WO2002072770A2 - Novel human proteins, polynucleotides encoding them and methods of using the same - Google Patents

Abstract

Disclosed are polypeptides and nucleic acids encoding same. Also disclosed are vectors, host cells, antibodies and recombinant methods for producing the polypeptides and polynucleotides, as well as methods for using same.

Description

NOVEL HUMAN PROTEINS, POLYNUCLEOTIDES ENCODING THEM AND METHODS OF USING THE SAME

FIELD OF THE INVENTION

The present invention is based in part on nucleic acids encoding proteins that are new members ofthe following protein families: RET finger-like proteins, RNA Polymerase I transcription factor Rrn3-like proteins, CG122292-like proteins, myosin NIIA-like proteins, cytoplasmic protein-like proteins, ankyrin repeat-containing protein-like proteins, WD40 repeat-containing protein-like proteins, zinc finger-containing protein-like proteins, nuclear protein ΝOP2-like proteins, intracellular protein-like proteins, HBV PX-associated protein 8-like proteins, SM-20-like proteins, synaptonemal complex protein 3-like proteins, paraneoplastic cancer-testis-brain antigen-like proteins, adenylate cyclase associated protein- like proteins, mitochondrial protein-like proteins, PRO2032-like proteins, Leman coiled-coil protein-like proteins, Pax8-like proteins, GTPase activating protein-like proteins, and F-box leucine rich protein-like proteins.

The invention relates to polynucleotides and the polypeptides encoded by such polynucleotides, as well as vectors, host cells, antibodies and recombinant methods for producing the polypeptides and polynucleotides, as well as methods for using the same.

BACKGROUND OF THE INVENTION

The invention generally relates to nucleic acids and polypeptides encoded therefrom. More specifically, the invention relates to nucleic acids encoding cytoplasmic, nuclear, membrane bound, and secreted polypeptides, as well as vectors, host cells, antibodies, and recombinant methods for producing these nucleic acids and polypeptides.

SUMMARY OF THE INVENTION

The present invention is based in part on nucleic acids encoding proteins that are members ofthe following protein families: RET finger-like proteins, RNA Polymerase I transcription factor Rrn3-like proteins, CG122292-like proteins, myosin VIIA-like proteins, cytoplasmic protein-like proteins, ankyrin repeat-containing protein-like proteins, WD40 repeat-containing protein-like proteins, zinc finger-containing protein-like proteins, nuclear protein NOP2-like proteins, intracellular protein-like proteins, HBV PX-associated protein

8-like proteins, SM-20-like proteins, synaptonemal complex protein 3-like proteins, paraneoplastic cancer-testis-brain antigen-like proteins, adenylate cyclase associated protein- like proteins, mitochondrial protein-like proteins, PRO2032-like proteins, Leman coiled-coil protein-like proteins, Pax8-like proteins, GTPase activating protein-like proteins, and F-box leucine rich protein-like proteins. The novel polynucleotides and polypeptides are referred to herein as NONla, ΝONlb, ΝON2, ΝOV3, NOV4, NOV5, NOV6a, NOVόb, NOW, NOV8, NOV9, NOVlOa, NOVlOb, NOV11, NOV12, NOV13a, NOV13b, NOV14, NOV15,

NOV16, NOV17a, NOV17b, NOV18, NOV19, NOV20, NOV21, NOV22, NOV23, NOV24, NOV25, NOV26, NOV27 and NOV28. These nucleic acids and polypeptides, as well as derivatives, homologs, analogs and fragments thereof, will hereinafter be collectively designated as "NOVX" nucleic acid or polypeptide sequences. In one aspect, the invention provides an isolated NOVX nucleic acid molecule encoding a NOVX polypeptide that includes a nucleic acid sequence that has identity to the nucleic acids disclosed in SEQ ID NO:2n-l, wherein n is an integer between 1 and 33. In some embodiments, the NOVX nucleic acid molecule will hybridize under stringent conditions to a nucleic acid sequence complementary to a nucleic acid molecule that includes a protein-coding sequence of a NOVX nucleic acid sequence. The invention also includes an isolated nucleic acid that encodes a NOVX polypeptide, or a fragment, homolog, analog or derivative thereof. For example, the nucleic acid can encode a polypeptide at least 80% identical to a polypeptide comprising the amino acid sequences of SEQ ID NO:2n, wherein n is an integer between 1 and 33. The nucleic acid can be, for example, a genomic DNA fragment or a cDNA molecule that includes the nucleic acid sequence of any of SEQ ID NO:2n-l, wherein n is an integer between 1 and 33.

Also included in the invention is an oligonucleotide, e.g., an oligonucleotide which includes at least 6 contiguous nucleotides of a NOVX nucleic acid (e.g., SEQ ID NO:2n-l, wherein n is an integer between 1 and 33) or a complement of said oligonucleotide. Also included in the invention are substantially purified NOVX polypeptides (SEQ ID NO:2n, wherem n is an integer between 1 and 33). In certain embodiments, the NOVX polypeptides include an amino acid sequence that is substantially identical to the amino acid sequence of a human NOVX polypeptide.

The invention also features antibodies that immunoselectively bind to NOVX polypeptides, or fragments, homologs, analogs or derivatives thereof.

In another aspect, the invention includes pharmaceutical compositions that include therapeutically- or prophylactically-effective amounts of a therapeutic and a pharmaceutically-acceptable carrier. The therapeutic can be, e.g., a NOVX nucleic acid, a NOVX polypeptide, or an antibody specific for a NOVX polypeptide. In a further aspect, the invention includes, in one or more containers, a therapeutically- or prophylactically-effective amount of this pharmaceutical composition.

In a further aspect, the invention includes a method of producing a polypeptide by culturing a cell that includes a NOVX nucleic acid, under conditions allowing for expression of the NOVX polypeptide encoded by the DNA. If desired, the NOVX polypeptide can then be recovered.

In another aspect, the invention includes a method of detecting the presence of a NOVX polypeptide in a sample. In the method, a sample is contacted with a compound that selectively binds to the polypeptide under conditions allowing for formation of a complex between the polypeptide and the compound. The complex is detected, if present, thereby identifying the NOVX polypeptide within the sample.

The invention also includes methods to identify specific cell or tissue types based on their expression of a NOVX.

Also mcluded in the invention is a method of detecting the presence of a NOVX nucleic acid molecule in a sample by contacting the sample with a NOVX nucleic acid probe or primer, and detecting whether the nucleic acid probe or primer bound to a NOVX nucleic acid molecule in the sample.

In a further aspect, the invention provides a method for modulating the activity of a NOVX polypeptide by contacting a cell sample that includes the NOVX polypeptide with a compound that binds to the NOVX polypeptide in an amount sufficient to modulate the activity of said polypeptide. The compound can be, e.g., a small molecule, such as a nucleic acid, peptide, polypeptide, peptidomimetic, carbohydrate, lipid or other organic (carbon containing) or inorganic molecule, as further described herein.

Also within the scope ofthe invention is the use of a therapeutic in the manufacture of a medicament for treating or preventing disorders or syndromes including, e.g., adrenoleukodystrophy, congenital adrenal hyperplasia, hemophilia, hypercoagulation, idiopathic thrombocytopenic purpura, autoimmune disease, allergies, immunodeficiencies, transplantation, graft versus host, Von Hippel-Lindau (VHL) syndrome, Alzheimer's disease, stroke, tuberous sclerosis, hypercalcemia, Parkinson's disease, Huntington's disease, cerebral palsy, epilepsy, Lesch-Nyhan syndrome, multiple sclerosis, ataxia-telangiectasia, leukodystrophies, behavioral disorders, addiction, anxiety, pain, neuroprotection, diabetes, autoimmune disease, renal artery stenosis, interstitial nephritis, glomerulonephritis, polycystic kidney disease, systemic lupus erythematosus, renal tubular acidosis, IgA nephropathy, hypercalcemia, Lesch-Nyhan syndrome, cirrhosis, transplantation, systemic lupus erythematosus, autoimmune disease, asthma, emphysema, scleroderma, allergy, adult respiratory distress syndrome (ARDS), lymphedema, allergies, hemophilia, hypercoagulation, idiopathic thrombocytopenic purpura, autoimmune disease, allergies, immunodeficiencies, transplantation, graft versus host disease (GVHD), lymphedema, fertility, diabetes, pancreatitis, obesity, hemophilia, hypercoagulation, idiopathic thrombocytopenic purpura, immunodeficiencies, graft versus host, hypercalcemia, ulcers, anemia, ataxia-telangiectasia, autoimmune disease, immunodeficiencies, cancer, trauma, regeneration (in vitro and in vivo), viral infections, bacterial infections, parasitic infections and/or other pathologies and disorders ofthe like. The therapeutic can be, e.g, a NOVX nucleic acid, a NOVX polypeptide, or a

NOVX-specific antibody, or biologically-active derivatives or fragments thereof.

For example, the compositions ofthe present invention will have efficacy for treatment of patients suffering from the diseases and disorders disclosed above and/or other pathologies and disorders ofthe like. The polypeptides can be used as immunogens to produce antibodies specific for the invention, and as vaccines. They can also be used to screen for potential agonist and antagonist compounds. For example, a cDNA encoding NOVX may be useful in gene therapy, and NOVX may be useful when administered to a subject in need thereof. By way of non-limiting example, the compositions ofthe present invention will have efficacy for treatment of patients suffering from the diseases and disorders disclosed above and/or other pathologies and disorders of the like.

The invention further includes a method for screening for a modulator of disorders or syndromes including, e.g., the diseases and disorders disclosed above and/or other pathologies and disorders ofthe like. The method includes contacting a test compound with a NOVX polypeptide and determining if the test compound binds to said NOVX polypeptide. Binding of the test compound to the NOVX polypeptide indicates the test compound is a modulator of activity, or of latency or predisposition to the aforementioned disorders or syndromes.

Also within the scope ofthe invention is a method for screening for a modulator of activity, or of latency or predisposition to disorders or syndromes including, e.g., the diseases and disorders disclosed above and/or other pathologies and disorders ofthe like by administering a test compound to a test animal at increased risk for the aforementioned disorders or syndromes. The test animal expresses a recombinant polypeptide encoded by a NOVX nucleic acid. Expression or activity of NOVX polypeptide is then measured in the test animal, as is expression or activity ofthe protein in a control animal which recombinantly-expresses NOVX polypeptide and is not at increased risk for the disorder or syndrome. Next, the expression of NOVX polypeptide in both the test animal and the control animal is compared. A change in the activity of NOVX polypeptide in the test animal relative to the control animal indicates the test compound is a modulator of latency ofthe disorder or syndrome.

In yet another aspect, the invention includes a method for determining the presence of or predisposition to a disease associated with altered levels of a NOVX polypeptide, a NOVX nucleic acid, or both, in a subject (e.g., a human subject). The method includes measuring the amount of the NOVX polypeptide in a test sample from the subject and comparing the amount ofthe polypeptide in the test sample to the amount ofthe NOVX polypeptide present in a control sample. An alteration in the level ofthe NOVX polypeptide in the test sample as compared to the control sample indicates the presence of or predisposition to a disease in the subject. Preferably, the predisposition includes, e.g., the diseases and disorders disclosed above and/or other pathologies and disorders ofthe like. Also, the expression levels ofthe new polypeptides of the invention can be used in a method to screen for various cancers as well as to determine the stage of cancers.

In a further aspect, the invention includes a method of treating or preventing a pathological condition associated with a disorder in a mammal by administering to the subject a NOVX polypeptide, a NOVX nucleic acid, or a NOVX-specifϊc antibody to a subject (e.g. , a human subject), in an amount sufficient to alleviate or prevent the pathological condition. In preferred embodiments, the disorder, includes, e.g., the diseases and disorders disclosed above and/or other pathologies and disorders ofthe like.

In yet another aspect, the invention can be used in a method to identity the cellular receptors and downstream effectors ofthe invention by any one of a number of techniques commonly employed in the art. These include but are not limited to the two-hybrid system, affinity purification, co-precipitation with antibodies or other specific-interacting molecules.

NOVX nucleic acids and polypeptides are further useful in the generation of antibodies that bind immuno-specifically to the novel NOVX substances for use in therapeutic or diagnostic methods. These NOVX antibodies may be generated according to methods known in the art, using prediction from hydrophobicity charts, as described in the "Anti-NOVX Antibodies" section below. The disclosed NOVX proteins have multiple hydrophilic regions, each of which can be used as an immunogen. These NOVX proteins can be used in assay systems for functional analysis of various human disorders, which will help in understanding of pathology ofthe disease and development of new drug targets for various disorders.

The NOVX nucleic acids and proteins identified here may be useful in potential therapeutic applications implicated in (but not limited to) various pathologies and disorders as indicated below. The potential therapeutic applications for this invention include, but are not limited to: protein therapeutic, small molecule drug target, antibody target (therapeutic, diagnostic, drug targeting/cytotoxic antibody), diagnostic and/or prognostic marker, gene therapy (gene delivery/gene ablation), research tools, tissue regeneration in vivo and in vitro of all tissues and cell types composing (but not limited to) those defined here.

Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing ofthe present invention, suitable methods and materials are described below. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety. In the case of conflict, the present specification, including definitions, will control. In addition, the materials, methods, and examples are illustrative only and not intended to be limiting.

Other features and advantages ofthe invention will be apparent from the following detailed description and claims.

DETAILED DESCRIPTION OF THE INVENTION

The present invention provides novel nucleotides and polypeptides encoded thereby. Included in the invention are the novel nucleic acid sequences, their encoded polypeptides, antibodies, and other related compounds. The sequences are collectively referred to herein as "NOVX nucleic acids" or "NOVX polynucleotides" and the corresponding encoded polypeptides are referred to as "NOVX polypeptides" or "NOVX proteins." Unless indicated otherwise, "NOVX" is meant to refer to any ofthe novel sequences disclosed herein. Table 1 provides a summary ofthe NOVX nucleic acids and their encoded polypeptides.

TABLE 1. Sequences and Corresponding SEQ ID Numbers

Table 1 indicates homology of NOVX nucleic acids to known protein families. Thus, the nucleic acids and polypeptides, antibodies and related compounds according to the invention corresponding to a NOVX as identified in column 1 of Table 1 will be useful in therapeutic and diagnostic applications implicated in, for example, pathologies and disorders associated with the known protein families identified in column 5 of Table 1.

NOVX nucleic acids and their encoded polypeptides are useful in a variety of applications and contexts. The various NOVX nucleic acids and polypeptides according to the invention are useful as novel members ofthe protein families according to the presence of domains and sequence relatedness to previously described proteins. Additionally, NOVX nucleic acids and polypeptides can also be used to identify proteins that are members ofthe family to which the NOVX polypeptides belong.

Consistent with other known members ofthe family of proteins, identified in column 5 of Table 1, the NOVX polypeptides ofthe present invention show homology to, and contain domains that are characteristic of, other members of such protein families. Details ofthe sequence relatedness and domain analysis for each NOVX are presented in Example A.

The NOVX nucleic acids and polypeptides can also be used to screen for molecules, which inhibit or enhance NOVX activity or function. Specifically, the nucleic acids and polypeptides according to the invention may be used as targets for the identification of small molecules that modulate or inhibit diseases associated with the protein families listed in Table 1.

The NOVX nucleic acids and polypeptides are also useful for detecting specific cell types. Details ofthe expression analysis for each NOVX are presented in Example C. Accordingly, the NOVX nucleic acids, polypeptides, antibodies and related compounds according to the invention will have diagnostic and therapeutic applications in the detection of a variety of diseases with differential expression in normal vs. diseased tissues, e.g., a variety of cancers. , Additional utilities for NOVX nucleic acids and polypeptides according to the invention are disclosed herein.

NOVl and NOV2: RET finger-like

NOVl and NOV2 are homologous to the Ring finger protein family. Thus, NOVl and NOV2 will function similarly to other members ofthe Ring finger protein family. This family has transcriptional regulatory activity and is also found in the cytoplasm. Members of this family (e.g., RET finger proteins) can also regulate protein degradation through the ubiquitin pathway. A number of RET finger proteins are involved in oncogenic transformation as well as normal developmental processes. Thus, NOVl and NOV2 proteins will play a role in normal development as well as dedifferentiation. Specifically, the NOV 1 and NOV2 nucleic acids and proteins ofthe invention have applications in the diagnosis and/or treatment of various diseases and disorders. For example, the compositions ofthe present invention will have efficacy for the treatment of patients suffering from: cancer, trauma, regeneration (in vitro and in vivo), viral infections, bacterial infections, parasitic infections, obesity, diabetes and other metabolic diseases as well as other diseases, disorders and conditions.

NO 3: RNA Polymerase I transcription factor Rrn3-like

NOV3 is homologous to the RNA polymerase I transcription factor Rrn3 protein family. Thus, NOV3 will function similarly to other members ofthe Rrn3 protein family. This family mediates transcription of rDNA by RNA polymerase I (Pol I) and is also found in the nucleus. Members of this family are essential genes. Thus, NOV3 nucleic acids and proteins ofthe invention have applications in the diagnosis and/or treatment of various diseases and disorders. For example, the compositions ofthe present invention will have efficacy for the treatment of patients suffering from: neurodegenerative disease, Alzheimer, Hodgkin's disease, perinatal asphyxia, systemic sclerosis (scleroderma), pituitary tumor, hepatocellular carcinoma and other malignancies, systemic lupus erythematosus, rheumatic autoimmune diseases, chronic leukemia as well as other diseases, disorders and conditions.

NOV4: CG122292-like NOV4 is homologous to IPR000620 CG12292 protein, a member of integral membrane protein DUF6 family. Thus, NOV4 will function similarly to other members of the DUF6 subfamily of CG122229-like proteins. The DUF6 family includes the Erwinia PecM protein, which is involved in pectinase, cellulase and blue pigment regulation. NOV4 nucleic acids and proteins ofthe invention have applications in the diagnosis and/or treatment of various diseases and disorders such as cancer (e.g., pancreatic adenocarcinoma).

NOV5: Myosin VHA-like

NOV5 is homologous to the Myosin VII protein family. Thus, NOV5 will function similarly to other members ofthe Myosin VII protein family. Among the members ofthe Myosin VII protein family are the microtubule-based kinesin motors and actin-based myosin motors generate motions, associated with intracellular trafficking, cell division, and muscle contraction. Thus, NOV5 proteins will play a role in muscle function. NOV5 nucleic acids and proteins ofthe invention have applications in the diagnosis and or treatment of various diseases and disorders. For example, the compositions ofthe present invention will have efficacy for the treatment of patients suffering from: Duchenne's Muscular Dystrophy, deafness, blindness, Inflammatory bowel disease, Diverticular disease, Hyperthyroidism and Hypothyroidism, Lymphedema, Allergies as well as other diseases, disorders and conditions.

