340   676       32   5018641T6   368   996       32   g2933440   374   696       32   1876772T6   379   940       32   g2703395   564   979       32   2652012T6   566   930       32   g1858336   575   1000       32   3124683H1   622   881       32   g1313829   631   867       32   4200739H1   671   956       32   2244549H1   671   882       32   2684460H1   696   909       33   g3232386   1   362       33   g3181700   7   419       33   6109493H1   322   613       34   3081155F6   1   417       34   3081155H1   2   315       34   3081155T6   31   463       34   g1648354   58   455       34   5800009H1   103   522       35   3517533R6   332   653       35   3517533H1   332   621       35   g5178468   190   667       35   3504115H1   162   477       35   3082646H1   123   415       35   259700H1   1   278       35   7169509H1   1   477       35   6768733H1   58   292       35   3176620H1   66   320       35   3176620F6   66   551       35   4613367H1   72   326       36   6753724J1   1   550       36   6769683J1   1   595       36   6772986J1   3   579       36   6769153J1   1   529       36   6764824J1   3   589       36   g6048166   3   423       37   653157H1   82   310       37   509350H1   107   307       37   3386816H1   1   183       37   3888124H1   4   268       37   6779840H1   9   549       37   3159561H1   58   220       37   6250084H1   67   593       37   2809974H1   67   324       37   3386816F6   1   622       37   3672749H1   81   364       37   4005442H1   2   285       37   70821167V1   627   1066       37   70822737V1   643   949       37   70832705V1   715   823       37   412478T6   835   1013       37   6779840J1   894   1315       37   g2631213   986   1049       37   5501816H1   1053   1261       37   5501816F6   1053   1293       37   4005442T6   487   1008       37   70818940V1   506   1068       37   059074H1   601   765       37   2862291H1   375   652       37   g3934153   376   841       37   g3900180   404   840       37   391612F1   432   1051       37   g895347   1   84       37   4005442F6   1   326       37   391612R1   353   898       37   412478R6   373   699       37   412478H1   373   600       37   3386816T6   337   818       37   1951349T6   296   802       37   6947424H1   302   813       38   6980110H1   1   501       38   g1897936   277   634       38   6264019H1   282   417       38   3099905H1   300   614       38   g2329399   438   689       38   2821248F6   452   766       38   2821248H1   452   755       38   g954493   494   556       38   6291249H1   507   718       38   5636134H1   640   912       38   6758395H1   658   1132       38   1903638H1   701   954       38   4991026H1   847   1075       38   4991026F6   847   1335       38   2821248T6   894   1434       38   6208066H1   893   1387       38   4453533H2   942   1149       38   5621950H1   1017   1297       38   6827666H1   1089   1614       38   3165907H1   1298   1569       38   4982360H1   1350   1616       39   5998240H1   1   484       39   458061H1   20   259       39   455162H1   20   272       39   460544R6   20   326       39   455966H1   20   255       39   455162R6   20   521       39   460544H1   20   245       39   3206542H1   25   202       39   4663964H1   37   289       39   g1807165   81   278       39   g2401959   129   486       39   3614680H1   155   272       39   g5101146   180   486       40   6927079H1   1   400       40   g922215   84   386       40   g922268   84   327       40   g991321   109   483       40   4991883T6   268   799       40   1958936H1   672   848       41   6033171H1   1   434       41   g1648352   23   428       41   3023715H1   184   448       41   2071327F6   285   584       41   2071327H1   285   475       41   6912958J1   351   944       41   4583753H1   378   666       41   2071327T6   676   1111       41   g4176194   713   1156       41   g3178308   731   1151       41   g4149782   824   1142       41   6912958H1   837   1307       41   g3797002   974   1088       41   g3180193   1135   1214       42   6937854H1   1927   2414       42   7032292H1   1837   2263       42   7002267H1   1385   1996       42   7153478H1   1986   2456       42   7100968H1   1404   1874       42   7035269H1   1372   1791       42   7166089H1   1147   1690       42   5637088H1   1339   1516       42   6778731H1   949   1515       42   2689778H1   1187   1439       42   4293860F6   818   1413       42   5975002H1   2124   2431       42   g1423371   869   1290       42   g3424813   874   1290       42   g1859062   1009   1290       42   1809216F6   809   1271       42   1809216H1   1109   1271       42   4290553H1   911   1161       42   6932930H1   733   1101       42   g1858731   690   1070       42   4293860H1   818   1065       42   4042230H1   673   849       42   g1423374   236   722       42   6132451H1   275   568       42   6781622H1   1   541       42   5966128H1   192   357       43   117874R6   1   438       43   4229003F6   171   674       43   4229003T6   271   738       43   g3895952   404   823       44   g3803941   113   583       44   g3805589   75   524       44   g3917967   73   520       44   3817246H1   28   320       44   2895718H1   1   283       45   4295967F8   1   404       45   g5811741   166   582       46   6708077H1   547   1179       46   5352502H1   578   664       46   6819988J1   1   605       46   6292367H1   493   609       46   g4900516   542   664       47   2607525T6   1   557       48   5120882F6   1   353       48   g4072524   256   619       49   4936033R6   591   1027       49   5387945T6   1   633       50   5401502T6   1   364       51   2205615T6   377   901       51   4150193F6   1   509       52   g2005559   1   239       52   1399832H1   1   227       52   1398471H1   1   238       52   g2027783   1   329       52   1322674H1   2   254       52   g2278640   13   442       52   g2159473   86   551       52   g2162158   123   594       52   g2156015   131   366       52   g2156002   208   366       52   g1954430   228   425       53   2737435F6   1   479       53   2737435T6   398   597       54   4654776T6   126   637       54   3796761F6   1   403       54   3796761H1   1   306       54   4654776H1   34   285       54   4654776F6   34   415       54   1729735H1   52   280       54   g895564   59   388       55   2920075R6   1   427       55   784499T6   20   470       55   2920075H1   169   427       55   269219H1   211   414       55   269219R1   212   754       55   g1978467   356   665       56   6183766H1   1   267       56   g1897936   1   354       56   3099905H1   19   334       56   g2329399   158   409       56   2821248F6   172   486       56   2821248H1   172   475       56   g954493   214   276       56   6291249H1   227   438       56   5636134H1   360   632       56   1903638H1   421   674       56   4991026F6   567   1055       56   4991026H1   567   795       56   6208066H1   659   1163       56   2821248T6   614   1154       56   4453533H2   662   869       56   5621950H1   737   1017       56   3165907H1   1018   1289       56   4982360H1   1070   1335       56   6417723H1   1325   1610       57   6061626H1   753   1059       57   5425067H1   798   1041       57   6331039H1   50   618       57   6132218H1   689   951       57   3499278F6   482   891       57   3499278H1   645   891       57   4657265H1   310   411       57   6309037H1   233   724       57   6538713H1   284   682       57   g1962414   304   564       57   g1376321   264   526       57   g4110945   3   417       57   g4310623   1   416       57   g2733078   2   384       57   g2835164   3   383       57   g3241486   1   382       57   g1376322   2   296       57   3351668F6   1   267       57   3351668H1   1   264       57   1785191H1   10   194       57   g2198029   1   129       57   4050289H1   28   127       57   4144289H1   20   125       58   g1979821   1   188       58   g3835204   1   325       58   g4072180   19   468       58   3519808H1   81   367       58   g843784   232   499       58   1614746F6   288   732       58   1614746H1   288   505       58   1575232H1   365   577       58   g2209653   403   912       58   6322462H1   520   718       58   2892611T6   645   1136       58   1614746T6   733   1148       58   g3644249   766   1189       58   670448R6   800   1186       58   677184R6   800   1152       58   670448T6   800   1149       58   677184H1   800   1071       58   670448H1   800   1055       58   g2810503   840   1186       58   g2336018   907   1187       58   2892611H1   945   1178       58   1289991H1   1081   1187       59   g3838936   83   488       59   g4085429   126   588       59   g1148294   133   524       59   g3057535   162   618       59   6429507H1   305   589       59   2865771F6   460   851       59   2865771H1   460   746       59   3639153H1   501   824       59   g1258980   584   839       59   3569914F6   601   1111       59   3569914H1   601   894       59   3569914T6   962   1569       59   5327014H1   1048   1291       59   1320957H1   1120   1353       59   2971828F6   1216   1745       59   g3049125   1226   1757       59   3077923H1   1284   1572       59   g4069747   1301   1758       59   g3417824   1307   1758       59   5732379H1   1398   1646       59   5732479H1   1399   1592       59   2971828H2   1452   1745       59   g1201083   1580   1748       59   4459657H1   1655   1775       60   60206922U1   296   954       60   368741T6   399   902       60   368741R6   1   397       60   368741H1   1   308       60   g4187853   375   750       60   368741R1   1   490       60   g3838805   409   751       61   5963989H1   1   502       61   3593809H1   1   298       61   g1059739   1   137       61   1850713F6   1   298       61   1850713H1   1   241       62   2872408F6   1   397       62   2872408H1   367   653       62   2872408T6   33   629       63   2862618T6   1   569       63   2838402H2   1   252       63   287364H1   1723   2056       63   2867867H1   198   481       63   2828380H1   522   792       63   2881835T6   606   1167       64   2882345T6   1809   2320       64   2820130T6   1421   1994       64   2837371H1   450   548       65   2222227T6   809   1284       66   270908H1   1   102       66   2698411H1   102   402       66   269967H1   55   354       66   2778926H1   24   260       66   2737435H1   1   207       66   2737435F6   19   501       66   2753160H1   1   119       66   2718845H1   1   245       66   2718845F6   1   395       66   2729146H1   107   344       66   2690266H1   126   385       66   2782984H1   438   571       66   2737435T6   420   617       67   2666231T6   1   523       68   1729868T6   1414   1975       69   3256037T6   1397   1940       70   3394349T6   33   600       71   5387945T6   685   1317       72   394892F1   793   1443       72   402386T6   930   1401       73   3318809F6   1   438       73   3318809H1   3   276       73   3318809T6   227   754       73   3255863H1   438   688       74   3685359H1   1   303       74   3685359F6   42   501       74   3685359T6   1   279       75   3637162H1   1   292       75   3637162F6   38   490       75   3699637H1   22   182       75   3637162T6   192   740       76   2007652H1   225   411       76   2007652T6   225   449       76   2007652R6   225   480       77   3325402T7   1764   2221       78   4654776F6   34   415       78   4654776H1   34   285       78   4654776T6   126   637       79   1476477F6   1   548       79   1476477H1   1   205       79   1476477H6   1   207       79   1476485H1   1   85       79   1476477T6   120   639       79   1500139H1   86   292       79   1496323H1   591   830       80   5200862H1   1   220       80   5200862F6   1   508       80   5200862T6   371   989       81   450278R6   619   1125       81   448904R6   1   390       81   450278R7   619   1046       81   448904H1   1   216       81   446282H1   1   256       81   450278H1   1   198       81   448904T6   721   1298       81   450278T7   788   1297       82   807409T6   210   590       82   807409R6   210   629       82   807409H1   1   78       83   3844814F6   147   721       83   3844814H1   433   721       83   3779417H1   1   204       83   3769201H1   63   208       83   3844814T6   63   317       84   1224910R6   1   543       84   1224910H1   881   1024       84   1224910T6   98   446       85   5296190T6   1523   1959       86   70375800D1   365   741       86   5884784H1   1617   1830       86   60203053V1   286   730       86   60203050V1   304   692       86   70374630D1   304   668       86   60209436U1   1   656       86   70373679D1   378   626       86   70373094D1   400   786       86   70375597D1   303   605       86   70373898D1   304   437       86   70373514D1   304   437       86   819073H1   1256   1510       86   2831185H1   1233   1479       86   70375782D1   380   772       86   70374947D1   304   772       86   g824801   1579   1852       86   70376281D1   472   766       86   60209435U1   102   747       86   g2780043   1733   2066       86   2751856R6   1651   2039       86   70374895D1   404   947       86   70376064D1   393   947       86   70374077D1   472   947       86   70373091D1   669   947       86   70373401D1   782   947       86   70376523D1   789   947       86   70375900D1   868   946       86   3999158H1   1844   1992       86   70376383D1   606   867       86   2751856H1   1651   1923       86   70374800D1   304   837       86   2751062H1   1651   1919       86   70373252D1   300   800       86   70375702D1   340   800       86   60203054V1   353   791       86   70373230D1   359   787       86   5822141H1   1871   2175       86   6332621H1   626   1098       86   1689922F6   1942   2175       86   5820119H1   1871   2166       86   g4222747   1715   2166       86   g3594875   1710   2160       86   g3751911   1700   2160       86   g3847457   1754   2160       86   g2675135   1863   2160       86   g4524280   1891   2160       86   g3422353   1857   2153       86   1689922H1   1942   2151       86   g1501747   1770   2146       86   g2342389   1928   2145       86   g846897   1832   2132       86   g4264802   1694   2117       86   g3693061   1788   2117       86   g4269694   1799   2117       86   g2269848   1756   2117       86   g824800   1834   2117       86   g3069508   1651   2117       86   g2055928   1783   2117       86   g2539104   1673   2117       86   g2901266   2005   2113       86   70375731D1   590   1018       86   g4390710   1751   2184       87   5059054H1   1   257       87   5059022F9   1   581       87   179544R6   84   529       87   1227952T6   125   595       87   064418H1   286   457       87   179544H1   329   529       87   g1011391   482   807       87   4667922H1   539   759       88   g4189831   12   63       88   4721701H1   360   439       88   3123901H1   1   66       88   6785315H1   12   481       88   7201126H2   51   570       89   2327449H1   15   248       89   2327457T6   1   364       89   2327449T6   1   288       89   2327449R6   13   408       89   2327457R6   13   402       90   g3694145   26   408       90   6779728J1   1   370       90   g3250058   1   408       90   6245903H1   120   532       90   g4333202   140   407       91   1895443H1   19   272       91   4729485H1   86   235       91   5217490H1   1   198       91   1542833H1   1   137       91   3836068F6   7   546       91   1891884H1   19   264       91   3052342H1   46   335       91   5735738H1   78   348       91   g5661058   155   347       91   3525414H1   171   494       91   3836068H1   7   289       92   1335071H1   515   737       92   1353732H1   535   775       92   6166536H1   736   1266       92   6483617F9   808   1320       92   6483617F8   828   1386       92   4326477F6   1   381       92   4326477H1   2   160       92   4140846T9   26   550       92   3888314H1   52   314       92   7073510H1   119   689       92   7081835H1   125   678       92   2079383H1   301   563       92   6405368H1   315   599       92   7254119H1   491   935       92   6420281H1   653   1159       92   6759433J1   668   1118       92   6483617H1   808   1319       93   257941R6   1   252       93   2764170H1   19   259       93   7040554H1   149   692       93   6477351H1   541   1039       93   g3163349   590   1022       93   3209591H1   1   149       93   5511002H1   628   880       93   5767313H1   1   246       93   000413H1   1   241       93   257941H1   1   237       93   2727658H1   1   173       93   5511002F6   628   905       93   g1977359   11   218       94   894136H1   1   167       94   893591H1   1   280       94   3685359F6   21   480       94   3685359H1   21   323       94   3685359T6   243   521       95   g1925745   1   143       95   g2752333   16   387       95   g2881366   16   370       95   3257022H1   28   287       95   2380565F6   186   681       95   2380565H1   186   401       95   6354028H1   192   471       95   2809746H1   248   487       95   5110009H1   249   500       95   5110002H1   249   441       95   6354128H1   326   480       95   1546477H1   378   584       95   5055767H1   565   845       95   3413877H1   572   817       95   2360256R6   719   1123       95   2360256H1   719   976       95   g1688388   781   1110       95   1330559H1   784   1027       95   g844802   968   1283       95   g1640803   1007   1243       96   4895030H1   2484   2784       96   g1941651   2486   2951       96   70657184V1   2503   3092       96   g658182   2512   2776       96   g2718978   2525   2989       96   g715365   2538   2875       96   5298571H1   2539   2819       96   4749511H1   2540   2838       96   5298771H1   2539   2804       96   5298612H1   2540   2664       96   6041370H1   2550   3199       96   g1471133   2556   2990       96   6555238H1   2565   3127       96   6556278H1   2565   3092       96   2913641H1   2604   2878       96   5825362H1   2609   3122       96   6400248H1   2624   2806       96   g1719353   2634   3091       96   g1960091   2671   3185       96   6829782H1   2683   3131       96   6829782J1   2683   3131       96   g6139947   2710   2989       96   70839657V1   2744   3193       96   70657566V1   2815   3435       96   2367903F6   2842   3294       96   2367903H1   2842   3079       96   2598088T6   2849   3428       96   2370164T6   2863   3400       96   7339911H1   2891   3434       96   494011F1   2890   3445       96   70658592V1   2911   3534       96   1398685H1   2935   3187       96   70656953V1   2938   3555       96   1380634H1   2939   3189       96   5138789H1   2949   3229       96   70655570V1   2957   3552       96   5067594H1   2958   3147       96   494011T6   2971   3405       96   g2051891   2978   3448       96   6850945H1   2986   3519       96   g4738083   2987   3447       96   6489656H1   2997   3108       96   g3419253   3004   3445       96   g5656805   3025   3445       96   g1444847   3030   3446       96   g3416160   3047   3449       96   g1719354   3058   3454       96   g1780421   3068   3450       96   6937801H1   3118   3682       96   6500625H1   3113   3727       96   g654349   3148   3441       96   g5747781   3156   3445       96   g758975   3157   3485       96   g564984   3156   3445       96   g1357788   3163   3465       96   g2931035   3178   3445       96   155875T6   3180   3849       96   3874011H1   3185   3455       96   g1190099   3218   3440       96   3117949H1   3251   3435       96   2756752H1   3308   3580       96   70837859V1   3306   3896       96   495759F1   3326   3893       96   1332986T6   3341   3399       96   3297117H1   3428   3687       96   g2959267   3429   3904       96   g3836196   3462   3898       96   154612T6   3493   3858       96   g2877501   3517   3898       96   g3838447   3541   3898       96   g1187596   3595   3893       96   7004590H1   3601   3894       96   6192804H1   3620   3893       96   6194735H1   3620   3893       96   6194703H1   3619   3877       96   495759T6   3642   3849       96   g4737879   3648   3893       96   g758939   3648   3873       96   g3895731   3648   3898       96   g1079906   3666   3944       96   767355H1   3801   4056       96   g1115031   3911   4343       96   7337250H1   1   574       96   2485552H1   46   257       96   7235980H1   61   574       96   g3869258   202   4338       96   g3869256   202   4338       96   2693067H1   559   813       96   6945783H1   613   1122       96   2535607H1   838   1099       96   70837439V1   956   1519       96   3502278H1   1050   1359       96   g1779649   1215   1673       96   60209482U1   1264   1763       96   495759R1   1304   1782       96   495759R6   1305   1682       96   495759H1   1305   1559       96   70375665D1   1438   2033       96   60207458U1   1487   2000       96   60209492U1   1489   2005       96   60209480U1   1489   1845       96   7006928H1   1535   1890       96   70375676D1   1644   2122       96   70375660D1   1684   1921       96   494011R1   1689   2115       96   494011R6   1689   2062       96   494011H1   1689   1921       96   g3896433   1698   2078       96   994895R1   1733   2255       96   994895H1   1733   2006       96   680754H1   1778   2047       96   70374292D1   1783   2008       96   70375422D1   1783   2241       96   70375842D1   1789   2222       96   3488220H1   1791   2075       96   71220762V1   1851   2480       96   1332986F6   1863   2182       96   71221685V1   1859   2529       96   1332986H1   1863   2097       96   70838337V1   1962   2525       96   1964279H1   1965   2251       96   71221831V1   2034   2530       96   70375269D1   2043   2527       96   g1357787   2095   2714       96   5874572H1   2106   2365       96   71221729V1   2117   2580       96   70838208V1   2146   2490       96   2598088F6   2168   2790       96   2598088H1   2168   2286       96   g573567   2168   2536       96   6409839H1   2218   2783       96   5035552H1   2282   2539       96   70658011V1   2283   2794       96   70656808V1   2299   2887       96   1997937R6   2308   2826       96   1997937H1   2308   2587       96   g3887783   2344   2793       96   1997937T6   2397   2962       96   g1280977   2404   2950       96   155875H1   2404   2627       96   155875R6   2404   2906       96   70839485V1   2403   2962       96   6096636H1   2466   2679       96   7126727H1   2484   3045       96   5876310H1   2484   2804       97   4719538H1   925   1185       97   4911350H1   964   1239       97   1903472H1   984   1228       97   3639862H1   1027   1306       97   4541862F6   1034   1422       97   6907459H1   1061   1564       97   g5657760   1076   1506       97   6908516J1   1077   1642       97   4697720H1   1125   1372       97   4238385H1   1   283       97   3403365H1   73   315       97   3403365F6   74   334       97   6907459J1   151   705       97   4551324H1   169   418       97   3374712H1   211   466       97   60264809D1   228   424       97   1954023H1   257   493       97   g792057   641   899       97   60264821D1   680   1235       97   60264804D1   680   1220       97   60264819D1   680   1192       97   g2011093   754   943       97   069791H1   807   940       97   4916612H1   819   1108       97   4916612F7   828   1360       97   4719538F6   925   1431       97   6337692H1   300   461       97   3371311H1   544   811       97   3091243H1   1164   1445       97   3915431H1   1199   1503       97   1954644H1   1145   1429       97   2596935H1   1269   1503       97   4719538T6   1597   1835       97   g812717   1598   1774       97   60100951B1   1279   1689       97   2410012H1   1308   1528       97   5781437H1   1353   1643       97   1499912H1   1387   1582       97   2777329H1   1412   1639       97   60100952B1   1415   1827       97   g2577280   1497   1671       97   g3648252   1574   1704       97   g1128730   1628   1854       97   g2837364   1650   1832       97   60100950D1   860   1177       97   60100952D1   860   1116       97   1264982H1   895   1131       97   6120149H1   908   1402       97   6119056H1   908   1530       97   60264817D1   625   1184       97   60100951D1   637   929       98   6197337H1   1   377       98   6198216T8   1   273       98   6197337F8   18   368       98   6331250H1   213   825       99   3016435F6   1   256       99   3016435H1   1   138       99   5964071H1   28   318       99   3016435T6   183   759       99   2094922H1   268   318       99   4587201H1   585   849       99   4762091H1   613   811       99   1495146R6   616   1100       99   1495146H1   616   775       99   5326946H1   671   928       99   1495146T6   681   1055       99   3703757H1   702   1004       99   2948181T6   719   1062       99   2948181F6   726   1100       99   2948181H1   726   983       99   g4620028   745   1100       99   g928615   748   1102       99   677653H1   1004   1100       100   5901489T6   1   584       101   2025051T6   1192   1583       102   g2932743   1   465       102   7165328H1   224   722       102   g5664446   237   597       102   2863343H1   346   643       102   4030732H1   346   455       102   2863343F6   346   769       102   3323244H1   597   856       103   4987160F6   1   601       103   4986076H1   251   398       103   2753343R6   273   678       103   2753343H1   273   523       103   4793432H1   351   482       103   4793224H1   353   635       103   2753343T6   394   811       103   4987160H1   1   271       103   3343341T6   430   814       103   4228058H1   434   708       103   864677H1   614   856       103   2755913T6   618   813       103   2755913H1   618   848       103   2755913R6   618   847       103   1971855T6   627   812       103   6161690H1   661   856       104   1006355H1   6597   6859       104   3957372H2   6610   6875       104   186558R6   7598   8102       104   g4838144   307   6193       104   g3847721   8369   8524       104   g3861906   8369   8524       104   185105H1   7598   7794       104   186558H1   7598   7745       104   186558T6   7859   8487       104   g1635936   7988   8197       104   4619766H1   8062   8337       104   7356046H1   8121   8524       104   g3797365   8183   8524       104   6432853H1   8263   8530       104   6434079H1   8263   8530       104   756968R6   8287   8524       104   756968H1   8287   8518       104   g184038   1   8521       104   7000401H1   343   428       104   1608059F6   445   836       104   1608059H1   445   664       104   2169635F6   1141   1544       104   2169635H1   1141   1373       104   2169635T6   1716   2134       104   7229203H1   2112   2612       104   g314259   4291   4813       104   3960183H1   6436   6705       105   70524885V1   1630   2395       105   70524843V1   1639   2253       105   6643169V1   1689   1909       105   70646269V1   1689   1909       105   70525307V1   1716   2188       105   g880693   1746   2259       105   70533384V1   2383   2603       105   2707682T6   2410   2903       105   70523481V1   2420   2965       105   70525826V1   2426   2939       105   70525835V1   2426   2939       105   70523321V1   2540   2944       105   g875595   2611   2954       105   70523241V1   2671   2938       105   g561027   2772   2938       105   g875594   1746   2106       105   g669502   1745   1996       105   70525213V1   1780   2434       105   70522841V1   1864   2410       105   70524066V1   1885   2366       105   70524543V1   1916   2434       105   7238311H1   1931   2473       105   70522425V1   1952   2725       105   70525244V1   2005   2625       105   70526939V1   2013   2610       105   70526388V1   2192   2409       105   70524889V1   2198   2767       105   70526612V1   2200   2358       105   70522907V1   2239   2973       105   70522785V1   2248   2927       105   70526373V1   2260   2416       105   7086069H1   2358   2898       105   7071063H1   1   547       105   g913241   159   2181       105   g6299529   164   599       105   7091369H1   245   847       105   7347105H1   266   460       105   5312260H1   357   578       105   7090770H1   1143   1683       105   70525682V1   1204   1896       105   70526748V1   1205   1699       105   2707682H1   1213   1477       105   2707682F6   1213   1439       105   70522990V1   1214   1837       105   1569986H1   1252   1374       105   70529862V1   1437   1903       105   70522200V1   1481   2029       105   70525480V1   1546   2185       105   6449560H1   1564   2157       106   3408296H1   1562   1812       106   3173959T6   1584   2175       106   2298374R6   1623   2077       106   2298374H1   1623   1895       106   1361272F1   1657   2216       106   1361366H1   1657   1834       106   1683557F6   1671   2227       106   1683573F6   1671   2086       106   1683573H1   1671   1907       106   1336611H1   1698   1941       106   71265365V1   1447   1892       106   70062844V1   1469   1865       106   2273538H1   1494   1763       106   3022474H1   1540   1833       106   71120676V1   1546   1932       106   70524882V1   1559   1725       106   70528634V1   558   1205       106   2408847H1   1882   2132       106   70059242V1   1888   2222       106   g5855887   1901   2223       106   g5858285   1901   2224       106   3409241T6   1911   2367       106   5669576H1   1931   2121       106   70062956V1   1793   2090       106   70061871V1   1800   2151       106   4778510H1   1802   2080       106   70061698V1   1811   2222       106   g4901915   1816   2222       106   70060889V1   1815   2268       106   1623591T6   1819   2177       106   1915736T6   1818   2176       106   g3155476   1821   2208       106   g5392637   1826   2222       106   70061837V1   1835   2222       106   1623591F6   1853   2222       106   1623591H1   1853   2074       106   g2198272   1858   2220       106   960516R1   1859   2222       106   70062540V1   1860   2222       106   960516H1   1859   2160       106   960387T1   1859   2185       106   2086141H1   1864   2127       106   4239442T8   1884   2294       106   5376913H1   683   950       106   2557738H1   650   892       106   2560627H1   650   911       106   1853191F6   1706   2218       106   1853191H1   1706   1960       106   1632340H1   1705   1922       106   614199H1   1706   1933       106   70059326V1   1719   2222       106   1683573T6   1725   2175       106   3751266T6   1725   2187       106   3321818T6   1730   2185       106   70062398V1   1341   1862       106   71118139V1   1346   1909       106   1865168H1   1348   1605       106   71294927V1   1403   1816       106   71120506V1   1404   1857       106   71118652V1   1408   2023       106   71266623V1   1448   1751       106   71265047V1   731   1261       106   71119759V1   731   1234       106   71266124V1   732   1366       106   71118641V1   731   1311       106   71118731V1   731   1292       106   71265792V1   731   1260       106   1915736R6   731   1137       106   2681128H1   696   984       106   70531848V1   706   1260       106   71264971V1   725   1481       106   71266413V1   922   1608       106   71117117V1   921   1580       106   3672409H1   937   1168       106   71118560V1   968   1537       106   71119741V1   973   1452       106   3964084H1   1006   1294       106   3628624H1   1936   2251       106   g2969311   1938   2222       106   g3151378   1960   2226       106   g1980460   1995   2313       106   223027R1   2064   2222       106   223027H1   2063   2222       106   223027F1   2064   2222       106   70529734V1   461   579       106   g850466   493   824       106   70530256V1   467   1041       106   3751266F6   1   358       106   