NOV6: Predicted cytoplasmic protein

NOV6 is an expressed human cytoplasmic protein with homology to an uncharacterized but expressed mouse protein. This human homolog is expressed in numerous human tissues, and will have implications in human diseases. NOV6 nucleic acids and proteins ofthe invention have applications in the diagnosis and/or treatment of various diseases and disorders. For example, the compositions ofthe present invention will have efficacy for the treatment of patients suffering from: adrenoleukodystrophy, congenital adrenal hyperplasia, hemophilia, hypercoagulation, idiopathic thrombocytopenic purpura, autoimmune disease, allergies, immunodeficiencies, transplantation, graft versus host, Von Hippel-Lindau (VHL) syndrome, Alzheimer's disease, stroke, tuberous sclerosis, hypercalcemia, Parkinson's disease, Huntington's disease, cerebral palsy, epilepsy, Lesch- Nyhan syndrome, multiple sclerosis, ataxia-telangiectasia, leukodystrophies, behavioral disorders, addiction, anxiety, pain, neuroprotection, diabetes, autoimmune disease, renal artery stenosis, interstitial nephritis, glomerulonephritis, polycystic kidney disease, systemic lupus erythematosus, renal tubular acidosis, IgA nephropathy, hypercalcemia, Lesch-Nyhan syndrome, cirrhosis, transplantation, systemic lupus erythematosus, autoimmune disease, asthma, emphysema, scleroderma, allergy, adult respiratory distress syndrome (ARDS), lymphedema, allergies, hemophilia, hypercoagulation, idiopathic thrombocytopenic purpura, autoimmune disease, allergies, immunodeficiencies, transplantation, graft versus host disease (GVHD), lymphedema, fertility, diabetes, pancreatitis, obesity, hemophilia, hypercoagulation, idiopathic thrombocytopenic purpura, immunodeficiencies, graft versus host, hypercalcemia, ulcers, anemia, ataxia-telangiectasia, autoimmune disease, immunodeficiencies, cancer, trauma, regeneration (in vitro and in vivo), viral infections, bacterial infections, parasitic infections, as well as other diseases, disorders and conditions.

NOV7: Ankyrin repeat-containing protein-like NOV7 is homologous to the Ankyrin repeat-containing protein family. Thus, NOV7 will function similarly to other members ofthe Ankyrin repeat-containing protein family. NOV7 protein ofthe present invention contains ankyrin repeats involved in protein-protein interaction and, thus, will regulate intracellular signal transduction. This domain can also regulate expression/activity of other proteins and /domains thereby affecting cell growth, proliferation, differentiation and survival. NOV7 nucleic acids and proteins ofthe invention, therefore, have applications in the diagnosis and/or treatment of various diseases and disorders. For example, the compositions ofthe present invention will have efficacy for the treatment of patients suffering from: cancer, trauma, tissue regeneration (in vitro and in vivo), viral infections, bacterial infections, parasitic infections, immunological disease, respiratory disease, gastro-intestinal diseases, reproductive health, neurological and neurodegenerative diseases, bone marrow transplantation, metabolic and endocrine diseases, allergy and inflammation, nephrological disorders, cardiovascular diseases, muscle, bone, joint and skeletal disorders, hematopoietic disorders, urinary system disorders as well as other diseases, disorders and conditions.

NOV8: WD40 repeat-containing protein-like

NOV8 is homologous to the WD40 repeat-containing protein family. Thus, NOV8 will function similarly to other members ofthe WD40 repeat-containing protein family. NOV7 ofthe present invention contains WD40 repeats, a domain has been shown to be involved in protein-protein interactions and signal transduction. WD-repeat proteins are found in all eukaryotes and are implicated in a variety of regulatory functions as a result of protein-protein interactions (e.g., detoxification). Thus, NOV7 nucleic acids and proteins of the invention have applications in the diagnosis and/or treatment of various diseases and disorders. For example, the compositions ofthe present invention will have efficacy for the treatment of patients suffering from: cancer, trauma, tissue regeneration (in vitro and in vivo), viral infections, bacterial infections, parasitic infections, immunological disease, respiratory disease, gastro-intestinal diseases, reproductive health, neurological and neurodegenerative diseases, bone marrow transplantation, metabolic and endocrine diseases, allergy and inflammation, nephrological disorders, cardiovascular diseases, muscle, bone, joint and skeletal disorders, hematopoietic disorders, urinary system disorders as well as other diseases, disorders and conditions.

NOV9: Zinc finger-containing protein-like NOV9 is homologous to the Zinc finger-containing protein family. Thus, NOV9 will function similarly to other members ofthe Zinc finger-containing protein family. NOV9 polypeptide ofthe present invention contains multiple zinc finger domains ofthe C2H2 type. These domains are mostly found in transcription factors that regulate gene expression in specific tissues. NOV9 nucleic acids and proteins ofthe invention have applications in the diagnosis and/or treatment of various diseases and disorders. NOV9 nucleic acid and protein can be used for targeting genes expressed in specific tissues because ofthe specificity ofthe members of this family. The compositions ofthe present invention will also have efficacy for the treatment of patients suffering from: cancer, trauma, regeneration (in vitro and in vivo), viral infections, bacterial infections, parasitic infections, cardiomyopathy, atherosclerosis, hypertension, congenital heart defects, aortic stenosis, atrial septal defect (ASD), atrioventricular (A-V) canal defect, ductus arteriosus, pulmonary stenosis, subaortic stenosis, ventricular septal defect (VSD), valve diseases, tuberous sclerosis, scleroderma, obesity, transplantation, Von Hippel-Lindau (VHL) syndrome, Alzheimer's disease, stroke, tuberous sclerosis, hypercalcemia, Parkinson's disease, Huntington's disease, cerebral palsy, epilepsy, Lesch-Nyhan syndrome, multiple sclerosis, ataxia-telangiectasia, leukodystrophies, behavioral disorders, addiction, anxiety, pain, neurodegeneration, hemophilia, hypercoagulation, idiopathic thrombocytopenic puφura, autoimmune disease, allergies, immunodeficiencies, transplantation, graft versus host disease, hypercalcemia, ulcers, pancreatitis, Hirschsprung's disease, Crohn's Disease, appendicitis, fertility, systemic lupus erythematosus, autoimmune disease, asthma, emphysema, scleroderma, allergy, adult respiratory distress syndrome (ARDS), adrenoleukodystrophy, congenital adrenal hypeφlasia as well as other diseases, disorders and conditions.

NO 10: Nuclear protein NOP2-like proteins

NOV10 is homologous to the NOP2 protein family and the pi 20 protein family. In yeast, NOP2 protein is a nucleolar protein that is plays an important role in maintaining the structure ofthe nucleolus and is essential for yeast cell proliferation. In humans, protein pi 20, is a cell proliferation marker that has been used as prognostic marker for stages of cancer. Thus, NOV10 ofthe present invention will play a role in cell proliferation and/or loss of expression associated with lack of cell integrity and apoptosis. NOV10 nucleic acids and proteins ofthe invention have applications in the diagnosis and/or treatment of various diseases and disorders. For example, the compositions ofthe present invention will have efficacy for the treatment of patients suffering from: cancer, trauma, regeneration (in vitro and in vivo), viral infections, bacterial infections, parasitic infections, autoimmune disease, asthma, emphysema, scleroderma, allergy, adult respiratory distress syndrome (ARDS), endocrine dysfunctions, diabetes, obesity, growth and reproductive disorders, Von Hippel- Lindau (VHL) syndrome, pancreatitis as well as other diseases, disorders and conditions. NOV11: Intracellular protein-like

NOVl 1 is homologous to bacterial intracellular proteins. NOVl 1 nucleic acids and proteins ofthe invention have applications in the diagnosis and/or treatment of various diseases and disorders. For example, the compositions ofthe present invention will have efficacy for the treatment of patients suffering from various disorders and conditions. NOV12: HBV PX-associated protein 8-like

•NOV12 is homologous to the HBV PX-associated protein family. Thus, NOV12 will function similarly to other members ofthe HBV PX-associated protein family. HBx/pX is implicated in the development of hepatocellular carcinoma (HCC) in chronic HBV-infected patients. NOVl 2 nucleic acids and proteins ofthe invention have applications in the diagnosis and/or treatment of various diseases and disorders. For example, measurement of NOV12 expression will be useful in the diagnosis of cancer. Also, the compositions ofthe present invention will have efficacy for the treatment of patients suffering from: liver diseases such as acute hepatitis, hepatitis B, liver transplantation, liver cirrhosis, Von Hippel- Lindau (VHL) syndrome and hepatocarcinoma as well as other diseases, disorders and conditions.

NOV13: SM-20-like NOV13 is homologous to the SM-20 protein family. Thus, NOV13 will function similarly to other members ofthe SM-20 protein family. SM-20 protein is a growth factor responsive protein that was first found expressed in blood vessels. SM-20 is also hypothesized to be a mitochondrial protein that promotes cell death through a caspase- dependent mechanism in neurons. NOVl 3 nucleic acids and proteins ofthe invention have applications in the diagnosis and/or treatment of various diseases and disorders. For example, the compositions ofthe present invention will have efficacy for the treatment of patients suffering from: cancer, trauma, tissue regeneration (in vitro and in vivo), viral infections, bacterial infections, parasitic infections, immunological disease, respiratory disease, gastrointestinal diseases, reproductive health, neurological and neurodegenerative diseases, bone marrow transplantation, metabolic and endocrine diseases, allergy and inflammation, nephrological disorders, cardiovascular diseases, muscle, bone, joint and skeletal disorders, hematopoietic disorders, urinary system disorders as well as other diseases, disorders and conditions.

NOV14: Synaptonemal complex protein 3-like NOVl 4 is homologous to the Synaptonemal complex protein 3 (SCP3) family. Thus,

NOV14 will function similarly to other members ofthe SCP3 family. SCP3 is responsible for synapsis of homologous chromosomes during meiosis. NOV14 nucleic acids and proteins ofthe invention have applications in the diagnosis and/or treatment of various diseases and disorders. For example, measurement of NOV14 expression will be useful in the diagnosis of cancer (e.g., testicular cancer) or infertility. Also, the compositions ofthe present invention will have efficacy for the treatment of patients suffering from: Von Hippel-Lindau (VHL) syndrome, Alzheimer's disease, stroke, tuberous sclerosis, hypercalcemia, Parkinson's disease, Huntington's disease, cerebral palsy, epilepsy, Lesch-Nyhan syndrome, multiple sclerosis, ataxia-telangiectasia, leukodystrophies, behavioral disorders, addiction, anxiety, pain, neurodegeneration, fertility disorders, Hirschsprung's disease, Crohn's disease, appendicitis, diabetes, autoimmune disease, renal artery stenosis, interstitial nephritis, glomerulonephritis, polycystic kidney disease, systemic lupus erythematosus, renal tubular acidosis, IgA nephropathy, cancer, tissue degeneration, bacterial/viral/parasitic infections as well as other diseases, disorders and conditions.

NOV15: paraneoplastic cancer-testis-brain antigen-like

NOVl 5 is homologous to the paraneoplastic cancer-testis-brain antigen-like protein family. Thus, NOVl 5 will function similarly to other members ofthe paraneoplastic cancer- testis-brain antigen-like protein family. Paraneoplastic syndromes are associated with certain types of cancer, such as cancer ofthe brain or testis. NOVl 5 nucleic acids and proteins of the invention have applications in the diagnosis and/or treatment of various diseases and disorders. For example, measurement of NOVl 15 expression will be useful in the diagnosis of neurological syndromes associated with different cancers, such as cancer ofthe brain or testis. Also, the compositions ofthe present invention will have efficacy for the treatment of patients suffering from: cancer, trauma, regeneration, viral/bacterial/parasitic infection, systemic lupis erythematosus, autoimmune disease, asthma, emphysema, scleroderma, allergies, and adult respiratory distress syndrome (ARDS), as well as other diseases, disorders and conditions. NOV16: adenylate cyclase associated protein-like

NOVl 6 is homologous to the adenylate cyclase associated protein family. Thus, NOVl 6 will function similarly to other members ofthe adenylate cyclase associated protein family. Adenylate cyclase associated proteins regulate actin cytoskeletal organization and are components ofthe ras signaling pathway. NOVl 6 nucleic acids and proteins ofthe invention have applications in the diagnosis and/or treatment of various diseases and disorders. For example, measurement of NOVl 6 expression will be useful in the diagnosis of cancer, neurological disorders and other diseases involving cell proliferation and/or differentiation. Also, the compositions ofthe present invention will have efficacy for the treatment of patients suffering from: brain disorders including epilepsy, eating disorders, schizophrenia, ADD, and cancer; heart disease; inflammation and autoimmune disorders including Crohn's disease, IBD, allergies, rheumatoid and osteoarthritis, inflammatory skin disorders, allergies, blood disorders; psoriasis colon cancer, leukemia AIDS; thalamus disorders; metabolic disorders including diabetes and obesity; lung diseases such as asthma, emphysema, polycystic kidney disease, cystic fibrosis, and cancer; pancreatic disorders including pancreatic insufficiency and cancer; and prostate disorders including prostate cancer as well as other diseases, disorders and conditions.

NOV17: expressed mitochondrial protein-like NOV17 is homologous to the expressed mitochondrial protein family. Thus, NOV17 will function similarly to other members ofthe expressed mitochondrial protein family. Mitochondrial proteins are important in metabolism, aging and apoptosis. NOVl 7 nucleic acids and proteins ofthe invention have applications in the diagnosis and/or treatment of various diseases and disorders. For example, measurement of NOVl 7 expression will be useful in the diagnosis of metabolic diseases and or mitochondrial storage diseases. Also, the compositions ofthe present invention will have efficacy for the treatment of patients suffering from: cardiomyopathy, atherosclerosis, hypertension, congenital heart defects, aortic stenosis, atrial septal defect (ASD), atrioventricular (A-V) canal defect, ductus arteriosus, pulmonary stenosis, subaortic stenosis, ventricular septal defect (VSD), valve diseases, tuberous sclerosis, scleroderma, obesity, transplantation, inflammatory bowel disease, diverticular disease, fertility, systemic lupus erythematosus, autoimmune disease, asthma, emphysema, scleroderma, allergy, adult respiratory distress syndrome (ARDS), Von Hippel-Lindau (VHL) syndrome, cirrhosis, transplantation, myasthenia gravis, leukodystrophies, pain, neuroprotection, cancer, trauma, regeneration (in vitro and in vivo), viral infections, bacterial infections, parasitic infections as well as other diseases, disorders and conditions.

NOV18: expressed cytoplasmic protein-like

NOVl 8 is homologous to the expressed cytoplasmic protein family. Thus, NOVl 8 will function similarly to other members ofthe expressed cytoplasmic protein family. Cytoplasmic proteins are important in cell metabolism, signaling, proliferation and differentiation. NOVl 8 nucleic acids and proteins ofthe invention have applications in the diagnosis and/or treatment of various diseases and disorders. For example, measurement of NOVl 8 expression will be useful in the diagnosis of diseases associated with altered expression of cytoplasmic proteins. Also, the compositions ofthe present invention will have efficacy for the treatment of patients suffering from: anemia, ataxia-telangiectasia, autoimmune disease, immunodeficiencies, hemophilia, hypercoagulation, idiopathic thrombocytopenic puφura, autoimmune disease, allergies, immunodeficiencies, transplantation, graft versus host, Von Hippel-Lindau (VHL) syndrome, Alzheimer's disease, stroke, tuberous sclerosis, hypercalcemia, Parkinson's disease, Huntington's disease, cerebral palsy, epilepsy, Lesch-Nyhan syndrome, multiple sclerosis, ataxia-telangiectasia, leukodystrophies, behavioral disorders, addiction, anxiety, pain, neuroprotection, diabetes, autoimmune disease, renal artery stenosis, interstitial nephritis, glomerulonephritis, polycystic kidney disease, systemic lupus erythematosus, renal tubular acidosis, IgA nephropafhy, hypercalcemia, Lesch-Nyhan syndrome, cirrhosis, transplantation, systemic lupus erythematosus, autoimmune disease, asthma, emphysema, scleroderma, allergy, adult respiratory distress syndrome (ARDS), endometriosis, fertility, diabetes, pancreatitis, obesity, hypercalcemia, ulcers, tonsillitis, endometriosis, cancer, trauma, regeneration (in vitro and in vivo), viral infections, bacterial infections, parasitic infections as well as other diseases, disorders and conditions.

NO 19: mitochondrial protein-like

NOV 19 is homologous to the mitochondrial protein family. Thus, NOV 19 will function similarly to other members ofthe mitochondrial protein family. Mitochondrial proteins are important in metabolism, aging and apoptosis. NOVl 9 nucleic acids and proteins ofthe invention have applications in the diagnosis and/or treatment of various diseases and disorders. For example, measurement of NOVl 9 expression will be useful in the diagnosis of mitochondrial storage diseases or metabolic disorders. Also, the compositions ofthe present invention will have efficacy for the treatment of patients suffering from: brain disorders including epilepsy, eating disorders, schizophrenia, ADD, and cancer; heart disease; inflammation and autoimmune disorders including Crohn's disease, IBD, allergies, rheumatoid and osteoarthritis, inflammatory skin disorders, allergies, blood disorders; psoriasis colon cancer, leukemia AIDS; thalamus disorders; metabolic disorders including diabetes and obesity; lung diseases such as asthma, emphysema, polycystic kidney disease, cystic fibrosis, and cancer; pancreatic disorders including pancreatic insufficiency and cancer; and prostate disorders including prostate cancer as well as other diseases, disorders and conditions.

NOV20: PRO2032-like

NOV20 is homologous to the PRO2032 family. Thus, NOV20 will function similarly to other members of the PRO2032 family. The gene encoding the PRO2032 protein maps to human chromosome 13q31-32, a locus associated with congenital microcoria. NOV20 nucleic acids and proteins ofthe invention have applications in the diagnosis and/or treatment of various diseases and disorders. For example, measurement of NOV20 expression will be useful in the diagnosis of congenital microcoria. Also, the compositions ofthe present invention will have efficacy for the treatment of patients suffering from: adrenoleukodystrophy, congenital adrenal hypeφlasia, osteoporosis, hypercalcemia, arthritis, ankylosing spondylitis, scoliosis, hemophilia, hypercoagulation, idiopathic thrombocytopenic puφura, autoimmune disease, allergies, immunodeficiencies, transplantation, graft versus host disease, Von Hippel-Lindau (VHL) syndrome, Alzheimer's disease, stroke, tuberous sclerosis, Parkinson's disease, Huntington's disease, cerebral palsy, epilepsy, Lesch-Nyhan syndrome, multiple sclerosis, ataxia-telangiectasia, leukodystrophies, behavioral disorders, addiction, anxiety, pain, neurodegeneration, renal artery stenosis, interstitial nephritis, glomerulonephritis, polycystic kidney disease, systemic lupus erythematosus, renal tubular acidosis, IgA nephropathy, cirrhosis, asthma, emphysema, scleroderma, adult respiratory distress syndrome (ARDS), lymphedema, hypeφarathyroidism, hypoparathyroidism, xerostomia, endocrine dysfunctions, diabetes, obesity, growth and reproductive disorders, hyperthyroidism, hypothyroidism, tonsillitis, endometriosis, fertility, cancer as well as other diseases, disorders and conditions.

NOV21: Leman coiled-coil protein-like

NOV21 is homologous to the Leman coiled-coil protein family. Thus, NOV21 will function similarly to other members ofthe Leman coiled-coil protein family. Extracellular matrix proteins such as keratins may belong to the superfamily of intermediate filament proteins that form alpha-helical coiled-coil dimers which associate laterally and end-to-end to form multimeric filaments. Mutations that perturb keratin filament assembly in vitro can cause blistering human skin disorders in vivo. NOV21 nucleic acids and proteins ofthe invention have applications in the diagnosis and/or treatment of various diseases and disorders. For example, measurement of NOV21 expression will be useful in the diagnosis of human skin diseases. Also, the compositions ofthe present invention will have efficacy for the treatment of patients suffering from: diseases ofthe musculoskeletal system, cancer, connective tissue disorders, heart diseases, Alzheimer's disease, abnormal wound healing, disorders ofthe skin as well as other diseases, disorders and conditions.

NOV22: Pax8-like NOV22 is homologous to the Pax8 family. Thus, NOV22 will function similarly to other members ofthe Pax8 family. Pax8 is a member of a family of transcription factors that are essentially required for the formation of several tissues from all germ layers in the mammalian embryo. In the thyroid gland, PAX8 is essential for the formation of thyroxine- producing follicular cells. NOV22 nucleic acids and proteins ofthe invention have applications in the diagnosis and/or treatment of various diseases and disorders. For example, measurement of NOV22 expression will be useful in the diagnosis of hyper/hypothyroidism. Also, the compositions ofthe present invention will have efficacy for the treatment of patients suffering from: thyroid diseases, developmental defects, cancer, especially thyroid carcinomas as well as other diseases, disorders and conditions.

NOV23 and NOV24: GTPase activating protein-like

NOV23 and NOV24 are homologous to the GTPase activating protein (GAP) family. Thus, NOV23 and NOV24 will function similarly to other members ofthe GAP family. The signaling pathway including the guanine nucleotide-binding proteins and the GAPs regulate a variety of processes, including sensual perception, protein synthesis, various transport processes, and cell growth and differentiation. NOV23 and NOV24 nucleic acids and proteins ofthe invention have applications in the diagnosis and/or treatment of various diseases and disorders. For example, measurement of NOV23 or NOV24 expression will be useful in the diagnosis of cancer. Also, the compositions ofthe present invention will have efficacy for the treatment of patients suffering from: adrenoleukodystrophy, congenital adrenal hypeφlasia, hemophilia, hypercoagulation, idiopathic thrombocytopenic puφura, autoimmune disease, allergies, immunodeficiencies, transplantation, graft versus host, Von Hippel-Lindau (VHL) syndrome, Alzheimer's disease, stroke, tuberous sclerosis, hypercalcemia, Parkinson's disease, Huntington's disease, cerebral palsy, epilepsy, Lesch- Nyhan syndrome, multiple sclerosis, ataxia-telangiectasia, leukodystrophies, behavioral disorders, addiction, anxiety, pain, neuroprotection, arthritis, tendinitis, diabetes, autoimmune disease, renal artery stenosis, interstitial nephritis, glomerulonephritis, polycystic kidney disease, systemic lupus erythematosus, renal tubular acidosis, IgA nephropathy, hypercalcemia, Lesch-Nyhan syndrome, cirrhosis, transplantation, systemic lupus erythematosus, autoimmune disease, asthma, emphysema, scleroderma, allergy, adult respiratory distress syndrome (ARDS), lymphedema, allergies, hemophilia, hypercoagulation, idiopathic thrombocytopenic puφura, autoimmune disease, allergies, immunodeficiencies, transplantation, graft versus host disease (GVHD), lymphedema, fertility, endometriosis, pancreatitis, obesity, hypeφarathyroidism, hypoparathyroidism, endocrine dysfunctions, diabetes, obesity, growth and reproductive disorders, inflammatory bowel disease, diverticular disease, hemophilia, hypercoagulation, idiopathic thrombocytopenic puφura, immunodeficiencies, graft versus host, hypercalcemia, ulcers, tonsillitis, cancer, trauma, regeneration (in vitro and in vivo), viral infections, bacterial infections, parasitic infections as well as other diseases, disorders and conditions.

NON25: F-box leucine rich protein-like

ΝOV25 is homologous to the F-box leucine rich protein family. Thus, NOV25 will function similarly to other members ofthe F-box leucine rich protein family. F-box proteins are critical components ofthe SCF ubiquitin-protein ligase complex and are involved in substrate recognition and recruitment for ubiquitination and consequent degradation by the proteasome. NOV25 nucleic acids and proteins ofthe invention have applications in the diagnosis and/or treatment of various diseases and disorders. For example, measurement of NOV25 expression will be useful in the diagnosis of lysosomal and other organelle storage diseases. Also, the compositions ofthe present invention will have efficacy for the treatment of patients suffering from: Adrenoleukodystrophy, Congenital Adrenal Hypeφlasia, Hemophilia, hypercoagulation, idiopathic thrombocytopenic pmpura, autoimmune disease, allergies, immunodeficiencies, transplantation, Graft versus host, Von Hippel-Lindau (VHL) syndrome, Alzheimer's disease, Stroke, Tuberous sclerosis, hypercalcemia, Parkinson's disease, Huntington's disease, Cerebral palsy, Epilepsy, Lesch-Nyhan syndrome, Multiple sclerosis, Ataxia-telangiectasia, Leukodystrophies, Behavioral disorders, Addiction, Anxiety, Pain, Neuroprotection, Cardiomyopathy, Atherosclerosis, Hypertension, Congenital heart defects, Aortic stenosis, Atrial septal defect (ASD), Atrioventricular (A-V) canal defect, Ductus arteriosus, Pulmonary stenosis, Subaortic stenosis, Ventricular septal defect (VSD), valve diseases, Tuberous sclerosis, Scleroderma, Obesity, Transplantation, Lymphedema, Allergies, Psoriasis, Actinic keratosis, Acne, Hair growth, allopecia, pigmentation disorders, endocrine disorders, Endocrine dysfunctions, Diabetes, obesity, Growth and reproductive disorders, Fertility, Endometriosis, Hemophilia, Hypercoagulation, idiopathic thrombocytopenic puφura, Immunodeficiencies, Graft versus host, Hyperthyroidism and Hypothyroidism as well as other diseases, disorders and conditions.

NOV26: GTPase activating protein-like

NOV26 is homologous to the GTPase activating protein (GAP) family. Thus, NOV26 will function similarly to other members ofthe GAP family. The signaling pathway including the guanine nucleotide-binding proteins and the GAPs regulate a variety of processes, including sensual perception, protein synthesis, various transport processes, and cell growth and differentiation. NOV26 nucleic acids and proteins ofthe invention have applications in the diagnosis and/or treatment of various diseases and disorders. For example, measurement of NOV26 expression will be useful in the diagnosis of cancer. Also, the compositions ofthe present invention will have efficacy for the treatment of patients suffering from: adrenoleukodystrophy, congenital adrenal hypeφlasia, hemophilia, hypercoagulation, idiopathic thrombocytopenic puφura, autoimmune disease, allergies, immunodeficiencies, transplantation, graft versus host, Von Hippel-Lindau (VHL) syndrome, Alzheimer's disease, stroke, tuberous sclerosis, hypercalcemia, Parkinson's disease, Huntington's disease, cerebral palsy, epilepsy, Lesch-Nyhan syndrome, multiple sclerosis, ataxia-telangiectasia, leukodystrophies, behavioral disorders, addiction, anxiety, pain, neuroprotection, arthritis, tendinitis, diabetes, autoimmune disease, renal artery stenosis, interstitial nephritis, glomerulonephritis, polycystic kidney disease, systemic lupus erythematosus, renal tubular acidosis, IgA nephropathy, hypercalcemia, Lesch-Nyhan syndrome, cirrhosis, transplantation, systemic lupus erythematosus, autoimmune disease, asthma, emphysema, scleroderma, allergy, ARDS, lymphedema, allergies, hemophilia, hypercoagulation, idiopathic thrombocytopenic puφura, autoimmune disease, allergies, immunodeficiencies, transplantation, graft versus host disease (GVHD), lymphedema, fertility, endometriosis, pancreatitis, obesity, hypeφarathyroidism, hypoparathyroidism, endocrine dysfunctions, diabetes, obesity, growth and reproductive disorders, inflammatory bowel disease, diverticular disease, hemophilia, hypercoagulation, idiopathic thrombocytopenic puφura, immunodeficiencies, graft versus host, hypercalcemia, ulcers, tonsillitis, cancer, trauma, regeneration (in vitro and in vivo), viral infections, bacterial infections, parasitic infections as well as other diseases, disorders and conditions.

NOV27: nuclear protein-like

NOV27 is homologous to the nuclear protein family. Thus, NOV27 will function similarly to other members ofthe nuclear protein family. Nuclear proteins are important in gene expression, cell cycle regulation, intracellular trafficking and cell shape. NOV27 nucleic acids and proteins ofthe invention have applications in the diagnosis and/or treatment of various diseases and disorders. The compositions ofthe present invention will have efficacy for the treatment of patients suffering from: adrenoleukodystrophy, congenital adrenal hypeφlasia, anemia, ataxia-telangiectasia, autoimmune disease, immunodeficiencies, hemophilia, hypercoagulation, idiopathic thrombocytopenic puφura, autoimmune disease, allergies, immunodeficiencies, transplantation, graft versus host, Von Hippel-Lindau (VHL) syndrome, Alzheimer's disease, stroke, tuberous sclerosis, hypercalcemia, Parkinson's disease, Huntington's disease, cerebral palsy, epilepsy, Lesch-Nyhan syndrome, multiple sclerosis, ataxia-telangiectasia, leukodystrophies, behavioral disorders, addiction, anxiety, pain, neuroprotection, diabetes, autoimmune disease, renal artery stenosis, interstitial nephritis, glomerulonephritis, polycystic kidney disease, systemic lupus erythematosus, renal tubular acidosis, IgA nephropathy, hypercalcemia, Lesch-Nyhan syndrome, systemic lupus erythematosus, autoimmune disease, asthma, emphysema, scleroderma, allergy, ARDS, lymphedema, allergies, hemophilia, hypercoagulation, idiopathic thrombocytopenic puφura, autoimmune disease, allergies, immunodeficiencies, transplantation, graft versus host disease (GVHD), lymphedema, fertility, endometriosis, fertility, diabetes, Von Hippel-Lindau (VHL) syndrome, pancreatitis, obesity, endocrine dysfunctions, diabetes, obesity, growth and reproductive disorders, hemophilia, hypercoagulation, idiopathic thrombocytopenic puφura, immunodeficiencies, graft versus host, hemophilia, hypercoagulation, idiopathic thrombocytopenic puφura, immunodeficiencies, hyperthyroidism and hypothyroidism, cystitis, incontinence, endometriosis, cancer, trauma, regeneration (in vitro and in vivo), viral infections, bacterial infections, parasitic infections as well as other diseases, disorders and conditions.

NOV28: WD40 repeat protein-like

NOV28 is homologous to the WD40 repeat protein family. Thus, NOV28 will function similarly to other members ofthe WD40 repeat protein family. The WD40 repeat consists of about 40 residues, each containing a central Tφ-Asp motif. The WD40 repeat is found in beta-transducin (G-beta) and G-beta-like peptides, yeast STE4, MSI1 , CDC4,

CDC20, MAKl 1, PRP4, PWP1 and TUP1, slime-mould AAC3 and coronin, and Drosophila Groucho protein. WD40 repeat-containing proteins are involved in cell cycle progression and cell differentiation. NOV28 nucleic acids and proteins ofthe invention have applications in the diagnosis and/or treatment of various diseases and disorders. For example, measurement of NOV28 expression will be useful in the diagnosis of defective cell differentiation such as in cancer. Also, the compositions ofthe present invention will have efficacy for the treatment of patients suffering from: adrenoleukodystrophy, congenital adrenal hypeφlasia, anemia, ataxia-telangiectasia, autoimmune disease, immunodeficiencies, hemophilia, hypercoagulation, idiopathic thrombocytopenic puφura, autoimmune disease, allergies, immunodeficiencies, transplantation, graft versus host, Von Hippel-Lindau (VHL) syndrome, Alzheimer's disease, stroke, tuberous sclerosis, hypercalcemia, Parkinson's disease, Huntington's disease, cerebral palsy, epilepsy, Lesch-Nyhan syndrome, multiple sclerosis, ataxia-telangiectasia, leukodystrophies, behavioral disorders, addiction, anxiety, pain, neuroprotection, fertility, atherosclerosis, aneurysm, hypertension, fibromuscular dysplasia, stroke, scleroderma, obesity, transplantation, myocardial infarction, embolism, cardiovascular disorders, bypass surgery, diabetes, tuberous sclerosis, cirrhosis, cardiomyopathy, atherosclerosis, hypertension, congenital heart defects, aortic stenosis, atrial septal defect (ASD), atrioventricular (A-V) canal defect, ductus arteriosus, pulmonary stenosis, subaortic stenosis, ventricular septal defect (VSD), valve diseases, tuberous sclerosis, scleroderma, obesity, transplantation, diabetes, autoimmune disease, renal artery stenosis, interstitial nephritis, glomerulonephritis, polycystic kidney disease, systemic lupus erythematosus, renal tubular acidosis, IgA nephropathy, hypercalcemia, Lesch-Nyhan syndrome, Von Hippel-Lindau (VHL) syndrome, cirrhosis, transplantation, systemic lupus erythematosus, autoimmune disease, asthma, emphysema, scleroderma, allergy, ARDS, lymphedema, allergies, hemophilia, hypercoagulation, idiopathic thrombocytopenic puφura, autoimmune disease, allergies, immunodeficiencies, transplantation, graft versus host disease (GVHD), lymphedema, muscular dystrophy, Lesch-Nyhan syndrome, myasthenia gravis, endometriosis, diabetes, Von Hippel-Lindau (VHL) syndrome, pancreatitis, obesity, endocrine dysfunctions, diabetes, obesity, growth and reproductive disorders, psoriasis, actinic keratosis, tuberous sclerosis, acne, hair growth, allopecia, pigmentation disorders, endocrine disorders, hemophilia, hypercoagulation, idiopathic thrombocytopenic puφura, immunodeficiencies, graft versus host, hypercalcemia, ulcers, hemophilia, hypercoagulation, idiopathic thrombocytopenic puφura, immunodeficiencies, hyperthyroidism and hypothyroidism, cancer, trauma, regeneration (in vitro and in vivo), viral¹ infections, bacterial infections, parasitic infections as well as other diseases, disorders and conditions.

NOVX clones

NOVX nucleic acids and their encoded polypeptides are useful in a variety of applications and contexts. The various NOVX nucleic acids and polypeptides according to the invention are useful as novel members ofthe protein families according to the presence of domains and sequence relatedness to previously described proteins. Additionally, NOVX nucleic acids and polypeptides can also be used to identify proteins that are members ofthe family to which the NOVX polypeptides belong. The NOVX genes and their corresponding encoded proteins are useful for preventing, treating or ameliorating medical conditions, e.g., by protein or gene therapy. Pathological conditions can be diagnosed by determining the amount ofthe new protein in a sample or by determming the presence of mutations in the new genes. Specific uses are described for each ofthe NOVX genes, based on the tissues in which they are most highly expressed. Uses include developing products for the diagnosis or treatment of a variety of diseases and disorders.

The NOVX nucleic acids and proteins ofthe invention are useful in potential diagnostic and therapeutic applications and as a research tool. These include serving as a specific or selective nucleic acid or protein diagnostic and/or prognostic marker, wherein the presence or amount ofthe nucleic acid or the protein are to be assessed, as well as potential therapeutic applications such as the following: (i) a protein therapeutic, (ii) a small molecule drug target, (iii) an antibody target (therapeutic, diagnostic, drug targeting/cytotoxic antibody), (iv) a nucleic acid useful in gene therapy (gene delivery/gene ablation), and (v) a composition promoting tissue regeneration in vitro and in vivo (vi) biological defense weapon.

In one specific embodiment, the invention includes an isolated polypeptide comprising an amino acid sequence selected from the group consisting of: (a) a mature form of the amino acid sequence selected from the group consisting of SEQ ID NO:2n, wherem n is an integer between 1 and 33; (b) a variant of a mature form ofthe amino acid sequence selected from the group consisting of SEQ ID NO:2n, wherein n is an integer between 1 and 33, wherein any amino acid in the mature form is changed to a different amino acid, provided that no more than 15% ofthe amino acid residues in the sequence ofthe mature form are so changed; (c) an amino acid sequence selected from the group consisting of SEQ ID NO:2n, wherein n is an integer between 1 and 33; (d) a variant ofthe amino acid sequence selected from the group consisting of SEQ ID NO:2n, wherein n is an integer between 1 and 33, wherein any amino acid specified in the chosen sequence is changed to a different amino acid, provided that no more than 15% ofthe amino acid residues in the sequence are so changed; and (e) a fragment of any of (a) through (d).