3751266H1   1   296       106   3321818F6   103   497       106   3321818H1   104   350       106   6351045H2   329   660       106   71265775V1   801   1368       106   4177726H1   808   1103       106   5296714H1   766   1037       106   7059995H1   799   1271       106   096309H1   1101   1323       106   71118507V1   1118   1700       106   71120385V1   1121   1448       106   g2198304   1133   1220       106   71119740V1   1134   1424       106   4174878H1   1137   1420       106   71117317V1   1153   1724       106   71119085V1   1197   1825       106   3792653H1   1224   1433       106   6329373H1   1235   1780       106   71119545V1   1267   1954       106   71120838V1   1275   1741       106   4717156H1   1277   1501       106   5391806H1   1282   1481       106   71118747V1   1295   1960       106   70527304V1   1301   1969       106   71120713V1   1306   1885       106   71118117V1   1311   1871       106   5290693H1   1314   1610       106   5287845H1   1314   1441       106   70530468V1   1320   1851       106   3173959F6   1322   1886       106   70061612V1   1322   1882       106   70059581V1   1322   1815       106   70058920V1   1322   1762       106   70060883V1   1322   1729       106   2298374T6   1735   2182       106   70061519V1   1747   2333       106   3021149H1   1758   2047       106   g2879046   1775   2225       106   1853191T6   1781   2376       106   g2444573   1794   2221       106   3934758F6   895   1411       106   70527491V1   846   1499       106   71119165V1   884   1492       106   71265279V1   889   1575       106   71264808V1   753   1263       106   71265724V1   731   1013       106   1915736H1   731   969       106   71265549V1   731   1202       106   71266530V1   732   1246       106   2681128F6   695   1180       106   g2541182   2106   2424       106   g3039868   2147   2225       106   3931985H1   896   1182       106   3934538H1   895   1200       106   3934758H1   895   1195       106   3934357H1   895   1200       106   3662984H1   1006   1254       106   70529227V1   1009   1601       106   71265619V1   1031   1689       106   5946829H1   1044   1325       106   71266345V1   1097   1749       106   71120487V1   1102   1594       106   71266010V1   1097   1765       106   5947885H1   1100   1363       106   3173959H1   1322   1589       106   70059533V1   1322   1585       106   g1981676   1322   1654       106   70062034V1   1326   1794       107   5913661H1   1   283       107   5913661F8   1   569       107   5913661F6   1   575       107   5913661T6   213   821       108   6796436H1   1   435       108   g5837313   111   559       109   2536867F6   918   1430       109   2536867H2   918   1187       109   g1926046   932   1379       109   g1925836   933   1201       109   3244923F6   993   1220       109   3244923H1   993   1213       109   2878889H1   1010   1304       109   2878889F6   1010   1349       109   g858504   1080   1411       109   5567006H1   1081   1257       109   1896278H1   1106   1341       109   g784668   1209   1286       109   3364252F6   1   432       109   3364252H1   1   226       109   096666H1   66   233       109   096675H1   67   240       109   7132087H1   255   624       109   4030547F8   461   1001       109   4030547H1   462   715       109   5077091H1   800   1072       110   2717953H1   1   259       110   2806157F6   27   606       110   2806157H1   26   323       110   2724233T6   367   954       111   166942F1   1   624       111   g3755789   134   505       111   g3109437   134   208       111   g3037830   135   509       111   g3180013   139   583       111   g4270829   142   429       111   g3594985   142   562       111   5282615T6   168   751       111   g2942533   248   563       111   g2953832   248   509       111   g3040122   248   503       111   g3051904   251   501       111   g3804542   368   750       111   5282615F6   635   1106       111   5282615H1   884   1106       112   g2563121   1   166       112   2792728F6   1   440       112   2792728T6   1   417       112   g3040744   1   338       112   g2714186   1   401       112   7157205H1   1   461       112   g4371924   71   205       112   1394888T6   90   304       112   1624877H1   94   288       112   2792728H1   147   439       113   g2943715   1   1450       113   6487571H1   657   1161       113   6487571F9   657   1207       113   70681361V1   692   1244       113   70681601V1   692   1178       113   1544823R6   692   1181       113   70681277V1   692   1164       113   1544823H1   692   898       113   g4686743   879   1327       113   70680264V1   950   1079       113   6476403H1   998   1525       114   6272292H2   1   507       114   5910821T8   365   636       114   5910821T9   365   662       114   5910821F8   365   793       114   5910821H1   365   676       115   1551035H1   1625   1845       115   2716914H1   1625   1873       115   4876737H1   1631   1915       115   7091270H1   1631   1955       115   674945H1   1636   1903       115   670546H1   1636   1756       115   2119143H1   1637   1891       115   2815231H1   1643   1910       115   g990246   1645   1912       115   957795H1   1653   1903       115   472488H1   1653   1882       115   472488R1   1654   2125       115   302216H1   1655   1877       115   1876833H1   1654   1913       115   2697195H1   1656   1898       115   5377444H1   1666   1918       115   2431879H1   1668   1914       115   777024H1   1669   1901       115   3236114H1   1675   1928       115   1641892H1   1677   1880       115   5569350H1   1678   1920       115   3857806H1   1686   1976       115   g2751499   1694   2029       115   g876527   1697   2020       115   4069649H1   1708   1834       115   4307481H1   1724   1900       115   031299H1   1750   1920       115   3623362H1   1779   2032       115   g5036497   1858   2136       115   3624213H1   1872   2081       115   g2411009   1907   2079       115   4721623H1   1918   2011       115   2839772H2   1922   2209       115   6056374H1   1933   2132       115   6056674H1   1933   2136       115   3802178H1   1933   2135       115   4371036H1   1940   2136       115   412662H1   1939   2136       115   1907478H1   1950   2136       115   5050475H1   1966   2136       115   6447704H1   1966   2106       115   g3888474   1988   2321       115   5379172H1   1989   2238       115   5015647H1   1994   2136       115   3327673H1   1995   2251       115   3567634H1   2021   2137       115   1739210H1   2025   2215       115   880124H1   2032   2132       115   2354054H1   2037   2132       115   1235532H1   2060   2132       115   2350867H1   2062   2132       115   g983285   2072   2421       115   4190021H1   2084   2132       115   g2017399   2116   2329       115   6550613H1   2153   2494       115   5195955H1   2155   2420       115   3907635H1   2185   2464       115   g3166884   2193   2356       115   g994523   2232   2476       115   g1099949   2244   2491       115   g757333   2245   2513       115   4534796H1   2255   2463       115   1543657H1   2262   2468       115   4754806H1   2295   2496       115   1947635H1   2336   2483       115   3930538H1   2360   2496       115   2760689H1   2377   2496       115   5563236H1   2404   2496       115   4310883H1   2408   2496       115   2778679H1   2439   2496       115   725638H1   1   243       115   6733284H1   96   640       115   3096672H1   98   410       115   7034622H1   100   606       115   g777461   120   197       115   5198834H1   146   385       115   2394482H2   157   372       115   4970513H1   161   431       115   4969579H1   160   361       115   377654H1   163   374       115   305879H1   187   444       115   307158H1   189   433       115   3319063H1   215   496       115   5504343H1   219   449       115   3513751H1   230   473       115   4775727H1   230   504       115   4696530H1   245   433       115   928643H1   289   554       115   4529248H1   370   618       115   3942806H1   418   694       115   7280921H1   466   677       115   g2013891   480   698       115   g2013455   480   709       115   5422141H1   481   726       115   3110608H1   525   776       115   5659913H1   620   896       115   5545664F8   647   982       115   5545664F6   647   1087       115   3163564H1   751   1011       115   761584H1   768   982       115   6887167J1   837   1454       115   1852303H1   1011   1084       115   6948478H1   1087   1504       115   4158605H1   1206   1469       115   5270074H1   1238   1440       115   2365846H1   1239   1469       115   2444605H1   1244   1464       115   3115706H1   1255   1482       115   6728257H1   1349   1938       115   5717548H1   1401   1922       115   5796003H1   1427   1849       115   779492H1   1436   1699       115   2279382H1   1436   1702       115   5370473H1   1439   1656       115   4152676H1   1450   1712       115   60123909B1   1458   2086       115   6960268H1   1466   1902       115   3449619H1   1461   1706       115   7039654H1   1461   1970       115   3890456H1   1461   1758       115   6966324H1   1466   2051       115   765511H1   1469   1812       115   4940482T9   1481   2031       115   6409578H1   1486   1978       115   6734637H1   1495   1912       115   2886241H1   1493   1744       115   1222871H1   1495   1739       115   4193226H1   1498   1707       115   7065251H1   1500   2097       115   2152140H1   1501   1772       115   3763167H1   1502   1559       115   3571390H1   1504   1790       115   942026H1   1509   1756       115   4533306T1   1513   2067       115   2124615H1   1517   1816       115   3684042H1   1522   1808       115   6715567H1   1523   2096       115   3322645H1   1523   1782       115   g1873672   1524   2006       115   3590863H1   1530   1809       115   60123902B1   1538   2104       115   1002539H1   1538   1639       115   6077425H1   1541   1858       115   6513731H1   1548   2079       115   3785763H1   1548   1837       115   538349H1   1550   1775       115   1832140H1   1551   1753       115   1669989H1   1553   1767       115   450252H1   1553   1773       115   737082H1   1556   1778       115   7065802H1   1560   2096       115   6741001H1   1563   2068       115   835332H1   1566   1869       115   1599938T6   1569   2099       115   4226362H1   1573   1848       115   529930H1   1578   1721       115   4771124H1   1577   1846       115   4715169H1   1583   1859       115   4533768T1   1584   2101       115   4895425H1   1591   1861       115   4348930H1   1593   1852       115   4348733H1   1594   1854       115   g2616416   1596   1833       115   5698825H1   1596   1847       115   2288814H1   1597   1844       115   5762293H1   1598   2136       115   3082229T6   1601   1987       115   3737781H1   1598   1897       115   883098H1   1600   1835       115   5336847H1   1601   1837       115   2741220H1   1601   1860       115   880546H1   1600   1841       115   6398288H1   1601   1744       115   3252954H1   1608   1863       115   3779658H1   1618   1921       116   g1891130   669   957       116   7333578H1   1   523       116   6545439H1   141   676       116   g3805536   534   966       116   g3322110   534   772       116   5610773H1   537   788       117   6929774H1   1   513       117   6052078J1   72   520       117   6052078H1   72   520       117   4970577H1   120   381       117   4970577F6   120   483       117   6292129H1   423   637       117   6294687H1   423   647       117   2807905H1   555   863       117   g2540618   597   871       117   4401727H1   650   916       117   5729803H1   731   1236       117   g1301257   787   1245       117   026879H1   838   1092       117   g1303063   897   1111       117   522135H1   1025   1160       117   522228H1   1025   1269       118   587588R6   1   336       118   587588T6   1   512       118   g1069975   229   539       119   g2809760   1   443       119   g2934256   68   387       119   2785236H2   220   481       119   3437984H1   274   530       119   2544176H1   332   520       120   2807456F6   1   508       120   2807456H1   1   249       120   2807456T6   122   671       121   g3595066   1   357       121   4665764H1   1   257       122   70151773V1   587   917       122   60203477U1   266   822       122   522228H1   955   1199       122   522135H1   955   1090       122   026879H1   768   1022       122   60203621U1   1   548       122   60203622U1   118   507       122   4401727H1   579   846       122   g2540618   526   801       122   2807905H1   484   793       122   70152547V1   797   1219       122   3812508H1   140   438       122   g1265991   198   338       122   3860472H1   256   546       122   3520754H1   334   621       122   70152228V1   391   1018       122   4350225H1   364   638       122   70155823V1   419   986       122   g3919706   1   424       122   2512390F6   1   316       122   g1792877   1   365       122   g4187765   4   446       122   g2905531   4   102       122   70156040V1   960   1356       122   70151954V1   828   1353       122   2512390H1   14   316       122   60202389B1   858   1335       122   60202388B1   870   1329       122   60202388B2   924   1329       122   999391H1   37   270       122   2512390T6   38   315       122   4970577T6   45   618       122   60110854B2   1164   1287       122   g2884782   1   452       122   g2220423   1   398       122   g1267721   1   282       122   g3918260   1   411       123   2807456F6   194   701       123   2807456H1   1   249       123   2807456T6   31   580       123   270567H1   78   161       123   269931H1   374   499       123   269626H1   1824   2053       123   269080H1   128   352       123   270403H1   85   349       123   270910R1   1824   2022       124   587588R6   220   555       124   587588T6   44   555       124   587588H1   1   165       125   3321035F6   317   786       125   3321035H1   330   595       125   g1319620   414   927       125   g2741801   414   556       125   g2841030   1354   1422       125   g2933104   421   906       126   5259815H1   1   206       126   3568526H1   71   366       126   1289824F6   181   734       126   g928730   693   889       126   764159H1   693   849       126   6620992H1   718   1297       126   839936R1   825   1369       126   1289824H1   181   349       126   839936H1   825   1066       126   3869224H1   996   1286       126   1685280F6   522   955       126   3223525H1   1137   1457       126   3843717H1   1   293       126   g1395923   1198   1531       126   1685280H1   522   754       126   5028090H1   1333   1598       126   4216695H1   1404   1656       126   1289824T6   249   852       126   g2540596   600   897       126   5724304H1   664   1233       126   1947742T6   688   858       126   g1225270   688   889       126   3438058F6   269   562       126   3438058H1   318   562       126   3438058T6   45   516       126   g3804916   6   436       126   g3254781   1   379       127   3504571H1   910   1216       127   g2055741   1038   1358       127   g1270278   1025   1374       127   2733544H1   1137   1405       127   g1162686   1144   1490       127   g1109059   1197   1484       127   g1774715   1212   1519       127   g3897241   1316   1718       127   5854467H1   1401   1557       127   g1898243   1492   1691       128   g2358498   616   997       128   183176H1   747   971       128   183176R6   507   971       128   183176R1   359   971       128   2733388H1   659   888       128   5616358H1   602   878       128   g2824012   433   792       128   g4762579   167   600       128   7104793H1   1   520       128   4004284H1   207   474       129   3928775H1   1   192       129   2562126T6   40   182       130   g4902006   709   895       130   2562126T6   708   852       130   839936R1   828   1385       130   839936H1   828   1073       130   1685280F6   1040   1477       130   3869224H1   1001   1301       130   4032140H1   1443   1700       130   1597096H1   1442   1628       130   3438058T6   1483   1957       130   4032240T9   1491   1900       130   g3804916   1564   1997       130   g3432508   1574   2002       130   g3254781   1621   2002       130   1947742T6   689   861       130   g928730   693   892       130   3223525H1   1152   1474       130   5259815H1   1   206       130   3568526H1   71   366       130   1289824F6   181   737       130   1289824H1   181   349       130   g2540596   600   900       130   4216695H1   1421   1676       130   3438058F6   1437   1733       130   3438058H1   1437   1682       130   1597096F6   1442   2036       130   1685280H1   1243   1477       130   5028090H1   1348   1616       130   7213258H1   1225   1803       130   g1395923   1213   1549       130   3928775H1   696   891       130   1289824T6   249   855       130   3843717H1   1153   1448       130   764159H1   693   852       130   5724304H1   664   1213       130   6040888H1   665   891       130   g1225270   689   892       130   7153412H1   656   1189       130   3640283T9   1701   1932       130   3640283T8   1701   1908       130   g3538751   1690   2005       130   3640283F8   1701   1949       130   6620992H1   717   1277       131   3404480H1   1921   2103       131   5665320H1   2177   2358       131   872922H1   7624   7881       131   6881955J1   2681   3266       131   2825680F6   7992   8265       131   g5036098   8059   8265       131   4027974T6   6903   7418       131   5717756H1   6914   7382       131   2717848T6   6942   7421       131   4722802H1   6965   7069       131   g3594812   7015   7468       131   6403421H1   7053   7317       131   g3163456   7067   7469       131   2649733F6   7123   7465       131   2649733H1   7123   7369       131   7003170H1   7124   7465       131   g683322   7127   7465       131   2649733T6   7128   7424       131   2484677H1   7142   7369       131   g6075627   7147   7469       131   g3048962   7149   7468       131   212391H1   7200   7437       131   g6506945   7080   7465       131   g6073337   7082   7469       131   g2630574   7111   7470       131   5207126H2   2813   2981       131   2825680H1   7994   8265       131   g560960   8006   8268       131   g796025   8011   8277       131   1415181H1   8034   8265       131   3336518H1   3434   3669       131   4723427H1   3458   3606       131   70390598D1   3509   4085       131   264846H1   2524   2856       131   6357277H1   2531   2839       131   5544294H1   2708   2828       131   2101112T6   7755   8220       131   7162079H1   218   671       131   g2599501   1   4447       131   6044517J1   7709   8237       131   810518H1   8140   8262       131   2101112H1   7763   8008       131   2101112R6   7764   8137       131   6559424H1   7774   8284       131   g2140984   7774   8165       131   3781662H1   7815   8126       131   744157H1   7818   8046       131   4774224T9   7821   8181       131   g2324704   7831   8266       131   g3796940   7834   8265       131   g5839127   7843   8265       131   g3932489   7870   8265       131   g683058   7964   8268       131   g1225252   7984   8268       131   2825680T6   7985   8226       131   3278368H1   7719   7959       131   4726587H1   7619   7844       131   2108075H1   7405   7654       131   g1241115   7445   7723       131   4632120H1   7488   7765       131   4632247H1   7488   7765       131   7154510H1   7499   7643       131   6162751H1   7497   8049       131   1593082F6   3583   4007       131   1593043H1   3583   3803       131   1593082H1   3583   3803       131   7236349H1   3664   4161       131   4004354H1   3665   3798       131   439020H1   3680   3902       131   3480603H1   3698   3865       131   g1678362   3734   3890       131   g4691014   3746   4204       131   g6132554   3779   4207       131   g3756265   3780   4206       131   g1199039   3835   4151       131   g819518   3859   4212       131   2155889F6   3867   4265       131   2155889H1   3867   4106       131   3513325H1   3880   4124       131   70391721D1   3896   4321       131   2155889T6   3964   4405       131   g5675561   3974   4452       131   6618788H1   3991   4472       131   g3144228   4048   4452       131   g2751511   4080   4441       131   1863153F6   4273   4666       131   1863161F6   4273   4526       131   1863153H1   4273   4520       131   385245H1   4275   4498       131   g4970402   4364   4819       131   5857654H1   4368   4636       131   g1624652   4395   4466       131   g1023422   4479   4773       131   g1023318   4511   4801       131   3341807F6   4527   4888       131   3341807H1   4527   4766       131   4027974F6   4592   4881       131   4027926H1   4592   4822       131   6024929H1   4601   4923       131   2203255H1   4755   5006       131   4184071H1   4839   5084       131   3322055F6   4994   5518       131   3322055H1   4994   5271       131   1863161T6   5057   5459       131   811828H1   5105   5352       131   789137H1   5167   5226       131   g3077295   5179   5635       131   3242918H1   5346   5610       131   4578433H1   5545   5787       131   5694130H1   5702   5886       131   4255728H1   5783   6052       131   5673677H1   5815   5974       131   g1880292   5855   6068       131   5768386H1   5903   6448       131   g1442274   5974   6178       131   g1678263   6029   6205       131   3607889H1   6044   6335       131   g709099   6063   6392       131   g769480   6063   6274       131   g692094   6065   6419       131   5036970H1   7696   7961       131   g2212423   7200   7465       131   3282024T6   7255   7409       131   3323350H1   7271   7524       131   g2768029   7301   7465       131   3322677H1   7313   7591       131   g1515911   7317   7464       131   3865622H1   7330   7526       131   g317850   7332   7592       131   g3174898   7357   7469       131   4253321H1   7391   7470       131   4244366H1   7390   7465       131   3334656H1   7394   7606       131   6355568H1   3040   3236       131   g1881143   3040   3347       131   1372008H1   7499   7667       131   2292865H1   7499   7721       131   g915845   7561   7784       131   2289132H1   7571   7802       131   5832896H1   7593   7874       131   4774272H1   7612   7881       131   1393295H1   7499   7692       131   1772773H1   7522   7784       131   6269862H1   6136   6658       131   g1975380   6238   6568       131   3389337H1   6336   6619       131   6706684H1   6370   6913       131   3282024F6   6416   6946       131   3282024H1   6416   6662       131   3854583H1   6428   6705       131   2509302H1   6442   6670       131   7225908H1   6547   7110       131   3147875H1   6581   6852       131   3797563H1   6583   6873       131   g2398342   1058   1351       131   6881955H1   1921   2185       131   2717848F6   6584   7049       131   2717848H1   6584   6832       131   4058694H1   6629   6895       131   025240H1   6665   6932       131   4270965H1   6782   6955       131   3341807T6   6814   7417       131   3322055T6   6820   7418       131   4027974T9   6822   7361       131   g691854   6874   7235       131   4029613H1   6873   7102       131   g708808   6874   7171       131   g565754   6874   7098       131   124772H1   6888   7030       132   1630826T6   1136   1410       132   g5813217   1172   1255       132   1631555T6   1194   1410       132   3878121H1   1210   1271       132   1986029H1   1210   1281       132   2956806H1   1210   1284       132   1372584H1   1214   1417       132   854216H1   1   247       132   854312R6   1   445       132   854312H1   1   151       132   2465049H1   150   358       132   2465049F6   150   697       132   2271395H1   192   455       132   2271395R6   192   704       132   g2184123   196   582       132   g1692736   348   740       132   2465049T6   519   1047       132   1601994H1   535   677       132   5274102T6   569   1060       132   g3413112   652   1088       132   g1692707   687   1094       132   g3428202   689   1088       132   g1686359   836   967       132   g2714128   860   1089       132   g3843958   950   1089       132   g4394372   950   1088       132   g3844127   950   1089       132   g661412   957   1252       133   g5438299   717   1143       133   g5109774   443   863       133   g5526440   420   841       133   g4599202   303   725       133   6306770H1   1   450       134   4186114H1   683   1024       134   4186114F6   683   1091       134   4186114T6   921   1587       134   4165023H1   674   955       135   2268189H1   1   231       135   2268189R6   1   379       135   1470335H1   37   230       135   g2505442   142   353       135   g2505398   157   521       135   g2458763   157   579       135   1445774H1   262   507       135   2268189T6   334   885       135   368103H1   520   763       135   g2328398   608   929       135   2322571H1   690   923       136   6114201H1   1   292       136   3287273H1   260   505       136   g779441   375   522       136   g879517   78   440       136   g870148   135   450       136   g883117   77   465       136   2120815H1   450   565       136   1631555H1   450   565       136   1631555F6   450   565       136   g705961   450   565       136   1547031H1   452   565       136   4796579H1   455   563       136   5081193H1   457   508       136   1630826F6   450   565       136   1630826H1   398   546       136   g2023471   491   696       137   6864812H1   1   502       137   292419H1   194   469       137   292419R6   196   486       137   3960046H2   333   473       137   3960046F8   333   926       137   g4114677   609   1055       137   g3674688   744   1053       137   2705604T6   744   1011       137   g3429159   744   1052       137   1871586H1   744   869       137   1998855H1   744   926       137   2135293H1   763   1036       137   2135293F6   763   1227       137   2100630H1   865   1055       137   292419T6   874   1001       137   3960046T8   887   1009       138   6100950H1   1   213       138   6302544H1   151   468       138   6144369H1   166   755       138   6144369F8   166   749       138   6099148H1   208   488       138   6281896H1   383   528       138   6111359H1   570   864       138   6028709H1   650   932       138   6144369T8   846   1259       139   5056523H1   2466   2604       139   2271032H1   893   1152       139   4243388H1   919   1251       139   4552488H1   958   1120       139   g1953334   885   1086       139   g1953371   885   986       139   g713204   1029   1191       139   g1967839   1091   1429       139   3737077H1   1295   1453       139   2271032R6   893   1370       139   6326476H1   1313   1610       139   1845337T6   2212   2566       139   1845337R6   2212   2562       139   1845337H1   2212   2433       139   g4288591   2215   2599       139   g5659132   2219   2551       139   913114H1   2228   2509       139   g1960980   2253   2604       139   g5887689   2244   2601       139   911966H1   2249   2503       139   g5838009   2250   2611       139   g5913083   2251   2604       139   g3741346   2254   2611       139   g3785022   2297   2602       139   g2463906   2312   2607       139   g4312734   2347   2599       139   g2100983   2358   2591       139   4519262H1   2360   2599       139   1272843T6   2368   2562       139   6104909H1   2413   2604       139   4180584H1   1872   2122       139   g1968950   1886   2368       139   5038396H1   1888   2146       139   6576957H1   1907   2455       139   3254138H1   1908   2115       139   5946104H1   1910   2145       139   2700155F6   1922   2433       139   2700155H1   1922   2121       139   2061567T6   1951   2554       139   3365965H1   1983   2108       139   g5591869   2006   2220       139   1749882T6   2039   2571       139   1752020H1   2049   2258       139   2700155T6   2063   2563       139   1889107T6   2099   2562       139   3236592H2   2125   2340       139   5291302H1   2152   2398       139   g5038163   2168   2608       139   g2355286   2200   2589       139   g3146568   2200   2604       139   5307668H1   1567   1691       139   g1968949   1345   1812       139   5840503H2   1578   1835       139   g2229641   1360   1792       139   6725031H1   1675   2299       139   476160H1   1389   1655       139   2061567R6   1698   2177       139   2061567H1   1698   1970       139   6158833H1   1433   1523       139   794690H1   1496   1698       139   6576416H1   1513   1850       139   1889107H1   1542   1812       139   2250985H1   1734   1964       139   1889107F6   1542   1892       139   4399135H1   1734   1991       139   4399331H1   1734   2000       139   1501428H1   1737   1930       139   6823952J1   1549   2162       139   6018675H1   1812   2378       139   4516041H1   1817   2066       139   1272843H1   1   245       139   1272843F6   1   593       139   1272843F1   1   331       139   1274319H1   21   218       139   2598183H1   24   263       139   3256051H1   25   268       139   6826541J1   33   663       139   6823903H1   34   188       139   6820526H1   422   747       139   6820526J1   422   747       139   6826541H1   672   1240       139   3857549H1   797   1106       139   1749882F6   811   1171       139   1749882H1   811   1085       139   6829715H1   860   1357       140   1272843F1   1   331       140   1272843F6   1   587       140   1272843H1   1   245       140   1274319H1   21   218       140   2598183H1   24   263       140   3256051H1   25   268       140   3857549H1   306   615       140   1749882F6   320   680       140   1749882H1   320   594       140   g1953334   394   595       140   g1953371   394   495       140   2271032R6   402   880       140   2271032H1   402   661       140   4243388H1   428   761       140   4552488H1   467   629       140   g713204   538   701       140   g1967839   600   939       140   3737077H1   805   963       140   6326476H1   823   1120       140   g1968949   855   1322       140   g2229641   870   1302       140   476160H1   899   1165       140   6158833H1   943   1033       140   794690H1   1006   1208       140   1889107F6   1052   1402       140   1889107H1   1052   1322       140   5840503H2   1088   1345       140   2061567H1   1208   1480       140   2061567R6   1208   1687       140   4399331H1   1244   1510       140   4399135H1   1244   1501       140   2250985H1   1244   1474       140   1501428H1   1247   1440       140   4516041H1   1327   1576       140   4180584H1   1382   1632       140   g1968950   1396   1878       140   5038396H1   1398   1656       140   3254138H1   1418   1625       140   5946104H1   1420   1655       140   2700155F6   1432   1943       140   2700155H1   1432   1631       140   2061567T6   1461   2064       140   3365965H1   1493   1618       140   1749882T6   1549   2081       140   70366789D1   1569   1883       140   1752020H1   1559   1768       140   2700155T6   1573   2073       140   1889107T6   1609   2072       140   3236592H2   1635   1850       140   5291302H1   1662   1908       140   g2355286   1710   2099       140   g3146568   1710   2114       140   1845337R6   1722   2072       140   1845337T6   1722   2076       140   1845337H1   1722   1943       140   g4288591   1725   2109       140   913114H1   1738   2019       140   g1960980   1763   2114       140   911966H1   1759   2013       140   g3741346   1764   2121       140   g3785022   1807   2112       140   g2463906   1822   2117       140   g4312734   1857   2109       140   g2100983   1868   2101       140   4519262H1   1870   2109       140   1272843T6   1878   2072       140   6104909H1   1923   2114       140   5056523H1   1976   2114       141   2211487F6   1   511       141   2211487T6   195   778       141   2412148H1   263   497       141   2564905H1   540   809       141   125647H1   559   768       141   2242295T6   625   773       141   2211487H1   1   241       142   3404890H1   1   218       142   3387263H1   44   343       142   g1225131   45   275       142   2745513H1   52   296       143   3330364H1   1   264       143   2532555H1   1   245       143   2532555F6   1   202       143   3452732H1   7   223       143   6206478H1   8   551       143   2240790H1   7   246       143   2240790F6   7   455       143   4052530H1   9   302       143   2211943H1   38   294       143   2207141H1   1   161       143   6338721H1   60   530       143   3722062H1   63   355       143   3564688H1   78   370       143   g1390898   122   415       143   5154455F6   261   738       143   5154455H1   489   738       143   5155056H1   492   738       143   5440206H1   376   645       143   6206972H1   638   1158       143   2240790T6   39   495       143   g2155919   45   468       143   3219860T6   39   451       143   3219860H1   135   451       143   3219860R6   1   451       143   3221716H1   149   451       143   2412827H1   233   450       143   g3932128   5   420       143   g2155881   1   335       143   g2775288   1   329       143   g3277838   1   325       143   g1390787   5   313       143   2532555T6   50   283       143   g1422648   1   217       143   g1783871   2   194       144   60210582U1   1   374       144   g857093   1   226       144   60124509B1   198   336       145   g2324899   1   329       145   g4525985   12   329       145   3516332H1   36   278       145   3012696F6   162   329       145   3012696H1   162   337       145   3012696T6   169   288       145   561748H1   231   329       146   6623203J1   1   573       146   3538176T6   590   927       147   5643301H1   1   266       147   5643301F6   1   412       147   7007819H1   12   557       147   7008530H1   19   463       147   7161666H1   34   435       147   7017571H1   42   211       147   3277305H1   128   393       147   3565182H1   492   782       147   g4293857   672   946       147   2992317H1   816   1097       147   5643301R6   823   1114       147   5643301R8   939   1115       148   g3742856   2099   2440       148   g1903217   1   2439       148   7183853H1   1   319       148   g3665358   2048   2440       148   6282319H1   2330   2439       148   g3931095   2387   2439       148   g1487041   2284   2448       148   g1365146   1897   2439       148   g1487088   1978   2188       148   7183071H1   2   532       148   7184162H1   6   440       148   7182312H1   265   747       148   g1365200   1875   2451       149   6807937J1   1   617       149   934453R6   169   650       149   934453T6   169   578       149   934453H1   169   430       150   g2356671   1   436       150   3878737H1   255   511       151   3613937H1   1   286       151   3613937F6   1   551       151   3356375H1   14   272       151   4365653H1   26   279       151   3133682F6   168   468       151   3133682H1   168   446       151   2784431H2   322   410       151   4760428H1   378   661       151   g1987179   426   713       151   3985930H1   427   714       151   g3840682   469   880       151   6619463H1   506   1081       151   g3594561   524   876       151   3613937T6   562   835       151   6148958H1   732   1299       151   5067091H1   1   243       151   2583002H1   33   298       151   2583002F6   33   533       151   g273554   104   290       152   70255820V1   1   485       152   70249242V1   70   324       152   70254597V1   114   627       152   70254754V1   114   607       152   70254688V1   137   517       152   70255790V1   244   565       152   70255539V1   507   565       152   70255201V1   514   565       153   g4194301   1   458       153   g3244448   3   471       153   g3804087   5   421       153   2383032T6   46   518       153   70046632V1   217   736       153   70046986V1   102   652       153   70047134V1   98   553       153   6208108H1   1   540       153   70046575V1   96   533       153   70047390V1   97   515       153   2383032F6   1   492       153   4938245H1   119   406       153   70047509V1   11   414       153   2383032H1   1   228       153   1952540H1   1   157       153   70046695V1   11   94       154   750543H1   1   183       154   g1474315   42   323       154   3722596H1   61   333       154   4875149F6   70   195       154   4875149H1   70   257       154   6561179H1   71   633       154   2029094H1   121   372       154   5950630H1   125   405       154   5950410H1   125   401       154   4933727H1   239   384       154   3533254H1   273   564       154   4875149T6   344   737       154   g3202357   457   784       154   748032H1   574   776       155   3751233H1   577   867       155   5812189H1   644   840       155   5812190H1   644   845       155   311752H1   657   744       155   373638H1   663   880       155   6333709H1   705   1240       155   6329688H1   705   1325       155   7080559H1   760   1170       155   6532643H1   803   1384       155   6141236T8   989   1579       155   4068325T6   1018   1641       155   7068876H1   1069   1483       155   3256362H1   1308   1554       155   g1400213   1337   1682       155   g3254782   1346   1682       155   g1383466   1395   1696       155   6412487H1   1494   1931       155   5534143H1   1578   1820       155   6869946H1   1717   2275       155   g4510725   1831   2272       155   6298042H1   1871   2146       155   g1998847   1973   2275       155   3541104H1   1991   2272       155   5499626H1   1   255       155   5500309H1   1   227       155   5500026H1   1   161       155   5499909H1   1   206       155   2723940H1   1   243       155   2723940F6   1   333       155   5643224H1   22   276       155   3126704H1   22   298       155   g1998848   22   296       155   3256088H1   24   259       155   1728945H1   26   246       155   6141236F8   28   633       155   6141236H1   28   370       155   6328822H1   47   592       155   3388023H1   89   361       155   6134014H1   110   401       155   3323733H1   192   461       155   3409854H1   357   618       155   4068325F6   515   1108       155   4068325H1   517   804       155   3629455H1   560   835       156   7006557H1   1   488       157   6459173H2   4960   5300       157   71034189V1   3224   3885       157   71033929V1   3308   3912       157   g2166710   3325   3492       157   71239210V1   3365   4028       157   71035047V1   3406   3932       157   71035847V1   3407   3932       157   71239003V1   3407   3934       157   7252515J2   3410   3964       157   617587H1   3462   3716       157   617587R6   3464   3954       157   71035452V1   3487   4142       157   71239589V1   3548   4177       157   71034494V1   3565   4148       157   71036467V1   3574   4249       157   71019079V1   3590   4079       157   71034628V1   3600   4175       157   71033566V1   3603   3776       157   71033563V1   3605   4306       157   71033740V1   3642   4142       157   71034052V1   3692   4322       157   2852347H1   3699   3792       157   71034523V1   3759   4400       157   71021626V1   3780   4091       157   71033276V1   3784   4394       157   71035439V1   3832   4224       157   71239794V1   3841   4296       157   71036851V1   3933   4532       157   2902436H1   4020   4325       157   71035620V1   4052   4596       157   617587T6   4094   4694       157   71240672V1   4243   4499       157   g2839119   4251   4718       157   71239718V1   4264   4729       157   71240638V1   4263   4415       157   g2716554   4274   4723       157   2839872F6   4473   4873       157   2839872H2   4473   4724       157   6701187H1   4517   5071       157   71033989V1   4552   4713       157   6700916H1   4758   5106       157   g1353781   1   3523       157   7183461H1   8   510       157   5878868F8   38   589       157   5878868T9   38   467       157   5878868H1   40   300       157   6621194J1   465   1063       157   7044151H1   1537   2130       157   7115836H2   1721   2207       157   6891967H1   1744   2304       157   7112010H2   2069   2273       157   7115993H2   2217   2833       157   71036402V1   2382   2971       157   71036528V1   2382   2844       157   71036302V1   2382   2836       157   5682881F6   2382   2755       157   5682881H1   2382   2556       157   7121747H1   2421   3009       157   6554467H1   2459   2877       157   71035730V1   2859   3427       157   71034930V1   2861   3427       157   5693974H1   2871   3141       157   71033786V1   2880   3524       157   71239770V1   2904   3495       157   71036410V1   2922   3457       157   71036110V1   2922   3457       157   71036841V1   2925   3050       157   71238907V1   3000   3542       157   71033235V1   3023   3512       157   71239855V1   3037   3682       157   71035104V1   3060   3655       157   71240132V1   3064   3600       157   71022089V1   3064   3174       157   71020694V1   3064   3173       157   71033750V1   3068   3576       157   71036780V1   3070   3575       157   71240343V1   3074   3590       157   71035904V1   3096   3656       157   71239106V1   3098   3535       157   71239593V1   3107   3663       157   71036268V1   3116   3650       157   71033766V1   3130   3794       157   71036727V1   3136   3763       157   71036255V1   3150   3690       157   3246035H1   3163   3414       157   71236782V1   3177   3581       157   71034117V1   3225   3899       158   2702437H1   3382   3692       158   2028546H1   3382   3674       158   70469621V1   3384   3974       158   70477624V1   3362   3551       158   70464393V1   3358   3664       158   2028546R6   3382   3697       158   897912H1   3532   3695       158   2657383H1   3542   3682       158   70466432V1   3600   4154       158   3405032H1   2336   2592       158   3099417H1   2407   2687       158   70467835V1   2535   3117       158   3936618H1   2535   2799       158   70468637V1   2538   3232       158   70465732V1   3029   3708       158   g4568142   3029   3550       158   70477055V1   3050   3634       158   70479313V1   3061   3562       158   70480477V1   3075   3701       158   70480157V1   3098   3662       158   4564168H1   3087   3365       158   70466004V1   3018   3732       158   70478380V1   3019   3638       158   4557976H1   3015   3240       158   3618747H1   1905   2177       158   6510691H1   1907   2199       158   6510634H1   1907   2192       158   6510739H1   1907   2183       158   4554621H1   2044   2312       158   70464785V1   2168   2732       158   4554554F6   2168   2649       158   4554554H1   2168   2409       158   70466681V1   2298   2736       158   70467583V1   2823   3476       158   70480720V1   2857   3511       158   70479363V1   2850   3206       158   70476260V1   2859   3447       158   70481657V1   2790   3146       158   70466450V1   2775   3513       158   70476879V1   2765   3450       158   70477920V1   2806   3426       158   70466934V1   2704   3432       158   70465560V1   3293   3609       158   70476467V1   3297   3594       158   70479516V1   3309   3697       158   4792962H1   3311   3605       158   70467134V1   3311   3697       158   4554470H1   3317   3600       158   70466403V1   3271   3697       158   70480199V1   3270   3697       158   70469816V1   3293   3654       158   70468353V1   3282   3697       158   70481137V1   3205   3697       158   70466975V1   3210   3697       158   70466049V1   3236   3697       158   70478359V1   3235   3666       158   6515587H1   3242   3671       158   70478211V1   3217   3682       158   4558722H1   1289   1534       158   3618747F6   1904   2401       158   g190751   50   3497       158   g1736195   979   1234       158   2624295R6   1016   1412       158   2624295H1   1016   1225       158   70466944V1   3104   3692       158   70480113V1   3129   3556       158   70468718V1   3142   3695       158   70477732V1   3142   3772       158   70469007V1   3149   3697       158   70469207V1   3322   3657       158   70480324V1   3326   3682       158   70466503V1   3328   4068       158   70479148V1   3328   3697       158   70464268V1   3329   3697       158   70466743V1   3331   3487       158   70467082V1   3389   3682       158   70481575V1   3408   3682       158   3381962H1   3429   3697       158   70468987V1   3435   3682       158   70465226V1   3440   3697       158   70466253V1   2697   3264       158   70469171V1   2699   3374       158   70469220V1   2765   3324       158   70481022V1   2759   3486       158   2624295T6   2762   3351       158   70469934V1   2769   3329       158   70480048V1   2644   3023       158   70478224V1   2650   3136       158   70467878V1   2660   3107       158   70481284V1   2587   3175       158   70467582V1   2607   3072       158   70465805V1   2630   3318       158   70481125V1   2624   3464       158   70469427V1   2642   3286       158   6287762H2   1   475       158   70478180V1   2901   3570       158   70465864V1   2900   3534       158   70479279V1   2923   3570       158   70476782V1   2927   3466       158   70469166V1   2935   3628       158   70476816V1   2992   3529       158   70480392V1   2746   3208       158   70478255V1   2751   3368       158   70478182V1   3467   4041       158   70498607V1   3496   3697       158   70465130V1   3440   3697       158   2301520H1   3457   3705       158   70467663V1   3171   3800       158   70479112V1   3208   3697       158   70467111V1   2890   3430       158   4562810H1   2883   3182       158   70473226V1   2882   3252       158   2620793H1   2886   3178       158   70464811V1   2708   3359       158   70481798V1   2721   3378       158   70478794V1   2722   3354       158   70467700V1   2732   3434       158   70481589V1   2742   3428       158   70466101V1   3337   3682       158   70468914V1   3348   3697       158   70479545V1   3359   3667       159   3874759H1   1   296       159   3685395H1   3   271       159   3692640H1   4   270       159   3692640F6   4   342       159   g3116688   4   443       159   3445829H2   6   264       159   306745H1   13   386       159   945423H1   11   293       159   988248R1   179   383       159   988248H1   179   291       159   4454887H1   250   519       159   3877090H1   350   619       159   6901148H1   368   825       159   3028782F6   505   841       159   3028782H1   505   807       159   1005592H1   599   889       159   4646393H1   801   1061       159   5278286H1   912   1125       159   2521969F6   950   1354       159   983403H1   1063   1335       159   983403R6   1064   1315       159   188949H1   1106   1306       159   983403T6   1123   1347       159   988925H1   1126   1349       159   2521969H1   1129   1354       159   2639257H1   1177   1339       159   983273H1   1241   1544       159   3692640T6   1247   1832       159   4013213H1   1308   1598       159   3877453H1   1331   1614       159   4010336H1   1328   1611       159   2324266H1   1337   1579       159   3011865F6   1340   1761       159   3576519H1   1339   1495       159   5176829H1   1348   1600       159   3873606H1   1351   1661       159   4852614H1   1363   1587       159   3016113F6   1376   1647       159   3011865H1   1376   1459       159   058529H1   1376   1522       159   3958442H1   1377   1474       159   5276870H1   1376   1535       159   3578346H1   1382   1634       159   3016113H1   1383   1669       159   6337413H1   1390   1507       159   6338013H1   1390   1881       159   6335720H1   1390   1883       159   3688195H1   1398   1695       159   4151882H1   1401   1641       159   3874704H1   1439   1714       159   5169119H1   1476   1610       159   5278359H1   1475   1705       159   3685458H1   1486   1783       159   6332978H1   1487   1982       159   5531678H1   1495   1624       159   g5395484   1513   1854       159   3693626H1   1512   1804       159   4152946H1   1520   1792       159   3875504H1   1549   1851       159   3890704H1   1548   1842       159   4466716H1   1582   1839       159   g395449   1591   1923       159   5167337H1   1592   1800       159   920392H1   1611   1923       159   3011865T6   1618   1972       159   3016113T6   1674   2155       159   5277911H1   1697   1935       159   2636153H1   1700   1929       159   1567405H1   1706   1889       159   462513H1   1706   1931       159   462513R6   1706   2044       159   3877471H1   1721   1990       159   5277625H1   1723   1973       159   3016674H1   1739   2024       159   3045188H1   1739   2025       159   977605T1   1764   2179       159   5299216H1   1761   2001       159   977605H1   1764   2075       159   977605R1   1764   2143       159   3028782T6   1767   2194       159   3487465H1   1765   2054       159   462261H1   1817   2014       159   5170457H1   1823   2011       159   3683935H1   1823   2109       159   5803204H1   1826   2024       159   979691H1   1836   2120       159   g434193   1852   2026       159   3013979H1   1877   2169       159   921802H1   1888   2214       159   920885H1   1888   2204       159   5278931H1   1996   2214       159   058768H1   1998   2201       159   3045908H1   2013   2281       159   g2787073   2048   2207       160   g6086623   4494   4929       160   g2569620   4498   4936       160   g2754538   4500   4930       160   3022465H1   4515   4743       160   2448511H1   4519   4765       160   3760535H1   4524   4835       160   2640174H1   4524   4780       160   517085T6   4532   4894       160   5338680H1   4533   4719       160   735966R1   4536   4961       160   735966H1   4536   4780       160   1592447H1   4536   4749       160   1591916H1   4536   4740       160   5022872H1   4538   4830       160   g3253612   4546   4951       160   g3843392   4553   4929       160   g3843398   4554   4929       160   g4080952   4558   4936       160   3814494H1   4561   4861       160   2009809H1   4568   4749       160   g1271129   4575   4962       160   g1068131   4586   4910       160   2096906H1   4584   4854       160   2603246H1   4589   4881       160   g829798   4598   4943       160   g1068126   4598   4905       160   2006525H1   4597   4764       160   628342H1   4598   4863       160   1943588H1   4599   4852       160   g1044543   4602   4916       160   g777642   4610   4930       160   2685217H1   4607   4864       160   1365826H1   4615   4876       160   1365826R1   4615   4941       160   1365826T1   4615   4910       160   6371841H1   4621   4912       160   g1525497   4622   4933       160   5704503H1   4643   4918       160   4060183H1   4644   4927       160   1416579H1   4644   4915       160   1416547H1   4644   4908       160   4569760H2   4658   4921       160   3721940H1   4651   4962       160   g852574   4663   4906       160   g1023637   4669   4890       160   3712504H1   4671   4971       160   4825195H1   4672   4924       160   g985536   4678   4902       160   3568055H1   4680   4986       160   g2819448   4690   4932       160   g1024915   4694   4924       160   g4125247   4702   4939       160   4254485T8   4705   4940       160   5714026H1   4707   4991       160   770816R1   4713   5202       160   770816H1   4713   4968       160   1901241H1   4719   4976       160   1898941H1   4719   4974       160   g2903316   4724   4936       160   6715734H1   4726   4923       160   2897118H1   4729   4947       160   986309R1   4747   5145       160   986309H1   4747   5067       160   986309T1   4747   5241       160   g2837150   4762   5278       160   2027901H1   4764   5060       160   2654529H1   4773   4942       160   g3057209   4773   5234       160   5871918H1   4775   5080       160   g3735780   4799   5287       160   g5424599   4804   5282       160   g4598235   4806   5285       160   g4684860   4807   5285       160   g5367615   4807   5287       160   g4437224   4810   5285       160   4790610H1   4810   4940       160   g3412956   4812   5285       160   1212364H1   4813   5002       160   g4153588   4815   5285       160   g3050313   4816   5285       160   4340133H1   4817   5098       160   g4071897   4821   5290       160   g4762931   4823   5287       160   g4393968   4823   5287       160   6872074H1   4828   5313       160   g3308744   4829   5293       160   g3887256   4843   5287       160   g5811461   4843   5285       160   g4372712   4842   5285       160   g3597624   4846   5291       160   g3869879   4848   5286       160   4201581H1   4848   4946       160   g4081955   4853   5290       160   g3840562   4854   5286       160   g5809990   4854   5270       160   4129277H2   4854   5127       160   g5904237   4855   5278       160   g4969558   4856   5276       160   g4292330   4857   5288       160   g5849107   4857   5291       160   3621721H1   4856   5149       160   g4987769   4863   4936       160   347302H1   4867   5086       160   5408358H1   4872   5129       160   g3896761   4879   5287       160   g3279962   4892   5285       160   g4244340   4895   5285       160   g3658851   4903   5285       160   g4983854   4906   5285       160   5388769H1   4911   5186       160   6215533H1   4912   5285       160   5388768H1   4911   5186       160   g2816194   4920   5285       160   g5547017   4930   5287       160   g4089434   4932   5285       160   g1484540   4934   5285       160   1739863R6   4940   5237       160   1739863H1   4940   5170       160   4207604H1   4940   5116       160   5578672H1   4950   5211       160   1689363H1   4952   5138       160   2606160H1   4957   5207       160   2007185H1   4999   5202       160   g795782   5014   5285       160   6613821H1   5033   5165       160   4772611H1   5036   5285       160   4923819H1   5037   5285       160   g1188096   5040   5232       160   g2540711   5047   5403       160   g2022546   5046   5285       160   3445038H1   5046   5257       160   g5447532   5058   5345       160   3486818H1   5071   5285       160   g1479409   5084   5403       160   g1064422   5084   5269       160   g2821180   5087   5406       160   g1543936   5098   5408       160   2769102H1   5102   5271       160   g815309   5131   5403       160   g4893445   5128   5293       160   1923809H1   5134   5289       160   g29237   5139   5403       160   g1548590   5143   5406       160   g1202222   5146   5319       160   g3202374   5145   5288       160   g1665338   5149   5271       160   g1228664   5151   5285       160   7336808H1   5161   5339       160   2807655H1   5176   5279       160   1741769R6   5216   5354       160   1741769H1   5216   5339       160   1741195H1   5216   5339       160   1741769T6   5216   5302       160   g810674   5258   5404       160   g944127   5261   5394       160   4125568H1   4959   5163       160   g6464630   4967   5403       160   g1501886   4969   5294       160   4537948H1   4971   5252       160   4699937H1   4974   5159       160   g3038275   4981   5352       160   g1664143   4988   5339       160   1283369T6   4991   5295       160   1283369F6   4997   5350       160   1283369H1   4997   5277       160   056712H1   3755   4005       160   g829797   3765   4195       160   g852673   3767   4118       160   g775984   3767   4023       160   2967706H1   3805   4117       160   2539208H1   3812   4049       160   2443012H1   3817   4068       160   988892H1   3821   4063       160   2291992H1   3821   4048       160   4178688H1   3822   4086       160   1833818H1   3831   4088       160   6368013H1   3838   4152       160   2134837H1   3858   4104       160   5657126H1   3862   4145       160   g1665244   3871   4196       160   2288056H1   3873   4122       160   3740644H1   3903   4101       160   4714119H1   3904   4202       160   5352624H1   3913   4159       160   g1272802   3939   4460       160   6388078H1   3935   4238       160   3056718H1   3936   4205       160   g1044542   3941   4227       160   g1525554   3942   4231       160   g1625857   3970   4363       160   g1977647   3972   4318       160   056202H1   3982   4165       160   1259638H1   4000   4255       160   1259479F1   4000   4527       160   1259638F1   4000   4492       160   4383783H1   4010   4274       160   g1440113   4032   4192       160   6856341H1   4048   4572       160   g944630   4051   4388       160   5950009H1   4064   4381       160   g1240400   4066   4202       160   5387777H1   4076   4340       160   4342232H1   4088   4243       160   3022534H1   4093   4335       160   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4488   4941       160   g6086615   4491   4929       160   g1469867   1   5285       160   7086887H1   1   516       160   3287254H1   8   248       160   g273451   81   456       160   g1057697   186   523       160   3903676H1   189   440       160   g434410   279   603       160   3698379H1   455   747       160   5643561H1   480   749       160   4960333H1   502   761       160   g2277376   522   931       160   5090209H1   584   831       160   525014H1   602   841       160   525014R6   602   845       160   162678H1   741   981       160   6332784H1   1028   1600       160   3872857H1   1053   1308       160   525014T6   1109   1587       160   g395681   1123   1460       160   g6086016   1169   1629       160   g2779861   1232   1536       160   g1057696   1305   1626       160   4806634H1   1367   1619       160   5864502H1   1415   1701       160   156350R6   1439   1717       160   156350H1   1439   1616       160   5502785H1   1447   1672       160   4696595H1   1488   1665       160   g3701554   1534   1625       160   3825811H1   1546   1852       160   3825859H1   1546   1803       160   g1880691   1643   1799       160   g691032   1722   2124       160   g573544   1722   2062       160   g708679   1722   2043       160   6610185H1   1732   2259       160   1257342H1   1775   2018       160   2532881H1   1859   2182       160   g3416512   1887   2323       160   2722041H1   1915   2155       160   7191660H2   2044   2659       160   1804357H1   2065   2340       160   1806920H1   2065   2330       160   5602907H1   2082   2351       160   2830493H1   2090   2352       160   5542070H1   2153   2364       160   7171003H1   2174   2627       160   3109807H1   2202   2486       160   1439519H1   2327   2604       160   086846H1   2351   2417       160   6743069H1   2379   2880       160   5092027H1   2401   2674       160   3595466H1   2452   2678       160   3120055H1   2600   2875       160   4028493H1   2623   2861       160   6918376H1   2649   3021       160   833106R1   2689   3287       160   833106H1   2689   2941       160   g1068214   2708   3062       160   g1068219   2719   3068       160   386166H1   2777   3061       160   3163260H1   2798   3086       160   6490409H1   2830   3338       160   2122144H1   2851   3109       160   g1484539   2902   3404       160   3120466H1   2924   3191       160   g1062289   2966   3116       160   4178517H1   3003   3265       160   2505067H1   3016   3259       160   2505367H1   3016   3245       160   7034903H1   3023   3630       160   g1259693   3042   3222       160   7315867H1   3070   3471       160   5370265H1   3126   3359       160   g2022547   3176   3463       160   2707316H1   3197   3502       160   2894887H1   3197   3446       160   g796865   3229   3473       160   3523324H1   3238   3589       160   g1479499   3257   3560       160   g1548639   3257   3626       160   g943258   3257   3559       160   g1067965   3257   3571       160   5371320H1   3264   3396       160   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2772059H1   3629   3889       160   2960117H1   3636   3937       160   g1969950   3641   4014       160   g1947603   3654   3817       160   5002748H1   3687   3844       160   4062722H1   3721   4010       160   7107841H1   3740   4101       160   g1024914   3749   4082       160   5546516H1   3750   3928       161   g1219848   2061   2389       161   5406570H1   2061   2269       161   5451646H1   2063   2243       161   g2000931   2063   2316       161   6517527H1   2068   2140       161   4426132H1   2214   2389       161   3468616H1   2068   2331       161   5451867H1   2073   2323       161   6441810H1   2085   2530       161   7076574H1   2068   2555       161   70346502D1   2070   2384       161   4772876H1   2073   2352       161   805234H1   2073   2269       161   6763871H1   2078   2569       161   7079082H1   2081   2565       161   6996593H1   2081   2575       161   5693746H1   2096   2319       161   4760746H1   2081   2371       161   4760754H1   2082   2371       161   3466673H1   2085   2180       161   3955495H1   2106   2385       161   3945275H1   2213   2490       161   6891449H1   2214   2316       161   4195061H1   2215   2518       161   3781801H1   2096   2379       161   2570710H1   2219   2467       161   4690839H1   2097   2352       161   g683009   2100   2389       161   5682533H1   2099   2378       161   5575001H1   2106   2253       161   1505114H1   2127   2344       161   4091605H1   2114   2422       161   5832027H1   2123   2377       161   5401121H1   2219   2429       161   5388353H1   2219   2494       161   2704568H1   2127   2350       161   5834524H1   2132   2405       161   7103141H1   2155   2265       161   4315855H1   2139   2389       161   5834443H1   2139   2343       161   664952H1   2142   2367       161   1992383H1   2143   2344       161   3468549H1   2219   2487       161   4256959H1   2161   2426       161   6993692H1   2148   2573       161   534218H1   2156   2414       161   664384H1   2156   2421       161   4713170H1   2177   2420       161   663504H1   2219   2347       161   2247226H1   2184   2445       161   2245218H1   2184   2452       161   531779H1   2184   2426       161   2042975H1   2184   2450       161   6741472H1   2204   2750       161   4490622H1   2204   2747       161   4063891H1   2241   2390       161   5960583H1   2242   2800       161   3781135H1   2190   2495       161   7071563H1   2208   2569       161   6891449J1   2214   2315       161   554022H1   2202   2438       161   4532533H1   2273   2435       161   6739828H1   2219   2805       161   4691246H1   2208   2424       161   3470773H1   2209   2454       161   5967249H1   2224   2744       161   6765073J1   2275   2692       161   666644H1   2212   2384       161   5579450H1   2212   2365       161   1618034H1   2212   2418       161   7088449H1   1   552       161   7073146H1   170   726       161   6784888H1   439   960       161   6882093H1   479   990       161   6882161H1   479   992       161   6766193H1   480   961       161   6977268H1   504   925       161   7230035H1   585   1180       161   7075220H1   629   1180       161   1485846F6   646   910       161   g1975476   664   884       161   1486578H1   676   912       161   5968971H1   685   1143       161   1476202H1   762   924       161   865333H1   780   910       161   7193726H2   786   1230       161   6891594J1   877   1507       161   4031012H1   981   1202       161   6446990H1   988   1498       161   g6133077   1024   1478       161   3941692H1   1056   1337       161   4772703H1   1105   1361       161   g668477   1117   1416       161   g870488   1118   1438       161   g698735   1117   1456       161   g669863   1117   1378       161   g768138   1143   1468       161   g1062711   1172   1485       161   g875401   1174   1475       161   1617044H1   1178   1392       161   5781411H1   1182   1459       161   532473H1   1193   1433       161   3945042H1   1194   1480       161   4758039H1   