In another specific embodiment, the invention includes an isolated nucleic acid molecule comprising a nucleic acid sequence encoding a polypeptide comprising an amino acid sequence selected from the group consisting of: (a) a mature form ofthe amino acid sequence given SEQ ID NO:2n, wherein n is an integer between 1 and 33; (b) a variant of a mature form ofthe amino acid sequence selected from the group consisting of SEQ ID

NO:2n, wherein n is an integer between 1 and 33, wherein any amino acid in the mature form ofthe chosen sequence is changed to a different amino acid, provided that no more than 15% ofthe amino acid residues in the sequence ofthe mature form are so changed; (c) the amino acid sequence selected from the group consisting of SEQ ID NO:2n, wherein n is an integer between 1 and 33; (d) a variant ofthe amino acid sequence selected from the group consisting of SEQ ID NO:2n, wherein n is an integer between 1 and 33, in which any amino acid specified in the chosen sequence is changed to a different amino acid, provided that no more than 15%) ofthe amino acid residues in the sequence are so changed; (e) a nucleic acid fragment encoding at least a portion of a polypeptide comprising the amino acid sequence selected from the group consisting of SEQ ID NO:2n, wherein n is an integer between 1 and 33, or any variant of said polypeptide wherein any amino acid ofthe chosen sequence is changed to a different amino acid, provided that no more than 10%> ofthe amino acid residues in the sequence are so changed; and (f) the complement of any of said nucleic acid molecules. In yet another specific embodiment, the invention includes an isolated nucleic acid molecule, wherein said nucleic acid molecule comprises a nucleotide sequence selected from the group consisting of: (a) the nucleotide sequence selected from the group consisting of SEQ ID NO:2n-l, wherein n is an integer between 1 and 33; (b) a nucleotide sequence wherein one or more nucleotides in the nucleotide sequence selected from the group consisting of SEQ ID NO:2n-l, wherein n is an integer between 1 and 33, is changed from that selected from the group consisting ofthe chosen sequence to a different nucleotide provided that no more than 15% ofthe nucleotides are so changed; (c) a nucleic acid fragment ofthe sequence selected from the group consisting of SEQ ID NO:2n-l, wherein n is an integer between 1 and 33; and (d) a nucleic acid fragment wherein one or more nucleotides in the nucleotide sequence selected from the group consisting of SEQ ID NO:2n- 1, wherein n is an integer between 1 and 33, is changed from that selected from the group consisting ofthe chosen sequence to a different nucleotide provided that no more than 15% ofthe nucleotides are so changed.

NOVX Nucleic Acids and Polypeptides One aspect ofthe invention pertains to isolated nucleic acid molecules that encode

NOVX polypeptides or biologically active portions thereof. Also included in the invention are nucleic acid fragments sufficient for use as hybridization probes to identify NOVX- encoding nucleic acids (e.g., NOVX mRNAs) and fragments for use as PCR primers for the amplification and/or mutation of NOVX nucleic acid molecules. As used herein, the term "nucleic acid molecule" is intended to include DNA molecules (e.g., cDNA or genomic DNA), RNA molecules (e.g., mRNA), analogs ofthe DNA or RNA generated using nucleotide analogs, and derivatives, fragments and homologs thereof. The nucleic acid molecule may be single-stranded or double-stranded, but preferably is comprised double- stranded DNA.

An NOVX nucleic acid can encode a mature NOVX polypeptide. As used herein, a "mature" form of a polypeptide or protein disclosed in the present invention is the product of a naturally occurring polypeptide or precursor form or proprotein. The naturally occurring polypeptide, precursor or proprotein includes, by way of nonlimitmg example, the full-length gene product, encoded by the corresponding gene. Alternatively, it may be defined as the polypeptide, precursor or proprotein encoded by an ORF described herein. The product "mature" form arises, again by way of nonlimiting example, as a result of one or more naturally occurring processing steps as they may take place within the cell, or host cell, in which the gene product arises. Examples of such processing steps leading to a "mature" form of a polypeptide or protein include the cleavage ofthe N-terminal methionine residue encoded by the initiation codon of an ORF, or the proteolytic cleavage of a signal peptide or leader sequence. Thus a mature form arising from a precursor polypeptide or protein that has residues 1 to N, where residue 1 is the N-terminal methionine, would have residues 2 through N remaining after removal ofthe N-terminal methionine. Alternatively, a mature form arising from a precursor polypeptide or protein having residues 1 to N, in which an N- terminal signal sequence from residue 1 to residue M is cleaved, would have the residues from residue M+1 to residue N remaining. Further as used herein, a "mature" form of a polypeptide or protein may arise from a step of post-translational modification other than a proteolytic cleavage event. Such additional processes include, by way of non-limiting example, glycosylation, myristoylation or phosphorylation. In general, a mature polypeptide or protein may result from the operation of only one of these processes, or a combination of any of them. The term "probes", as utilized herein, refers to nucleic acid sequences of variable length, preferably between at least about 10 nucleotides (nt), 100 nt, or as many as approximately, e.g., 6,000 nt, depending upon the specific use. Probes are used in the detection of identical, similar, or complementary nucleic acid sequences. Longer length probes are generally obtained from a natural or recombinant source, are highly specific, and much slower to hybridize than shorter-length oligomer probes. Probes may be single- or double-stranded and designed to have specificity in PCR, membrane-based hybridization technologies, or ELISA-like technologies.

The term "isolated" nucleic acid molecule, as utilized herein, is one, which is separated from other nucleic acid molecules which are present in the natural source ofthe nucleic acid. Preferably, an "isolated" nucleic acid is free of sequences which naturally flank the nucleic acid (i.e., sequences located at the 5'- and 3'-termini ofthe nucleic acid) in the genomic DNA ofthe organism from which the nucleic acid is derived. For example, in various embodiments, the isolated NOVX nucleic acid molecules can contain less than about 5 kb, 4 kb, 3 kb, 2 kb, 1 kb, 0.5 kb or 0.1 kb of nucleotide sequences which naturally flank the nucleic acid molecule in genomic DNA ofthe cell/tissue from which the nucleic acid is derived (e.g., brain, heart, liver, spleen, etc.). Moreover, an "isolated" nucleic acid molecule, such as a cDNA molecule, can be substantially free of other cellular material or culture medium when produced by recombinant techniques, or of chemical precursors or other chemicals when chemically synthesized.

A nucleic acid molecule ofthe invention, e.g., a nucleic acid molecule having the nucleotide sequence SEQ ID NO:2n-l, wherein n is an integer between 1 and 33, or a complement of this aforementioned nucleotide sequence, can be isolated using standard molecular biology techniques and the sequence information provided herein. Using all or a portion of the nucleic acid sequence of SEQ ID NO :2n- 1 , wherein n is an integer between 1 and 33, as a hybridization probe, NOVX molecules can be isolated using standard hybridization and cloning techniques (e.g., as described in Sambrook, et al., (eds.), MOLECULAR CLONING: A LABORATORY MANUAL 2^nd Ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, 1989; and Ausubel, et al., (eds.), CURRENT PROTOCOLS IN MOLECULAR BIOLOGY, John Wiley & Sons, New York, NY, 1993.)

A nucleic acid ofthe invention can be amplified using cDNA, mRNA or alternatively, genomic DNA, as a template and appropriate oligonucleotide primers according to standard PCR amplification techniques. The nucleic acid so amplified can be cloned into an appropriate vector and characterized by DNA sequence analysis. Furthermore, oligonucleotides corresponding to NOVX nucleotide sequences can be prepared by standard synthetic techniques, e.g., using an automated DNA synthesizer.

As used herein, the term "oligonucleotide" refers to a series of linked nucleotide residues, which oligonucleotide has a sufficient number of nucleotide bases to be used in a PCR reaction. A short oligonucleotide sequence may be based on, or designed from, a genomic or cDNA sequence and is used to amplify, confirm, or reveal the presence of an identical, similar or complementary DNA or RNA in a particular cell or tissue. Oligonucleotides comprise portions of a nucleic acid sequence having about 10 nt, 50 nt, or 100 nt in length, preferably about 15 nt to 30 nt in length. In one embodiment of the invention, an oligonucleotide comprising a nucleic acid molecule less than 100 nt in length would further comprise at least 6 contiguous nucleotides SEQ ID NO:2n-l, wherein n is an mteger between 1 and 33, or a complement thereof. Oligonucleotides may be chemically synthesized and may also be used as probes.

In another embodiment, an isolated nucleic acid molecule ofthe invention comprises a nucleic acid molecule that is a complement ofthe nucleotide sequence shown in SEQ ID NO:2n-l, wherein n is an integer between 1 and 33, or a portion of this nucleotide sequence (e.g. , a fragment that can be used as a probe or primer or a fragment encoding a biologically- active portion of an NOVX polypeptide). A nucleic acid molecule that is complementary to the nucleotide sequence shown SEQ ID NO:2n-l, wherein n is an integer between 1 and 33is one that is sufficiently complementary to the nucleotide sequence shown SEQ ID NO:2n-l, wherein n is an integer between 1 and 33, that it can hydrogen bond with little or no mismatches to the nucleotide sequence shown SEQ ID NO:2n-l, wherein n is an integer between 1 and 33, thereby forming a stable duplex.

As used herein, the term "complementary" refers to Watson-Crick or Hoogsteen base pairing between nucleotides units of a nucleic acid molecule, and the term "binding" means the physical or chemical interaction between two polypeptides or compounds or associated polypeptides or compounds or combinations thereof. Binding includes ionic, non-ionic, van der Waals, hydrophobic interactions, and the like. A physical interaction can be either direct or indirect. Indirect interactions may be through or due to the effects of another polypeptide or compound. Direct binding refers to interactions that do not take place through, or due to, the effect of another polypeptide or compound, but instead are without other substantial chemical intermediates.

Fragments provided herein are defined as sequences of at least 6 (contiguous) nucleic acids or at least 4 (contiguous) amino acids, a length sufficient to allow for specific hybridization in the case of nucleic acids or for specific recognition of an epitope in the case of amino acids, respectively, and are at most some portion less than a full length sequence. Fragments may be derived from any contiguous portion of a nucleic acid or amino acid sequence of choice. Derivatives are nucleic acid sequences or amino acid sequences formed from the native compounds either directly or by modification or partial substitution. Analogs are nucleic acid sequences or amino acid sequences that have a structure similar to, but not identical to, the native compound but differs from it in respect to certain components or side chains. Analogs may be synthetic or from a different evolutionary origin and may have a similar or opposite metabolic activity compared to wild type. Homologs are nucleic acid sequences or amino acid sequences of a particular gene that are derived from different species.

A full-length NOVX clone is identified as containing an ATG translation start codon and an in-frame stop codon. Any disclosed NOVX nucleotide sequence lacking an ATG start codon therefore encodes a truncated C-terminal fragment ofthe respective NOVX polypeptide, and requires that the corresponding full-length cDNA extend in the 5' direction ofthe disclosed sequence. Any disclosed NOVX nucleotide sequence lacking an in-frame stop codon similarly encodes a truncated N-terminal fragment ofthe respective NOVX polypeptide, and requires that the corresponding full-length cDNA extend in the 3' direction of the disclosed sequence.

Derivatives and analogs may be full length or other than full length, if the derivative or analog contains a modified nucleic acid or amino acid, as described below. Derivatives or analogs ofthe nucleic acids or proteins ofthe invention include, but are not limited to, molecules comprising regions that are substantially homologous to the nucleic acids or proteins ofthe invention, in various embodiments, by at least about 70%, 80%, or 95% identity (with a preferred identity of 80-95%) over a nucleic acid or amino acid sequence of identical size or when compared to an aligned sequence in which the alignment is done by a computer homology program known in the art, or whose encoding nucleic acid is capable of hybridizing to the complement of a sequence encoding the aforementioned proteins under stringent, moderately stringent, or low stringent conditions. See e.g. Ausubel, et al.,

CURRENT PROTOCOLS IN MOLECULAR BIOLOGY, John Wiley & Sons, New York, NY, 1993, and below.

A "homologous nucleic acid sequence" or "homologous amino acid sequence," or variations thereof, refer to sequences characterized by a homology at the nucleotide level or amino acid level as discussed above. Homologous nucleotide sequences encode those sequences coding for isoforms of NOVX polypeptides. Isoforms can be expressed in different tissues ofthe same organism as a result of, for example, alternative splicing of RNA. Alternatively, isoforms can be encoded by different genes. In the invention, homologous nucleotide sequences include nucleotide sequences encoding for an NOVX polypeptide of species other than humans, including, but not limited to: vertebrates, and thus can include, e.g., frog, mouse, rat, rabbit, dog, cat cow, horse, and other organisms. Homologous nucleotide sequences also include, but are not limited to, naturally occurring allelic variations and mutations ofthe nucleotide sequences set forth herein. A homologous nucleotide sequence does not, however, include the exact nucleotide sequence encoding human NOVX protein. Homologous nucleic acid sequences include those nucleic acid sequences that encode conservative amino acid substitutions (see below) in SEQ ID NO:2n-l, wherein n is an integer between 1 and 33, as well as a polypeptide possessing NOVX biological activity. Various biological activities ofthe NOVX proteins are described below. An NOVX polypeptide is encoded by the open reading frame ("ORF") of an NOVX nucleic acid. An ORF corresponds to a nucleotide sequence that could potentially be translated into a polypeptide. A stretch of nucleic acids comprising an ORF is uninterrupted by a stop codon. An ORF that represents the coding sequence for a full protein begins with an ATG "start" codon and terminates with one ofthe three "stop" codons, namely, TAA, TAG, or TGA. For the puφoses of this invention, an ORF may be any part of a coding sequence, with or without a start codon, a stop codon, or both. For an ORF to be considered as a good candidate for coding for a bonafide cellular protein, a minimum size requirement is often set, e.g., a stretch of DNA that would encode a protein of 50 amino acids or more.

The nucleotide sequences determined from the cloning ofthe human NOVX genes allows for the generation of probes and primers designed for use in identifying and/or cloning NOVX homologues in other cell types, e.g. from other tissues, as well as NOVX homologues from other vertebrates. The probe/primer typically comprises substantially purified oligonucleotide. The oligonucleotide typically comprises a region of nucleotide sequence that hybridizes under stringent conditions to at least about 12, 25, 50, 100, 150, 200, 250, 300, 350 or 400 consecutive sense strand nucleotide sequence SEQ ID NO:2n-l, wherein n is an integer between 1 and 33; or an anti-sense strand nucleotide sequence of SEQ ID NO:2n- 1, wherein n is an integer between 1 and 33; or of a naturally occurring mutant of SEQ ID NO:2n-l, wherein n is an integer between 1 and 33.

Probes based on the human NOVX nucleotide sequences can be used to detect transcripts or genomic sequences encoding the same or homologous proteins. In various embodiments, the probe further comprises a label group attached thereto, e.g. the label group can be a radioisotope, a fluorescent compound, an enzyme, or an enzyme co-factor. Such probes can be used as a part of a diagnostic test kit for identifying cells or tissues which mis- express an NOVX protein, such as by measuring a level of an NOVX-encoding nucleic acid in a sample of cells from a subject e.g. , detecting NOVX mRNA levels or determining whether a genomic NOVX gene has been mutated or deleted.

"A polypeptide having a biologically-active portion of an NOVX polypeptide" refers to polypeptides exhibiting activity similar, but not necessarily identical to, an activity of a polypeptide ofthe invention, including mature forms, as measured in a particular biological assay, with or without dose dependency. A nucleic acid fragment encoding a "biologically- active portion of NOVX" can be prepared by isolating a portion SEQ ID NO:2n-l, wherein n is an integer between 1 and 33, that encodes a polypeptide having an NOVX biological activity (the biological activities ofthe NOVX proteins are described below), expressing the encoded portion of NOVX protein (e.g., by recombinant expression in vitro) and assessing the activity ofthe encoded portion of NOVX.

NOVX Nucleic Acid and Polypeptide Variants

The invention further encompasses nucleic acid molecules that differ from the nucleotide sequences shown in SEQ ID NO:2n-l, wherein n is an integer between 1 and 33, due to degeneracy ofthe genetic code and thus encode the same NOVX proteins as that encoded by the nucleotide sequences shown in SEQ ID NO:2n-l, wherein n is an integer between 1 and 33. In another embodiment, an isolated nucleic acid molecule ofthe invention has a nucleotide sequence encoding a protein having an amino acid sequence shown in SEQ ID NO:2n, wherein n is an integer between 1 and 33. In addition to the human NOVX nucleotide sequences shown in SEQ ID NO:2n-l , wherein n is an integer between 1 and 33, it will be appreciated by those skilled in the art that DNA sequence polymoφhisms that lead to changes in the amino acid sequences ofthe NOVX polypeptides may exist within a population (e.g., the human population). Such genetic polymoφhism in the NOVX genes may exist among individuals within a population due to natural allelic variation. As used herein, the terms "gene" and "recombinant gene" refer to nucleic acid molecules comprising an open reading frame (ORF) encoding an NOVX protein, preferably a vertebrate NOVX protein. Such natural allelic variations can typically result in 1-5% variance in the nucleotide sequence ofthe NOVX genes. Any and all such nucleotide variations and resulting amino acid polymoφhisms in the NOVX polypeptides, which are the result of natural allelic variation and that do not alter the functional activity of the NOVX polypeptides, are intended to be within the scope ofthe invention.

Moreover, nucleic acid molecules encoding NOVX proteins from other species, and thus that have a nucleotide sequence that differs from the human SEQ ID NO:2n-l, wherein n is an integer between 1 and 3.3, are intended to be within the scope ofthe invention. Nucleic acid molecules corresponding to natural allelic variants and homologues ofthe NOVX cDNAs ofthe invention can be isolated based on their homology to the human NOVX nucleic acids disclosed herein using the human cDNAs, or a portion thereof, as a hybridization probe according to standard hybridization techniques under stringent hybridization conditions.

Accordingly, in another embodiment, an isolated nucleic acid molecule ofthe invention is at least 6 nucleotides in length and hybridizes under stringent conditions to the nucleic acid molecule comprising the nucleotide sequence of SEQ ID NO:2n-l, wherein n is an integer between 1 and 33. In another embodiment, the nucleic acid is at least 10, 25, 50, 100, 250, 500, 750, 1000, 1500, or 2000 or more nucleotides in length. In yet another embodiment, an isolated nucleic acid molecule ofthe invention hybridizes to the coding region. As used herein, the term "hybridizes under stringent conditions" is intended to describe conditions for hybridization and washing under which nucleotide sequences at least 60% homologous to each other typically remain hybridized to each other.

Homologs (i.e., nucleic acids encoding NOVX proteins derived from species other than human) or other related sequences (e.g., paralogs) can be obtained by low, moderate or high stringency hybridization with all or a portion ofthe particular human sequence as a probe using methods well known in the art for nucleic acid hybridization and cloning.

As used herein, the phrase "stringent hybridization conditions" refers to conditions under which a probe, primer or oligonucleotide will hybridize to its target sequence, but to no other sequences. Stringent conditions are sequence-dependent and will be different in different circumstances. Longer sequences hybridize specifically at higher temperatures than shorter sequences. Generally, stringent conditions are selected to be about 5 °C lower than the thermal melting point (Tm) for the specific sequence at a defined ionic strength and pH. The Tm is the temperature (under defined ionic strength, pH and nucleic acid concentration) at which 50%) ofthe probes complementary to the target sequence hybridize to the target sequence at equilibrium. Since the target sequences are generally present at excess, at Tm, 50% ofthe probes are occupied at equilibrium. Typically, stringent conditions will be those in which the salt concentration is less than about 1.0 M sodium ion, typically about 0.01 to 1.0 M sodium ion (or other salts) at pH 7.0 to 8.3 and the temperature is at least about 30°C for short probes, primers or oligonucleotides (e.g., 10 nt to 50 nt) and at least about 60°C for longer probes, primers and oligonucleotides. Stringent conditions may also be achieved with the addition of destabilizing agents, such as formamide.

Stringent conditions are known to those skilled in the art and can be found in Ausubel, et al., (eds.), CURRENT PROTOCOLS IN MOLECULAR BIOLOGY, John Wiley & Sons, N.Y. (1989), 6.3.1-6.3.6. Preferably, the conditions are such that sequences at least about 65%, 70%, 75%, 85%, 90%, 95%, 98%, or 99% homologous to each other typically remain hybridized to each other. A non-limiting example of stringent hybridization conditions are hybridization in a high salt buffer comprising 6X SSC, 50 mM Tris-HCl (pH 7.5), 1 mM EDTA, 0.02% PVP, 0.02% Ficoll, 0.02% BSA, and 500 mg/ml denatured salmon sperm DNA at 65°C, followed by one or more washes in 0.2X SSC, 0.01% BSA at 50°C. An isolated nucleic acid molecule of the invention that hybridizes under stringent conditions to the sequences SEQ ID NO:2n-l, wherein n is an integer between 1 and 33, corresponds to a naturally-occurring nucleic acid molecule. As used herein, a "naturally-occurring" nucleic acid molecule refers to an RNA or DNA molecule having a nucleotide sequence that occurs in nature (e.g., encodes a natural protein). In a second embodiment, a nucleic acid sequence that is hybridizable to the nucleic acid molecule comprising the nucleotide sequence of SEQ ID NO:2n-l, wherein n is an integer between 1 and 33, or fragments, analogs or derivatives thereof, under conditions of moderate stringency is provided. A non-limiting example of moderate stringency hybridization conditions are hybridization in 6X SSC, 5X Denhardt's solution, 0.5% SDS and 100 mg/ml denatured salmon sperm DNA at 55°C, followed by one or more washes in

IX SSC, 0.1% SDS at 37°C. Other conditions of moderate stringency that may be used are well-known within the art. See, e.g., Ausubel, et al. (eds.), 1993, CURRENT PROTOCOLS IN MOLECULAR BIOLOGY, John Wiley & Sons, NY, and Kriegler, 1990; GENE TRANSFER AND EXPRESSION, A LABORATORY MANUAL, Stockton Press, NY. In a third embodiment, a nucleic acid that is hybridizable to the nucleic acid molecule comprising the nucleotide sequences SEQ ID NO:2n-l, wherein n is an integer between 1 and 33, or fragments, analogs or derivatives thereof, under conditions of low stringency, is provided. A non-limiting example of low stringency hybridization conditions are hybridization in 35% formamide, 5X SSC, 50 mM Tris-HCl (pH 7.5), 5 mM EDTA, 0.02% PVP, 0.02% Ficoll, 0.2% BSA, 100 mg/ml denatured salmon sperm DNA, 10% (wt/vol) dextran sulfate at 40°C, followed by one or more washes in 2X SSC, 25 mM Tris-HCl (pH 7.4), 5 mM EDTA, and 0.1% SDS at 50°C. Other conditions of low stringency that may be used are well known in the art (e.g., as employed for cross-species hybridizations). See, e.g., Ausubel, et at. (eds.), 1993, CURRENT PROTOCOLS IN MOLECULAR BIOLOGY, John Wiley & Sons, NY, and Kriegler, 1990, GENE TRANSFER AND EXPRESSION, A LABORATORY MANUAL, Stockton Press, NY; Shilo and Weinberg, 1981. Proc Natl Acad Sci USA 78: 6789-6792. Conservative Mutations

In addition to naturally-occurring allelic variants of NOVX sequences that may exist in the population, the skilled artisan will further appreciate that changes can be introduced by mutation into the nucleotide sequences SEQ ID NO:2n-l, wherein n is an integer between 1 and 33, thereby leading to changes in the amino acid sequences ofthe encoded NOVX proteins, without altering the functional ability of said NOVX proteins. For example, nucleotide substitutions leading to amino acid substitutions at "non-essential" amino acid residues can be made in the sequence SEQ ID NO:2n, wherein n is an integer between 1 and 33. A "non-essential" amino acid residue is a residue that can be altered from the wild-type sequences ofthe NOVX proteins without altering their biological activity, whereas an "essential" amino acid residue is required for such biological activity. For example, amino acid residues that are conserved among the NOVX proteins ofthe invention are predicted to be particularly non-amenable to alteration. Amino acids for which conservative substitutions can be made are well-known within the art.

Another aspect ofthe invention pertains to nucleic acid molecules encoding NOVX proteins that contain changes in amino acid residues that are not essential for activity. Such NOVX proteins differ in amino acid sequence from SEQ ID NO:2n, wherein n is an integer between 1 and 33, yet retain biological activity. In one embodiment, the isolated nucleic acid molecule comprises a nucleotide sequence encoding a protein, wherein the protein comprises an amino acid sequence at least about 45% homologous to the amino acid sequences SEQ ID NO:2n, wherein n is an integer between 1 and 33. Preferably, the protein encoded by the nucleic acid molecule is at least about 60% homologous to SEQ ID NO:2n, wherein n is an integer between 1 and 33; more preferably at least about 70% homologous SEQ ID NO:2n, wherein n is an integer between 1 and 33; still more preferably at least about 80% homologous to SEQ ID NO:2n, wherein n is an integer between 1 and 33; even more preferably at least about 90% homologous to SEQ ID NO:2n, wherein n is an integer between 1 and 33; and most preferably at least about 95% homologous to SEQ ID NO:2n, wherein n is an integer between 1 and 33.

An isolated nucleic acid molecule encoding an NOVX protein homologous to the protein of SEQ ID NO:2n, wherein n is an integer between 1 and 33, can be created by introducing one or more nucleotide substitutions, additions or deletions into the nucleotide sequence of SEQ ID NO:2n-l, wherein n is an integer between 1 and 33, such that one or more amino acid substitutions, additions or deletions are introduced into the encoded protein. Mutations can be introduced into SEQ ID NO:2n-l, wherein n is an integer between 1 and 33, by standard techniques, such as site-directed mutagenesis and PCR-mediated mutagenesis. Preferably, conservative amino acid substitutions are made at one or more predicted, non-essential amino acid residues. A "conservative amino acid substitution" is one in which the amino acid residue is replaced with an amino acid residue having a similar side chain. Families of amino acid residues having similar side chains have been defined within the art. These families include amino acids with basic side chains (e.g., lysine, arginine, histidine), acidic side chains (e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine), nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan), beta-branched side chains (e.g., threonine, valine, isoleucine) and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, histidine). Thus, a predicted non-essential amino acid residue in the NOVX protein is replaced with another amino acid residue from the same side chain family. Alternatively, in another embodiment, mutations can be introduced randomly along all or part of an NOVX coding sequence, such as by saturation mutagenesis, and the resultant mutants can be screened for NOVX biological activity to identify mutants that retain activity. Following mutagenesis of SEQ ID NO:2n-l, wherein n is an integer between 1 and 33, the encoded protein can be expressed by any recombinant technology known in the art and the activity of the protein can be determined.

The relatedness of amino acid families may also be determined based on side chain interactions. Substituted amino acids may be fully conserved "strong" residues or fully conserved "weak" residues. The "strong" group of conserved amino acid residues may be any one ofthe following groups: STA, NEQK, NHQK, NDEQ, QHRK, MILV, MILF, HY, FYW, wherein the single letter amino acid codes are grouped by those amino acids that may be substituted for each other. Likewise, the "weak" group of conserved residues may be any one ofthe following: CSA, ATV, SAG, STNK, STPA, SGND, SNDEQK, NDEQHK, NEQHRK, HFY, wherein the letters within each group represent the single letter amino acid code. In one embodiment, a mutant NOVX protein can be assayed for (j) the ability to form proteimprotein interactions with other NOVX proteins, other cell-surface proteins, or biologically-active portions thereof, (ii) complex formation between a mutant NOVX protein and an NOVX ligand; or (in) the ability of a mutant NOVX protein to bind to an intracellular target protein or biologically-active portion thereof; (e.g. avidin proteins). In yet another embodiment, a mutant NOVX protein can be assayed for the ability to regulate a specific biological function (e.g., regulation of insulin release). Antisense Nucleic Acids

Another aspect ofthe invention pertains to isolated antisense nucleic acid molecules that are hybridizable to or complementary to the nucleic acid molecule comprising the nucleotide sequence of SEQ ID NO:2n-l, wherein n is an integer between 1 and 33, or fragments, analogs or derivatives thereof. An "antisense" nucleic acid comprises a nucleotide sequence that is complementary to a "sense" nucleic acid encoding a protein (e.g., complementary to the coding strand of a double-stranded cDNA molecule or complementary to an mRNA sequence). In specific aspects, antisense nucleic acid molecules are provided that comprise a sequence complementary to at least about 10, 25, 50, 100, 250 or 500 nucleotides or an entire NOVX coding strand, or to only a portion thereof. Nucleic acid molecules encoding fragments, homologs, derivatives and analogs of an NOVX protein of SEQ ID NO:2n, wherein n is an integer between 1 and 33, or antisense nucleic acids complementary to an NOVX nucleic acid sequence of SEQ ID NO:2n-l, wherein n is an integer between 1 and 33, are additionally provided. In one embodiment, an antisense nucleic acid molecule is antisense to a "coding region" ofthe coding strand of a nucleotide sequence encoding an NOVX protein. The term "coding region" refers to the region ofthe nucleotide sequence comprising codons which are translated into amino acid residues. In another embodiment, the antisense nucleic acid molecule is antisense to a "noncoding region" ofthe coding strand of a nucleotide sequence encoding the NOVX protein. The term "noncoding region" refers to 5' and 3' sequences which flank the coding region that are not translated into amino acids (i.e., also referred to as 5' and 3' untranslated regions).

Given the coding strand sequences encoding the NOVX protein disclosed herein, antisense nucleic acids ofthe invention can be designed according to the rules of Watson and Crick or Hoogsteen base pairing. The antisense nucleic acid molecule can be complementary to the entire coding region of NOVX mRNA, but more preferably is an oligonucleotide that is antisense to only a portion ofthe coding or noncoding region of NOVX mRNA. For example, the antisense oligonucleotide can be complementary to the region surrounding the translation start site of NOVX mRNA. An antisense oligonucleotide can be, for example, about 5, 10, 15, 20, 25, 30, 35, 40, 45 or 50 nucleotides in length. An antisense nucleic acid ofthe invention can be constructed using chemical synthesis or enzymatic ligation reactions using procedures known in the art. For example, an antisense nucleic acid (e.g., an antisense oligonucleotide) can be chemically synthesized using naturally-occurring nucleotides or variously modified nucleotides designed to increase the biological stability ofthe molecules or to increase the physical stability ofthe duplex formed between the antisense and sense nucleic acids (e.g., phosphorothioate derivatives and acridine substituted nucleotides can be used).

Examples of modified nucleotides that can be used to generate the antisense nucleic acid include: 5-fluorouracil, 5-bromouracil, 5-chlorouracil, 5-iodouracil, hypoxanthine, xanthine, 4-acetylcytosine, 5-(carboxyhydroxylmethyl) uracil, 5-carboxymethylammomethyl- 2-thiouridme, 5-carboxymethylaminomethyluracil, dihydrouracil, beta-D-galactosylqueosine, inosine, N6-isopentenyladenine, 1-methylguanine, 1-methylinosine, 2,2-dimethylguanine, 2-methyladenine, 2-methylguanine, 3-methylcytosine, 5-methylcytosine, N6-adenine, 7-methylguanine, 5-methylaminomethyluracil, 5-methoxyaminomethyl-2-thiouracil, beta-D-mannosylqueosine, 5'-methoxycarboxymethyluracil, 5-methoxyuracil, 2-methylthio-N6-isopentenyladenine, uracil-5-oxyacetic acid (v), wybutoxosine, pseudouracil, queosine, 2-thiocytosine, 5-methyl-2-thiouracil, 2-thiouracil, 4-thiouracil, 5-methyluracil, uracil-5-oxyacetic acid methylester, uracil-5-oxyacetic acid (v), 5-methyl-2-thiouracil, 3-(3-amino-3-N-2-carboxypropyl) uracil, (acp3)w, and

2,6-diaminopurine. Alternatively, the antisense nucleic acid can be produced biologically using an expression vector into which a nucleic acid has been subcloned in an antisense orientation (t.e., RNA transcribed from the inserted nucleic acid will be of an antisense orientation to a target nucleic acid of interest, described further in the following subsection). The antisense nucleic acid molecules of the invention are typically administered to a subject or generated in situ such that they hybridize with or bind to cellular mRNA and/or genomic DNA encoding an NOVX protein to thereby inhibit expression ofthe protein (e.g., by inhibiting transcription and/or translation). The hybridization can be by conventional nucleotide complementarity to form a stable duplex, or, for example, in the case of an antisense nucleic acid molecule that binds to DNA duplexes, through specific interactions in the major groove ofthe double helix. An example of a route of administration of antisense nucleic acid molecules ofthe invention includes direct injection at a tissue site. Alternatively, antisense nucleic acid molecules can be modified to target selected cells and then administered systemically. For example, for systemic administration, antisense molecules can be modified such that they specifically bind to receptors or antigens expressed on a selected cell surface (e.g., by linking the antisense nucleic acid molecules to peptides or antibodies that bind to cell surface receptors or antigens). The antisense nucleic acid molecules can also be delivered to cells using the vectors described herein. To achieve sufficient nucleic acid molecules, vector constructs in which the antisense nucleic acid molecule is placed under the control of a strong pol II or pol UI promoter are preferred.

In yet another embodiment, the antisense nucleic acid molecule ofthe invention is an α-anomeric nucleic acid molecule. An α-anomeric nucleic acid molecule forms specific double-stranded hybrids with complementary RNA in which, contrary to the usual β-units, the strands run parallel to each other. See, e.g., Gaultier, et al., 1987. Nucl. Acids Res. 15: 6625-6641. The antisense nucleic acid molecule can also comprise a 2'-o-methylribonucleotide (See, e.g., Inoue, et al. 1987. Nucl. Acids Res. 15: 6131-6148) or a chimeric RNA-DNA analogue (See, e.g., Inoue, et al., 1987. FEBS Lett. 215: 327-330.

Ribozymes and PNA Moieties

Nucleic acid modifications include, by way of non-limiting example, modified bases, and nucleic acids whose sugar phosphate backbones are modified or derivatized. These modifications are carried out at least in part to enhance the chemical stability ofthe modified nucleic acid, such that they may be used, for example, as antisense binding nucleic acids in therapeutic applications in a subject.

In one embodiment, an antisense nucleic acid ofthe invention is a ribozyme. Ribozymes are catalytic RNA molecules with ribonuclease activity that are capable of cleaving a single-stranded nucleic acid, such as an mRNA, to which they have a complementary region. Thus, ribozymes (e.g., hammerhead ribozymes as described in

Haselhoff and Gerlach 1988. Nature 334: 585-591) can be used to catalytically cleave NOVX mRNA transcripts to thereby inhibit translation of NOVX mRNA. A ribozyme having specificity for an NOVX-encoding nucleic acid can be designed based upon the nucleotide sequence of an NOVX cDNA disclosed herein (i.e., SEQ ID NO:2n-l, wherein n is an integer between 1 and 33). For example, a derivative of a Tetrahymena L-19 IVS RNA can be constructed in which the nucleotide sequence ofthe active site is complementary to the nucleotide sequence to be cleaved in an NOVX-encoding mRNA. See, e.g., U.S. Patent 4,987,071 to Cech, et al. and U.S. Patent 5,116,742 to Cech, et al. NOVX mRNA can also be used to select a catalytic RNA having a specific ribonuclease activity from a pool of RNA molecules. See, e.g., Bartel et al., (1993) Science 261:1411-1418.

Alternatively, NOVX gene expression can be inhibited by targeting nucleotide sequences complementary to the regulatory region ofthe NOVX nucleic acid (e.g., the NOVX promoter and/or enhancers) to form triple helical structures that prevent transcription ofthe NOVX gene in target cells. See, e.g., Helene, 1991. Anticancer Drug Des. 6: 569-84; Helene, et al. 1992. Ann. N.Y. Acad. Sci. 660:27-36; Maher, 1992. Bioassays 14: 807-15.

In various embodiments, the NOVX nucleic acids can be modified at the base moiety, sugar moiety or phosphate backbone to improve, e.g., the stability, hybridization, or solubility ofthe molecule. For example, the deoxyribose phosphate backbone ofthe nucleic acids can be modified to generate peptide nucleic acids. See, e.g., Hyrup, et al., 1996. Bioorg Med Chem 4: 5-23. As used herein, the terms "peptide nucleic acids" or "PNAs" refer to nucleic acid mimics (e.g., DNA mimics) in which the deoxyribose phosphate backbone is replaced by a pseudopeptide backbone and only the four natural nucleobases are retained. The neutral backbone of PNAs has been shown to allow for specific hybridization to DNA and RNA under conditions of low ionic strength. The synthesis of PNA oligomers can be performed using standard solid phase peptide synthesis protocols as described in Hyrup, et al., 1996. supra; Perry-O'Keefe, et al., 1996. Proc. Natl. Acad. Sci. USA 93: 14670-14675.

PNAs of NOVX can be used in therapeutic and diagnostic applications. For example, PNAs can be used as antisense or antigene agents for sequence-specific modulation of gene expression by, e.g., inducing transcription or translation arrest or inhibiting replication. PNAs of NOVX can also be used, for example, in the analysis of single base pair mutations in a gene (e.g., PNA directed PCR clamping; as artificial restriction enzymes when used in combination with other enzymes, e.g., S_\ nucleases (See, Hyrup, et al, 1996. supra); or as probes or primers for DNA sequence and hybridization (See, Hyrup, et al., 1996, supra; Perry-O'Keefe, et al, 1996. supra).