1193   1440       161   3466710H1   1203   1464       161   5860513H1   1221   1490       161   6770719H1   1238   1355       161   5656957H1   1238   1487       161   5204013H1   1238   1505       161   4254309H1   1257   1459       161   5578708H1   1259   1489       161   5013751H1   1264   1491       161   4019484H1   1278   1543       161   6063870H1   1281   1564       161   3471004H1   1298   1570       161   5206590H1   1304   1546       161   6746950H1   1304   1831       161   5857309H1   1303   1576       161   5968610H1   1308   1851       161   g2012398   1308   1663       161   3469688H1   1317   1580       161   5857943H1   1354   1638       161   7070664H1   1508   1855       161   6416952H1   1519   1822       161   6894069H1   1526   2069       161   5013816H1   1533   1783       161   533363H1   1538   1775       161   4761822H1   1540   1812       161   2904083H1   1554   1850       161   4306790H1   1577   1687       161   1991775H1   1578   1797       161   4337965H1   1580   1827       161   5833237H1   1585   1750       161   3469095H1   1597   1855       161   4316943H1   1635   1803       161   5876761H1   1647   1931       161   5015603H1   1648   1813       161   2963466H1   1724   2028       161   4316745H1   1724   2020       161   4315891H1   1744   2022       161   4091142H1   1749   2023       161   4753020H1   1751   2007       161   6518022H1   1757   1838       161   4316639H1   1772   2046       161   2705655H1   1800   2083       161   5961924H1   1790   2298       161   4695643H1   1820   2036       161   2707689H1   1896   2192       161   4342587H1   1979   2088       161   6770516H1   1988   2569       161   4342620H1   1992   2246       161   6882093J1   1996   2568       161   6886296J1   2001   2481       161   7253724H1   2024   2531       161   2423349H1   2027   2272       161   4316734H1   2032   2342       161   4256579H1   2036   2265       161   6763709H1   2023   2568       161   6771870J1   2031   2578       161   4772777H1   2046   2149       161   6740527H1   2041   2522       161   4691683H1   2058   2293       161   5205527H1   2058   2293       161   5575884H1   2060   2251       161   3466389H1   2061   2306       161   5955966H1   2315   2492       161   5205250H2   2316   2479       161   4776444H1   2311   2566       161   6517485H1   2347   2825       161   2292420H1   2408   2647       161   6552809H1   2494   2887       161   6552988H1   2494   2887       161   6552963H1   2494   2887       161   4619412H1   2571   2791       161   4775811H1   2566   2801       161   g1194667   2595   2887       161   5405506H1   2604   2840       161   4314575H1   2614   2895       161   1289406H1   2619   2877       161   g1190424   2629   2887       161   g1071637   2650   2858       161   4773458H1   2681   2887       161   4756793H1   2681   2826       161   4341024H1   2694   2874       161   4298642H1   2694   2887       161   1596024H1   2694   2887       161   552947H1   2699   2887       161   5371715H1   2713   2886       161   5447990H2   2737   2906       161   667160H1   2739   2887       162   668331H1   2789   2883       161   5657894H1   2793   2887       162   6127609T8   1   306       162   6959915H1   108   561       163   5914133H1   1   282       163   5914133F6   1   630       163   5321108F9   1   537       163   5914133F8   1   391       163   5914133T6   52   609       164   5911540F8   1   460       164   5911540H1   1   250       164   5911540T8   78   570       165   5905252F8   1   497       165   5905252F6   35   576       165   5905252H1   35   313       165   5905252T6   376   821       166   4020439F8   1   391       166   2773907F6   1   173       166   2773907H1   1   146       166   4020439H1   1   115       166   4020439T8   6   503       166   2773907T6   129   435       166   g820143   313   435       167   6919815H1   1   89       167   6100456H1   12   272       167   5840353H1   19   294       167   3055393H1   1   77       167   4528621H1   3   250       167   482608H1   28   255       167   3477717H1   3   119       167   2472906H1   3   240       167   2444808H1   4   234       167   3167552H1   6   74       167   2615139H1   9   226       167   3199504H1   13   106       167   2455307H1   10   197       167   2428111H1   11   248       167   3458104H1   12   270       167   3596416H1   12   327       167   2453021H1   12   213       167   4298232H1   12   271       167   g1956274   35   272       167   g1880655   12   256       167   3458004H1   12   279       167   5694361H1   12   286       167   g2026069   15   306       167   477069H1   15   278       167   1918196H1   15   293       167   5587272H1   18   282       167   4997069H1   18   284       167   1551008H1   18   211       167   5347226H1   19   275       167   6819761H1   19   616       167   5117110H1   20   289       167   3447366H2   19   275       167   2445594H1   19   249       167   4622780H1   19   288       167   2111981H1   20   282       167   552401H1   20   258       167   2461616H1   19   197       167   5379341H1   19   275       167   3820792H1   20   310       167   4347282H1   21   264       167   1322148H1   22   257       167   4222393H1   23   304       167   4223817H1   22   336       167   5843163H1   20   289       167   3657588H1   23   218       167   4721051H1   25   271       167   4790883H1   21   278       167   5377733H1   24   280       167   g1638522   24   353       167   3596245H1   25   318       167   781633H1   25   270       167   5374490H1   26   286       167   2161371H1   31   277       167   3597248H1   32   267       167   2934534H1   36   172       167   4386768H1   37   326       167   3399780H1   126   191       167   6399947H1   126   272       167   6897251H1   166   634       167   4431669H2   173   312       167   g1779781   330   692       167   g826488   330   717       167   4794163H1   346   595       167   4880181H1   349   609       167   g889083   366   638       167   6819761J1   396   1013       167   g916666   408   513       167   4249123H1   452   539       167   6481276H1   584   800       167   4191747H1   624   709       167   638004H1   648   753       167   1452178H1   676   753       167   g2615681   692   753       167   g1665347   697   753       167   g2558364   697   753       168   6795278H1   184   698       168   6796542H1   1   546       168   6796380H1   8   563       168   6795463H1   8   523       168   3941984H1   26   315       168   g1260435   112   287       168   6798249H1   184   635       168   6791366H1   238   804       168   6790685H1   239   699       168   1242854H1   698   885       169   2904954T6   1   522       169   4739603H1   11   303       169   5614905H1   213   492       169   g2006850   277   557       169   2008385H1   464   562       169   2014576T6   464   523       169   2014576H1   464   586       169   3294227T6   223   557       169   2014576R6   464   553       169   1716729H1   501   556       170   3966795F6   1   365       170   3966795H1   1   267       170   3274864H1   14   268       170   3966795T6   23   659       171   3033193F6   1   272       171   3033193H1   1   216       171   3033193T6   129   445       172   5908301F8   1   519       172   5908301H1   1   311       172   6271267H2   24   492       172   5908301T9   248   586       173   5907939T9   1   517       173   5907939F8   1   551       173   5907939H1   1   310       173   6271008H2   10   483       174   5912415F8   1   376       174   5912415H1   1   299       174   5912415F6   12   565       174   5912415T9   66   535       175   4119207F6   1   336       175   4119207T6   1   336       175   4119207H1   1   175       176   5905477F6   1   564       176   5905477H1   1   271       176   5905477T9   411   932       177   5907791F8   1   360       177   5907791H1   1   280       177   5907791F6   1   305       177   5907791T9   155   682       177   5907791T6   248   733       178   4770137H1   1   144       178   5564253H1   2   235       178   606101H1   1   169       178   4650476H1   1   271       178   592893H1   2   130       178   781453H1   17   276       178   2793564H1   28   317       178   888943H1   151   287       178   6589872H1   194   698       178   2190828T6   207   699       178   1503654H1   225   491       178   4200384H1   388   662       179   5911540F8   1   460       179   5911540H1   1   250       179   5911540T9   27   568       179   5911540T8   78   569       180   g4325750   1   103       180   4290049F6   1   353       180   4290049H1   1   124       180   5493752H1   172   444       181   6729842H1   1   412       181   5401350H1   1   105       181   6057617H1   56   643       181   5401350T9   82   666       181   g3214092   406   782       181   3524102H1   479   779       182   7030475H1   1   524       182   7030327H1   3   376       183   5271230H1   1485   1748       183   240641H1   1485   1657       183   2948213H1   1486   1768       183   g1291432   1492   1896       183   353018H1   1496   1708       183   349497H1   1496   1680       183   g2276981   1533   1859       183   6804414J1   1537   2080       183   3499213H1   1537   1841       183   g2537462   1542   1981       183   1551014H1   1549   1763       183   808076H1   1554   1769       183   2307944H1   1557   1762       183   g2222952   1559   1851       183   2804178H1   1565   1852       183   6801505J1   1566   1881       183   g888315   1569   1927       183   g2056786   1570   2035       183   6901949H1   1   518       183   4183549H1   91   259       183   6980581H1   104   452       183   5839778H1   107   359       183   808102H1   172   406       183   4323252H1   224   469       183   g2003585   258   517       183   g900863   361   447       183   6979927H1   437   785       183   582110H1   487   739       183   3323870H1   499   778       183   2115479R6   515   913       183   4779981H1   538   793       183   1818066H1   552   808       183   1818066F6   552   893       183   g4438745   584   797       183   5450643H1   614   851       183   7066292H1   636   1078       183   3538459H1   648   855       183   2115479H1   655   913       183   4090518H1   669   864       183   6077621H1   686   797       183   4947628H1   702   832       183   4759655H1   713   993       183   1699317H1   715   931       183   4303659H1   740   991       183   591324R6   752   1180       183   591324H1   752   975       183   591174H1   752   955       183   5998787H1   772   1179       183   1672160H1   778   991       183   1672152H1   778   991       183   4694794H1   784   1045       183   2972664H2   782   1078       183   1718469H1   795   1006       183   1718480H1   795   998       183   3270027H1   802   1059       183   927251R1   806   1419       183   927251H1   806   1058       183   g1986696   809   992       183   5102583H1   821   1068       183   7094153H1   822   1103       183   5948962H1   843   1137       183   1571253H1   845   1013       183   5371617H1   860   1101       183   3784982H1   895   1195       183   6980693H1   897   1257       183   4770011H1   901   1166       183   1695218H1   903   1119       183   3697260H1   962   1179       183   1331860H1   981   1179       183   g1644929   1018   1179       183   g1880762   1046   1163       183   6804414H1   1065   1601       183   1722938H1   1074   1327       183   5451478H1   1104   1348       183   1500938H1   1108   1304       183   5950551H1   1120   1413       183   3376840H1   1121   1367       183   5950519H1   1121   1320       183   4193013H1   1121   1438       183   4556920H1   1124   1261       183   g1046489   1143   1470       183   g1046497   1143   1455       183   1283213H1   1202   1390       183   1283213F6   1202   1658       183   4225341H1   1206   1487       183   6714888H1   1214   1778       183   1346996H1   1259   1485       183   1348304H1   1259   1481       183   1348303H1   1259   1481       183   6954001H1   1261   1764       183   g2107139   1290   1761       183   4720048H1   1308   1583       183   1720725H1   1328   1538       183   1722139H1   1328   1553       183   5970873H1   1334   1887       183   4693221H1   1344   1601       183   2571078H1   1343   1591       183   6121921H1   1351   1904       183   4215895H1   1363   1625       183   g2188904   1372   1857       183   g1646485   1374   1468       183   004844H1   1386   1669       183   6313521H1   1386   1950       183   2427675H1   1392   1622       183   5478565H1   1401   1608       183   5480821H1   1402   1635       183   5483319H1   1402   1501       183   5322481H1   1403   1641       183   5948083H1   1409   1714       183   1996549H1   1415   1554       183   1996549R6   1415   1751       183   5405753H1   1434   1586       183   5293687H2   1434   1680       183   935623R1   1465   1989       183   936611H1   1465   1767       183   935623H1   1465   1708       183   3409340H1   1473   1629       183   1819126H1   1480   1748       183   1819126F6   1480   1979       183   g2053565   1480   1888       183   g2209615   1481   1929       183   5879274H1   1482   1754       183   4570795H1   2032   2292       183   g1046490   2044   2349       183   1416514H1   2046   2291       183   4193606H1   2048   2324       183   g1858344   2051   2338       183   5021880T1   2055   2300       183   1281333H1   2058   2294       183   2222539H1   2060   2298       183   2222539F6   2060   2338       183   2222539T6   2061   2300       183   5341229H1   2066   2265       183   2659133H1   2067   2280       183   g3871000   2069   2338       183   g4524292   2077   2340       183   g1858279   2081   2342       183   1301182H1   2082   2343       183   2256984H1   2105   2338       183   6498361H1   2106   2338       183   2574115H1   2106   2337       183   2273027H1   2107   2310       183   g4899838   2119   2346       183   g3051313   2128   2338       183   3142955H1   2158   2340       183   2878176H1   2164   2339       183   3271618H1   2181   2337       183   g2958193   2190   2340       183   2247168H1   2197   2344       183   2246357H1   2225   2339       183   2247256H1   2254   2339       183   3812939H1   2274   2338       183   5103254H1   2283   2339       183   7064872H1   1575   2105       183   6818666H1   1579   1827       183   4726069H1   1581   1854       183   g4662834   1586   2036       183   g1182379   1589   1804       183   g4874675   1588   2033       183   5661656H1   1589   1849       183   6868292H1   1591   2060       183   3706916H1   1605   1904       183   g2208265   1633   2033       183   2424009H1   1649   1907       183   1470490H1   1663   1849       183   797359H1   1664   1903       183   3245252H1   1669   1936       183   4781187H1   1671   1939       183   g1688385   1690   2024       183   1413737H1   1712   1978       183   g2279273   1737   2033       183   g1146486   1741   1926       183   g1644873   1745   2038       183   g888316   1749   2033       183   099594H1   1754   1972       183   3996904H1   1754   1913       183   3916961H1   1759   2048       183   3917411H1   1759   2036       183   6801505H1   1759   2125       183   910557H1   1785   1850       183   2921975H1   1788   2061       183   5003193H1   1790   2060       183   g1442965   1791   2033       183   5018209H1   1797   1957       183   3246162H1   1797   2036       183   1344364H1   1798   2013       183   1344388H1   1798   2036       183   5928305H1   1801   2111       183   683816H1   1805   2040       183   5451469H1   1808   2048       183   2402750H1   1811   1919       183   1851976H1   1816   2034       183   3702341H1   1825   2066       183   2963989H1   1819   2096       183   g1441785   1826   2091       183   5777701H1   1828   2105       183   g3898324   1831   2042       183   1819126T6   1843   2298       183   1484988H1   1847   2095       183   2106229T6   1851   2298       183   g4762523   1859   2033       183   g4620507   1860   2039       183   1964264H1   1872   2033       183   1964264R6   1872   2040       183   1964264T6   1872   2002       183   g2945963   1876   2340       183   4696529H1   1885   2095       183   g4564438   1886   2339       183   g3741810   1886   2341       183   2913361H1   1889   1977       183   6862628H1   1894   2033       183   g3742173   1905   2340       183   g4083420   1910   2338       183   g3678703   1912   2342       183   6327536H1   1912   2340       183   g2910125   1916   2341       183   g3308355   1919   2342       183   2556534H1   1933   2178       183   g3922126   1935   2338       183   g4083267   1936   2338       183   g4083582   1951   2338       183   g1646486   1958   2340       183   g2902884   1998   2339       183   953800T1   2000   2301       183   g895530   2006   2338       183   953800H1   2012   2296       183   953800R1   2012   2338       183   4977128H1   2022   2281       183   g1046498   2026   2338       184   3497339T6   1   492       184   g1060678   211   436       184   6258709H1   213   444       184   2570554H1   239   484       184   2570554R6   239   634       184   6200064H1   459   921       184   g734035   473   792       184   1400373F6   474   1007       184   1400373H1   474   733       184   g2167556   488   897       184   6389638H1   542   823       184   6344656H1   598   885       184   6344688H1   606   888       184   6328470H1   664   1018       184   g4148622   723   1072       184   6338470H1   793   1018       184   3784456H1   876   1200       184   3781165H1   943   1049       184   g733951   1031   1403       184   g734304   1149   1333       184   3554877H1   1   294       185   4111213H1   1   243       185   5624259R8   1   350       185   3139315H1   126   427       185   5545642H1   130   363       185   3141553H1   139   412       185   g564381   139   369       185   987690H1   154   424       185   4833664H1   169   319       185   116132H1   214   403       185   3691576H1   244   399       185   1667502F6   265   644       185   3283023H1   267   528       185   3959293H2   290   564       185   2675859H1   347   596       185   2676108H1   347   594       185   2676122H1   347   591       185   5301231H1   387   569       185   1598950H1   387   567       185   2352332H1   456   626       185   5038795H1   459   689       185   2534205H1   459   705       185   3252993H1   464   711       185   1742040H1   490   721       185   4592168H1   498   648       185   g29244   1   255       185   2635196H1   1   249       186   6428735H1   867   1184       187   2525961F6   1   449       187   2525961H1   2   243       187   6560586H1   64   613       187   g1357976   235   523       187   2525961T6   253   677       187   g3245391   532   894       188   g1277774   1   322       188   g2186366   1   442       188   5533987H1   1   268       188   5320656H1   23   287       188   985175H1   22   276       188   5478163H1   23   303       188   5476045H1   23   301       188   5325512H1   23   291       188   5479637H1   23   159       188   7289635H1   196   762       188   6559813H1   603   1192       188   6858544H1   732   1210       188   5929387H1   845   1124       188   5778630H1   845   1114       188   4017917H1   851   1115       188   5206602H1   905   1185       188   4776841H1   1078   1295       188   6314480H1   1093   1213       189   70395867D1   1   542       189   2525961F6   4   452       189   2525961H1   5   246       189   70395557D1   40   642       189   70397658D1   59   589       189   6560586H1   67   616       189   70396453D1   72   478       189   70396167D1   119   654       189   70396459D1   118   585       189   70396121D1   192   481       189   g1357976   238   526       189   2525961T6   256   680       189   70396053D1   260   679       189   70396654D1   278   768       189   70395799D1   279   680       189   70395826D1   279   680       189   70397446D1   279   676       189   70397464D1   292   676       189   70394880D1   293   746       189   70397538D1   293   768       189   70396886D1   292   564       189   70395257D1   293   751       189   70395475D1   298   676       189   70395110D1   307   676       189   70395982D1   312   680       189   70397165D1   352   676       189   70396476D1   483   676       189   g3245391   535   897       190   2402420H1   4083   4349       190   1340154H1   4091   4319       190   2791792R6   4108   4475       190   2079255T6   4053   4414       190   g2884989   4111   4451       190   g1189027   4060   4458       190   2412005H1   4120   4352       190   g2354871   4123   4450       190   g2350487   4133   4457       190   g5662773   4156   4466       190   g5639034   4061   4453       190   g816766   4165   4460       190   3041690H1   4157   4451       190   977648R1   4063   4431       190   g1193159   4182   4458       190   977648T1   4063   4415       190   977648H1   4063   4386       190   978306H1   4063   4319       190   g2051455   4184   4460       190   5280171H1   4187   4442       190   g3886427   4204   4347       190   1381539H1   4210   4457       190   552969H1   4210   4458       190   2760603H1   4210   4453       190   g3647431   4212   4454       190   1917113H1   4245   4457       190   4787118H1   4063   4343       190   g892648   4259   4458       190   g2229339   4272   4456       190   1942308H1   4292   4461       190   1942315H1   4292   4461       190   1941519H1   4327   4457       190   5007945H1   4389   4457       190   1979718T6   4069   4419       190   240941H1   4066   4182       190   1979718R6   4069   4452       190   1979718H1   4069   4174       190   g682911   4083   4457       190   4465894H1   3788   4048       190   2819748T6   3815   4417       190   3445036H1   3816   4085       190   6735779H1   3822   4433       190   2279717T6   3824   4418       190   3720714H1   3832   4020       190   4828820H1   3844   4119       190   1369089H1   3847   4077       190   1833781H1   3850   4130       190   1965941T6   3865   4422       190   2559627H1   3868   3984       190   2285422H1   3881   4083       190   5483595H1   3904   4084       190   5322590H1   3904   4157       190   g892647   3925   4226       190   4030718T6   3927   4433       190   6476518H1   3939   4460       190   5013604H1   3940   4221       190   2245864H1   3949   4223       190   2301055T6   3955   4416       190   2292517T6   3967   4419       190   g5444044   3982   4452       190   g5436138   3984   4465       190   g3427279   3990   4459       190   6612393H1   4008   4444       190   g6044228   4017   4457       190   g4650722   4020   4460       190   g5109970   4020   4459       190   g2656315   4021   4456       190   g3277136   4026   4453       190   615565H1   4040   4314       190   g1192291   4042   4126       190   g4002109   4047   4457       190   933913R1   3411   3813       190   933913H1   3411   3683       190   3056157H1   3442   3663       190   6853558H1   3446   3911       190   1267149F1   3446   3913       190   1267149H1   3446   3659       190   1413302H1   3453   3663       190   4092170H1   3473   3749       190   1961533H1   3491   3760       190   1961533R6   3491   3663       190   464481H1   3491   3663       190   g2141846   3498   3817       190   1373705H1   3514   3748       190   3961925H1   3584   3641       190   5285788H1   3610   3739       190   2301055R6   3663   3918       190   1965941R6   3692   4177       190   3991277H1   3692   3981       190   1923669H1   3692   3923       190   2301055H1   3692   3919       190   1965941H1   3692   3916       190   1833543H1   3692   3890       190   927374H1   3696   3966       190   4511590H1   3696   3931       190   6517421H1   3703   4233       190   1551235T6   3716   4417       190   2674692H1   3731   3965       190   2288745H1   3734   3969       190   4311971H1   3740   4037       190   6118751H1   3753   4264       190   6119992H1   3753   4329       190   6126908H1   3753   4255       190   g4629955   3755   3976       190   2157206H1   3772   4029       190   71226132V1   2110   2748       190   g3092783   2129   2371       190   70863741V1   2130   2716       190   2079255F6   2130   2513       190   2079255H1   2130   2367       190   6167128H1   2173   2701       190   70856205V1   2176   2782       190   6482958H1   2180   2277       190   70149143V1   2191   2726       190   5372491H1   2218   2355       190   71226268V1   2225   2707       190   g5234288   2245   2718       190   g389953   2249   2637       190   g769298   2249   2567       190   579131H1   2257   2504       190   579198H1   2257   2503       190   g3308511   2277   2721       190   083837H1   2276   2489       190   g4534587   2287   2674       190   g4188225   2296   2712       190   3380281H1   2315   2563       190   g3870352   2316   2713       190   3086136H1   2326   2531       190   g3228885   2328   2714       190   g2541163   2344   2721       190   g3884137   2348   2721       190   1819282F6   2363   2713       190   1819282H1   2363   2618       190   6492522H1   2369   2978       190   g875856   2373   2721       190   4176829H1   2371   2638       190   4176860H1   2371   2634       190   70864219V1   2391   2984       190   1819282T6   2405   2666       190   g819854   2421   2722       190   g561301   2418   2717       190   70145373V1   2426   2719       190   4670647H1   2442   2720       190   70855556V1   2465   3045       190   6844792H1   2498   2713       190   3724780H1   2530   2833       190   4822074H1   2542   2658       190   g794631   2547   2721       190   g708639   2550   2858       190   70146025V1   2559   3033       190   g691143   2573   2932       190   g3096354   2577   2716       190   70855455V1   2586   3160       190   4259987H1   2619   2702       190   6545104H1   2633   2713       190   g874560   2650   2722       190   4612918H1   2655   2837       190   4144852H1   2661   2942       190   985756R1   2688   3124       190   985756H1   2688   2979       190   5570469H1   2726   2925       190   6323423H1   2731   2988       190   985756R6   2731   2808       190   6835880H1   2740   3320       190   6742687H1   2750   3268       190   5350646H1   2785   3035       190   71227966V1   2797   3000       190   3091359H1   2803   3085       190   985756T6   2829   3266       190   g1198696   2825   3127       190   g3416884   2871   3306       190   3695945H1   2881   3163       190   5377540H1   2893   3153       190   g5363556   2895   3301       190   g618368   2909   3252       190   1305496T6   2927   3263       190   1551235R6   2945   3460       190   g530325   2945   3279       190   1551235H1   2945   3153       190   4311884H1   2976   3299       190   3659340H1   2985   3090       190   6125263H1   2989   3523       190   1984710H1   2986   3241       190   6124418H1   2989   3449       190   1008121H1   2998   3264       190   4402539H1   3002   3244       190   3518554H1   3018   3268       190   3345944H1   3032   3288       190   2819748F6   3075   3394       190   2819748H1   3077   3231       190   5660765H1   3096   3330       190   71225736V1   3132   3768       190   70857262V1   3133   3301       190   3785435H1   3154   3448       190   2279717H1   3183   3424       190   2279717R6   3183   3373       190   3469571H1   3207   3464       190   3356238H1   3209   3470       190   1805536H1   3212   3479       190   4160048H1   3214   3272       190   4942785H1   3234   3464       190   5585444H1   3235   3448       190   6269078H1   3252   3752       190   127072H1   3265   3456       190   7290084H1   3269   3683       190   7011256H1   3273   3657       190   g1893973   3278   3669       190   3992283H1   3291   3405       190   3793161H1   3292   3493       190   3325588H1   3301   3515       190   1008331H1   3310   3628       190   2637310F6   3313   3559       190   2637310H1   3313   3583       190   4623479H1   3314   3577       190   3769624H1   3325   3624       190   3725948H1   3328   3615       190   3433892H1   3345   3591       190   3919666H1   3349   3447       190   5732787H1   3359   3579       190   3741626H1   3360   3641       190   5874821H1   3380   3628       190   2292517H1   3382   3639       190   3559065H1   3393   3574       190   3778532H1   3404   3653       190   6520495H1   3409   3864       190   5510632F6   1   402       190   5510632H1   1   201       190   6780915H1   41   512       190   4782751H1   127   371       190   4782751F6   127   187       190   2987413H1   192   477       190   6064208H1   203   464       190   5336504H1   219   435       190   5337416H1   219   473       190   g1984325   271   504       190   7245264H1   286   854       190   g3239921   293   648       190   g2026718   299   586       190   7245164H1   302   823       190   g2816869   320   565       190   5510632R6   364   855       190   4760384H1   491   575       190   4760384F6   491   961       190   1305496F6   798   1267       190   70145686V1   798   1046       190   1305496H1   798   941       190   3617505H1   819   1021       190   70861843V1   870   1297       190   g669800   873   1196       190   g875546   874   1129       190   g874649   874   1176       190   70862573V1   891   1352       190   70856733V1   964   1627       190   g771213   1008   1060       190   7048824H1   1036   1629       190   71225244V1   1056   1612       190   3211592H1   1054   1289       190   093259H1   1079   1319       190   926880H1   1120   1361       190   6163034H1   1127   1456       190   70860860V1   1134   1758       190   1366443R6   1134   1468       190   1366443H1   1134   1383       190   70145966V1   1141   1461       190   6448764H1   1151   1743       190   4960778H1   1157   1430       190   71226715V1   1203   1824       190   71228090V1   1215   1654       190   70857707V1   1220   1774       190   4625867H1   1221   1490       190   71225380V1   1274   1804       190   70858188V1   1290   1935       190   70858768V1   1303   1903       190   70864189V1   1307   1926       190   6911209H1   1323   1505       190   70858579V1   1330   1996       190   70858582V1   1366   2008       190   71224853V1   1396   1993       190   5640688H1   1416   1664       190   71226033V1   1421   2078       190   7359924H1   1437   2064       190   70861230V1   1450   2100       190   71226687V1   1457   2020       190   70864162V1   1463   2044       190   70855640V1   1491   2187       190   71225393V1   1507   2076       190   70858387V1   1539   2062       190   70855984V1   1520   1993       190   70857482V1   1520   2169       190   70855452V1   1526   2133       190   70861672V1   1541   2227       190   70861119V1   1548   2117       190   71225746V1   1558   2147       190   70856590V1   1568   2109       190   4030718F6   1573   1963       190   4030718H1   1573   1845       190   70856815V1   1611   2119       190   2742284H1   1626   1887       190   71228652V1   1651   2284       190   70855093V1   1689   2167       190   70863689V1   1722   2354       190   6412316H1   1730   1956       190   70857818V1   1740   2409       190   70860827V1   1745   2302       190   70864783V1   1755   2382       190   70863459V1   1752   2053       190   70858650V1   1822   2413       190   6911209J1   1830   2415       190   70146804V1   1869   2439       190   70146516V1   1869   2409       190   70863645V1   1877   2491       190   71225144V1   1889   2474       190   71224913V1   1929   2422       190   70146682V1   