In another embodiment, PNAs of NOVX can be modified, e.g., to enhance their stability or cellular uptake, by attaching lipophilic or other helper groups to PNA, by the formation of PNA-DNA chimeras, or by the use of liposomes or other techniques of drug delivery known in the art. For example, PNA-DNA chimeras of NOVX can be generated that may combine the advantageous properties of PNA and DNA. Such chimeras allow DNA recognition enzymes (e.g., RNase H and DNA polymerases) to interact with the DNA portion while the PNA portion would provide high binding affinity and specificity. PNA-DNA chimeras can be linked using linkers of appropriate lengths selected in terms of base stacking, number of bonds between the nucleobases, and orientation (see, Hyrup, et al., 1996. supra). The synthesis of PNA-DNA chimeras can be performed as described in Hyrup, et al, 1996. supra and Finn, et al., 1996. Nucl Acids Res 24: 3357-3363. For example, a DNA chain can be synthesized on a solid support using standard phosphoramidite coupling chemistry, and modified nucleoside analogs, e.g., 5'-(4-methoxytrityl)amino-5'-deoxy-thymidine phosphoramidite, can be used between the PNA and the 5' end of DNA. See, e.g., Mag, et al, 1989. Nucl Acid Res 17: 5973-5988. PNA monomers are then coupled in a stepwise manner to produce a chimeric molecule with a 5' PNA segment and a 3' DNA segment. See, e.g., Finn, et al, 1996. supra. Alternatively, chimeric molecules can be synthesized with a 5' DNA segment and a 3' PNA segment. See, e.g., Petersen, et al, 1975. Bioorg. Med. Chem. Lett. 5: 1119-11124.

In other embodiments, the oligonucleotide may include other appended groups such as peptides (e.g., for targeting host cell receptors in vivo), or agents facilitating transport across the cell membrane (see, e.g., Letsinger, et al, 1989. Proc. Natl Acad. Sci. U.S.A. 86: 6553-6556; Lemaitre, et al., 1987. Proc. Natl. Acad. Sci. 84: 648-652; PCT Publication No. WO88/09810) or the blood-brain barrier (see, e.g., PCT Publication No. WO 89/10134). In addition, oligonucleotides can be modified with hybridization triggered cleavage agents ( ee, e.g., Krol, et al, 1988. BioTechniques 6:958-976) or intercalating agents (see, e.g., Zsm, 1988. Pharm. Res. 5: 539-549). To this end, the oligonucleotide may be conjugated to another molecule, e.g. , a peptide, a hybridization triggered cross-linking agent, a transport agent, a hybridization-triggered cleavage agent, and the like.

NOVX Polypeptides

A polypeptide accordmg to the invention includes a polypeptide including the amino acid sequence of NOVX polypeptides whose sequences are provided in SEQ ID NO:2n, wherein n is an integer between 1 and 33. The invention also includes a mutant or variant protein any of whose residues may be changed from the corresponding residues shown in SEQ ID NO:2n, wherein n is an integer between 1 and 33, while still encoding a protein that maintains its NOVX activities and physiological functions, or a functional fragment thereof. In general, an NOVX variant that preserves NOVX-like function includes any variant in which residues at a particular position in the sequence have been substituted by other amino acids, and further include the possibility of inserting an additional residue or residues between two residues ofthe parent protein as well as the possibility of deleting one or more residues from the parent sequence. Any amino acid substitution, insertion, or deletion is encompassed by the invention. In favorable circumstances, the substitution is a conservative substitution as defined above.

One aspect ofthe invention pertains to isolated NOVX proteins, and biologically- active portions thereof, or derivatives, fragments, analogs or homologs thereof. Also provided are polypeptide fragments suitable for use as immunogens to raise anti-NOVX antibodies. In one embodiment, native NOVX proteins can be isolated from cells or tissue sources by an appropriate purification scheme using standard protein purification techniques. In another embodiment, NOVX proteins are produced by recombinant DNA techniques. Alternative to recombinant expression, an NOVX protein or polypeptide can be synthesized chemically using standard peptide synthesis techniques.

An "isolated" or "purified" polypeptide or protein or biologically-active portion thereof is substantially free of cellular material or other contaminating proteins from the cell or tissue source from which the NOVX protein is derived, or substantially free from chemical precursors or other chemicals when chemically synthesized. The language "substantially free of cellular material" includes preparations of NOVX proteins in which the protein is separated from cellular components ofthe cells from which it is isolated or recombinantly- produced. In one embodiment, the language "substantially free of cellular material" includes preparations of NOVX proteins having less than about 30%> (by dry weight) of non-NOVX proteins (also referred to herein as a "contaminating protein"), more preferably less than about 20% of non-NOVX proteins, still more preferably less than about 10% of non-NOVX proteins, and most preferably less than about 5% of non-NOVX proteins. When the NOVX protein or biologically-active portion thereof is recombinantly-produced, it is also preferably substantially free of culture medium, i.e., culture medium represents less than about 20%, more preferably less than about 10%, and most preferably less than about 5% ofthe volume of the NOVX protein preparation.

The language "substantially free of chemical precursors or other chemicals" includes preparations of NOVX proteins in which the protein is separated from chemical precursors or other chemicals that are involved in the synthesis ofthe protein. In one embodiment, the language "substantially free of chemical precursors or other chemicals" includes preparations of NOVX proteins having less than about 30% (by dry weight) of chemical precursors or non-NOVX chemicals, more preferably less than about 20% chemical precursors or non-NOVX chemicals, still more preferably less than about 10% chemical precursors or non-NOVX chemicals, and most preferably less than about 5% chemical precursors or non-NOVX chemicals. Biologically-active portions of NOVX proteins include peptides comprising amino acid sequences sufficiently homologous to or derived from the amino acid sequences ofthe NOVX proteins (e.g., the amino acid sequence shown in SEQ ID NO:2n, wherein n is an integer between 1 and 33) that include fewer amino acids than the full-length NOVX proteins, and exhibit at least one activity of an NOVX protein. Typically, biologically-active portions comprise a domain or motif with at least one activity ofthe NOVX protein. A biologically-active portion of an NOVX protein can be a polypeptide which is, for example, 10, 25, 50, 100 or more amino acid residues in length.

Moreover, other biologically-active portions, in which other regions ofthe protein are deleted, can be prepared by recombinant techniques and evaluated for one or more of the functional activities of a native NOVX protein.

In an embodiment, the NOVX protein has an amino acid sequence shown SEQ ID NO:2n, wherein n is an integer between 1 and 33. In other embodiments, the NOVX protein is substantially homologous to SEQ ID NO:2n, wherein n is an integer between 1 and 33, and retains the functional activity of the protein of SEQ ID NO:2n, wherein n is an integer between 1 and 33, yet differs in amino acid sequence due to natural allelic variation or mutagenesis, as described in detail, below. Accordingly, in another embodiment, the NOVX protein is a protein that comprises an amino acid sequence at least about 45% homologous to the amino acid sequence SEQ ID NO:2n, wherein n is an integer between 1 and 33, and retains the functional activity of the NOVX proteins of SEQ ID NO:2n, wherein n is an integer between 1 and 33.

Determining Homology Between Two or More Sequences

To determine the percent homology of two amino acid sequences or of two nucleic acids, the sequences are aligned for optimal comparison puφoses (e.g., gaps can be introduced in the sequence of a first amino acid or nucleic acid sequence for optimal alignment with a second amino or nucleic acid sequence). The amino acid residues or nucleotides at corresponding amino acid positions or nucleotide positions are then compared. When a position in the first sequence is occupied by the same amino acid residue or nucleotide as the corresponding position in the second sequence, then the molecules are homologous at that position (t.e., as used herein amino acid or nucleic acid "homology" is equivalent to amino acid or nucleic acid "identity").

The nucleic acid sequence homology may be determined as the degree of identity between two sequences. The homology may be determined using computer programs known in the art, such as GAP software provided in the GCG program package. See, Needleman and Wunsch, 1970. JMol Biol 48: 443-453. Using GCG GAP software with the following settings for nucleic acid sequence comparison: GAP creation penalty of 5.0 and GAP extension penalty of 0.3, the coding region ofthe analogous nucleic acid sequences referred to above exhibits a degree of identity preferably of at least 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99%o, with the CDS (encoding) part ofthe DNA sequence shown in SEQ ID NO:2n- 1, wherein n is an mteger between 1 and 33.

The term "sequence identity" refers to the degree to which two polynucleotide or polypeptide sequences are identical on a residue-by-residue basis over a particular region of comparison. The term "percentage of sequence identity" is calculated by comparing two optimally aligned sequences over that region of comparison, determining the number of positions at which the identical nucleic acid base (e.g., A, T, C, G, U, or I, in the case of nucleic acids) occurs in both sequences to yield the number of matched positions, dividing the number of matched positions by the total number of positions in the region of comparison (i.e., the window size), and multiplying the result by 100 to yield the percentage of sequence identity. The term "substantial identity" as used herein denotes a characteristic of a polynucleotide sequence, wherein the polynucleotide comprises a sequence that has at least 80 percent sequence identity, preferably at least 85 percent identity and often 90 to 95 percent sequence identity, more usually at least 99 percent sequence identity as compared to a reference sequence over a comparison region.

Chimeric and Fusion Proteins

The invention also provides NOVX chimeric or fusion proteins. As used herein, an NOVX "chimeric protein" or "fusion protein" comprises an NOVX polypeptide operatively- linked to a non-NOVX polypeptide. An "NOVX polypeptide" refers to a polypeptide having an amino acid sequence corresponding to an NOVX protein SEQ ID NO:2n, wherein n is an integer between 1 and 33), whereas a "non-NOVX polypeptide" refers to a polypeptide having an amino acid sequence corresponding to a protein that is not substantially homologous to the NOVX protein, e.g., a protein that is different from the NOVX protein and that is derived from the same or a different organism. Within an NOVX fusion protein the NOVX polypeptide can correspond to all or a portion of an NOVX protein. In one embodiment, an NOVX fusion protein comprises at least one biologically-active portion of an NOVX protein. In another embodiment, an NOVX fusion protein comprises at least two biologically-active portions of an NOVX protein. In yet another embodiment, an NOVX fusion protein comprises at least three biologically-active portions of an NOVX protein. Within the fusion protein, the term "operatively-linked" is intended to indicate that the NOVX polypeptide and the non-NOVX polypeptide are fused in-frame with one another. The non-NOVX polypeptide can be fused to the N-terminus or C-terminus ofthe NOVX polypeptide. In one embodiment, the fusion protein is a GST-NO VX fusion protein in which the NOVX sequences are fused to the C-terminus ofthe GST (glutathione S-transferase) sequences. Such fusion proteins can facilitate the purification of recombinant NOVX polypeptides. In another embodiment, the fusion protein is an NOVX protein containing a heterologous signal sequence at its N-terminus. In certain host cells (e.g., mammalian host cells), expression and/or secretion of NOVX can be increased through use of a heterologous signal sequence.

In yet another embodiment, the fusion protein is an NOVX-immunoglobulin fusion protein in which the NOVX sequences are fused to sequences derived from a member ofthe immunoglobulin protein family. The NOVX-immunoglobulin fusion proteins ofthe invention can be incoφorated into pharmaceutical compositions and administered to a subject to inhibit an interaction between an NOVX ligand and an NOVX protein on the surface of a cell, to thereby suppress NOVX-mediated signal transduction in vivo. The NOVX- immunoglobulin fusion proteins can be used to affect the bioavailability of an NOVX cognate ligand. Inhibition ofthe NOVX ligand/NOVX interaction may be useful therapeutically for both the treatment of proliferative and differentiative disorders, as well as modulating (e.g. promoting or inhibiting) cell survival. Moreover, the NOVX-immunoglobulin fusion proteins ofthe invention can be used as immunogens to produce anti-NOVX antibodies in a subject, to purify NOVX ligands, and in screening assays to identify molecules that inhibit the interaction of NOVX with an NOVX ligand.

An NOVX chimeric or fusion protein ofthe invention can be produced by standard recombinant DNA techniques. For example, DNA fragments coding for the different polypeptide sequences are ligated together in-frame in accordance with conventional techniques, e.g., by employing blunt-ended or stagger-ended termini for ligation, restriction enzyme digestion to provide for appropriate termini, filling-in of cohesive ends as appropriate, alkaline phosphatase treatment to avoid undesirable joining, and enzymatic ligation. In another embodiment, the fusion gene can be synthesized by conventional techniques including automated DNA synthesizers. Alternatively, PCR amplification of gene fragments can be carried out using anchor primers that give rise to complementary overhangs between two consecutive gene fragments that can subsequently be annealed and reamplified to generate a chimeric gene sequence (see, e.g., Ausubel, et al. (eds.) CURRENT PROTOCOLS IN MOLECULAR BIOLOGY, John Wiley & Sons, 1992). Moreover, many expression vectors are commercially available that already encode a fusion moiety (e.g., a GST polypeptide). An NOVX-encoding nucleic acid can be cloned into such an expression vector such that the fusion moiety is linked in-frame to the NOVX protein.

NOVX Agonists and Antagonists

The invention also pertains to variants ofthe NOVX proteins that function as either NOVX agonists (i.e., mimetics) or as NOVX antagonists. Variants ofthe NOVX protein can be generated by mutagenesis (e.g., discrete point mutation or truncation ofthe NOVX protein). An agonist ofthe NOVX protein can retain substantially the same, or a subset of, the biological activities ofthe naturally occurring form ofthe NOVX protein. An antagonist ofthe NOVX protein can inhibit one or more ofthe activities ofthe naturally occurring form of the NOVX protein by, for example, competitively binding to a downstream or upstream member of a cellular signaling cascade which includes the NOVX protein. Thus, specific biological effects can be elicited by treatment with a variant of limited function. In one embodiment, treatment of a subject with a variant having a subset ofthe biological activities ofthe naturally occurring form ofthe protein has fewer side effects in a subject relative to treatment with the naturally occurring form of the NOVX proteins.

Variants ofthe NOVX proteins that function as either NOVX agonists (i.e., mimetics) or as NOVX antagonists can be identified by screening combinatorial libraries of mutants (e.g., truncation mutants) ofthe NOVX proteins for NOVX protein agonist or antagonist activity. In one embodiment, a variegated library of NOVX variants is generated by combinatorial mutagenesis at the nucleic acid level and is encoded by a variegated gene library. A variegated library of NOVX variants can be produced by, for example, enzymatically ligating a mixture of synthetic oligonucleotides into gene sequences such that a degenerate set of potential NOVX sequences is expressible as individual polypeptides, or alternatively, as a set of larger fusion proteins (e.g. , for phage display) containing the set of NOVX sequences therein. There are a variety of methods which can be used to produce libraries of potential NOVX variants from a degenerate oligonucleotide sequence. Chemical synthesis of a degenerate gene sequence can be performed in an automatic DNA synthesizer, and the synthetic gene then ligated into an appropriate expression vector. Use of a degenerate set of genes allows for the provision, in one mixture, of all ofthe sequences encoding the desired set of potential NOVX sequences. Methods for synthesizing degenerate oligonucleotides are well-known within the art. See, e.g., Narang, 1983. Tetrahedron 39: 3; Itakura, et al, 1984. Annu. Rev. Biochem. 53: 323; Itakura, et al, 1984. Science 198: 1056; Ike, et al, 1983. Nucl. Acids Res. 11: 477. Polypeptide Libraries

In addition, libraries of fragments ofthe NOVX protein coding sequences can be used to generate a variegated population of NOVX fragments for screening and subsequent selection of variants of an NOVX protein. In one embodiment, a library of coding sequence fragments can be generated by treating a double stranded PCR fragment of an NOVX coding sequence with a nuclease under conditions wherein nicking occurs only about once per molecule, denaturing the double stranded DNA, renaturing the DNA to form double-stranded DNA that can include sense/antisense pairs from different nicked products, removing single stranded portions from reformed duplexes by treatment with S₁ nuclease, and ligating the resulting fragment library into an expression vector. By this method, expression libraries can be derived which encodes N-terminal and internal fragments of various sizes ofthe NOVX proteins.

Various techniques are known in the art for screening gene products of combinatorial libraries made by point mutations or truncation, and for screening cDNA libraries for gene products having a selected property. Such techniques are adaptable for rapid screening of the gene libraries generated by the combinatorial mutagenesis of NOVX proteins. The most widely used techniques, which are amenable to high throughput analysis, for screening large gene libraries typically include cloning the gene library into replicable expression vectors, transforming appropriate cells with the resulting library of vectors, and expressing the combinatorial genes under conditions in which detection of a desired activity facilitates isolation ofthe vector encoding the gene whose product was detected. Recursive ensemble mutagenesis (REM), a new technique that enhances the frequency of functional mutants in the libraries, can be used in combination with the screening assays to identify NOVX variants. See, e.g., Arkin and Yourvan, 1992. Proc. Natl. Acad. Sci. USA 89: 7811-7815; Delgrave, et al, 1993. Protein Engineering 6:327-331.

Anti-NOVX Antibodies

Also included in the invention are antibodies to NOVX proteins, or fragments of NOVX proteins. The term "antibody" as used herein refers to immunoglobulin molecules and immunologically active portions of immunoglobulin (Ig) molecules, i.e., molecules that contain an antigen binding site that specifically binds (immunoreacts with) an antigen. Such antibodies include, but are not limited to, polyclonal, monoclonal, chimeric, single chain, F_ab, F_ab- and F(_ab_')₂ fragments, and an F_ab expression library. In general, an antibody molecule obtained from humans relates to any ofthe classes IgG, IgM, IgA, IgE and IgD, which differ from one another by the nature ofthe heavy chain present in the molecule. Certain classes have subclasses as well, such as IgG_1; IgG₂, and others. Furthermore, in humans, the light chain may be a kappa chain or a lambda chain. Reference herein to antibodies includes a reference to all such classes, subclasses and types of human antibody species. An isolated NOVX-related protein ofthe invention may be intended to serve as an antigen, or a portion or fragment thereof, and additionally can be used as an immunogen to generate antibodies that immunospecifically bind the antigen, using standard techniques for polyclonal and monoclonal antibody preparation. The full-length protein can be used or, alternatively, the invention provides antigenic peptide fragments ofthe antigen for use as immunogens. An antigenic peptide fragment comprises at least 6 amino acid residues ofthe amino acid sequence ofthe full length protein and encompasses an epitope thereof such that an antibody raised against the peptide forms a specific immune complex with the full length protein or with any fragment that contains the epitope. Preferably, the antigenic peptide comprises at least 10 amino acid residues, or at least 15 amino acid residues, or at least 20 amino acid residues, or at least 30 amino acid residues. Preferred epitopes encompassed by the antigenic peptide are regions ofthe protein that are located on its surface; commonly these are hydrophilic regions.

In certain embodiments ofthe invention, at least one epitope encompassed by the antigenic peptide is a region of NOVX-related protein that is located on the surface ofthe protein, e.g., a hydrophilic region. A hydrophobicity analysis ofthe human NOVX-related protein sequence will indicate which regions of a NOVX-related protein are particularly hydrophilic and, therefore, are likely to encode surface residues useful for targeting antibody production. As a means for targeting antibody production, hydropathy plots showing regions of hydrophilicity and hydrophobicity may be generated by any method well known in the art, including, for example, the Kyte Doolittle or the Hopp Woods methods, either with or without Fourier transformation. See, e.g., Hopp and Woods, 1981, Proc. Nat. Acad. Sci. USA 78: 3824-3828; Kyte and Doolittle 1982, J Mol Biol. 157: 105-142, each of which is incoφorated herein by reference in its entirety. Antibodies that are specific for one or more domains within an antigenic protein, or derivatives, fragments, analogs or homologs thereof, are also provided herein.

A protein ofthe invention, or a derivative, fragment, analog, homolog or ortholog thereof, may be utilized as an immunogen in the generation of antibodies that immunospecifically bind these protein components. Various procedures known within the art may be used for the production of polyclonal or monoclonal antibodies directed against a protein ofthe invention, or against derivatives, fragments, analogs homologs or orthologs thereof (see, for example, Antibodies: A Laboratory Manual, Harlow and Lane, 1988, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, incoφorated herein by reference). Some of these antibodies are discussed below.

Polyclonal Antibodies

For the production of polyclonal antibodies, various suitable host animals (e.g., rabbit, goat, mouse or other mammal) may be immunized by one or more injections with the native protein, a synthetic variant thereof, or a derivative ofthe foregoing. An appropriate immunogenic preparation can contain, for example, the naturally occurring immunogenic protein, a chemically synthesized polypeptide representing the immunogenic protein, or a recombinantly expressed immunogenic protein. Furthermore, the protein may be conjugated to a second protein known to be immunogenic in the mammal being immunized. Examples of such immunogenic proteins include but are not limited to keyhole limpet hemocyanin, serum albumin, bovine thyroglobulin, and soybean trypsin inhibitor. The preparation can further include an adjuvant. Various adjuvants used to increase the immunological response include, but are not limited to, Freund's (complete and incomplete), mineral gels (e.g., aluminum hydroxide), surface active substances (e.g., lysolecithin, pluronic polyols, polyanions, peptides, oil emulsions, dinitiophenol, etc.), adjuvants usable in humans such as Bacille Calmette-Guerin and Corynebacterium parvum, or similar immunostimulatory agents. Additional examples of adjuvants which can be employed include MPL-TDM adjuvant (monophosphoryl Lipid A, synthetic trehalose dicorynomycolate).

The polyclonal antibody molecules directed against the immunogenic protein can be isolated from the mammal (e.g., from the blood) and further purified by well known techniques, such as affinity chromatography using protein A or protein G, which provide primarily the IgG fraction of immune serum. Subsequently, or alternatively, the specific antigen which is the target ofthe immunoglobulin sought, or an epitope thereof, may be immobilized on a column to purify the immune specific antibody by immunoaffinity chromatography. Purification of immunoglobulins is discussed, for example, by D.

Wilkinson (The Scientist, published by The Scientist, Inc., Philadelphia PA, Vol. 14, No. 8 (April 17, 2000), pp. 25-28). Monoclonal Antibodies

The term "monoclonal antibody" (MAb) or "monoclonal antibody composition", as used herein, refers to a population of antibody molecules that contain only one molecular species of antibody molecule consisting of a unique light chain gene product and a unique heavy chain gene product. In particular, the complementarity determining regions (CDRs) of the monoclonal antibody are identical in all the molecules ofthe population. MAbs thus contain an antigen binding site capable of immunoreacting with a particular epitope ofthe antigen characterized by a unique binding affinity for it.

Monoclonal antibodies can be prepared using hybridoma methods, such as those described by Kohler and Milstein, Nature, 256:495 (1975). In a hybridoma method, a mouse, hamster, or other appropriate host animal, is typically immunized with an immunizing agent to elicit lymphocytes that produce or are capable of producing antibodies that will specifically bind to the immunizing agent. Alternatively, the lymphocytes can be immunized in vitro. The immunizing agent will typically include the protein antigen, a fragment thereof or a fusion protein thereof. Generally, either peripheral blood lymphocytes are used if cells of human origin are desired, or spleen cells or lymph node cells are used if non-human mammalian sources are desired. The lymphocytes are then fused with an immortalized cell line using a suitable fusing agent, such as polyethylene glycol, to form a hybridoma cell (Goding, MONOCLONAL ANTIBODIES: PRINCIPLES AND PRACTICE, Academic Press, (1986) pp. 59-103). Immortalized cell lines are usually transformed mammalian cells, particularly myeloma cells of rodent, bovine and human origin. Usually, rat or mouse myeloma cell lines are employed. The hybridoma cells can be cultured in a suitable culture medium that preferably contains one or more substances that inhibit the growth or survival ofthe unfused, immortalized cells. For example, if the parental cells lack the enzyme hypoxanthine guanine phosphoribosyl transferase (HGPRT or HPRT), the culture medium for the hybridomas typically will include hypoxanthine, aminopterin, and thymidine ("HAT medium"), which substances prevent the growth of HGPRT-deficient cells.

Preferred immortalized cell lines are those that fuse efficiently, support stable high level expression of antibody by the selected antibody-producing cells, and are sensitive to a medium such as HAT medium. More preferred immortalized cell lines are murine myeloma lines, which can be obtained, for instance, from the Salk Institute Cell Distribution Center, San Diego, California and the American Type Culture Collection, Manassas, Virginia. Human myeloma and mouse-human heteromyeloma cell lines also have been described for the production of human monoclonal antibodies (Kozbor, J. Immunol., 133:3001 (1984); Brodeur et al, MONOCLONAL ANTIBODY PRODUCTION TECHNIQUES AND APPLICATIONS, Marcel Dekker, Inc., New York, (1987) pp. 51-63).

The culture medium in which the hybridoma cells are cultured can then be assayed for the presence of monoclonal antibodies directed against the antigen. Preferably, the binding specificity of monoclonal antibodies produced by the hybridoma cells is determined by immunoprecipitation or by an in vitro binding assay, such as radioimmunoassay (RIA) or enzyme-linked immunoabsorbent assay (ELISA). Such techniques and assays are known in the art. The binding affinity ofthe monoclonal antibody can, for example, be determined by the Scatchard analysis of Munson and Pollard, Anal. Biochem., 107:220 (1980). Preferably, antibodies having a high degree of specificity and a high binding affinity for the target antigen are isolated.

After the desired hybridoma cells are identified, the clones can be subcloned by limiting dilution procedures and grown by standard methods. Suitable culture media for this puφose include, for example, Dulbecco's Modified Eagle's Medium and RPMI- 1640 medium. Alternatively, the hybridoma cells can be grown in vivo as ascites in a mammal.

The monoclonal antibodies secreted by the subclones can be isolated or purified from the culture medium or ascites fluid by conventional immunoglobulin purification procedures such as, for example, protein A-Sepharose, hydroxylapatite chromatography, gel electrophoresis, dialysis, or affinity chromatography.

The monoclonal antibodies can also be made by recombinant DNA methods, such as those described in U.S. Patent No. 4,816,567. DNA encoding the monoclonal antibodies of the invention can be readily isolated and sequenced using conventional procedures (e.g., by using oligonucleotide probes that are capable of binding specifically to genes encoding the heavy and light chains of murine antibodies). The hybridoma cells ofthe invention serve as a preferred source of such DNA. Once isolated, the DNA can be placed into expression vectors, which are then transfected into host cells such as simian COS cells, Chinese hamster ovary (CHO) cells, or myeloma cells that do not otherwise produce immunoglobulin protein, to obtain the synthesis of monoclonal antibodies in the recombinant host cells. The DNA also can be modified, for example, by substituting the coding sequence for human heavy and light chain constant domains in place ofthe homologous murine sequences (U.S. Patent No. 4,816,567; Morrison, Nature 368, 812-13 (1994)) or by covalently joining to the immunoglobulin coding sequence all or part ofthe coding sequence for a non- immunoglobulin polypeptide. Such a non-immunoglobulin polypeptide can be substituted for the constant domains of an antibody of the invention, or can be substituted for the variable domains of one antigen-combining site of an antibody ofthe invention to create a chimeric bivalent antibody. Humanized Antibodies The antibodies directed against the protein antigens ofthe invention can further comprise humanized antibodies or human antibodies. These antibodies are suitable for administration to humans without engendering an immune response by the human against the administered immunoglobulin. Humanized forms of antibodies are chimeric immunoglobulins, immunoglobulin chains or fragments thereof (such as Fv, Fab, Fab', F(ab')₂ or other antigen-binding subsequences of antibodies) that are principally comprised ofthe sequence of a human immunoglobulin, and contain minimal sequence derived from a non- human immunoglobulin. Humanization can be performed following the method of Winter and co-workers (Jones et al, Nature, 321:522-525 (1986); Riechmann et al, Nature, 332:323-327 (1988); Verhoeyen et al, Science, 239:1534-1536 (1988)), by substituting rodent CDRs or CDR sequences for the corresponding sequences of a human antibody. (See also U.S. Patent No. 5,225,539.) In some instances, Fv framework residues ofthe human immunoglobulin are replaced by corresponding non-human residues. Humanized antibodies can also comprise residues which are found neither in the recipient antibody nor in the imported CDR or framework sequences. In general, the humanized antibody will comprise substantially all of at least one, and typically two, variable domains, in which all or substantially all ofthe CDR regions correspond to those of a non-human immunoglobulin and all or substantially all ofthe framework regions are those of a human immunoglobulin consensus sequence. The humanized antibody optimally also will comprise at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin (Jones et al, 1986; Riechmann et al, 1988; and Presta, Curr. Op. Struct. Biol, 2:593-596 (1992)). Human Antibodies

Fully human antibodies relate to antibody molecules in which essentially the entire sequences of both the light chain and the heavy chain, including the CDRs, arise from human genes. Such antibodies are termed "human antibodies", or "fully human antibodies" herein. Human monoclonal antibodies can be prepared by the trioma technique; the human B-cell hybridoma technique (see Kozbor, et al, 1983 Immunol Today 4: 72) and the EBV hybridoma technique to produce human monoclonal antibodies (see Cole, et al, 1985 In: MONOCLONAL ANTIBODIES AND CANCER THERAPY, Alan R. Liss, Inc., pp. 77-96). Human monoclonal antibodies may be utilized in the practice ofthe present invention and may be produced by using human hybridomas (see Cote, et al, 1983. Proc Natl Acad Sci USA 80:2026-2030) or by transforming human B-cells with Epstein Barr Virus in vitro (see Cole, et al, 1985 In: MONOCLONAL ANTIBODIES AND CANCER THERAPY, Alan R. Liss, Inc., pp. 77-96). In addition, human antibodies can also be produced using additional techniques, including phage display libraries (Hoogenboom and Winter, J. Mol. Biol, 227:381 (1991); Marks et al, J. Mol. Biol, 222:581 (1991)). Similarly, human antibodies can be made by introducing human immunoglobulin loci into transgenic animals, e.g., mice in which the endogenous immunoglobulin genes have been partially or completely inactivated. Upon challenge, human antibody production is observed, which closely resembles that seen in humans in all respects, including gene rearrangement, assembly, and antibody repertoire. This approach is described, for example, in U.S. Patent Nos. 5,545,807; 5,545,806; 5,569,825; 5,625,126; 5,633,425; 5,661,016, and in Marks et al. (Bio/Technology 10, 779- 783 (1992)); Lonberg et al. (Nature 368 856-859 (1994)); Morrison ( Nature 368, 812-13 (1994)); Fishwild et al,( Nature Biotechnology 14, 845-51 (1996)); Neuberger (Nature Biotechnology 14, 826 (1996)); and Lonberg and Huszar (Intern. Rev. Immunol 13 65-93 (1995)).

Human antibodies may additionally be produced using transgenic nonhuman animals which are modified so as to produce fully human antibodies rather than the animal's endogenous antibodies in response to challenge by an antigen. (See PCT publication

WO94/02602). The endogenous genes encoding the heavy and light immunoglobulin chains in the nonhuman host have been incapacitated, and active loci encoding human heavy and light chain immunoglobulins are inserted into the host's genome. The human genes are incoφorated, for example, using yeast artificial chromosomes containing the requisite human DNA segments. An animal which provides all the desired modifications is then obtained as progeny by crossbreeding intermediate transgenic animals containing fewer than the full complement ofthe modifications. The preferred embodiment of such a nonhuman animal is a mouse, and is termed the Xenomouse™ as disclosed in PCT publications WO 96/33735 and WO 96/34096. This animal produces B cells which secrete fully human immunoglobulins. The antibodies can be obtained directly from the animal after immunization with an immunogen of interest, as, for example, a preparation of a polyclonal antibody, or alternatively from immortalized B cells derived from the animal, such as hybridomas producing monoclonal antibodies. Additionally, the genes encoding the immunoglobulins with human variable regions can be recovered and expressed to obtain the antibodies directly, or can be further modified to obtain analogs of antibodies such as, for example, single chain Fv molecules.

An example of a method of producing a nonhuman host, exemplified as a mouse, lacking expression of an endogenous immunoglobulin heavy chain is disclosed in U.S. Patent No. 5,939,598. It can be obtained by a method including deleting the J segment genes from at least one endogenous heavy chain locus in an embryonic stem cell to prevent rearrangement of the locus and to prevent formation of a transcript of a rearranged immunoglobulin heavy chain locus, the deletion being effected by a targeting vector containing a gene encoding a selectable marker; and producing from the embryonic stem cell a transgenic mouse whose somatic and germ cells contain the gene encoding the selectable marker.

A method for producing an antibody of interest, such as a human antibody, is disclosed in U.S. Patent No. 5,916,771. It includes introducing an expression vector that contains a nucleotide sequence encoding a heavy chain into one mammalian host cell in culture, introducing an expression vector containing a nucleotide sequence encoding a light chain into another mammalian host cell, and fusing the two cells to form a hybrid cell. The hybrid cell expresses an antibody containing the heavy chain and the light chain.

In a further improvement on this procedure, a method for identifying a clinically relevant epitope on an immunogen, and a correlative method for selecting an antibody that binds immunospecifically to the relevant epitope with high affinity, are disclosed in PCT publication WO 99/53049.

F_ab Fragments and Single Chain Antibodies

According to the invention, techniques can be adapted for the production of single-chain antibodies specific to an antigenic protein ofthe invention (see e.g., U.S. Patent No. 4,946,778). In addition, methods can be adapted for the construction of F_ab expression libraries (see e.g., Huse, et al, 1989 Science 246: 1275-1281) to allow rapid and effective identification of monoclonal F_ab fragments with the desired specificity for a protein or derivatives, fragments, analogs or homologs thereof. Antibody fragments that contain the idiotypes to a protein antigen may be produced by techniques known in the art including, but not limited to: (i) an F(_ab_')₂ fragment produced by pepsin digestion of an antibody molecule; (ii) an F_ab fragment generated by reducing the disulfide bridges of an F(_{a '})₂ fragment; (iii) an F_ab fragment generated by the treatment ofthe antibody molecule with papain and a reducing agent and (iv) F_v fragments. Bispecific Antibodies

Bispecific antibodies are monoclonal, preferably human or humanized, antibodies that have binding specificities for at least two different antigens. In the present case, one ofthe binding specificities is for an antigenic protein ofthe invention. The second binding target is any other antigen, and advantageously is a cell-surface protein or receptor or receptor subunit.

Methods for making bispecific antibodies are known in the art. Traditionally, the recombinant production of bispecific antibodies is based on the co-expression of two immunoglobulin heavy-chain/light-chain pairs, where the two heavy chains have different specificities (Milstein and Cuello, Nature, 305:537-539 (1983)). Because ofthe random assortment of immunoglobulin heavy and light chains, these hybridomas (quadromas) produce a potential mixture often different antibody molecules, of which only one has the correct bispecific structure. The purification ofthe correct molecule is usually accomplished by affinity chromatography steps. Similar procedures are disclosed in WO 93/08829, published 13 May 1993, and in Traunecker et al, 1991 EMBO J, 10:3655-3659.

Antibody variable domains with the desired binding specificities (antibody-antigen combining sites) can be fused to immunoglobulin constant domain sequences. The fusion preferably is with an immunoglobulin heavy-chain constant domain, comprising at least part ofthe hinge, CH2, and CH3 regions. It is preferred to have the first heavy-chain constant region (CHI) containing the site necessary for light-chain binding present in at least one of the fusions. DNAs encoding the immunoglobulin heavy-chain fusions and, if desired, the immunoglobulin light chain, are inserted into separate expression vectors, and are co- transfected into a suitable host organism. For further details of generating bispecific antibodies see, for example, Suresh et al, Methods in Enzymology, 121:210 (1986). According to another approach described in WO 96/27011 , the interface between a pair of antibody molecules can be engineered to maximize the percentage of heterodimers which are recovered from recombinant cell culture. The preferred interface comprises at least a part ofthe CH3 region of an antibody constant domain. In this method, one or more small amino acid side chains ftom the interface ofthe first antibody molecule are replaced with larger side chains (e.g. tyrosine or tryptophan). Compensatory "cavities" of identical or similar size to the large side chain(s) are created on the interface ofthe second antibody molecule by replacing large amino acid side chains with smaller ones (e.g. alanine or threonine). This provides a mechanism for increasing the yield ofthe heterodimer over other unwanted end-products such as homodimers. Bispecific antibodies can be prepared as full length antibodies or antibody fragments (e.g. F(ab')₂ bispecific antibodies). Techniques for generating bispecific antibodies from antibody fragments have been described in the literature. For example, bispecific antibodies can be prepared using chemical linkage. Brennan et al, Science 229:81 (1985) describe a procedure wherein intact antibodies are proteolytically cleaved to generate F(ab')₂ fragments. These fragments are reduced in the presence ofthe dithiol complexing agent sodium arsenite to stabilize vicinal dithiols and prevent intermolecular disulfide formation. The Fab' fragments generated are then converted to thionitrobenzoate (TNB) derivatives. One ofthe Fab'-TNB derivatives is then reconverted to the Fab '-thiol by reduction with mercaptoethylamine and is mixed with an equimolar amount ofthe other Fab'-TNB derivative to form the bispecific antibody. The bispecific antibodies produced can be used as agents for the selective immobilization of enzymes.