1936   2394       190   060994H1   1936   2131       190   70861917V1   1973   2563       190   2791792F6   2000   2315       190   2791792H1   2000   2297       190   70857602V1   2001   2402       190   71225364V1   2013   2660       190   70858585V1   2015   2639       190   70854964V1   2015   2644       190   70860984V1   2016   2573       190   70855603V1   2033   2704       190   70858486V1   2050   2280       190   70858723V1   2050   2280       190   70861733V1   2048   2785       190   70855437V1   2059   2683       190   70858374V1   2060   2596       190   449622H1   2069   2240       190   71228093V1   2070   2326       190   71227893V1   2070   2351       190   71227986V1   2072   2687       190   71225556V1   2086   2744       190   70864357V1   2095   2685       190   70855127V1   2110   2729       191   2106710R6   1204   1492       191   71032896V1   417   928       191   2004771H1   381   441       191   71134281V1   928   1489       191   71130579V1   1003   1490       191   71129556V1   1016   1488       191   71133640V1   1088   1509       191   g3427925   591   1011       191   71133255V1   605   931       191   70643569V1   613   810       191   70639731V1   613   829       191   71133577V1   653   1365       191   71134103V1   657   1276       191   968289H1   143   416       191   7212584H1   176   467       191   g4735242   212   473       191   g4533052   255   473       191   2705278H1   1   210       191   2427258H1   8   163       191   991952H1   127   401       191   3070166H1   399   700       191   71131960V1   399   833       191   71130961V1   406   930       191   71133228V1   779   1472       191   2301872R6   503   813       191   g1263124   505   671       191   1965957R6   512   843       191   6742448H1   398   474       191   6843692H1   413   692       191   g3433649   563   1009       191   6883312H1   565   1039       191   70747238V1   585   712       191   1965957H1   512   685       191   2301872H1   512   686       191   6202014H1   534   931       191   g4113683   562   1004       191   70749633V1   562   749       191   71129118V1   690   1233       191   5769401H1   682   1308       191   g611097   808   1182       191   70641000V1   873   1297       191   2106710H1   1204   1488       191   2106710T6   1204   1448       191   861904H1   1343   1488       191   2709045F6   1441   1798       191   2709045H1   1442   1757       192   7345973H1   862   1457       192   7322101H1   921   1460       192   6850665H1   1366   1818       192   2654475F6   826   1310       192   2654475H1   826   1121       192   g831331   1984   2353       192   1787542H1   2005   2223       192   6930205H1   2259   2725       192   4020229H1   1952   2228       192   6894957J1   1752   2259       192   4152694H1   1653   1911       192   3246645H1   1698   1866       192   g4901556   1740   2205       192   2740754F6   2311   2784       192   2740754H1   2311   2564       192   1834220H1   2425   2687       192   4941620H1   2432   2695       192   4941983H1   2432   2711       192   2072075H1   2522   2772       192   5373319H1   2564   2779       192   3619907H1   2644   2921       192   4543335H1   2703   2966       192   6894957H1   2731   3286       192   4694271H1   2766   3009       192   4852731H1   2880   3138       192   2154473H1   2918   3190       192   2284114H1   2944   3172       192   4878579H1   2942   3086       192   4775770H1   2975   3235       192   2548439H1   3011   3245       192   2740754T6   3136   3642       192   7214289H1   3146   3371       192   3732858F6   3172   3639       192   3732858H1   3173   3398       192   4136158H1   3193   3478       192   2654475T6   3195   3653       192   3732858T6   3217   3639       192   g4189007   3240   3680       192   856846H1   3327   3543       192   g4076116   3329   3683       192   g440877   1   3690       192   g5544790   141   538       192   7106835H1   489   657       192   2313806H1   3487   3683       192   2841392H1   3499   3686       192   2097796H1   3545   3686       192   666777H1   3347   3550       192   815360H1   3387   3620       192   2104253H1   3439   3675       192   g831278   3464   3690       193   5911845T6   1   432       193   5911845F8   1   588       193   5911845F6   1   484       193   5911845H1   1   254       193   5911845T8   22   443       194   5910555F8   1   587       194   5910555T8   85   489       194   5910555F6   1   640       194   5910555T6   1   575       194   5910555H1   1   194       195   6790675H1   1   380       195   g1688992   67   154       195   g4326739   68   512       196   g274447   2556   2884       196   g2051352   2558   2924       196   g1859536   2558   2909       196   g2817917   2567   2931       196   g1783714   2570   2878       196   g823824   2587   2923       196   g2782889   2604   2887       196   4840554T6   2612   2903       196   g565986   2639   2909       196   g3277818   2645   2926       196   g4149369   2649   2925       196   1575363H1   2664   2888       196   1575363F6   2664   2917       196   1575363T6   2666   2879       196   g788188   2690   2909       196   g1482370   2691   2910       196   g864606   2696   2879       196   2006905H1   2712   2899       196   806183H1   2716   2867       196   g842117   2730   2907       196   g3742246   2738   2909       196   4882821H1   2771   3047       196   4884359H1   2771   3033       196   5090054H1   2781   3025       196   2277831T6   2784   2878       196   2343515H1   2792   2906       196   2343515F6   2792   2906       196   g788214   2821   2899       196   g1995403   2857   3182       196   5119623H1   2995   3277       196   3369954H1   3012   3294       196   5346406H1   3028   3177       196   2675533H1   3208   3394       196   3077845H1   3217   3451       196   5592052H1   3236   3495       196   996009H1   3251   3493       196   5041507H1   3296   3548       196   7032868H1   3301   3854       196   1665916H1   3358   3611       196   1665916F6   3358   3807       196   5513048H1   3393   3621       196   2969909H2   3439   3736       196   3978484H1   3473   3755       196   g831015   3530   3895       196   g821280   3561   3895       196   g1885508   1   289       196   g1110043   1   305       196   3349547F6   1   411       196   3349547H1   1   271       196   2741917H1   5   251       196   6991627H1   203   727       196   1434004H1   241   528       196   5587446H1   272   506       196   g2880882   276   409       196   349280H1   375   614       196   g713456   380   729       196   g698683   403   727       196   6916554H1   603   1106       196   6719552H1   659   1266       196   4384945H1   704   965       196   4384979H1   704   882       196   g1885551   910   1284       196   5019503H1   953   1210       196   4876805H1   996   1272       196   2479328H1   1055   1293       196   037774H1   1083   1306       196   g1717439   1121   1557       196   4840554H1   1189   1453       196   4840554F6   1189   1633       196   3279387H1   1202   1456       196   5373713H1   1216   1468       196   4145650H1   1246   1331       196   4916687H1   1293   1450       196   4581558H1   1328   1543       196   g2307061   1330   1763       196   4386045H1   1371   1651       196   4700744H1   1373   1635       196   6812975J1   1426   2021       196   6484322H1   1438   1623       196   1912429H1   1473   1693       196   g1440342   1498   1770       196   g4524899   1511   1896       196   g768901   1513   1820       196   g571029   1512   1840       196   g880779   1513   1841       196   g832097   1513   1884       196   4623266H1   1566   1825       196   3519427H1   1590   1955       196   3680478H1   1608   1840       196   6332293H1   1668   2034       196   4832174H1   1735   1965       196   g1329683   1739   2321       196   g787502   1779   2009       196   g864605   1779   2053       196   g787457   1779   2024       196   g841915   1778   2047       196   5589418H1   1784   2030       196   5071250H1   1803   2062       196   3865563H1   1843   2217       196   1955962H1   1894   2151       196   4750673H2   1945   2009       196   442154H1   1966   2272       196   261197H1   1965   2281       196   444335H1   1966   2225       196   4515301H1   2028   2284       196   3780534H1   2055   2353       196   2191782F6   2063   2373       196   2191782H1   2063   2304       196   3735668H1   2072   2331       196   3038357H1   2117   2391       196   6812975H1   2124   2462       196   4633908H1   2138   2408       196   1339250F6   2140   2483       196   1339250H1   2140   2404       196   1339250T6   2142   2469       196   4324610H1   2150   2401       196   4266784H2   2156   2439       196   4834124H1   2174   2358       196   g1783896   2175   2516       196   g2207390   2175   2518       196   908344R2   2180   2516       196   908344H1   2180   2285       196   4748325H1   2199   2467       196   4746557H1   2200   2439       196   g1859648   2231   2520       196   5565332H1   2244   2501       196   g778446   2320   2632       196   g831074   2319   2636       196   g2053046   2330   2794       196   3031124H1   2353   2644       196   2277831H1   2391   2660       196   2277831R6   2391   2516       196   g2552923   2437   2906       196   g832055   2523   2923       196   g1329627   2545   2922       196   g4990052   2547   2834       196   g4311136   2547   2993       196   g4988520   2547   3007       196   g4740301   2547   2833       196   g3422370   2548   2923       196   g4289096   2548   2904       196   g3431513   2549   2876       196   000140H1   2555   2911       196   2543809H1   2555   2770       196   2435619H1   2555   2713       196   g5113593   2555   2917       196   g4189560   2555   2877       196   g3108551   2555   2916       196   g1476749   2557   2878       196   g5112875   2555   2909       196   g4565709   2555   2921       196   g3280248   2555   2910       196   g2208127   2555   2878       197   5965475H1   1   564       197   g1162029   1   146       197   g1648409   1   322       197   5833130H1   1   266       197   6151063H1   53   353       198   6764201J1   1   585       198   986476R6   5   479       198   986476H1   5   303       198   4180212H1   8   260       198   g3933038   75   566       198   4029248H1   122   377       198   4029227H1   122   370       199   g4196260   1   320       199   g2218495   1   313       199   3685061H1   1   293       199   6821713J1   1   518       199   5372078H1   25   226       199   4205860H1   169   437       199   4205654H1   169   416       200   6868907H1   1   542       200   1543453R6   24   440       200   1543453H1   24   217       200   5055563H1   38   307       200   5055563F9   38   607       200   4778273F8   126   752       200   4778273H1   126   397       200   2724784H1   137   372       200   2925738H1   339   605       200   4778273T9   559   1136       200   1543453T6   610   1221       200   1921258H1   832   1046       200   1921258F6   833   1233       201   g4664146   1   478       201   g6140998   2   458       201   g6144190   2   422       201   g6139959   2   400       201   6764092J1   10   589       201   g2910438   59   537       201   4570842H1   67   315       201   4921180H1   69   331       201   5955592H1   86   596       201   g2904695   89   474       201   g2841122   108   375       201   6982639H1   384   913       201   1570643T6   422   944       201   g5232046   430   870       201   7377338H1   537   1127       201   1570643F6   661   972       201   3119826H1   732   1002       201   6097252H1   753   954       201   1570316H1   810   972       201   1570643H1   814   972       202   5913683H1   1   281       202   5320619F9   1   372       202   5913683F6   4   358       202   5321234F9   6   484       202   5914061H1   6   264       202   5914061F8   6   444       202   5913683T6   28   431       202   5913683F8   27   200       202   6269343H1   36   444       202   6269670H1   59   444       203   71035493V1   961   1616       203   5771773H1   971   1635       203   5952931H1   971   1230       203   5989634H1   968   1168       203   5800985H1   971   1270       203   5800543H1   976   1477       203   5805534H1   976   1299       203   5831382H2   976   1193       203   6166604H1   984   1567       203   6153585H1   982   1221       203   5804655H1   1003   1323       203   7075719H1   1022   1600       203   5798865H1   1027   1604       203   6964153H1   1031   1629       203   6183568H1   1031   1322       203   6183681H1   1031   1308       203   71001091V1   1050   1610       203   6217476H1   1061   1617       203   6148902H1   1068   1615       203   6114310H1   1078   1353       203   5806522H1   1090   1418       203   6146547H1   1093   1723       203   7220801H1   1094   1676       203   6132328H1   1094   1386       203   5995785H1   1095   1395       203   7322507H1   1099   1470       203   6113383H1   1100   1461       203   5975102H1   1102   1685       203   6148050H1   1102   1633       203   5977862H1   1102   1618       203   6038505H1   1104   1743       203   7080569H1   1103   1454       203   6116490H1   1102   1394       203   71360704V1   1105   1709       203   5892681H1   1104   1351       203   6164075H1   1107   1725       203   6181324H1   1107   1428       203   5770720H1   1114   1752       203   5797766H1   1114   1712       203   5826374H1   1118   1687       203   5796423H1   1114   1401       203   6036824H1   1117   1719       203   6034194H1   1117   1678       203   5769794H1   1118   1656       203   6134213H1   1117   1460       203   5770890H1   1122   1760       203   5976811H1   1134   1770       203   6178910H1   1139   1429       203   5975668H1   1145   1778       203   5975687H1   1145   1729       203   6214924H1   1146   1720       203   6183602H1   1162   1505       203   6183689H1   1162   1461       203   5803566H1   1165   1465       203   6040276H1   1169   1782       203   5975156H1   1169   1687       203   5973594H1   1169   1675       203   5995226H1   1169   1499       203   5993875H1   1170   1487       203   6078894H1   1169   1465       203   5825616H1   1173   1541       203   5921387T8   1177   1760       203   6042666H1   1187   1479       203   7185125H1   1194   1778       203   5993778H1   1197   1463       203   5993775H1   1193   1513       203   6168554H1   1198   1502       203   60047572D4   1199   1757       203   5824125H1   1200   1655       203   5994991H1   1200   1515       203   6112929H1   1204   1502       203   7330358H1   1219   1909       203   5768301H1   1220   1806       203   5769260H1   1218   1756       203   7330481H1   1221   1868       203   5995163H1   1233   1569       203   6959960H1   1236   1747       203   60206936U1   1239   1912       203   6034683H1   1243   1803       203   6169613H1   1244   1582       203   4294314T9   1258   1713       203   6041063H1   1248   1905       203   71035949V1   1252   1885       203   5952448H1   1256   1571       203   5974564H1   1254   1859       203   5991486H1   1255   1599       203   7264840H1   1257   1915       203   5991328H1   1255   1611       203   6112291H1   1256   1594       203   7080084H1   1261   1885       203   5771870H1   1263   1827       203   6170079H1   1262   1582       203   6080872H1   1264   1862       203   5824738H1   1269   1837       203   3390106T6   1274   1662       203   60221621V1   1274   1677       203   7004345H1   1284   1909       203   60109238B1   1286   1912       203   7186303H1   1290   1881       203   7025213H1   1297   1900       203   7182261H1   1299   1807       203   7193486H2   1306   1929       203   6736012H1   1319   1907       203   5970654H1   1322   1912       203   5770017H1   1327   1919       203   5802957H1   1327   1561       203   5800770H1   1328   1601       203   5804521H1   1331   1550       203   5995788H1   1330   1639       203   7331927H1   1332   1935       203   5769517H1   1333   1918       203   5967562H1   1333   1856       203   6163438H1   1337   1896       203   5953896H1   1338   1640       203   5789044H1   1338   1633       203   5975734H1   1339   1916       203   6078714H1   1338   1668       203   5786342H1   1338   1646       203   6175953H1   1340   1636       203   5805592H1   1340   1639       203   5952736H1   1351   1684       203   3693154F6   1354   1935       203   6217933H1   1356   1756       203   7160128H1   628   1197       203   6117709H1   630   956       203   6180232H1   632   859       203   6173095H1   645   974       203   6132472H1   645   956       203   6150854H1   647   996       203   5804474H1   647   944       203   6043158H1   647   922       203   6150959H1   647   898       203   6162572H1   648   1211       203   5803707H1   648   950       203   6111936H1   653   955       203   6111235H1   656   969       203   5953419H1   659   1020       203   5995735H1   659   974       203   6002288H1   659   917       203   6144013H1   664   1176       203   6166779H1   664   1176       203   6052294H1   666   889       203   6053478J1   664   1250       203   6052294J1   666   1098       203   5995222H1   671   1001       203   5990146H1   671   994       203   6079085H1   671   956       203   5798802H1   672   1277       203   5975323H1   681   1238       203   7327981H2   684   1173       203   6155086H1   691   1046       203   6183751H1   699   1001       203   6183830H1   699   983       203   6215696H1   701   1298       203   6109481H1   701   1023       203   6030242H2   702   1001       203   6163760H1   706   1074       203   6153438H1   710   1236       203   6111179H1   710   982       203   6116630H1   723   1044       203   6151576H1   726   998       203   6111753H1   732   1001       203   5824855H1   736   1199       203   5991422H1   740   1085       203   6180759H1   740   1069       203   6135676H1   741   1031       203   5954296H1   741   983       203   6154930H1   741   1095       203   6145090H1   747   1336       203   7215869H1   750   1359       203   6110012H1   754   1069       203   6135068H1   757   1061       203   6035941H1   759   1259       203   6163478H1   764   1336       203   5968127H1   760   1255       203   6178490H1   763   1018       203   60204862U1   771   1320       203   6108986H1   773   1083       203   6097994H1   787   1118       203   6165174H1   793   1081       203   6115201H1   792   1129       203   5798369H1   797   1419       203   5801102H1   797   1105       203   6112951H1   797   997       203   6109295H1   803   1129       203   6110881H1   805   1081       203   6179893H1   804   1105       203   5955376H1   806   1170       203   7359669H1   815   1090       203   6176408H1   813   1095       203   7004683H1   820   1427       203   7004582H1   820   1396       203   71033163V1   820   1364       203   5974746H1   827   1459       203   7325468H1   829   1480       203   6035163H1   839   1457       203   6112093H1   835   1148       203   6177785H1   836   1124       203   5798782H1   836   917       203   6150235H1   848   1459       203   5952964H1   849   1130       203   6218804H1   851   1196       203   5804744H1   853   1162       203   5771556H1   858   1477       203   5994894H1   856   1166       203   71036246V1   861   1368       203   5915476H1   858   1096       203   6163363H1   863   1499       203   5796628H1   870   1046       203   6115353H1   872   1144       203   6115444H1   876   1097       203   6033285H1   884   1516       203   6145417H1   890   1443       203   6172878H1   890   1197       203   6172790H1   890   1176       203   5801236H1   902   1168       203   6113354H1   906   1275       203   6032732H1   906   1459       203   6130142H1   908   1245       203   6168522H1   913   1264       203   6037518H1   915   1525       203   6032546H1   914   1542       203   60104786D4   912   1222       203   6176164H1   924   1214       203   6133149H1   913   1026       203   7320972H1   920   1515       203   6033149H1   923   1523       203   5970892H1   924   1548       203   6036731H1   925   1529       203   5970884H1   924   1510       203   6113366H1   923   1026       203   7185073H1   932   1524       203   5827058H1   935   988       203   5770524H1   940   1589       203   6114346H1   936   1210       203   5990721H1   942   1263       203   5975145H1   947   1489       203   6044290H1   941   1226       203   5768644H1   957   1532       203   5971326H1   948   1554       203   6110988H1   946   1173       203   6099321H1   948   1236       203   6173091H1   62   390       203   6036155H1   65   613       203   6180780H1   65   387       203   5770305H1   66   615       203   7215805H1   67   610       203   7024381H1   67   430       203   6107796H1   67   255       203   7221886H1   69   646       203   5769589H1   70   647       203   7328380H1   73   643       203   4571391F8   75   628       203   5771842H1   79   676       203   6108158H1   79   418       203   6176812H1   79   395       203   6177958H1   79   393       203   6179512H1   79   393       203   6181431H1   79   384       203   7008218H1   79   694       203   6215727H1   79   590       203   6151732H1   84   317       203   5973312H1   84   615       203   60126922U1   86   662       203   5974449H1   88   675       203   6111184H1   90   170       203   6113115H1   90   429       203   6000095H1   91   474       203   6182051H1   91   394       203   7272693H1   91   677       203   7385648H1   96   287       203   6135054H1   99   403       203   6182449H1   104   467       203   5775693H1   104   719       203   6175825H1   106   381       203   5968167H1   118   728       203   60120598D1   108   278       203   6147691H1   122   616       203   6115311H1   133   425       203   5975411H1   137   630       203   5915392H1   144   416       203   6130193H1   148   465       203   7328558H1   147   830       203   5996192H1   166   467       203   5977403H1   168   658       203   5989334H1   177   480       203   6109928H1   179   509       203   6172501H1   186   392       203   6172633H1   186   479       203   7183712H1   187   769       203   7025064H1   194   618       203   7325279H1   201   812       203   6116494H1   204   493       203   5953527H1   210   543       203   5892842H1   214   481       203   5993811H1   218   546       203   6109818H1   226   448       203   6044864H1   228   751       203   6182368H1   242   578       203   7326560H2   250   893       203   6033805H1   253   837       203   6101869H1   257   545       203   6170505H1   265   596       203   6152125H1   265   596       203   6137458H1   265   568       203   5958352H1   265   856       203   5975525H1   294   899       203   5838053H1   296   585       203   5796171H1   304   858       203   6112187H1   310   639       203   6165284H1   313   858       203   5826529H1   320   764       203   6172048H1   336   644       203   6035764H1   326   932       203   6966778H1   329   425       203   6172080H1   328   576       203   7182401H1   354   954       203   7269240H1   357   994       203   6029089H1   360   667       203   6100553H1   367   674       203   5825841H1   387   1002       203   7214023H1   396   923       203   6109604H1   393   729       203   6143986H1   411   1031       203   6117765H1   421   543       203   6136677H1   423   744       203   6041789H1   430   1047       203   5989594H1   448   730       203   6053478H1   452   1086       203   6109628H1   453   711       203   6044864J1   455   1098       203   7180879H1   457   1063       203   5797850H1   461   930       203   6115841H1   461   756       203   6137438H1   468   772       203   6731970H1   474   1055       203   6108268H1   478   793       203   6115462H1   491   718       203   5891369H1   517   775       203   6001442H1   524   1096       203   7360854H1   525   694       203   5974453H1   539   1087       203   6109032H1   534   897       203   5892176H1   534   868       203   5953990H1   534   804       203   5831262H1   534   726       203   5955474H1   543   886       203   5955458H1   543   888       203   6111439H1   544   769       203   6111316H1   546   881       203   5797137H1   577   1094       203   6133207H1   586   897       203   7328606H1   589   1133       203   7261854H1   590   1225       203   6100514H1   596   898       203   6040074H1   599   1199       203   5824550H1   606   1060       203   2814920F7   612   1092       203   6181437H1   617   905       203   5771704H1   626   1260       203   6131361H1   624   937       203   71216557V1   628   1188       203   6141011H1   1   156       203   6108212H1   1   324       203   6177861H1   2   301       203   5976933H1   12   479       203   7024610H1   39   249       203   5973147H1   35   622       203   6735535H1   39   221       203   7279008H1   39   531       203   60264017D1   32   569       203   7079629H2   44   358       203   5972573H1   41   686       203   7160586H1   34   619       203   6784013H1   35   645       203   7083178H1   36   637       203   7182216H1   36   592       203   7184701H1   36   578       203   6170753H1   36   341       203   7261837H1   36   644       203   6180976H1   37   372       203   6170740H1   37   340       203   7262607H1   39   648       203   7220667H1   38   636       203   7182859H1   39   616       203   6099372H1   39   324       203   7260783H1   39   675       203   7316234H1   39   709       203   4625563F9   39   673       203   7160372H1   39   643       203   7160758H1   39   615       203   7216608H1   39   623       203   7219614H1   39   613       203   7213493H1   39   643       203   7160235H1   39   555       203   7317368H1   39   420       203   6097675H1   39   380       203   6098127H1   39   375       203   6171834H1   39   358       203   6169216H1   39   372       203   6131659H1   39   358       203   6177913H1   39   350       203   6109871H1   39   347       203   6132882H1   39   346       203   6136114H1   39   340       203   6132559H1   39   328       203   5989784H1   39   328       203   6135227H1   39   339       203   6028575H1   39   267       203   5914782H1   57   278       203   7359725H1   39   685       203   6084122H1   40   673       203   7083662H1   39   685       203   7358734H1   39   682       203   7181284H1   39   557       203   60264004D1   39   552       203   6080769H1   50   645       203   7212965H1   39   444       203   6174304H1   39   387       203   6117564H1   40   363       203   6132067H1   39   350       203   6099630H1   39   322       203   6134055H1   62   379       203   5994380H1   42   384       203   6098224H1   42   362       203   6132993H1   42   337       203   7213881H1   43   644       203   6081567H1   43   636       203   6132164H1   43   385       203   6101109H1   43   377       203   5992072H1   43   372       203   6078760H1   43   365       203   5995191H1   43   374       203   6171974H1   43   359       203   5989890H1   43   350       203   6181359H1   44   353       203   6110331H1   43   347       203   6111434H1   43   339       203   6168435H1   43   284       203   7268992H1   45   675       203   6081557H1   44   614       203   6110567H1   43   283       203   7272829H1   45   685       203   5992910H1   45   388       203   6131653H1   47   373       203   5796567H1   45   109       203   7263260H1   46   647       203   7158311H1   46   588       203   6097721H1   47   393       203   6136962H1   46   404       203   6170754H1   47   353       203   6107830H1   47   235       203   7275183H1   48   644       203   7182041H1   49   616       203   7182971H1   49   616       203   5991415H1   48   388       203   6112705H1   48   373       203   6098861H1   48   347       203   6099983H1   48   243       203   7283631H1   49   718       203   6039521H1   49   650       203   5992224H1   49   375       203   6170022H1   49   387       203   5992273H1   49   363       203   6114538H1   48   348       203   7272102H1   50   635       203   6136863H1   50   389       203   6085163H1   68   331       203   6115314H1   50   306       203   7270954H1   51   687       203   7190289H2   51   619       203   4624253F8   54   733       203   6037863H1   54   590       203   5973394H1   54   630       203   6168342H1   54   412       203   6733426H1   73   419       203   5770643H1   55   644       203   6733166H1   54   647       203   7321688H1   58   593       203   6038935H1   58   635       203   6134077H1   56   399       203   7185189H1   56   643       203   6034778H1   59   683       203   7330333H1   58   524       203   7184720H1   62   640       203   60104786B2   1354   1910       203   6177422H1   1353   1657       203   5824565H1   1360   1894       203   6110675H1   1364   1676       203   6042367H1   1364   1661       203   6035204H1   1366   1929       203   6170972H1   1365   1682       203   6181601H1   1365   1667       203   5802968H1   1367   1676       203   4362784F8   1372   1927       203   5996082H1   1377   1699       203   5996003H1   1375   1666       203   5995203H1   1382   1721       203   6133437H1   1383   1700       203   6178414H1   1393   1699       203   6179001H1   1401   1548       203   5769429H1   1403   1918       203   5785086H1   1404   1722       203   5801902H1   1405   1725       203   5792833H1   1404   1477       203   5949501H1   1415   1784       203   g6228962   1428   1937       203   g6228709   1432   1938       203   g6300732   1431   1938       203   60126922B1   1434   1887       203   6173439H1   1434   1747       203   5769804H1   1434   1920       203   g6301637   1436   1938       203   g6302015   1437   1936       203   5953730H1   1440   1530       203   5770049H1   1440   1927       203   7082145H1   1440   1907       203   g6301063   1443   1936       203   6133907H1   1444   1778       203   g6039672   1445   1932       203   g6301561   1453   1938       203   5990839H1   1456   1799       203   5768633H1   1459   1920       203   g6229211   1555   1933       203   g6132479   1555   1933       203   g6030521   1555   1935       203   g6197062   1561   1937       203   6152884H1   1564   1869       203   g6471957   1565   1928       203   5955189H1   1566   1854       203   6030104H2   1566   1861       203   5805947H1   1565   1840       203   5771690H1   1566   1928       203   g6030549   1568   1928       203   g6198562   1567   1928       203   6097965H1   1568   1889       203   6151677H1   1571   1866       203   5974518H1   1571   1926       203   g6229210   1574   1933       203   6028702H1   1579   1813       203   g6470581   1580   1920       203   g6198908   1587   1928       203   g6044672   1588   1928       203   g6470803   1511   1930       203   g6027906   1513   1934       203   g6074740   1524   1928       203   g6026164   1523   1928       203   g6133768   1528   1932       203   6179968H1   1526   1784       203   5991468H1   1528   1863       203   5786634H1   1530   1832       203   5786701H1   1530   1715       203   5786734H1   1532   1652       203   g6047869   1534   1921       203   6176458H1   1547   1825       203   5816770H1   1550   1883       203   5817968H1   1550   1746       203   g6471065   1550   1928       203   5817174H1   1550   1832       203   g6471718   1552   1937       203   g6464005   1552   1937       203   6108818H1   1554   1787       203   7181625H1   1553   1920       203   g6043984   1488   1938       203   g6476041   1488   1930       203   g6075390   1489   1937       203   g6028910   1490   1937       203   g6073495   1490   1930       203   g6401184   1490   1930       203   g6073672   1490   1937       203   g6036791   1489   1928       203   g6026147   1492   1935       203   g6036640   1492   1920       203   6157091H1   1492   1813       203   g6033899   1493   1928       203   g6198865   1493   1938       203   6219812H2   1500   1886       203   6192668H1   1501   1814       203   5768793H1   1505   1920       203   6191666H1   1501   1804       203   g6473184   1502   1934       203   g6476087   1502   1928       203   5993364H1   1503   1813       203   g6438645   1503   1928       203   g6076043   1504   1928       203   6043152H1   1519   1787       203   g6472040   1509   1937       203   6179110H1   1511   1806       203   g6400409   1476   1929       203   g6036079   1476   1928       203   g6086332   1477   1934       203   g6199417   1477   1933       203   g6044232   1478   1936       203   g6029466   1478   1935       203   g6439518   1479   1934       203   g6462956   1479   1934       203   g6439604   1480   1937       203   g6196631   1480   1938       203   g6439647   1479   1930       203   g6131743   1480   1935       203   g6044247   1480   1928       203   6034847H1   1482   1911       203   g6117455   1481   1933       203   g6464482   1481   1928       203   g6036141   1481   1928       203   g6047637   1481   1928       203   g6117632   1482   1930       203   g6463078   1482   1936       203   g6035703   1480   1936       203   g6076131   1482   1932       203   g6196780   1482   1932       203   g6031028   1483   1930       203   g6464375   1484   1928       203   g6043231   1484   1929       203   g6117300   1485   1933       203   g6200216   1484   1938       203   g6040103   1485   1932       203   g6399002   1485   1926       203   g6029144   1486   1934       203   g6043655   1487   1937       203   g6196472   1489   1934       203   g6034277   1815   1920       203   g6117025   1815   1920       203   g6199037   1815   1920       203   g6198912   1823   1928       203   g6473327   1833   1928       203   g6463164   1841   1930       203   5920960H1   1844   1925       203   g6198784   1856   1926       203   5787770H1   1722   1981       203   5789227H1   1722   1927       203   g6074506   1728   1938       203   6166065H1   1741   1956       203   g6029762   1746   1876       203   5786021H1   1754   1926       203   g6438899   1763   1928       203   5992585H1   1769   1920       203   6158235H1   1865   1940       203   5946071H1   1668   1917       203   5950118H1   1668   1905       203   6157445H1   1668   1850       203   5946973H1   1668   1920       203   5767935H1   1670   1920       203   5767973H1   1670   1912       203   5947508H1   1672   1934       203   5790266H1   1674   1920       203   g6400408   1676   1938       203   7338126H1   1680   1932       203   7350525H1   1686   1936       203   g6398110   1686   1938       203   g6402278   1686   1938       203   6157246H1   1686   1827       203   g6197883   1687   1928       203   g6046641   1691   1928       203   g6400764   1692   1928       203   g6076220   1693   1928       203   5794559H1   1695   1926       203   g6033633   1695   1926       203   5787563H1   1695   1925       203   g6117328   1708   1937       203   g6044253   1710   1929       203   g6398246   1721   1928       203   g6131945   1465   1929       203   6181714H1   1466   1806       203   g6048081   1466   1928       203   g6047077   1466   1932       203   g6132102   1468   1929       203   g6398647   1468   1929       203   g6400444   1468   1929       203   g6086266   1468   1928       203   g6132321   1469   1928       203   g6132239   1470   1930       203   g6074116   1468   1928       203   5990574H1   1470   1804       203   g6400620   1470   1930       203   g6074889   1470   1928       203   g6473566   1469   1928       203   g6400824   1472   1936       203   g6132812   1472   1930       203   g6400805   1472   1938       203   g6400717   1472   1928       203   g6030907   1473   1928       203   g6047234   1474   1930       203   7180844H1   1459   1928       203   g6035938   1464   1916       203   g6439666   1475   1928       203   g6036991   1475   1928       203   g6047968   1475   1930       203   g6036801   1476   1937       203   g6196212   1600   1928       203   6177877H1   1603   1888       203   g6039199   1605   1928       203   g6438679   1604   1920       203   g6400557   1611   1928       203   5817262H1   1615   1904       203   5817362H1   1615   1906       203   5820474H1   1615   1915       203   6102129H1   1619   1928       203   g6074067   1653   1932       203   g6132060   1656   1935       203   3775108F9   1656   1912       203   g6198405   1665   1929       203   6178153H1   1665   1928       203   g6045038   1666   1911       203   5770026H1   1668   1946       203   5792284H1   1668   1920       203   g6028253   1588   1930       203   g6475978   1593   1932       203   6175948H1   1593   1891       203   g6132639   1594   1928       203   g6463340   1597   1928       203   g6046756   1596   1928       204   6844571H1   664   1221       204   1299074T6   715   1264       204   1751671T6   860   1258       204   6941680H1   769   1245       204   1426502T6   979   1248       204   2825904H1   984   1241       204   6825511H1   594   1115       204   6827717H1   676   1155       204   6828967J1   813   1145       204   6825030H1   823   1116       204   6825030J1   822   1115       204   1470306H1   921   1114       204   6827717J1   434   1076       204   1945447H1   808   1051       204   5870352H1   729   1007       204   6826536J1   1157   1756       204   6822128J1   829   984       204   6826536H1   1143   1742       204   g3056033   1065   1305       204   g5837846   816   1289       204   g2986193   1027   1297       204   6831310H1   845   1278       204   1631532H1   1227   1297       204   g3150672   914   1287       204   g2236974   899   1287       204   g4889379   1021   1302       204   2255054R6   393   777       204   6498130H1   312   777       204   2255054H1   549   777       204   6826447H1   357   752       204   6826447J1   357   752       204   1579339H1   502   713       204   4443559H1   474   613       204   2042892H1   315   568       204   3358238H1   329   566       204   4750087H1   185   453       204   6824269H1   386   880       204   6824349H1   350   851       204   2184748H1   179   463       204   6756704J1   235   808       204   2184748F6   1   463       204   6756496H1   234   432       204   6756704H1   255   432       204   1299074H1   161   401       204   6824091H1   156   422       204   3698810H1   13   304       204   1751671H1   621   843       204   g1616031   448   783       204   6824349J1   147   796       204   4516155H1   541   775       204   4672305H1   94   298       204   6829925H1   1303   1619       204   1426502H1   1438   1679       204   6829925J1   1303   1618       204   6824269J1   875   1484       204   6824288J1   805   1385       204   6825511J1   786   1392       204   g5856181   886   1309       204   g824615   1112   1275       204   6802929J1   833   1289       204   g5127144   929   1299       204   g2344128   1075   1315       204   6458046H1   929   1285       204   g2397910   1064   1302       204   g2278427   1157   1301       204   g3058045   823   1284       204   6820991H1   747   1284       204   2255054T6   705   1267       204   6822128H1   514   951       204   6802929H1   535   920       204   6825384H1   424   928       204   6824288H1   498   991       204   6824091J1   335   879       204   3512547H1   1455   1695       204   6804257H1   1157   1691       204   3700061H1   1404   1683       204   1426502F6   1335   1679       204   755545H1   1439   1679       204   6498381H1   1158   1726       205   1858538T6   1354   1583       205   1810993T6   1360   1586       205   1810792H1   1281   1506       205   2625825H1   1367   1595       205   2228636T6   1368   1586       205   1724417T6   1386   1586       205   2689550T6   1333   1586       205   2689550F6   1   511       205   2689550H1   1   265       205   6476804H1   3   563       205   1545750H1   17   163       205   g2244465   68   341       205   1494653H1   351   402       205   1858538H1   400   682       205   2218749F6   474   916       205   2228636F6   474   908       205   2228636H1   474   714       205   1724288H1   642   852       205   1724417H1   642   852       205   1724417F6   642   998       205   1724064H1   642   841       205   1724064F6   642   1096       205   6549336H1   1002   1419       205   4822845H1   1024   1261       205   1810993F6   1281   1558       205   1810993H1   1281   1518       206   6045641H1   1   364       206   6045641J1   1   364       206   3179825H1   120   424       206   3179825F6   120   672       206   4758644H1   141   409       206   2966833H1   222   484       206   5372448H1   327   481       206   2686267H1   359   619       206   3722865H1   425   558       206   5036810H1   463   731       206   4164428H1   519   787       206   3799675H1   563   731       206   4708839H1   568   851       206   2952970H1   602   865       206   3561988H1   633   928       206   856538H1   642   846       206   2487001F6   672   1061       206   2487001H1   672   829       206   4980831H1   862   1147       206   2620607H1   875   1129       206   3873170H1   876   1028       206   4575663H1   912   1190       206   6155913H1   953   1276       206   4543658H1   1005   1249       206   5064775H1   1104   1345       206   g1014062   1144   1418       206   6249620H1   1173   1668       206   2881132F6   1223   1489       206   2881132H1   1223   1496       206   g1949025   1228   1546       206   4552262H1   1238   1500       206   950155H1   1278   1523       206   1705026H1   1316   1527       206   4398180H1   1331   1571       206   3179825T6   1418   1909       206   2487001T6   1441   2006       206   6097383H1   1482   1798       206   g2251371   1596   2047       206   g4072810   1631   2050       206   g3889774   1700   2050       206   g4087488   1749   2056       206   g1014063   1798   2030       206   2881132T6   1809   2005       206   g1195443   1885   2047       206   g2155148   1942   2053       207   6803514H1   503   984       207   6803514J1   242   823       207   6286909H2   316   799       207   g4568721   260   685       207   g3742689   209   684       207   g5365250   586   679       207   g5676607   451   679       207   g4111591   263   679       207   g2539496   299   678       207   g5769156   224   677       207   g5232991   277   675       207   g3090059   202   676       207   g3751723   261   676       207   g4299176   248   676       207   g5630519   273   675       207   5743674T7   354   545       207   5743674H1   1   297       207   4127943H1   1   246       207   5743674R7   1   183       208   1952854T6   1   565       209   2917244T6   1   567       209   2917244H1   1   156       209   2917244F6   1   567       210   5507875H1   1   219       210   5507875F6   1   393       210   3510289H1   1   276       210   6985373H1   20   382       210   6985355H1   20   540       210   4715879H1   20   238       210   645341H1   21   220       210   645341R6   21   432       210   4551228H1   20   212       210   3182009H1   24   326       210   3182022H1   24   321       210   5156573H1   27   272       210   4800844H1   28   277       210   5864355H1   31   324       210   6219781H1   32   251       210   3375196H1   35   280       210   6306192H1   38   512       210   g575091   38   316       210   2924014F6   38   385       210   2924014H1   38   247       210   2825826H1   56   355       210   3695750H1   62   182       210   941644R6   66   275       210   5310424H1   66   282       210   3580982H1   36   340       210   941644R1   66   365       210   5967756H1   70   570       210   g831059   85   445       210   5727850H1   131   666       210   2053435H1   218   446       210   g4682505   234   685       210   5507875R6   308   769       210   6874493H1   354   971       210   g5109730   543   998       210   g1391905   560   858       211   6409981H1   1   486                    
     [0901]                       TABLE 5                       SEQ ID NO:   Template ID   Tissue Distribution                                            1   LG:1040582.1:2000FEB18   Liver - 41%, Pancreas - 34%, Cardiovascular System -               14%       2   LG:453570.1:2000FEB18   Nervous System - 100%       3   LG:408751.3:2000FEB18   Sense Organs - 63%, Nervous System - 22%       4   LI:090574.1:2000FEB01   Nervous System - 46%, Unclassified/Mixed - 36%       5   LI:229932.2:2000FEB01   Musculoskeletal System - 80%       6   LI:332176.1:2000FEB01   Urinary Tract - 95%       7   LI:403248.2:2000FEB01   Respiratory System - 60%, Hemic and Immune               System - 40%       8   LG:220992.1:2000MAY19   Embryonic Structures - 17%, Male Genitalia - 12%       9   LG:1094571.1:2000MAY19   Liver - 19%, Embryonic Structures - 16%,               Cardiovascular System - 14%       10   LI:350754.4:2000MAY01   Skin - 47%, Stomatognathic System - 27%, Sense               Organs - 14%       11   LI:255828.29:2000MAY01   Musculoskeletal System - 100%       12   LI:1190263.1:2000MAY01   Urinary Tract - 80%, Urinary Tract - 15%       13   LG:270916.2:2000FEB18   Female Genitalia - 100%       14   LG:999414.3:2000FEB18   Embryonic Structures - 30%, Urinary Tract - 13%,               Digestive System - 11%, Musculoskeletal System -               11%       15   LG:429446.1:2000FEB18   Urinary Tract - 80%, Hemic and Immune System -               20%       16   LI:057229.1:2000FEB01   Male Genitalia - 71%, Hemic and Immune System -               29%       17   LI:351965.1:2000FEB01   Unclassified/Mixed - 53%, Male Genitalia - 12%       18   LG:068682.1:2000FEB18   Unclassified/Mixed - 49%, Male Genitalia - 27%       19   LG:242665.1:2000FEB18   Germ Cells - 47%, Female Genitalia - 13%, Male               Genitalia - 12%       20   LG:241743.1:2000FEB18   Liver - 27%, Urinary Tract - 27%, Respiratory System -               14%       21   LI:034212.1:2000FEB01   Digestive System - 24%, Musculoskeletal System -               22%, Nervous System - 11%       22   LG:344886.1:2000MAY19   Germ Cells - 24%, Nervous System - 12%       23   LG:228930.1:2000MAY19   Embryonic Structures - 43%, Nervous System - 29%,               Respiratory System - 14%, Hemic and Immune               System - 14%       24   LG:338927.1:2000MAY19   Digestive System - 23%, Unclassified/Mixed - 21%,               Embryonic Structures - 19%, Hemic and Immune               System - 19%       25   LG:898771.1:2000MAY19   Pancreas - 13%, Embryonic Structures - 11%,               Female Genitalia - 10%, Urinary Tract - 10%, Hemic               and Immune System - 10%, Cardiovascular System -               10%       26   LI:257664.67:2000MAY01   Hemic and Immune System - 100%       27   LI:001496.2:2000MAY01   Endocrine System - 27%, Female Genitalia - 25%,               Embryonic Structures - 25%       28   LI:1085273.2:2000MAY01   Digestive System - 29%, Skin - 24%, Endocrine               System - 16%       29   LI:333138.2:2000MAY01   Exocrine Glands - 61%, Nervous System - 13%,               Nervous System - 11%       30   LI:338927.1:2000MAY01   Embryonic Structures - 51%, Digestive System - 17%       31   LG:335558.1:2000FEB18   Endocrine System - 45%, Nervous System - 18%,               Exocrine Glands - 11%       32   LG:998283.7:2000FEB18   Sense Organs - 33%, Germ Cells - 18%       33   LI:402739.1:2000FEB01   Unclassified/Mixed - 78%, Mate Genitalia - 11%,               Hemic and Immune System - 11%       34   LI:175223.1:2000FEB01   Embryonic Structures - 99%       35   LG:981076.2:2000MAY19   Endocrine System - 28%, Nervous System - 22%,               Respiratory System - 17%, Female Genitalia - 17%,               Hemic and Immune System - 17%       36   LI:1008973.1:2000MAY01   Nervous System - 57%, Digestive System - 41%       37   LI:1190250.1:2000MAY01   Female Genitalia - 48%, Respiratory System - 25%       38   LG:021371.3:2000FEB18   Liver - 23%, Endocrine System - 17%, Hemic and               Immune System - 17%       39   LG:475404.1:2000FEB18   Skin - 82%       40   LG:979406.2:2000FEB18   Liver - 46%, Connective Tissue - 31%, Nervous               System - 15%       41   LG:410726.1:2000FEB18   Embryonic Structures - 52%, Endocrine System -               26%       42   LG:200005.1:2000FEB18   Unclassified/Mixed - 26%, Cardiovascular System -               14%, Female Genitalia - 13%       43   LG:1076828.1:2000FEB18   Unclassified/Mixed - 69%, Urinary Tract - 25%       44   LG:1076931.1:2000FEB18   Unclassified/Mixed - 63%, Musculoskeletal System -               20%, Urinary Tract - 11%       45   LG:1078121.1:2000FEB18   Female Genitalia - 75%, Nervous System - 25%       46   LG:1079203.1:2000FEB18   Female Genitalia - 42%, Cardiovascular System -               33%, Hemic and Immune System - 17%       47   LG:1082586.1:2000FEB18   Respiratory System - 100%       48   LG:1082774.1:2000FEB18   Respiratory System - 50%, Female Genitalia - 50%       49   LG:1082775.1:2000FEB18   Female Genitalia - 75%, Nervous System - 25%       50   LG:1083120.1:2000FEB18   Nervous System - 100%       51   LG:1087707.1:2000FEB18   Stomatognathic System - 98%       52   LG:1090915.1:2000FEB18   Embryonic Structures - 44%, Connective tissue -               19%       53   LG:1094230.1:2000FEB18   Female Genitalia - 100%       54   LG:474848.3:2000FEB18   Connective Tissue - 44%, Exocrine Glands - 44%,               Hemic and Immune System - 11%       55   LI:251656.1:2000FEB01   Nervous System - 38%, Digestive System - 38%, Male               Genitalia - 25%       56   LI:021371.1:2000FEB01   Hemic and Immune System - 69%, Endocrine               System - 14%       57   LI:133095.1:2000FEB01   Respiratory System - 67%, Nervous System - 13%       58   LI:236654.2:2000FEB01   Unclassified/Mixed - 30%, Respiratory System - 19%,               Nervous System - 13%, Digestive System - 13%       59   LI:200009.1:2000FEB01   Unclassified/Mixed - 37%, Urinary Tract - 16%,               Cardiovascular System - 15%       60   LI:758502.1:2000FEB01   Unclassified/Mixed - 78%, Musculoskeletal System -               22%       61   LI:344772.1:2000FEB01   Nervous System - 56%, Skin - 27%, Connective Tissue -               13%       62   LI:789445.1:2000FEB01   Endocrine System - 100%       63   LI:789657.1:2000FEB01   Urinary Tract - 31%, Female Genitalia - 19%,               Digestive System - 19%, Hemic and Immune System -               19%       64   LI:789808.1:2000FEB01   Exocrine Glands - 44%, Female Genitalia - 33%,               Nervous System - 22%       65   LI:792919.1:2000FEB01   Respiratory System - 100%       66   LI:793949.1:2000FEB01   Female Genitalia - 42%, Endocrine System - 19%,               Exocrine Glands - 13%       67   LI:794389.1:2000FEB01   Endocrine System - 100%       68   LI:796010.1:2000FEB01   Exocrine Glands - 100%       69   LI:796324.1:2000FEB01   Female Genitalia - 100%       70   LI:796373.1:2000FEB01   Respiratory System - 100%       71   LI:796415.1:2000FEB01   Nervous System - 100%       72   LI:798636.1:2000FEB01   Hemic and Immune System - 100%       73   LI:800045.1:2000FEB01   Female Genitalia - 60%, Male Genitalia - 40%       74   LI:800680.1:2000FEB01   Cardiovascular System - 100%       75   LI:800894.1:2000FEB01   Respiratory System - 50%, Digestive System - 50%       76   LI:801015.1:2000FEB01   Male Genitalia - 100%       77   LI:801236.1:2000FEB01   Endocrine System - 100%       78   LI:803335.1:2000FEB01   Connective Tissue - 100%       79   LI:803998.1:2000FEB01   Nervous System - 38%, Digestive System - 38%, Male               Genitalia - 25%       80   LI:478757.1:2000FEB01   Digestive System - 100%       81   LI:808532.1:2000FEB01   Hemic and Immune System - 100%       82   LI:443073.1:2000FEB01   Digestive System - 100%       83   LI:479671.1:2000FEB01   Exocrine Glands - 80%, Hemic and Immune System -               20%       84   LI:810078.1:2000FEB01   Digestive System - 100%       85   LI:810224.1:2000FEB01   Digestive System - 100%       86   LI:817052.2:2000FEB01   Nervous System - 24%, Unclassified/Mixed - 18%,               Exocrine Glands - 14%       87   LG:892274.1:2000MAY19   Embryonic Structures - 63%, Digestive System - 30%       88   LG:1080959.1:2000MAY19   Digestive System - 40%, Respiratory System - 30%,               Hemic and Immune System - 30%       89   LG:1054900.1:2000MAY19   Digestive System - 100%       90   LG:1077357.1:2000MAY19   Nervous System - 38%, Female Genitalia - 38%,               Male Genitalia - 25%       91   LG:1084051.1:2000MAY19   Pancreas - 31%, Digestive System - 22%, Hemic and               Immune System - 16%       92   LG:1076853.1:2000MAY19   Female Genitalia - 23%, Unclassified/Mixed - 23%,               Cardiovascular System - 18%, Exocrine Glands -               18%       93   LG:481631.10:2000MAY19   Female Genitalia - 22%, Nervous System - 17%,               Exocrine Glands - 17%, Urinary Tract - 17%       94   LG:1088431.2:2000MAY19   Exocrine Glands - 67%, Cardiovascular System -               33%       95   LI:401619.10:2000MAY01   Endocrine System - 18%, Embryonic Structures -               16%, Pancreas - 15%       96   LI:1144007.1:2000MAY01   Hemic and Immune System - 27%, Female               Genitalia - 13%       97   LI:331074.1:2000MAY01   Endocrine System - 28%, Sense Organs - 22%,               Connective Tissue - 10%       98   LI:1170349.1:2000MAY01   Endocrine System - 91%       99   LG:335097.1:2000FEB18   Embryonic Structures - 24%, Musculoskeletal System -               19%, Nervous System - 16%       100   LG:1076451.1:2000FEB18   Nervous System - 100%       101   LI:805478.1:2000FEB01   Skin - 100%       102   LG:101269.1:2000MAY19   Endocrine System - 33%, Embryonic Structures -               33%, Urinary Tract - 30%       103   LI:331087.1:2000MAY01   Liver - 82%, Hemic and Immune System - 13%       104   LI:410188.1:2000MAY01   Cardiovascular System - 81%, Cardiovascular               System - 12%       105   LI:1188288.1:2000MAY01   Nervous System - 73%       106   LI:427997.4:2000MAY01   Liver - 16%, Male Genitalia - 13%, Embryonic               Structures - 11%       107   LG:451682.1:2000FEB18   Nervous System - 100%       108   LG:1077283.1:2000FEB18   Liver - 86%, Hemic and Immune System - 14%       109   LG:481436.5:2000FEB18   Embryonic Structures - 41%, Endocrine System -               20%, Hemic and Immune System - 13%       110   LI:793701.1:2000FEB01   Endocrine System - 43%, Urinary Tract - 36%,               Respiratory System - 21%       111   LI:373637.1:2000FEB01   Germ Cells - 74%, Unclassified/Mixed - 16%       112   LG:239368.2:2000MAY19   Digestive System - 43%, Male Genitalia - 24%,               Endocrine System - 24%       113   LI:053826.1:2000MAY01   Germ Cells - 66%, Unclassified/Mixed - 22%, Male               Genitalia - 12%       114   LI:449393.1:2000MAY01   Nervous System - 100%       115   LI:1071427.96:2000MAY01   Stomatognathic System - 13%       116   LI:336338.8:2000MAY01   Unclassified/Mixed - 55%, Connective Tissue - 26%       117   LG:345527.1:2000FEB18   Urinary Tract - 24%, Hemic and Immune System -               24%, Respiratory System - 18%       118   LG:1089383.1:2000FEB18   Connective Tissue - 73%, Female Genitalia - 27%       119   LG:1092522.1:2000FEB18   Female Genitalia - 38%, Exocrine Glands - 31%,               Male Genitalia - 15%, Hemic and Immune System -               15%       120   LG:1093216.1:2000FEB18   Urinary Tract - 100%       121   LI:270318.3:2000FEB01   Embryonic Structures - 86%, Hemic and Immune               System - 14%       122   LI:335671.2:2000FEB01   Unclassified/Mixed - 34%, Hemic and Immune               System - 20%, Urinary Tract - 17%       123   LI:793758.1:2000FEB01   Nervous System - 62%, Urinary Tract - 38%       124   LI:803718.1:2000FEB01   Female Genitalia - 100%       125   LI:412179.1:2000FEB01   Endocrine System - 100%       126   LI:815679.1:2000FEB01   Digestive System - 75%       127   LI:481361.3:2000FEB01   Embryonic Structures - 28%, Skin - 20%,               Unclassified/Mixed - 16%       128   LG:247388.1:2000MAY19   Cardiovascular System - 33%, Endocrine System -               21%, Male Genitalia - 21%       129   LG:255789.10:2000MAY19   Endocrine System - 56%, Urinary Tract - 44%       130   LI:787618.1:2000MAY01   Endocrine System - 22%, Digestive System - 13%,               Endocrine System - 12%       139   LG:337818.2:2000FEB18   Sense Organs - 18%, Nervous System - 11%,               Digestive System - 34%, Liver - 17%, Female               Genitalia - 11%       140   LI:337818.1:2000FEB01   Digestive System - 27%, Liver - 19%, Female               Genitalia - 15%       141   LG:241577.4:2000MAY19   Pancreas - 48%, Endocrine System - 24%,               Respiratory System - 14%       142   LG:344786.4:2000MAY19   Respiratory System - 67%, Digestive System - 22%,               Nervous System - 11%       143   LI:414307.1:2000FEB01   Endocrine System - 44%, Unclassified/Mixed - 17%,               Nervous System - 11%       144   LI:202943.2:2000FEB01   Embryonic Structures - 100%       145   LI:246194.2:2000FEB01   Germ Cells - 75%, Pancreas - 13%       146   LI:815961.1:2000FEB01   Digestive System - 99%       147   LG:120744.1:2000MAY19   Skin - 33%, Embryonic Structures - 21%, Digestive               System - 21%       148   LI:757520.1:2000MAY01   Musculoskeletal System - 45%, Cardiovascular               System - 26%, Skin - 24%       149   LG:160570.1:2000FEB18   Skin - 84%, Female Genitalia - 16%       150   LI:350398.3:2000FEB01   Male Genitalia - 50%, Hemic and Immune System -               50%       151   LI:221285.1:2000FEB01   Endocrine System - 42%, Nervous System - 21%       153   LI:329017.1:2000FEB01   Endocrine System - 62%, Unclassified/Mixed - 24%       154   LI:401322.1:2000FEB01   Sense Organs - 44%, Liver - 22%, Skin - 14%       155   LG:403409.1:2000MAY19   Respiratory System - 18%, Female Genitalia - 16%,               Cardiovascular System - 13%       156   LG:233933.5:2000MAY19   Digestive System - 100%       157   LI:290344.1:2000MAY01   Connective Tissue - 40%, Nervous System - 19%,               Embryonic Structures - 12%       158   LI:410742.1:2000MAY01   Respiratory System - 47%, Skin - 42%       159   LG:406568.1:2000MAY19   Stomatognathic System - 57%, Musculoskeletal               System - 21%, Cardiovascular System - 16%       160   LI:283762.1:2000MAY01   Sense Organs - 25%       161   LI:347687.113:2000MAY01   Nervous System - 45%, Nervous System - 38%       162   LI:1146510.1:2000MAY01   Skin - 94%       163   LG:451710.1:2000FEB18   Connective Tissue - 89%, Nervous System - 11%       164   LG:455771.1:2000FEB18   Nervous System - 100%       165   LG:452089.1:2000FEB18   Nervous System - 100%       166   LG:246415.1:2000FEB18   Pancreas - 83%, Nervous System - 17%       167   LG:414144.10:2000FEB18   Cardiovascular System - 17%, Connective Tissue -               12%       168   LG:1101445.1:2000FEB18   Liver - 91%       169   LG:452134.1:2000FEB18   Hemic and Immune System - 64%, Male Genitalia -               36%       170   LI:903021.1:2000FEB01   Male Genitalia - 100%       171   LI:246422.1:2000FEB01   Hemic and Immune System - 100%       172   LG:449404.1:2000MAY19   Nervous System - 100%       173   LG:449413.1:2000MAY19   Nervous System - 100%       174   LG:450105.1:2000MAY19   Nervous System - 100%       175   LG:460809.1:2000MAY19   Exocrine Glands - 100%       176   LG:481781.1:2000MAY19   Nervous System - 100%       177   LG:1101153.1:2000MAY19   Nervous System - 100%       178   LI:257695.20:2000MAY01   Exocrine Glands - 28%, Endocrine System - 19%,               Nervous System - 16%, Digestive System - 16%       179   LI:455771.1:2000MAY01   Nervous System - 100%       180   LI:274551.1:2000MAY01   Nervous System - 60%, Hemic and Immune System -               40%       181   LI:035973.1:2000MAY01   Embryonic Structures - 58%, Digestive System - 26%,               Nervous System - 16%       182   LG:978427.5:2000FEB18   Nervous System - 100%       183   LG:247781.2:2000FEB18   Nervous System - 11%       184   LI:034583.1:2000FEB01   Nervous System - 35%, Endocrine System - 35%       185   LI:333307.2:2000FEB01   Cardiovascular System - 28%, Urinary Tract - 27%,               Musculoskeletal System - 17%       186   LI:814710.2:2000FEB01   Respiratory System - 100%       187   LG:414732.1:2000MAY19   Endocrine System - 82%, Nervous System - 18%       188   LG:413910.6:2000MAY19   Connective Tissue - 55%, Nervous System - 15%,               Embryonic Structures - 13%       189   LI:414732.2:2000MAY01   Endocrine System - 80%, Nervous System - 20%       190   LI:900264.2:2000MAY01   Urinary Tract - 15%, Male Genitalia - 12%       191   LI:335593.1:2000MAY01   Urinary Tract - 46%, Endocrine System - 17%, Germ               Cells - 14%       192   LI:1189543.1:2000MAY01   Stomatognathic System - 35%, Digestive System -               14%       193   LG:455450.1:2000FEB18   Nervous System - 100%       194   LG:1040978.1:2000FEB18   Nervous System - 100%       195   LG:446649.1:2000FEB18   Liver - 80%, Hemic and Immune System - 13%       196   LG:132147.3:2000FEB18   Unclassified/Mixed - 17%, Sense Organs - 16%,               Embryonic Structures - 10%       197   LI:036034.1:2000FEB01   Nervous System - 80%       198   LG:162161.1:2000MAY19   Unclassified/Mixed - 53%, Cardiovascular System -               21%, Nervous System - 16%       199   LG:407214.10:2000MAY19   Unclassified/Mixed - 40%, Respiratory System - 24%,               Cardiovascular System - 16%       200   LG:204626.1:2000MAY19   Digestive System - 41%, Exocrine Glands - 24%,               Female Genitalia - 18%       201   LI:007401.1:2000MAY01   Unclassified/Mixed - 31%, Nervous System - 25%,               Urinary Tract - 11%       202   LI:476342.1:2000MAY01   Connective Tissue - 77%, Nervous System - 23%       203   LI:1072759.1:2000MAY01   Hemic and Immune System - 27%, Musculoskeletal               System - 19%, Endocrine System - 11%       204   LG:998857.1:2000FEB18   Digestive System - 58%, Pancreas - 12%       205   LG:482261.1:2000FEB18   Male Genitalia - 85%, Respiratory System - 15%       206   LG:480328.1:2000FEB18   Skin - 20%, Germ Cells - 18%, Female Genitalia -               10%       207   LG:311197.1:2000MAY19   Germ Cells - 44%, Digestive System - 15%, Male               Genitalia - 11%       208   LG:1054883.1:2000MAY19   Endocrine System - 100%       209   LG:399395.1:2000MAY19   Hemic and Immune System - 100%       210   LG:380497.2:2000MAY19   Germ Cells - 23%, Exocrine Glands - 14%,               Connective Tissue - 13%       211   LI:272913.22:2000MAY01   Female Genitalia - 100%                    
     [0902]                                           TABLE 6                       SEQ                                   ID                       Probability       NO:   Frame   Length   Start   Stop   GI Number   Score   Annotation                                                                212   3   115   198   542   g399660   3.00E−51   aldehyde reductase [ Rattus norvegicus ]       212   3   115   198   542   g7677318   7.00E−51   aldehyde reductase [ Mus musculus ]       212   3   115   198   542   g6013149   2.00E−48   aldehyde reductase [ Homo sapiens ]       213   3   161   3   485   g2909424   2.00E−60   Glyoxalase I [ Cicer arietinum ]       213   3   161   3   485   g2113825   2.00E−58   Glyoxalase I [ Brassica juncea ]       213   3   161   3   485   g1177314   4.00E−57   glyoxalase-I [ Lycopersicon esculentum ]       214   2   332   2   997   g8671168   0   hypothetical protein [ Homo sapiens ]       214   2   332   2   997   g8886025   0   collapsin response mediator protein-5 [ Homo sapiens ]       214   2   332   2   997   g8671360   1.00E−179   Ulip-like protein [ Rattus norvegicus ]       215   3   274   12   833   g29600   2.00E−86   carbonic anhydrase I (AA 1-261) [ Homo sapiens ]       215   3   274   12   833   g179793   2.00E−86   carbonic anhydrase I (EC 4.2.1.1) [ Homo sapiens ]       215   3   274   12   833   g29587   4.00E−84   carbonic anhydrase II (AA 1-260) [ Homo sapiens ]       216   1   182   742   1287   g10438188   1.00E−102   unnamed protein product [ Homo sapiens ]       216   1   182   742   1287   g9949721   3.00E−49   probable acetyl-coa synthetase [ Pseudomonas aeruginosa ]       216   1   182   742   1287   g9655831   7.00E−46   prpE protein [ Vibrio cholerae ]       217   2   359   2   1078   g2104689   1.00E−111   alpha glucosidase II, alpha subunit [ Mus musculus ]       217   2   359   2   1078   g7672977   1.00E−111   glucosidase II alpha subunit [ Homo sapiens ]       217   2   359   2   1078   g577295   1.00E−110   The ha1225 gene product is related to human alpha-glucosidase. [ Homo                                       sapiens ]       218   2   110   161   490   g9653274   1.00E−26   ornithine decarboxylase-2 [ Xenopus laevis ]       218   2   110   161   490   g200124   5.00E−18   ornithine decarboxylase [ Mus pahari ]       218   2   110   161   490   g53518   1.00E−17   ornithine decarboxylase [ Mus musculus ]       219   3   549   36   1682   g10435462   0   unnamed protein product [ Homo sapiens ]       219   3   549   36   1682   g7023375   0   unnamed protein product [ Homo sapiens ]       219   3   549   36   1682   g10433608   1.00E−164   unnamed protein product [ Homo sapiens ]       220   1   264   1   792   g7023634   3.00E−92   unnamed protein product [ Homo sapiens ]       220   1   264   1   792   g3213202   3.00E−49   similarto  C. elegans  R10H10.6 and  S. cerevisiae  YD8419.03c [ Drosophila                                       melanogaster ]       220   1   264   1   792   g7298960   3.00E−49   CG2846 gene product [ Drosophila melanogaster ]       221   3   701   33   2135   g307504   0   transglutaminase E3 [ Homo sapiens ]       221   3   701   33   2135   g4467804   0   TGM3 (PROTEIN-GLUTAMINE GLUTAMYLTRANSFERASE E3                                   PRECURSOR (EC 2.3.2.13) (TGASE E3) (TRANSGLUTAMINASE 3).) [ Homo                                       sapiens ]       221   3   701   33   2135   g309521   0   transglutaminase E3 [ Mus musculus ]       222   2   150   2   451   g35505   7.00E−65   pyruvate kinase [ Homo sapiens ]       222   2   150   2   451   g189998   7.00E−65   M2-type pyruvate kinase [ Homo sapiens ]       222   2   150   2   451   g2623945   3.00E−64   pyruvate kinase; ATP: pyruvate 2-o-phosphotransferase [ Oryctolagus                                       cuniculus ]       223   2   234   866   1567   g2576305   1.00E−128   arylsulphatase [ Homo sapiens ]       223   2   234   866   1567   g791002   3.00E−82   ARSD [ Homo sapiens ]       223   2   234   866   1567   g791004   4.00E−75   ARSE [ Homo sapiens ]       224   2   86   2   259       225   2   173   1049   1567   g4092820   8.00E−62   BC319430_7 [ Homo sapiens ]       225   2   173   1049   1567   g2792016   2.00E−54   olfactory receptor [ Homo sapiens ]       225   2   173   1049   1567   g4092819   2.00E−54   BC319430_5 [ Homo sapiens ]       226   2   68   86   289   g8272468   4.00E−15   envelope protein [ Homo sapiens ]       226   2   68   86   289   g4773880   4.00E−15   envelope protein precursor [ Homo sapiens ]       226   2   68   86   289   g4262296   4.00E−15   envelope protein [ Homo sapiens ]       227   1   70   79   288   g11231093   1.00E−11   hypothetical protein [ Macaca fascicularis ]       227   1   70   79   288   g10435559   3.00E−10   unnamed protein product [ Homo sapiens ]       227   1   70   79   288   g7020625   2.00E−09   unnamed protein product [ Homo sapiens ]       228   2   117   836   1186   g5726235   3.00E−18   unknown protein U5/2 [multiple sclerosis associated retrovirus element]       229   2   294   2   883   g404634   1.00E−59   serine/threonine kinase [ Mus musculus ]       229   2   294   2   883   g2738898   3.00E−59   protein kinase [ Mus musculus ]       229   2   294   2   883   g8101585   2.00E−54   testis specific serine kinase-3 [ Mus musculus ]       230   1   326   1   978   g2117166   1.00E−160   Ras like GTPase [ Homo sapiens ]       230   1   326   1   978   g466271   1.00E−140   Rar protein [ Homo sapiens ]       230   1   326   1   978   g3036779   1.00E−102   match: multiple proteins; RAR (RAS like GTPASE) like [ Homo sapiens ]       231   1   182   40   585   g5763838   1.00E−66   dJ593C16.1 (ras GTPase activating protein) [ Homo sapiens ]       231   1   182   40   585   g4417207   1.00E−66   synGAP-d [ Rattus norvegicus ]       231   1   182   40   585   g4105589   1.00E−66   nGAP [ Homo sapiens ]       232   1   358   58   1131   g1469876   1.00E−103   The KIAA0147 gene product is related to adenylyl cyclase. [ Homo sapiens ]       232   1   358   58   1131   g6850952   1.00E−86   vartul-2 protein [ Drosophila melanogaster ]       232   1   358   58   1131   g6782322   1.00E−86   Vartul-1 protein [ Drosophila melanogaster ]       233   1   194   370   951   g7008402   1.00E−107   kappa B-ras 1 [ Homo sapiens ]       233   1   194   370   951   g7239257   1.00E−103   kappaB-Ras 1 [ Mus musculus ]       233   1   194   370   951   g7008404   8.00E−75   kappa B-ras 2 [ Homo sapiens ]       234   2   222   17   682   g9368448   1.00E−111   phospholipase C-beta-1a [ Homo sapiens ]       234   2   222   17   682   g9368450   1.00E−111   phospholipase C-beta-1b [ Homo sapiens ]       234   2   222   17   682   g206218   1.00E−110   phospholipase C-1 [Rattus sp.]       235   3   185   126   680   g3599940   1.00E−57   faciogenital dysplasia protein 2 [ Mus musculus ]       235   3   185   126   680   g10440426   8.00E−42   FLJ00048 protein [ Homo sapiens ]       235   3   185   126   680   g595425   4.00E−20   FGD1 [ Homo sapiens ]       236   2   192   707   1282       237   3   61   204   386       238   2   335   17   1021   g3005085   2.00E−92   hook1 protein [ Homo sapiens ]       238   2   335   17   1021   g5706448   2.00E−92   dJ782L23.1 (HOOK1) [ Homo sapiens ]       238   2   335   17   1021   g3005087   2.00E−70   hook2 protein [ Homo sapiens ]       239   1   346   1261   2298   g1109782   1.00E−105   protein-tyrosine phosphatase [ Homo sapiens ]       239   1   346   1261   2298   g1781037   1.00E−76   neuronal tyrosine threonine phosphatase 1 [ Mus musculus ]       239   1   346   1261   2298   g10241798   5.00E−11   hypothetical protein SCE41.24c [ Streptomyces coelicolor ]       240   3   298   147   1040   g4678722   1.00E−156   hypothetical protein [ Homo sapiens ]       240   3   298   147   1040   g4007153   1.00E−153   dJ272L16.1 (Rat Ca2+/Calmodulin dependent Protein Kinase LIKE protein)                                   [ Homo sapiens ]       240   3   298   147   1040   g2077934   1.00E−152   Protein Kinase [ Rattus norvegicus ]       241   1   133   133   531   g10440426   1.00E−34   FLJ00048 protein [ Homo sapiens ]       241   1   133   133   531   g3599940   2.00E−16   faciogenital dysplasia protein 2 [ Mus musculus ]       242   2   354   821   1882   g11907572   1.00E−143   TSC22-related inducible leucine zipper 1b [ Mus musculus ]       242   2   354   821   1882   g1181619   1.00E−106   a variant of TSC-22 [ Gallus gallus ]       242   2   354   821   1882   g3327152   9.00E−16   KIAA0669 protein [ Homo sapiens ]       243   1   237   1   711   g6683492   1.00E−105   bromodomain PHD finger transcription factor [ Homo sapiens ]       243   1   237   1   711   g3876452   9.00E−53   contains similarity to Pfam domain: PF00439 (Bromodomain), Score = 125.5, E-                                   value = 1.5e−35, N = 1; PF00628 (PHD-finger), Score = 102.0,                                   E-value =3.8e−27, N = 2 [ Caenorhabditis elegans ]       243   1   237   1   711   g3876449   9.00E−53   predicted using Genefinder˜contains similarity to Pfam domain: PF00439                                   (Bromodomain), Score = 125.5, E-value = 1.5e−35, N = 1; PF00628                                   (PHD-finger), Score = 102.0, E-value = 3.8e−27, N = 2 [ Caenorhabditis elegans ]       244   1   161   1   483   g6330736   1.00E−42   KIAA1234 protein [ Homo sapiens ]       244   1   161   1   483   g11244871   1.00E−40   dioxin receptor repressor [ Homo sapiens ]       244   1   161   1   483   g4164151   4.00E−35   AhR repressor [ Mus musculus ]       245   3   151   54   506   g10433955   9.00E−44   unnamed protein product [ Homo sapiens ]       245   3   151   54   506   g7295442   1.00E−16   CG17334 gene product [ Drosophila melanogaster ]       245   3   151   54   506   g2745892   1.00E−12   Y box transcription factor [ Mus musculus ]       246   2   160   173   652   g3924670   4.00E−68   supported by Genscan and several ESTs: C83049 (NID: g3062006),                                   AA823760 (NID: g2893628), AA215791 (NID: g1815572), AI095488                                   (NID: g3434464), and AA969095 (NID: g3144275) [ Homo sapiens ]       246   2   160   173   652   g5640105   2.00E−59   homeobox protein LSX [ Homo sapiens ]       246   2   160   173   652   g6523391   6.00E−59   phtf protein [ Mus musculus ]       247   3   160   108   587   g6939732   1.00E−52   transcription factor Elongin A2 [ Homo sapiens ]       247   3   160   108   587   g4581412   1.00E−29   dJ886K2.1 (elongin A; RNA polymerase; RNA polymerase II; RNA polymerase                                   II elongation factor.) [ Homo sapiens ]       247   3   160   108   587   g992563   1.00E−29   elongin A [ Homo sapiens ]       248   1   171   25   537   g11907923   4.00E−29   enhancer of polycomb [ Homo sapiens ]       248   1   171   25   537   g3757890   3.00E−18   enhancer of polycomb [ Drosophila melanogaster ]       248   1   171   25   537   g7303589   3.00E−18   E(Pc) gene product [ Drosophila melanogaster ]       249   2   449   266   1612   g10443047   0   bA465L10.2 (novel C2H2 type zinc finger protein similar to chicken FZF-1)                                   [ Homo sapiens ]       249   2   449   266   1612   g10438918   0   unnamed protein product [ Homo sapiens ]       249   2   449   266   1612   g984814   8.00E−98   zinc finger protein [ Gallus gallus ]       250   2   127   140   520   g10434195   2.00E−64   unnamed protein product [ Homo sapiens ]       250   2   127   140   520   g6467206   3.00E−36   gonadotropin inducible transcription repressor-4 [ Homo sapiens ]       250   2   127   140   520   g6330394   4.00E−34   KIAA1198 protein [ Homo sapiens ]       251   1   157   1   471   g340446   2.00E−17   zinc finger protein 7 (ZFP7) [ Homo sapiens ]       251   1   157   1   471   g4325310   2.00E−17   zinc-finger protein 7 [ Homo sapiens ]       251   1   157   1   471   g6007771   5.00E−17   KID2 [ Mus musculus ]       252   1   305   145   1059   g6002480   3.00E−49   BWSCR2 associated zinc-finger protein BAZ2 [ Homo sapiens ]       252   1   305   145   1059   g9963806   3.00E−47   zinc finger protein ZNF287 [ Homo sapiens ]       252   1   305   145   1059   g11527849   8.00E−43   zinc finger protein SKAT2 [ Mus musculus ]       253   2   717   305   2455   g10047335   0   KIAA1629 protein [ Homo sapiens ]       253   2   717   305   2455   g1504006   1.00E−96   similar to human ZFY protein. [ Homo sapiens ]       253   2   717   305   2455   g7243280   4.00E−66   KIAA1441 protein [ Homo sapiens ]       254   1   211   1   633   g10047183   3.00E−49   KIAA1559 protein [ Homo sapiens ]       254   1   211   1   633   g5080758   2.00E−45   BC331191_1 [ Homo sapiens ]       254   1   211   1   633   g498721   3.00E−44   zinc finger protein [ Homo sapiens ]       255   2   103   2   310   g498152   2.00E−20   ha0946 protein is Kruppel-related. [ Homo sapiens ]       255   2   103   2   310   g7576272   2.00E−20   bA393J16.1 (zinc finger protein 33a (KOX 31)) [ Homo sapiens ]       255   2   103   2   310   g10440081   2.00E−19   unnamed protein product [ Homo sapiens ]       256   3   84   135   386   g347906   2.00E−26   zinc finger protein [ Homo sapiens ]       256   3   84   135   386   g3342002   1.00E−25   hematopoietic cell derived zinc finger protein [ Homo sapiens ]       256   3   84   135   386   g8163824   5.00E−25   krueppel-like zinc finger protein HZF2 [ Homo sapiens ]       257   1   194   103   684   g10435738   4.00E−74   unnamed protein product [ Homo sapiens ]       257   1   194   103   684   g1017722   8.00E−73   repressor transcriptional factor [ Homo sapiens ]       257   1   194   103   684   g7959207   3.00E−71   KIAA1473 protein [ Homo sapiens ]       258   1   129   28   414   g2072955   6.00E−07   p40 [ Homo sapiens ]       258   1   129   28   414   g483915   8.00E−07   ORF1, encodes a 40 kDa product [ Homo sapiens ]       258   1   129   28   414   g339776   8.00E−07   ORF1 codes for a 40 kDa product [ Homo sapiens ]       259   3   93   75   353   g3329372   4.00E−36   DNA-binding protein [ Homo sapiens ]       259   3   93   75   353   g7959207   1.00E−33   KIAA1473 protein [ Homo sapiens ]       259   3   93   75   353   g184452   3.00E−33   Krueppel-related DNA-binding protein [ Homo sapiens ]       260   3   193   369   947   g8099348   1.00E−38   zinc finger protein [ Homo sapiens ]       260   3   193   369   947   g5730196   2.00E−38   Kruppel-type zinc finger [ Homo sapiens ]       260   3   193   369   947   g8050899   4.00E−38   ZNF180 [ Homo sapiens ]       261   3   111   3   335   g7023216   1.00E−14   unnamed protein product [ Homo sapiens ]       261   3   111   3   335   g3406676   6.00E−14   zinc finger protein 54 [ Mus musculus ]       261   3   111   3   335   g9802037   3.00E−13   zinc finger protein SBZF3 [ Homo sapiens ]       262   3   137   75   485   g186774   1.00E−26   zinc finger protein [ Homo sapiens ]       262   3   137   75   485   g2384653   6.00E−26   Krueppel family zinc finger protein [ Homo sapiens ]       262   3   137   75   485   g8163824   6.00E−26   krueppel-like zinc finger protein HZF2 [ Homo sapiens ]       263   3   68   51   254   g7239109   2.00E−15   HSPC059 [ Homo sapiens ]       263   3   68   51   254   g347906   4.00E−15   zinc finger protein [ Homo sapiens ]       263   3   68   51   254   g7023216   2.00E−14   unnamed protein product [ Homo sapiens ]       264   3   101   90   392   g3329372   8.00E−35   DNA-binding protein [ Homo sapiens ]       264   3   101   90   392   g4559318   7.00E−32   BC273239_1 [ Homo sapiens ]       264   3   101   90   392   g184452   9.00E−32   Krueppel-related DNA-binding protein [ Homo sapiens ]       265   1   96   184   471   g4589588   5.00E−22   KIAA0972 protein [ Homo sapiens ]       265   1   96   184   471   g4514561   6.00E−22   KRAB-containing zinc-finger protein KRAZ2 [ Mus musculus ]       265   1   96   184   471   g7576272   2.00E−21   bA393J16.1 (zinc finger protein 33a (KOX 31)) [ Homo sapiens ]       266   2   251   2   754   g55471   1.00E−134   Zfp-29 [ Mus musculus ]       266   2   251   2   754   g1020145   3.00E−73   DNA binding protein [ Homo sapiens ]       266   2   251   2   754   g6002478   3.00E−72   BWSCR2 associated zinc-finger protein BAZ1 [ Homo sapiens ]       267   3   522   36   1601   g10443047   0   bA465L10.2 (novel C2H2 type zinc finger protein similar to chicken FZF-1)                                   [ Homo sapiens ]       267   3   522   36   1601   g10438918   0   unnamed protein product [ Homo sapiens ]       267   3   522   36   1601   g984814   2.00E−97   zinc finger protein [ Gallus gallus ]       268   2   267   2   802   g9886891   4.00E−45   zinc finger protein 276 C2H2 type [ Mus musculus ]       268   2   267   2   802   g11611571   3.00E−43   hypothetical protein [ Macaca fascicularis ]       268   2   267   2   802   g453376   4.00E−43   zinc finger protein PZF [ Mus musculus ]       269   2   286   2   859   g2754696   9.00E−08   high molecular mass nuclear antigen [ Gallus gallus ]       269   2   286   2   859   g2078483   9.00E−06   antifreeze glycopeptide AFGP polyprotein precursor [ Boreogadus saida ]       270   3   194   270   851   g8575782   1.00E−112   PR-domain zinc finger protein 6 isoform A; PR-domain family protein 3 isoform                                   A; PRDM6A; PFM3A [ Homo sapiens ]       270   3   194   270   851   g10437767   1.00E−26   unnamed protein product [ Homo sapiens ]       270   3   194   270   851   g7295698   9.00E−26   CG15436 gene product [ Drosophila melanogaster ]       271   3   263   3   791   g6409345   1.00E−107   zinc finger protein ZNF180 [ Homo sapiens ]       271   3   263   3   791   g8050899   1.00E−107   ZNF180 [ Homo sapiens ]       271   3   263   3   791   g200407   1.00E−101   pMLZ-4 [ Mus musculus ]       272   2   142   290   715   g4062983   5.00E−65   Eos protein [ Mus musculus ]       272   2   142   290   715   g9408382   4.00E−46   eos [ Raja eglanteria ]       272   2   142   290   715   g11612390   3.00E−42   zinc finger transcription factor Eos [ Homo sapiens ]       273   2   164   2   493   g1049301   3.00E−25   KRAB zinc finger protein; Method: conceptual translation supplied by author                                   [ Homo sapiens ]       273   2   164   2   493   g10047251   9.00E−25   KIAA1588 protein [ Homo sapiens ]       273   2   164   2   493   g8809810   1.00E−19   KRAB zinc finger protein [ Mus musculus ]       274   2   107   509   829   g1237278   2.00E−36   zinc finger protein [ Cavia porcellus ]       274   2   107   509   829   g7023417   4.00E−36   unnamed protein product [ Homo sapiens ]       274   2   107   509   829   g11917507   5.00E−36   HPF1 protein [ Homo sapiens ]       275   3   105   336   650   g9801232   2.00E−51   bA508N22.2 (zinc finger protein 37a (KOX 21)) [ Homo sapiens ]       275   3   105   336   650   g829151   2.00E−51   ZNF37A [ Homo sapiens ]       275   3   105   336   650   g5730196   4.00E−36   Kruppel-type zinc finger [ Homo sapiens ]       276   1   149   1   447   g7656698   3.00E−91   Zinc finger protein 222 [ Homo sapiens ]       276   1   149   1   447   g6118381   3.00E−91   zinc finger protein ZNF222 [ Homo sapiens ]       276   1   149   1   447   g6118383   1.00E−81   zinc finger protein ZNF223 [ Homo sapiens ]       277   3   101   90   392   g3329372   1.00E−30   DNA-binding protein [ Homo sapiens ]       277   3   101   90   392   g4559318   3.00E−29   BC273239_1 [ Homo sapiens ]       277   3   101   90   392   g1124876   5.00E−29   Krueppel-related DNA-binding protein [ Homo sapiens ]       278   3   137   6   416   g11062533   2.00E−46   bA245E14.1 (novel zinc finger protein similar to ZFP47) [ Homo sapiens ]       278   3   137   6   416   g5640017   2.00E−46   zinc finger protein ZFP113 [ Mus musculus ]       278   3   137   6   416   g186774   5.00E−46   zinc finger protein [ Homo sapiens ]       279   3   97   165   455   g829151   2.00E−27   ZNF37A [ Homo sapiens ]       279   3   97   165   455   g9801232   2.00E−27   bA508N22.2 (zinc finger protein 37a (KOX 21)) [ Homo sapiens ]       279   3   97   165   455   g3702137   9.00E−20   dJ733D15.1 (Zinc-finger protein) [ Homo sapiens ]       280   2   97   182   472   g9801232   4.00E−29   bA508N22.2 (zinc finger protein 37a (KOX 21)) [ Homo sapiens ]       280   2   97   182   472   g829151   4.00E−29   ZNF37A [ Homo sapiens ]       280   2   97   182   472   g200407   4.00E−21   pMLZ-4 [ Mus musculus ]       281   1   179   31   567   g10442700   3.00E−61   zinc-finger protein ZBRK1 [ Homo sapiens ]       281   1   179   31   567   g10435411   3.00E−61   unnamed protein product [ Homo sapiens ]       281   1   179   31   567   g10954044   3.00E−61   KRAB zinc finger protein ZFQR [ Homo sapiens ]       282   3   87   369   629   g8099348   2.00E−14   zinc finger protein [ Homo sapiens ]       282   3   87   369   629   g498725   2.00E−14   zinc finger protein [ Homo sapiens ]       282   3   87   369   629   g495568   2.00E−13   zinc finger protein [ Homo sapiens ]       283   2   172   2   517   g6007771   4.00E−97   KID2 [ Mus musculus ]       283   2   172   2   517   g2970038   2.00E−93   HKL1 [ Homo sapiens ]       283   2   172   2   517   g205067   2.00E−93   zinc finger protein [ Rattus norvegicus ]       284   1   151   1   453   g1806134   5.00E−57   zinc finger protein [ Mus musculus ]       284   1   151   1   453   g538413   5.00E−57   zinc finger protein [ Mus musculus ]       284   1   151   1   453   g186774   3.00E−55   zinc finger protein [ Homo sapiens ]       285   2   89   83   349   g7023216   2.00E−18   unnamed protein product [ Homo sapiens ]       285   2   89   83   349   g9802037   4.00E−16   zinc finger protein SBZF3 [ Homo sapiens ]       285   2   89   83   349   g7239109   7.00E−15   HSPC059 [ Homo sapiens ]       286   2   146   62   499   g2739353   7.00E−56   ZNF91L [ Homo sapiens ]       286   2   146   62   499   g7959207   5.00E−50   KIAA1473 protein [ Homo sapiens ]       286   2   146   62   499   g3342002   7.00E−50   hematopoietic cell derived zinc finger protein [ Homo sapiens ]       287   1   78   1   234   g487785   4.00E−16   zinc finger protein ZNF136 [ Homo sapiens ]       287   1   78   1   234   g5262560   7.00E−15   hypothetical protein [ Homo sapiens ]       287   1   78   1   234   g10434856   9.00E−15   unnamed protein product [ Homo sapiens ]       288   3   126   78   455   g9963804   4.00E−47   zinc finger protein ZNF286 [ Homo sapiens ]       288   3   126   78   455   g5640017   2.00E−46   zinc finger protein ZFP113 [ Mus musculus ]       288   3   126   78   455   g7020166   4.00E−46   unnamed protein product [ Homo sapiens ]       289   1   96   151   438   g4589588   5.00E−22   KIAA0972 protein [ Homo sapiens ]       289   1   96   151   438   g4514561   6.00E−22   KRAB-containing zinc-finger protein KRAZ2 [ Mus musculus ]       289   1   96   151   438   g7576272   2.00E−21   bA393J16.1 (zinc finger protein 33a (KOX 31)) [ Homo sapiens ]       290   1   149   118   564   g7959207   1.00E−26   KIAA1473 protein [ Homo sapiens ]       290   1   149   118   564   g498736   3.00E−26   zinc finger protein [ Homo sapiens ]       290   1   149   118   564   g4454678   4.00E−23   zinc finger protein 4 [ Homo sapiens ]       291   2   134   152   553   g498152   1.00E−06   ha0946 protein is Kruppel-related. [ Homo sapiens ]       291   2   134   152   553   g10440081   1.00E−06   unnamed protein product [ Homo sapiens ]       291   2   134   152   553   g7576272   1.00E−06   bA393J16.1 (zinc finger protein 33a (KOX 31)) [ Homo sapiens ]       292   2   212   2   637   g7656698   1.00E−133   Zinc finger protein 222 [ Homo sapiens ]       292   2   212   2   637   g6118381   1.00E−133   zinc finger protein ZNF222 [ Homo sapiens ]       292   2   212   2   637   g6118383   1.00E−122   zinc finger protein ZNF223 [ Homo sapiens ]       293   2   108   2   325   g4567179   2.00E−33   BC37295_1 [ Homo sapiens ]       293   2   108   2   325   g10434142   9.00E−31   unnamed protein product [ Homo sapiens ]       293   2   108   2   325   g5817149   9.00E−31   hypothetical protein [ Homo sapiens ]       294   1   83   97   345   g930123   9.00E−24   zinc finger protein (583 AA) [ Homo sapiens ]       294   1   83   97   345   g487785   8.00E−23   zinc finger protein ZNF136 [ Homo sapiens ]       294   1   83   97   345   g5262560   1.00E−22   hypothetical protein [ Homo sapiens ]       295   1   180   1   540   g498719   2.00E−83   zinc finger protein [ Homo sapiens ]       295   1   180   1   540   g3953593   3.00E−69   Zinc finger protein s11-6 [ Mus musculus ]       295   1   180   1   540   g6467206   4.00E−68   gonadotropin inducible transcription repressor-4 [ Homo sapiens ]       296   3   97   57   347   g9801232   3.00E−28   bA508N22.2 (zinc finger protein 37a (KOX 21)) [ Homo sapiens ]       296   3   97   57   347   g829151   3.00E−28   ZNF37A [ Homo sapiens ]       296   3   97   57   347   g881564   4.00E−20   ZNF157 [ Homo sapiens ]       297   1   217   421   1071   g6331377   1.00E−131   KIAA1285 protein [ Homo sapiens ]       297   1   217   421   1071   g1020145   6.00E−53   DNA binding protein [ Homo sapiens ]       297   1   217   421   1071   g2224593   1.00E−52   KIAA0326 [ Homo sapiens ]       298   3   137   15   425   g4456989   4.00E−20   protease [ Homo sapiens ]       298   3   137   15   425   g9558703   4.00E−20   protease [ Homo sapiens ]       298   3   137   15   425   g1780976   5.00E−20   protease [Human endogenous retrovirus K]       299   2   169   59   565   g10434856   2.00E−40   unnamed protein product [ Homo sapiens ]       299   2   169   59   565   g5262560   2.00E−40   hypothetical protein [ Homo sapiens ]       299   2   169   59   565   g930123   1.00E−31   zinc finger protein (583 AA) [ Homo sapiens ]       300   3   135   3   407   g10434856   3.00E−35   unnamed protein product [ Homo sapiens ]       300   3   135   3   407   g5262560   3.00E−35   hypothetical protein [ Homo sapiens ]       300   3   135   3   407   g10434195   2.00E−27   unnamed protein product [ Homo sapiens ]       301   1   170   22   531   g10047297   2.00E−23   KIAA1611 protein [ Homo sapiens ]       301   1   170   22   531   g7023216   2.00E−22   unnamed protein product [ Homo sapiens ]       301   1   170   22   531   g347906   5.00E−16   zinc finger protein [ Homo sapiens ]       302   3   181   3   545   g5931821   8.00E−79   dJ228H13.3 (zinc finger protein) [ Homo sapiens ]       302   3   181   3   545   g6807587   8.00E−79   hypothetical protein [ Homo sapiens ]       302   3   181   3   545   g488555   2.00E−63   zinc finger protein ZNF135 [ Homo sapiens ]       303   1   263   1   789   g506502   1.00E−141   NK10 [ Mus musculus ]       303   1   263   1   789   g488555   1.00E−92   zinc finger protein ZNF135 [ Homo sapiens ]       303   1   263   1   789   g8453103   7.00E−88   zinc finger protein [ Homo sapiens ]       304   3   340   18   1037   g7023216   1.00E−142   unnamed protein product [ Homo sapiens ]       304   3   340   18   1037   g7023703   2.00E−89   unnamed protein product [ Homo sapiens ]       304   3   340   18   1037   g10436789   7.00E−54   unnamed protein product [ Homo sapiens ]       305   1   89   103   369   g7023216   2.00E−18   unnamed protein product [ Homo sapiens ]       305   1   89   103   369   g9802037   4.00E−16   zinc finger protein SBZF3 [ Homo sapiens ]       305   1   89   103   369   g7239109   7.00E−15   HSPC059 [ Homo sapiens ]       306   1   80   1   240   g7959865   9.00E−20   PRO2032 [ Homo sapiens ]       306   1   80   1   240   g8099520   6.00E−11   muscleblind [ Mus musculus ]       306   1   80   1   240   g8515711   2.00E−10   EXP35 [ Homo sapiens ]       307   2   386   176   1333   g3869259   0   ZNF202 beta [ Homo sapiens ]       307   2   386   176   1333   g7328045   0   hypothetical protein [ Homo sapiens ]       307   2   386   176   1333   g5360097   1.00E−123   putative kruppel-related zinc finger protein NY-REN-23 antigen [ Homo                                       sapiens ]       308   2   368   71   1174   g3882241   0   KIAA0760 protein [ Homo sapiens ]       308   2   368   71   1174   g6760445   0   Smad-and Olf-interacting zinc finger protein [ Homo sapiens ]       308   2   368   71   1174   g2149792   0   Roaz [ Rattus norvegicus ]       309   2   175   191   715   g487787   8.00E−15   zinc finger protein ZNF140 [ Homo sapiens ]       309   2   175   191   715   g10047183   9.00E−31   KIAA1559 protein [ Homo sapiens ]       309   2   175   191   715   g4567179   2.00E−29   BC37295_1 [ Homo sapiens ]       310   2   78   521   754       311   1   61   394   576       312   1   73   172   390   g2587027   4.00E−13   HERV-E envelope glycoprotein [ Homo sapiens ]       312   1   73   172   390   g2587024   4.00E−13   HERV-E envelope glycoprotein [ Homo sapiens ]       312   1   73   172   390   g1049232   2.00E−10   HERV-E envelope protein [Human endogenous retrovirus]       313   1   184   304   855   g8132311   2.00E−74   inwardly-rectifying potassium channel Kir5.1 [ Homo sapiens ]       313   1   184   304   855   g8132295   2.00E−74   inwardly-rectifying potassium channel Kir5.1 [ Homo sapiens ]       313   1   184   304   855   g8132293   2.00E−74   inwardly-rectifying potassium channel Kir5.1 [ Homo sapiens ]       314   2   219   164   820   g7105926   2.00E−22   calcium channel alpha2-delta3 subunit [ Homo sapiens ]       314   2   219   164   820   g4186073   2.00E−22   calcium channel alpha-2-delta-C subunit [ Mus musculus ]       314   2   219   164   820   g9929977   2.00E−22   hypothetical protein [ Macaca fascicularis ]       315   1   1603   1   4809   g184039   0   sodium channel alpha subunit [ Homo sapiens ]       315   1   1603   1   4809   g6782382   0   voltage-gated sodium channel [ Mus musculus ]       315   1   1603   1   4809   g206858   0   sodium channel alpha-subunit [ Rattus norvegicus ]       316   3   200   240   839   g913242   5.00E−71   gamma-aminobutyric acid transporter type 3, GABA transporter type 3, GAT-3                                   [human, fetal brain, Peptide, 632 aa] [ Homo sapiens ]       316   3   200   240   839   g204220   2.00E−69   beta-alanine-sensitive neuronal GABA transporter [ Rattus norvegicus ]       316   3   200   240   839   g202535   2.00E−69   GABA transporter [ Rattus norvegicus ]       317   3   329   3   989   g6996442   4.00E−61   CTL1 protein [ Homo sapiens ]       317   3   329   3   989   g6996589   1.00E−59   CTL1 protein [ Rattus norvegicus ]       317   3   329   3   989   g6996587   2.00E−51   CTL1 protein [ Torpedo marmorata ]       318   3   256   3   770   g5091520   1.00E−134   ESTs AU058081(E30812),AU058365(E50679), AU030138(E50679)                                   correspond to a region of the predicted gene.; Similar to  Spinacia oleracea                                     mRNA for proteasome 37 kD subunit.(X96974) [ Oryza sativa ]       318   3   256   3   770   g8096329   1.00E−134   ESTs AU058081(E3082),AU075427(E30384) correspond to a region of the                                   predicted gene.˜Similar to  Spinacia oleracea  proteasome 27 kD subunit                                   (P52427) [ Oryza sativa ]       318   3   256   3   770   g8096319   1.00E−134   ESTs AU058081(E3082),AU075427(E30384) correspond to a region of the                                   predicted gene. ˜Similar to  Spinacia oleracea  proteasome 27 kD subunit                                   (P52427) [ Oryza sativa ]       319   2   76   2   229   g951425   2.00E−07   housekeeping protein [ Rattus norvegicus ]       319   2   76   2   229   g5759144   2.00E−07   cyclophilin A [ Mus musculus ]       319   2   76   2   229   g50621   2.00E−07   cyclophilin (AA 1-164) [ Mus musculus ]       320   3   276   354   1181   g7019854   1.00E−84   unnamed protein product [ Homo sapiens ]       320   3   276   354   1181   g6567172   7.00E−84   mDj10 [ Mus musculus ]       320   3   276   354   1181   g10436329   5.00E−81   unnamed protein product [ Homo sapiens ]       321   1   115   328   672   g1049232   3.00E−24   HERV-E envelope protein [Human endogenous retrovirus]       321   1   115   328   672   g2587024   2.00E−23   HERV-E envelope glycoprotein [ Homo sapiens ]       321   1   115   328   672   g2587027   2.00E−23   HERV-E envelope glycoprotein [ Homo sapiens ]       322   3   227   3   683   g2286123   6.00E−33   testis specific DNAj-homolog [ Mus musculus ]       322   3   227   3   683   g6681592   1.00E−32   DnaJ homolog [ Homo sapiens ]       322   3   227   3   683   g6648623   1.00E−32   DNAJ homolog [ Homo sapiens ]       323   3   100   153   452       324   3   142   840   1265   g2943716   5.00E−81   25 kDa trypsin inhibitor [ Homo sapiens ]       324   3   142   840   1265   g9885193   5.00E−54   dJ881L22.3 (novel protein similar to a trypsin inhibitor) [ Homo sapiens ]       324   3   142   840   1265   g4324682   2.00E−52   late gestation lung protein 1 [ Rattus norvegicus ]       325   3   263   3   791   g6957716   1.00E−135   putative chaperonin [ Arabidopsis thaliana ]       325   3   263   3   791   g9755653   1.00E−132   TCP-1 chaperonin-like protein [ Arabidopsis thallana ]       325   3   263   3   791   g5295933   2.00E−93   chaperonin containing TCP-1 zeta-1 subunit [ Mus musculus ]       326   2   357   23   1093   g3882167   1.00E−171   KIAA0723 protein [ Homo sapiens ]       326   2   357   23   1093   g9956070   1.00E−171   similar to Homo sapiens mRNA for KIAA0723 protein with GenBank                                   Accession Number AB018266.1 []       326   2   357   23   1093   g6563246   1.00E−170   matrin 3 [ Homo sapiens ]       327   2   100   656   955       328   2   303   2   910   g8980660   1.00E−158   proliferation-associated SNF2-like protein [ Homo sapiens ]       328   2   303   2   910   g805296   1.00E−149   lymphocyte specific helicase [ Mus musculus ]       328   2   303   2   910   g9956001   8.00E−86   similar to  Mus musculus  lymphocyte specific helicase mRNA with GenBank                                   Accession Number U25691.1 [ Homo sapiens ]       329   2   72   167   382       330   2   76   80   307       331   2   74   446   667   g2104910   1.00E−29   ORF derived from D1 leader region and integrase coding region [ Homo                                       sapiens ]       331   2   74   446   667   g2104914   5.00E−21   ORF derived from protease and integrase coding regions [ Homo sapiens ]       331   2   74   446   667   g4959374   5.00E−21   pol protein [ Homo sapiens ]       332   3   67   57   257       333   2   192   302   877   g8980660   8.00E−92   proliferation-associated SNF2-like protein [ Homo sapiens ]       333   2   192   302   877   g9956001   8.00E−92   similar to  Mus musculus  lymphocyte specific helicase mRNA with GenBank                                   Accession Number U25691.1 [ Homo sapiens ]       333   2   192   302   877   g7022306   1.00E−89   unnamed protein product [ Homo sapiens ]       334   2   74   446   667   g2104910   1.00E−30   ORF derived from D1 leader region and integrase coding region [ Homo                                       sapiens ]       334   2   74   446   667   g2104914   5.00E−21   ORF derived from protease and integrase coding regions [ Homo sapiens ]       334   2   74   446   667   g4959374   5.00E−21   pol protein [ Homo sapiens ]       335   2   72   167   382       336   2   55   557   721   g2231380   8.00E−12   orf; encodes putative chimeric protein with SET domain in N-terminus with                                   similarity to several other human, Drosophlla, nematode and yeast proteins                                   [ Homo sapiens ]       336   2   55   557   721   g3005702   8.00E−12   unknown [ Homo sapiens ]       336   2   55   557   721   g1263081   1.00E−11   mariner transposase [ Homo sapiens ]       337   3   107   1614   1934       338   3   147   63   503   g10047265   7.00E−81   KIAA1595 protein [ Homo sapiens ]       338   3   147   63   503   g10176757   3.00E−26   ATP-dependent RNA helicase-like protein [ Arabidopsis thaliana ]       338   3   147   63   503   g3776011   3.00E−26   RNA helicase [ Arabidopsis thaliana ]       339   1   257   199   969   g10434055   1.00E−128   unnamed protein product [ Homo sapiens ]       339   1   257   199   969   g7243213   1.00E−126   KIAA1416 protein [ Homo sapiens ]       339   1   257   199   969   g11345539   1.00E−120   dJ620E11.1 (novel Helicase C-terminal domain and SNF2 N-terminal domains                                   containing protein, similar to KIAA0308) [ Homo sapiens ]       340   3   63   3   191       341   1   112   1639   1974       342   3   427   2097   3377   g2599502   0   protocadherin 68 [ Homo sapiens ]       342   3   427   2097   3377   g7243181   4.00E−49   KIAA1400 protein [ Homo sapiens ]       342   3   427   2097   3377   g4099551   5.00E−48   OL-protocadherin [ Mus musculus ]       343   2   144   635   1066   g10436424   1.00E−10   unnamed protein product [ Homo sapiens ]       344   2   97   557   847       345   3   75   675   899   g2587027   4.00E−13   HERV-E envelope glycoprotein [ Homo sapiens ]       345   3   75   675   899   g2587024   4.00E−13   HERV-E envelope glycoprotein [ Homo sapiens ]       345   3   75   675   899   g1049232   2.00E−10   HERV-E envelope protein [Human endogenous retrovirus]       346   3   135   399   803   g9368839   2.00E−71   hypothetical protein [ Homo sapiens ]       346   3   135   399   803   g2739452   6.00E−58   ribosomal protein L23A [ Homo sapiens ]       346   3   135   399   803   g1399086   6.00E−58   ribosomal protein L23a [ Homo sapiens ]       347   2   55   179   343       348   2   129   425   811   g11493463   2.00E−22   PRO2852 [ Homo sapiens ]       348   2   129   425   811   g9280152   5.00E−22   unnamed portein product [ Macaca fascicularis ]       348   2   129   425   811   g10437485   5.00E−21   unnamed protein product [ Homo sapiens ]       349   2   291   122   994   g673417   1.00E−152   class II antigen [ Homo sapiens ]       349   2   291   122   994   g703089   1.00E−152   MHC class II DP3-alpha [ Homo sapiens ]       349   2   291   122   994   g758100   1.00E−137   SB classII histocompatibility antigen alpha- chain [ Homo sapiens ]       350   1   517   1   1551   g402843   1.00E−144   cytochrome P450 2B-Bx = phenobarbital-inducible [rabbits, kidney, Peptide, 491                                   aa] [ Oryctolagus cuniculus ]       350   1   517   1   1551   g404777   1.00E−144   cytochrome P-450 2B-Bx [ Oryctolagus cuniculus ]       350   1   517   1   1551   g164959   1.00E−142   cytochrome P-450 [ Oryctolagus cuniculus ]       351   1   232   1300   1995       352   1   220   67   726   g11863734   2.00E−80   dJ857M17.2 (novel protein similar to cytochrome c oxidase subunit IV                                   (COX4)) [ Homo sapiens ]       352   1   220   67   726   g8809758   9.00E−42   cytochrome c oxidase subunit IV isoform 2 precursor [ Thunnus obesus ]       352   1   220   67   726   g2809498   3.00E−41   cytochrome c oxidase subunit IV [ Gorilla gorilla ]       353   1   95   1   285       354   2   331   2   994   g11229985   1.00E−176   unnamed protein product [ Homo sapiens ]       354   2   331   2   994   g11229992   6.00E−57   unnamed protein product [ Mus musculus ]       354   2   331   2   994   g30095   6.00E−49   collagen subunit (alpha-1 (X)) 3 [ Homo sapiens ]       355   3   93   54   332   g11177164   4.00E−12   polydom protein [ Mus musculus ]       355   3   93   54   332   g391669   4.00E−07   hikaru genki type4 product precursor [ Drosophila melanogaster ]       355   3   93   54   332   g391667   4.00E−07   hikaru genki type3 product precursor [ Drosophila melanogaster ]       356   1   112   1   336       357   3   73   192   410       358   1   239   181   897   g4582324   1.00E−129   dJ708F5.1 (PUTATIVE novel Collagen alpha 1 LIKE protein) [ Homo sapiens ]       358   1   239   181   897   g1732121   4.00E−36   cartilage matrix protein [ Homo sapiens ]       358   1   239   181   897   g180654   2.00E−35   cartilage matrix protein [ Homo sapiens ]       359   1   528   4   1587   g1903218   0   type II intermediate filament of hair keratin [ Homo sapiens ]       359   1   528   4   1587   g7161771   0   keratin [ Homo sapiens ]       359   1   528   4   1587   g4103156   0   hair keratin basic 5; keratin Hb5 [ Mus musculus ]       360   2   157   161   631   g11034725   2.00E−64   hNBL4 [ Homo sapiens ]       360   2   157   161   631   g466548   3.00E−63   NBL4 [ Mus musculus ]       360   2   157   161   631   g2822458   5.00E−54   band 4.1-like protein 4 [ Danio rerio ]       361   3   65   54   248   g3724141   6.00E−08   myosin I [ Rattus norvegicus ]       361   3   65   54   248   g3882175   6.00E−08   KIAA0727 protein [ Homo sapiens ]       362   3   517   3   1553   g6855339   1.00E−120   dJ111C20.1 (similar to Chlamydomonas radial spoke protein 3) [ Homo                                       sapiens ]       362   3   517   3   1553   g18218   1.00E−75   spoke protein [ Chlamydomonas reinhardtii ]       362   3   517   3   1553   g7295323   9.00E−47   CG10099 gene product [ Drosophila melanogaster ]       363   2   60   314   493       364   1   239   127   843   g1813638   9.00E−53   PF20 [ Chlamydomonas reinhardtii ]       364   1   239   127   843   g3983133   2.00E−47   pf20 homolog [ Trypanosoma brucei ]       364   1   239   127   843   g607003   1.00E−37   beta transducin-like protein [ Podospora anserina ]       365   1   160   1   480       366   3   757   3   2273   g8896164   0   kinesin-like protein GAKIN [ Homo sapiens ]       366   3   757   3   2273   g10697238   0   KIF13A [ Mus musculus ]       366   3   757   3   2273   g11761613   0   kinesin-like protein RBKIN2 [ Homo sapiens ]       367   3   162   3   488   g11231085   1.00E−56    hypothetical protein [ Macaca fascicularis ]       367   3   162   3   488   g7385113   2.00E−18   ankyrin 1 [ Bos taurus ]       367   3   162   3   488   g747710   2.00E−18   alt, ankyrin (variant 2.2) [ Homo sapiens ]       368   2   635   308   2212   g1353782   0   dystrophin-related protein 2 [ Homo sapiens ]       368   2   635   308   2212   g11066165   0   dystrophin-related protein 2 A-form splice variant [ Rattus norvegicus ]       368   2   635   308   2212   g11066167   0   dystrophin-related protein 2 B-form splice variant [ Rattus norvegicus ]       369   3   433   999   2297   g190752   0   pemphigus vulgaris antigen [ Homo sapiens ]       369   3   433   999   2297   g2290200   1.00E−176   desmoglein 3 [ Mus musculus ]       369   3   433   999   2297   g416178   2.00E−58   desmoglein 2 [ Homo sapiens ]       370   3   531   3   1595   g28969   7.00E−71   64 Kd autoantigen [ Homo sapiens ]       370   3   531   3   1595   g6934240   8.00E−61   tropomodulin 2 [ Homo sapiens ]       370   3   531   3   1595   g7288857   3.00E−60   neural tropomodulin N-Tmod [ Mus musculus ]       371   2   257   383   1153   g1469868   1.00E−124   The KIAA0143 gene product is related to a putative  C.elegans  gene encoded                                   on cosmid C32D5. [ Homo sapiens ]       371   2   257   383   1153   g4589550   4.00E−82   KIAA0953 protein [ Homo sapiens ]       371   2   257   383   1153   g7304005   1.00E−55   cmp44E gene product [alt 1] [ Drosophila melanogaster ]       372   1   242   139   864   g387514   1.00E−123   DM-20 protein [ Mus musculus ]       372   1   242   139   864   g190088   1.00E−123   DM-20 [ Homo sapiens ]       372   1   242   139   864   g200409   1.00E−122   proteolipid protein variant Dm-20 [ Mus musculus ]       373   2   60   380   559       374   1   157   22   492   g7268562   1.00E−59   ribosomal protein L32-like protein [ Arabidopsis thaliana ]       374   1   157   22   492   g5816996   1.00E−59   ribosomal protein L32-like protein [ Arabidopsis thaliana ]       374   1   157   22   492   g10177580   7.00E−59   ribosomal protein L32 [ Arabidopsis thaliana ]       375   3   158   3   476   g643074   4.00E−76   putative 40S ribosomal protein s12 [ Fragaria  x  ananassa ]       375   3   158   3   476   g6716785   1.00E−75   40s ribosomal protein S23 [ Euphorbia esula ]       375   3   158   3   476   g7413571   6.00E−75   putative protein [ Arabidopsis thaliana ]       376   2   238   14   727   g10799832   1.00E−93   ribosomal protein L11-like [ Nicotiana tabacum ]       376   2   238   14   727   g7630065   4.00E−93   ribosomal protein L11-like [ Arabidopsis thaliana ]       376   2   238   14   727   g11908058   4.00E−93   ribosomal protein L11, cytosolic [ Arabidopsis thaliana ]       377   3   102   3   308   g57131   7.00E−41   ribosomal protein S26 [ Rattus norvegicus ]       377   3   102   3   308   g296452   7.00E−41   ribosomal protein S26 [ Homo sapiens ]       377   3   102   3   308   g3335024   7.00E−41   ribosomal protein S26 [ Homo sapiens ]       378   1   102   316   621   g6969165   6.00E−53   dJ475N16.3 (novel protein similar to RPL7A (60S ribosomal protein L7A))                                   [ Homo sapiens ]       378   1   102   316   621   g6687301   2.00E−21   60S ribosomal protein L7 [ Cyanophora paradoxa ]       378   1   102   316   621   g200785   1.00E−20   ribosomal protein L7 [ Mus musculus ]       379   3   177   3   533   g206736   1.00E−82   ribosomal protein L7 [ Rattus norvegicus ]       379   3   177   3   533   g200785   2.00E−80   ribosomal protein L7 [ Mus musculus ]       379   3   177   3   533   g554269   2.00E−80   ribosomal protein L7 [ Mus musculus ]       380   2   86   257   514   g550025   2.00E−31   ribosomal protein S10 [ Homo sapiens ]       380   2   86   257   514   g57127   3.00E−30   ribosomal protein S10 (AA 1-165) [ Rattus norvegicus ]       380   2   86   257   514   g9581772   3.00E−29   bA371L19.2 (similar to ribosomal protein S10) [ Homo sapiens ]       381   1   97   286   576   g36140   2.00E−31   ribosomal protein L7 [ Homo sapiens ]       381   1   97   286   576   g307388   2.00E−31   ribosomal protein L7 [ Homo sapiens ]       381   1   97   286   576   g1335288   2.00E−31   ribosomal protein L7 [ Homo sapiens ]       382   1   82   70   315   g409074   2.00E−19   HBp15/L22 [ Sus scrofa ]       382   1   82   70   315   g409072   2.00E−19   HBp15/L22 [ Mus musculus ]       382   1   82   70   315   g409070   2.00E−19   HBp15/L22 [ Homo sapiens ]       383   1   180   46   585   g4886269   2.00E−75   putative ribosomal protein S14 [ Arabidopsis thaliana ]       383   1   180   46   585   g6006890   6.00E−75   putative 40S ribosomal protein s14; 67401-66292 [ Arabidopsis thaliana ]       383   1   180   46   585   g4678226   3.00E−74   40S ribosomal protein S14 [ Arabidopsis thaliana ]       384   3   118   21   374   g643074   2.00E−49   putative 40S ribosomal protein s12 [ Fragaria  x  ananassa ]       384   3   118   21   374   g6716785   6.00E−49   40s ribosomal protein S23 [ Euphorbia esula ]       384   3   118   21   374   g7413571   3.00E−48   putative protein [ Arabidopsis thaliana ]       385   2   164   2   493   g643074   4.00E−76   putative 40S ribosomal protein s12 [ Fragaria  x  ananassa ]       385   2   164   2   493   g6716785   1.00E−75   40s ribosomal protein S23 [ Euphorbia esula ]       385   2   164   2   493   g7413571   6.00E−75   putative protein [ Arabidopsis thaliana ]       386   3   101   3   305   g36130   1.00E−22   ribosomal protein L31 (AA 1-125) [ Homo sapiens ]       386   3   101   3   305   g1655596   1.00E−22   ribosomal protein L31 [ Homo sapiens ]       386   3   101   3   305   g57115   1.00E−22   ribosomal protein L31 (AA 1-125) [ Rattus norvegicus ]       387   3   259   3   779   g2331301   1.00E−122   ribosomal protein S4 type I [ Zea mays ]       387   3   259   3   779   g2345154   1.00E−120   ribsomal protein S4 [ Zea mays ]       387   3   259   3   779   g7546687   1.00E−116   ribosomal protein S4 [ Arabidopsis thaliana ]       388   2   184   2   553   g2668748   1.00E−95   ribosomal protein L17 [ Zea mays ]       388   2   184   2   553   g19104   8.00E−85   ribosomal protein L17-2 [ Hordeum vulgare ]       388   2   184   2   553   g19102   1.00E−82   ribosomal protein L17-1 [ Hordeum vulgare ]       389   2   152   2   457   g338447   5.00E−28   RPS16 [ Homo sapiens ]       389   2   152   2   457   g57714   5.00E−28   ribosomal protein S16 (AA 1-146) [Rattus rattus]       389   2   152   2   457   g200796   2.00E−27   16S ribosomal protein [ Mus musculus ]       390   3   158   3   476   g643074   4.00E−76   putative 40S ribosomal protein s12 [ Fragaria  x  ananassa ]       390   3   158   3   476   g6716785   1.00E−75   40s ribosomal protein S23 [ Euphorbia esula ]       390   3   158   3   476   g7413571   6.00E−75   putative protein [ Arabidopsis thaliana ]       391   1   94   34   315       392   3   83   303   551   g57121   3.00E−18   ribosomal protein L37 [ Rattus norvegicus ]       392   3   83   303   551   g292441   3.00E−18   ribosomal protein L37 [ Homo sapiens ]       392   3   83   303   551   g1839334   3.00E−18   ribosomal protein L37 {C2-C2 zinc-finger-like} [human, HeLa cells, Peptlde, 97                                   aa] [ Homo sapiens ]       393   2   174   2   523   g10433651   3.00E−80   unnamed protein product [ Homo sapiens ]       393   2   174   2   523   g10434617   3.00E−80   unnamed protein product [ Homo sapiens ]       393   2   174   2   523   g545998   6.00E−79   tricarboxylate carrier [Rattus sp.]       394   3   183   3   551       395   1   399   1   1197   g11907599   0   protein kinase HIPK2 [ Homo sapiens ]       395   1   399   1   1197   g5815141   0   nuclear body associated kinase 1b [ Mus musculus ]       395   1   399   1   1197   g5815139   0   nuclear body associated kinase 1a [ Mus musculus ]       396   1   301   109   1011   g7688667   1.00E−161   PC326 protein [ Homo sapiens ]       396   1   301   109   1011   g2734854   1.00E−08     Mus musculus  Dentin Matrix Protein 1 []       396   1   301   109   1011   g6137020   1.00E−08   dentin matrix protein-1 [ Mus musculus ]       397   2   105   2   316   g178281   1.00E−47   AHNAK nucleoprotein [ Homo sapiens ]       397   2   105   2   316   g50675   2.00E−47   desmoyokin [ Mus musculus ]       397   2   105   2   316   g897824   5.00E−47   AHNAK gene product [ Homo sapiens ]       398   1   153   202   660   g183233   1.00E−34   beta-glucuronidase precursor (EC 3.2.1.31) [ Homo sapiens ]       398   1   153   202   660   g3549609   2.00E−33   beta-glucuronidase [Cercopithecus aethiops]       398   1   153   202   660   g4102553   3.00E−29   mutant beta-glucuronidase [ Felis catus ]       399   1   161   106   588   g7022046   1.00E−36   unnamed protein product [ Homo sapiens ]       399   1   161   106   588   g7670456   5.00E−34   unnamed protein product [ Mus musculus ]       399   1   161   106   588   g8671586   1.00E−29   ataxin 2-binding protein [ Homo sapiens ]       400   1   153   205   663   g183233   1.00E−34   beta-glucuronidase precursor (EC 3.2.1.31) [ Homo sapiens ]       400   1   153   205   663   g3549609   2.00E−33   beta-glucuronidase [ Cercopithecus aethiops ]       400   1   153   205   663   g4102553   3.00E−29   mutant beta-glucuronidase [ Felis catus ]       401   3   135   651   1055   g414797   9.00E−58   pyruvate dehydrogenase phosphatase [ Bos taurus ]       401   3   135   651   1055   g3298607   3.00E−56   pyruvate dehydrogenase phosphatase isoenzyme 1 [ Rattus norvegicus ]       401   3   135   651   1055   g7688679   3.00E−53   pyruvate dehydrogenase [ Homo sapiens ]       402   3   129   30   416       403   1   299   1   897   g440878   1.00E−149   onconeural ventral antigen-1 [ Homo sapiens ]       403   1   299   1   897   g7025507   1.00E−137   ventral neuron-specific protein 1 NOVA1 [ Mus musculus ]       403   1   299   1   897   g2673961   9.00E−99   astrocytic NOVA-like RNA-binding protein [ Homo sapiens ]       404   1   142   1   426   g4105111   1.00E−43   dehydrin 6 [ Hordeum vulgare ]       404   1   142   1   426   g6017938   4.00E−43   dehydrin; DHN6 [ Hordeum vulgare ]       404   1   142   1   426   g5738195   1.00E−28   abscisic acid response protein [ Prunus dulcis ]       405   2   168   2   505   g453189   9.00E−59   acyl carrier protein [ Zea mays ]       405   2   168   2   505   g166971   4.00E−49   acyl carrier protein III [ Hordeum vulgare ]       405   2   168   2   505   g166969   6.00E−41   acyl carrier protein II [ Hordeum vulgare ]       406   2   117   2   352   g203923   1.00E−40   diazepam binding inhibitor [ Rattus norvegicus ]       406   2   117   2   352   g1228089   1.00E−40   multifunctional acyl-CoA-binding protein [ Rattus norvegicus ]       406   2   117   2   352   g203925   1.00E−40   diazepam binding inhibitor [ Rattus norvegicus ]       407   3   804   3   2414   g10953883   0   ubiquitin E3 ligase SMURF2 [ Homo sapiens ]       407   3   804   3   2414   g10047327   0   KIAA1625 protein [ Homo sapiens ]       407   3   804   3   2414   g6446606   0   E3 ubiquitin ligase SMURF1 [ Homo sapiens ]       408   1   220   244   903   g9622856   9.00E−24   sorting nexin 15A [ Homo sapiens ]       408   1   220   244   903   g2529709   1.00E−23   unknown [ Homo sapiens ]       408   1   220   244   903   g9622854   1.00E−23   sorting nexin 15 [ Homo sapiens ]       409   2   168   80   583   g5823961   2.00E−87   dJ20B11.1 (ortholog of rat RSEC5 (mammalian exocyst complex subunit))                                   [ Homo sapiens ]       409   2   168   80   583   g2827158   2.00E−84   rsec5 [ Rattus norvegicus ]       409   2   168   80   583   g7295804   8.00E−29   CG8843 gene product [ Drosophila melanogaster ]       410   2   108   194   517   g9963839   1.00E−50   lipase [ Homo sapiens ]       411   1   314   277   1218   g3243240   4.00E−56   syntaxin 11 [ Homo sapiens ]       411   1   314   277   1218   g4104685   1.00E−53   syntaxin 11 [ Homo sapiens ]       411   1   314   277   1218   g3248918   8.00E−46   syntaxin 11 [ Homo sapiens ]       412   2   143   212   640   g4512103   3.00E−57   rab11 binding protein [ Bos taurus ]       412   2   143   212   640   g6049150   8.00E−43   WD-containing protein [ Rattus norvegicus ]       413   1   122   1   366       414   2   86   623   880       415   3   213   183   821       416   1   263   40   828   g9558701   3.00E−31   gag [ Homo sapiens ]       416   1   263   40   828   g5802824   3.00E−31   Gag-Pro-Pol protein [ Homo sapiens ]       416   1   263   40   828   g5802821   3.00E−31   Gag-Pro-Pol protein [ Homo sapiens ]       417   1   175   940   1464   g246483   1.00E−63   prohibitin [human, Peptide, 272 aa] [ Homo sapiens ]       417   1   175   940   1464   g206384   2.00E−63   prohibitin [ Rattus norvegicus ]       417   1   175   940   1464   g541732   2.00E−63   prohibitin or B-cell receptor associated protein (BAP) 32 [ Mus musculus ]       418   2   272   167   982   g505033   6.00E−75   mitogen inducible gene mlg-2 [ Homo sapiens ]       418   2   272   167   982   g10727293   5.00E−33   CG14991 gene product [alt 2] [ Drosophila melanogaster ]       418   2   272   167   982   g7292434   5.00E−33   CG14991 gene product [alt 1] [ Drosophila melanogaster ]       419   1   167   16   516   g2587027   3.00E−34   HERV-E envelope glycoprotein [ Homo sapiens ]       419   1   167   16   516   g2587024   3.00E−34   HERV-E envelope glycoprotein [ Homo sapiens ]       419   1   167   16   516   g1049232   2.00E−31   HERV-E envelope protein [Human endogenous retrovirus]       420   2   59   227   403       421   1   216   1   648   g10504238   1.00E−101   hepatocellular carcinoma-related putative tumor suppressor [ Homo sapiens ]       421   1   216   1   648   g7020759   7.00E−75   unnamed protein product [ Homo sapiens ]       421   1   216   1   648   g3880143   1.00E−28   contains similarity to Pfam domain: PF01585 (G-patch domain), Score = 67.0, E                                   value = 1.3e−16, N = 1 [ Caenorhabditis elegans ]       422   1   162   1   486   g4982485   7.00E−55   apoptosis related protein APR-3 [ Homo sapiens ]       422   1   162   1   486   g4689122   3.00E−49   HSPC013 [ Homo sapiens ]                    
     [0903]                           TABLE 7                                   Parameter       Program   Description   Reference   Threshold                  ABIFACTURA   A program that removes vector sequences and   Applied Biosystems, Foster City, CA.               masks ambiguous bases in nucleic acid sequences.       ABI/   A Fast Data Finder useful in comparing and   Applied Biosystems, Foster City, CA;   Mismatch &lt;       PARACEL   annotating amino acid or nucleic acid sequences.   Paracel Inc., Pasadena, CA.   50%       FDF       ABI   A program that assembles nucleic acid sequences.   Applied Biosystems, Foster City, CA.       AutoAssembler       BLAST   A Basic Local Alignment Search Tool useful in   Altschul, S. F. et al. (1990) J. Mol. Biol.   ESTs:           sequence similarity search for amino acid and   215: 403-410; Altschul, S. F. et al. (1997)   Probability           nucleic acid sequences. BLAST includes five   Nucleic Acids Res. 25: 3389-3402.   value = 1.0E−8           functions: blastp, blastn, blastx, tblastn, and tblastx.       or less Full                   Length                   sequences:                   Probability                   value =                   1.0E−10 or less       FASTA   A Pearson and Lipman algorithm that searches for   Pearson, W. R. and D. J. Lipman (1988) Proc.   ESTs: fasta E           similarity between a query sequence and a group of   Natl. Acad Sci. USA 85: 2444-2448; Pearson,   value =           sequences of the same type. FASTA comprises as   W. R. (1990) Methods Enzymol. 183: 63-98;   1.06E−6           least five functions: fasta, tfasta, fastx, tfastx, and   and Smith, T. F. and M. S. Waterman (1981)   Assembled           ssearch.   Adv. Appl. Math. 2: 482-489.   ESTs: fasta                   Identity = 95%                   or greater and                   Match length =                   200 bases or                   greater; fastx E                   value = 1.0E−8                   or less Full                   Length                   sequences:                   fastx score =                   100 or greater       BLIMPS   A BLocks IMProved Searcher that matches a   Henikoff, S. and J. G. Henikoff (1991) Nucleic   Probability           sequence against those in BLOCKS, PRINTS,   Acids Res. 19: 6565-6572; Henikoff, J. G. and   value = 1.0E−3           DOMO, PRODOM, and PFAM databases to search   S. Henikoff (1996) Methods Enzymol.   or less           for gene families, sequence homology, and structural   266: 88-105; and Attwood, T. K. et al. (1997) J.           fingerprint regions.   Chem. Inf. Comput. Sci. 37: 417-424.       HMMER   An algorithm for searching a query sequence against   Krogh, A. et al. (1994) J. Mol. Biol.   PFAM hits:           hidden Markov model (HMM)-based databases of   235: 1501-1531; Sonnhammer, E. L. L. et al.   Probability           protein family consensus sequences, such as PFAM.   (1988) Nucleic Acids Res. 26: 320-322;   value = 1.0E−3               Durbin, R. et al. (1998) Our World View, in a   or less               Nutshell, Cambridge Univ. Press, pp. 1-350.   Signal peptide                   hits: Score = 0                   or greater       ProfileScan   An algorithm that searches for structural and sequence   Gribskov, M. et al. (1988) CABIOS 4: 61-66;   Normalized           motifs in protein sequences that match sequence patterns   Gribskov, M. et al. (1989) Methods Enzymol.   quality score ≧           defined in Prosite.   183: 146-159; Bairoch, A. et al. (1997)   GCG-specified               Nucleic Acids Res. 25: 217-221.   “HIGH” value                   for that                   particular                   Prosite motif.                   Generally,                   score =                   1.4-2.1.       Phred   A base-calling algorithm that examines automated   Ewing, B. et al. (1998) Genome Res.           sequencer traces with high sensitivity and probability.   8: 175-185; Ewing, B. and P. Green               (1998) Genome Res. 8: 186-194.       Phrap   A Phils Revised Assembly Program including SWAT and   Smith, T. F. and M. S. Waterman (1981) Adv.   Score = 120 or           CrossMatch, programs based on efficient implementation   Appl. Math. 2: 482-489; Smith, T.F. and M.S.   greater;           of the Smith-Waterman algorithm, useful in searching   Waterman (1981) J. Mol. Biol. 147: 195-197;   Match length =           sequence homology and assembling DNA sequences.   and Green, P., University of Washington,   56 or greater               Seattle, WA.       Consed   A graphical tool for viewing and editing Phrap assemblies.   Gordon, D. et al. (1998) Genome Res. 8: 195-202.       SPScan   A weight matrix analysis program that scans protein   Nielson, H. et al. (1997) Protein Engineering   Score = 3.5 or           sequences for the presence of secretory signal peptides.   10: 1-6; Claverie, J.M. and S. Audic (1997)   greater               CABIOS 12: 431-439.       TMAP   A program that uses weight matrices to delineate   Persson, B. and P. Argos (1994) J. Mol. Biol.           transmembrane segments on protein sequences and   237: 182-192; Persson, B. and P. Argos (1996)           determine orientation.   Protein Sci. 5: 363-371.       TMHMMER   A program that uses a hidden Markov model (HMM) to   Sonnhammer, E. L. et al. (1998) Proc. Sixth Intl.           delineate transmembrane segments on protein sequences   Conf. on Intelligent Systems for Mol. Biol.,           and determine orientation.   Glasgow et al., eds., The Am. Assoc. for Artificial               Intelligence Press, Menlo Park, CA, pp. 175-182.       Motifs   A program that searches amino acid sequences for patterns   Bairoch, A. et al. (1997) Nucleic Acids           that matched those defined in Prosite.   Res. 25: 217-221;               Wisconsin Package Program Manual, version 9, page               M51-59, Genetics Computer Group, Madison, WI.                    
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                 SEQUENCE LISTING 
               
            
           
           
               
            
               
                 The patent application contains a lengthy “Sequence Listing” section. A copy of the “Sequence Listing” is available in electronic form from the USPTO 
               
               
                 web site (http://seqdata.uspto.gov/sequence.html?DocID=20040048253). An electronic copy of the “Sequence Listing” will also be available from the 
               
               
                 USPTO upon request and payment of the fee set forth in 37 CFR 1.19(b)(3).