Additionally, Fab' fragments can be directly recovered from E. coli and chemically coupled to form bispecific antibodies. Shalaby et al., J. Exp. Med. 175:217-225 (1992) describe the production of a fully humanized bispecific antibody F(ab')₂ molecule. Each Fab' fragment was separately secreted from E. coli and subjected to directed chemical coupling in vitro to form the bispecific antibody. The bispecific antibody thus formed was able to bind to cells overexpressing the ErbB2 receptor and normal human T cells, as well as trigger the lytic activity of human cytotoxic lymphocytes against human breast tumor targets. Various techniques for making and isolating bispecific antibody fragments directly from recombinant cell culture have also been described. For example, bispecific antibodies have been produced using leucine zippers. Kostelny et al, J. Immunol. 148(5):1547-1553 (1992). The leucine zipper peptides from the Fos and Jun proteins were linked to the Fab' portions of two different antibodies by gene fusion. The antibody homodimers were reduced at the hinge region to form monomers and then re-oxidized to form the antibody heterodimers. This method can also be utilized for the production of antibody homodimers. The "diabody" technology described by Hollinger et al, Proc. Natl Acad. Sci. USA 90:6444-6448 (1993) has provided an alternative mechanism for making bispecific antibody fragments. The fragments comprise a heavy-chain variable domain (V_H) connected to a light-chain variable domain (VL) by a linker which is too short to allow pairing between the two domains on the same chain. Accordingly, the V_H and V_L domains of one fragment are forced to pair with the complementary V_L and V_H domains of another fragment, thereby forming two antigen-binding sites. Another strategy for making bispecific antibody fragments by the use of single-chain Fv (sFv) dimers has also been reported. See, Gruber et al., J. Immunol. 152:5368 (1994).

Antibodies with more than two valencies are contemplated. For example, trispecific antibodies can be prepared. Tutt et al, J. Immunol. 147:60 (1991). Exemplary bispecific antibodies can bind to two different epitopes, at least one of which originates in the protein antigen ofthe invention. Alternatively, an anti-antigenic arm of an immunoglobulin molecule can be combined with an arm which binds to a triggering molecule on a leukocyte such as a T-cell receptor molecule (e.g. CD2, CD3, CD28, or B7), or Fc receptors for IgG (FcγR), such as FcγRI (CD64), FcγRII (CD32) and FcγRIII (CD 16) so as to focus cellular defense mechanisms to the cell expressing the particular antigen.

Bispecific antibodies can also be used to direct cytotoxic agents to cells which express a particular antigen. These antibodies possess an antigen-binding arm and an arm which binds a cytotoxic agent or a radionuclide chelator, such as EOTUBE, DPT A, DOTA, or TET A. Another bispecific antibody of interest binds the protein antigen described herein and further binds tissue factor (TF).

Heteroconjugate Antibodies

Heteroconjugate antibodies are also within the scope ofthe present invention. Heteroconjugate antibodies are composed of two covalently joined antibodies. Such antibodies have, for example, been proposed to target immune system cells to unwanted cells (U.S. Patent No. 4,676,980), and for treatment of FUN infection (WO 91/00360; WO

92/200373; EP 03089). It is contemplated that the antibodies can be prepared in vitro using known methods in synthetic protein chemistry, including those involving crosslinking agents. For example, immunotoxins can be constructed using a disulfide exchange reaction or by forming a thioether bond. Examples of suitable reagents for this puφose include iminothiolate and methyl-4-mercaptobutyrimidate and those disclosed, for example, in U.S. Patent No. 4,676,980. Effector Function Engineering

It can be desirable to modify the antibody ofthe invention with respect to effector function, so as to enhance, e.g., the effectiveness ofthe antibody in treating cancer. For example, cysteine residue(s) can be introduced into the Fc region, thereby allowing interchain disulfide bond formation in this region. The homodimeric antibody thus generated can have improved internalization capability and/or increased complement-mediated cell killing and antibody-dependent cellular cytotoxicity (ADCC). See Caron et al, J. Exp Med., 176: 1191- 1195 (1992) and Shopes, J. Immunol., 148:2918-2922 (1992). Homodimeric antibodies with enhanced anti-tumor activity can also be prepared using heterobifunctional cross-linkers as described in Wolff et al. Cancer Research, 53:2560-2565 (1993). Alternatively, an antibody can be engineered that has dual Fc regions and can thereby have enhanced complement lysis and ADCC capabilities. See Stevenson et al, Anti-Cancer Drug Design, 3:219-230 (1989). Immunoconjugates

The invention also pertains to immunoconjugates comprising an antibody conjugated to a cytotoxic agent such as a chemotherapeutic agent, toxin (e.g., an enzymatically active toxin of bacterial, fungal, plant, or animal origin, or fragments thereof), or a radioactive isotope (i.e., a radioconjugate). Chemotherapeutic agents useful in the generation of such immunoconjugates have been described above. Enzymatically active toxins and fragments thereof that can be used include diphtheria A chain, nonbinding active fragments of diphtheria toxin, exotoxin A chain (from Pseudomonas aeruginosa), ricin A chain, abrin A chain, modeccin A chain, alpha-sarcin, Aleurites fordii proteins, dianthin proteins, Phytolaca americana proteins (PAPI, PAPII, and PAP-S), momordica charantia inhibitor, curcin, crotin, sapaonaria officinalis inhibitor, gelonin, mitogellin, restrictocin, phenomycin, enomycin, and the tricothecenes. A variety of radionuclides are available for the production of radioconjugated antibodies. Examples include ²¹²Bi, ¹³¹1, ¹³¹In, ⁹⁰Y, and ¹⁸⁶Re.

Conjugates ofthe antibody and cytotoxic agent are made using a variety of bifunctional protein-coupling agents such as N-succinimidyl-3-(2-pyridyldithiol) propionate (SPDP), iminothiolane (IT), bifunctional derivatives of imidoesters (such as dimethyl adipimidate HCL), active esters (such- as disuccinimidyl suberate), aldehydes (such as glutareldehyde), bis-azido compounds (such as bis (p-azidobenzoyl) hexanediamine), bis- diazonium derivatives (such as bis-(p-diazoniumbenzoyl)-ethylenediamine), diisocyanates (such as tolyene 2,6-diisocyanate), and bis-active fluorine compounds (such as 1,5-difluoro- 2,4-dinitrobenzene). For example, a ricin immunotoxin can be prepared as described in Vitetta et al, Science, 238: 1098 (1987). Carbon- 14-labeled l-isothiocyanatobenzyl-3- methyldiethylene triaminepentaacetic acid (MX-DTPA) is an exemplary chelating agent for conjugation of radionucleotide to the antibody. See W094/11026. In another embodiment, the antibody can be conjugated to a "receptor" (such streptavidin) for utilization in tumor pretargeting wherein the antibody-receptor conjugate is administered to the patient, followed by removal of unbound conjugate from the circulation using a clearing agent and then administration of a "ligand" (e.g., avidin) that is in turn conjugated to a cytotoxic agent. In one embodiment, methods for the screening of antibodies that possess the desired specificity include, but are not limited to, enzyme-linked immunosorbent assay (ELISA) and other immunologically-mediated techniques known within the art. In a specific embodiment, selection of antibodies that are specific to a particular domain of an NOVX protein is facilitated by generation of hybridomas that bind to the fragment of an NOVX protein possessing such a domain. Thus, antibodies that are specific for a desired domain within an NOVX protein, or derivatives, fragments, analogs or homologs thereof, are also provided herein.

Anti-NOVX antibodies may be used in methods known within the art relating to the localization and/or quantitation of an NOVX protein (e.g., for use in measuring levels of the NOVX protein within appropriate physiological samples, for use in diagnostic methods, for use in imaging the protein, and the like). In a given embodiment, antibodies for NOVX proteins, or derivatives, fragments, analogs or homologs thereof, that contain the antibody derived binding domain, are utilized as pharmacologically-active compounds (hereinafter "Therapeutics").

An anti-NOVX antibody (e.g., monoclonal antibody) can be used to isolate an NOVX polypeptide by standard techniques, such as affinity chromatography or immunoprecipitation. An anti-NOVX antibody can facilitate the purification of natural NOVX polypeptide from cells and of recombinantly-produced NOVX polypeptide expressed in host cells. Moreover, an anti-NOVX antibody can be used to detect NOVX protein (e.g. , in a cellular lysate or cell supernatant) in order to evaluate the abundance and pattern of expression ofthe NOVX protein. Anti-NOVX antibodies can be used diagnostically to monitor protein levels in tissue as part of a clinical testing procedure, e.g., to, for example, determine the efficacy of a given treatment regimen. Detection can be facilitated by coupling (i.e., physically linking) the antibody to a detectable substance. Examples of detectable substances include various enzymes, prosthetic groups, fluorescent materials, luminescent materials, bioluminescent materials, and radioactive materials. Examples of suitable enzymes include horseradish peroxidase, alkaline phosphatase, β-galactosidase, or acetylcholinesterase; examples of suitable prosthetic group complexes include streptavidin/biotin and avidin/biotin; examples of suitable fluorescent materials include umbelliferone, fluorescein, fluorescein isothiocyanate, rhodamine, dichlorotriazinylamine fluorescein, dansyl chloride or phycoerythrin; an example of a luminescent material includes luminol; examples of bioluminescent materials include luciferase, luciferin, and aequorin, and examples of suitable radioactive material include ¹²⁵1, ¹³¹1, ³⁵S or ³H. NOVX Recombinant Expression Vectors and Host Cells

Another aspect ofthe invention pertains to vectors, preferably expression vectors, containing a nucleic acid encoding an NOVX protein, or derivatives, fragments, analogs or homologs thereof. As used herein, the term "vector" refers to a nucleic acid molecule capable of transporting another nucleic acid to which it has been linked. One type of vector is a

"plasmid", which refers to a circular double stranded DNA loop into which additional DNA segments can be ligated. Another type of vector is a viral vector, wherein additional DNA segments can be ligated into the viral genome. Certain vectors are capable of autonomous replication in a host cell into which they are introduced (e.g., bacterial vectors having a bacterial origin of replication and episomal mammalian vectors) . Other vectors (e.g. , non-episomal mammalian vectors) are integrated into the genome of a host cell upon introduction into the host cell, and thereby are replicated along with the host genome. Moreover, certain vectors are capable of directing the expression of genes to which they are operatively-linked. Such vectors are referred to herein as "expression vectors". In general, expression vectors of utility in recombinant DNA techniques are often in the form of plasmids. In the present specification, "plasmid" and "vector" can be used interchangeably as the plasmid is the most commonly used form of vector. However, the invention is intended to include such other forms of expression vectors, such as viral vectors (e.g., replication defective retroviruses, adenoviruses and adeno-associated viruses), which serve equivalent functions.

The recombinant expression vectors ofthe invention comprise a nucleic acid ofthe invention in a form suitable for expression ofthe nucleic acid in a host cell, which means that the recombinant expression vectors include one or more regulatory sequences, selected on the basis ofthe host cells to be used for expression, that is operatively-linked to the nucleic acid sequence to be expressed. Within a recombinant expression vector, "operably-linked" is intended to mean that the nucleotide sequence of mterest is linked to the regulatory sequence(s) in a manner that allows for expression ofthe nucleotide sequence (e.g., in an in vitro transcription/translation system or in a host cell when the vector is introduced into the host cell). The term "regulatory sequence" is intended to includes promoters, enhancers and other expression control elements (e.g., polyadenylation signals). Such regulatory sequences are described, for example, in Goeddel, GENE EXPRESSION TECHNOLOGY: METHODS IN ENZYMOLOGY 185, Academic Press, San Diego, Calif. (1990). Regulatory sequences include those that direct constitutive expression of a nucleotide sequence in many types of host cell and those that direct expression ofthe nucleotide sequence only in certain host cells (e.g., tissue-specific regulatory sequences). It will be appreciated by those skilled in the art that the design ofthe expression vector can depend on such factors as the choice ofthe host cell to be transformed, the level of expression of protein desired, etc. The expression vectors ofthe invention can be introduced into host cells to thereby produce proteins or peptides, including fusion proteins or peptides, encoded by nucleic acids as described herein (e.g., NOVX proteins, mutant forms of NOVX proteins, fusion proteins, etc.).

The recombinant expression vectors ofthe invention can be designed for expression of NOVX proteins in prokaryotic or eukaryotic cells. For example, NOVX proteins can be expressed in bacterial cells such as Escherichia coli, insect cells (using baculovirus expression vectors) yeast cells or mammalian cells. Suitable host cells are discussed further in Goeddel, GENE EXPRESSION TECHNOLOGY: METHODS IN ENZYMOLOGY 185, Academic Press, San Diego, Calif. (1990). Alternatively, the recombinant expression vector can be transcribed and translated in vitro, for example using T7 promoter regulatory sequences and T7 polymerase.

Expression of proteins in prokaryotes is most often carried out in Escherichia coli with vectors containing constitutive or inducible promoters directing the expression of either fusion or non-fusion proteins. Fusion vectors add a number of amino acids to a protein encoded therein, usually to the amino terminus ofthe recombinant protein. Such fusion vectors typically serve three puφoses: ( ) to increase expression of recombinant protein; (ii) to increase the solubility ofthe recombinant protein; and (in) to aid in the purification ofthe recombinant protein by acting as a ligand in affinity purification. Often, in fusion expression vectors, a proteolytic cleavage site is introduced at the junction ofthe fusion moiety and the recombinant protein to enable separation ofthe recombinant protein from the fusion moiety subsequent to purification of the fusion protein. Such enzymes, and their cognate recognition sequences, include Factor Xa, thrombin and enterokinase. Typical fusion expression vectors include pGEX (Pharmacia Biotech Inc; Smith and Johnson, 1988. Gene 67: 31-40), pMAL (New England Biolabs, Beverly, Mass.) and pRIT5 (Pharmacia, Piscataway, NJ.) that fuse glutathione S-transferase (GST), maltose E binding protein, or protein A, respectively, to the target recombinant protein.

Examples of suitable inducible non-fusion E. coli expression vectors include pTrc (Amrann et al, (1988) Gene 69:301-315) and pET lid (Studier et al, GENE EXPRESSION TECHNOLOGY: METHODS IN ENZYMOLOGY 185, Academic Press, San Diego, Calif. (1990) 60-89). One strategy to maximize recombinant protein expression in E. coli is to express the protein in a host bacteria with an impaired capacity to proteolytically cleave the recombinant protein. See, e.g., Gottesman, GΕNΕ EXPRESSION TECHNOLOGY: METHODS IN ENZYMOLOGY 185, Academic Press, San Diego, Calif. (1990) 119-128. Another strategy is to alter the nucleic acid sequence ofthe nucleic acid to be inserted into an expression vector so that the individual codons for each amino acid are those preferentially utilized inE. coli (see, e.g., Wada, et al., 1992. Nucl. Acids Res. 20:2111-2118). Such alteration of nucleic acid sequences ofthe invention can be carried out by standard DNA synthesis techniques.

In another embodiment, the NOVX expression vector is a yeast expression vector. Examples of vectors for expression in yeast Saccharomyces cerivisae include pYepSecl (Baldari, et al, 1987. EMBO J. 6:229-234), pMFa (Kurjan and Herskowitz, 1982. Cell 30: 933-943), pJRY88 (Schultz et al, 1987. Gene 54: 113-123), pYES2 (Invitrogen Coφoration, San Diego, Calif), and picZ (InVitrogen Coφ, San Diego, Calif).

Alternatively, NOVX can be expressed in insect cells using baculovirus expression vectors. Baculovirus vectors available for expression of proteins in cultured insect cells (e.g., SF9 cells) include the pAc series (Smith, et al, 1983. Mol. Cell Biol. 3:2156-2165) and the pVL series (Lucklow and Summers, 1989. Virology 170: 31-39).

In yet another embodiment, a nucleic acid ofthe invention is expressed in mammalian cells using a mammalian expression vector. Examples of mammalian expression vectors include pCDM8 (Seed, 1987. Nαtwre 329: 840) and pMT2PC (Kaufman, et al, 1987. EMBO J. 6: 187-195). When used in mammalian cells, the expression vector's control functions are often provided by viral regulatory elements. For example, commonly used promoters are derived from polyoma, adenovirus 2, cytomegalovirus, and simian virus 40. For other suitable expression systems for both prokaryotic and eukaryotic cells see, e.g., Chapters 16 and 17 of Sambrook, et al, MOLECULAR CLONING: A LABORATORY MANUAL. 2nd ed., Cold Spring Harbor Laboratory, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1989.

In another embodiment, the recombinant mammalian expression vector is capable of directing expression ofthe nucleic acid preferentially in a particular cell type (e.g., tissue-specific regulatory elements are used to express the nucleic acid). Tissue-specific regulatory elements are known in the art. Non-limiting examples of suitable tissue-specific promoters include the albumin promoter (liver-specific; Pinkert, et al, 1987. Genes Dev. 1:268-277), lymphoid-specific promoters (Calame and Eaton, 1988. Adv. Immunol. 43:235-275), in particular promoters of T cell receptors (Winoto and Baltimore, 1989. EMBO J. 8: 729-733) and immunoglobulins (Banerji, et al, 1983. Cell 33: 729-740; Queen and Baltimore, 1983. Cell 33: 741-748), neuron-specific promoters (e.g., the neurofilament promoter; Byrne and Ruddle, 1989. Proc. Natl. Acad. Sci. USA 86: 5473-5477), pancreas-specific promoters (Edlund, et al, 1985. Science 230: 912-916), and mammary gland-specific promoters (e.g., milk whey promoter; U.S. Pat. No. 4,873,316 and European Application Publication No. 264,166). Developmentally-regulated promoters are also encompassed, e.g., the murine hox promoters (Kessel and Grass, 1990. Science 249: 374-379) and the α-fetoprotein promoter (Campes and Tilghman, 1989. Genes Dev. 3: 537-546). The invention further provides a recombinant expression vector comprising a DNA molecule ofthe invention cloned into the expression vector in an antisense orientation. That is, the DNA molecule is operatively-linked to a regulatory sequence in a manner that allows for expression (by transcription ofthe DNA molecule) of an RNA molecule that is antisense to NOVX mRNA. Regulatory sequences operatively linked to a nucleic acid cloned in the antisense orientation can be chosen that direct the continuous expression ofthe antisense RNA molecule in a variety of cell types, for instance viral promoters and/or enhancers, or regulatory sequences can be chosen that direct constitutive, tissue specific or cell type specific expression of antisense RNA. The antisense expression vector can be in the form of a recombinant plasmid, phagemid or attenuated virus in which antisense nucleic acids are produced under the control of a high efficiency regulatory region, the activity of which can be determined by the cell type into which the vector is introduced. For a discussion ofthe regulation of gene expression using antisense genes see, e.g., Weintraub, et al, "Antisense RNA as a molecular tool for genetic analysis," Reviews-Trends in Genetics, Vol. 1(1) 1986. Another aspect ofthe invention pertains to host cells into which a recombinant expression vector ofthe invention has been introduced. The terms "host cell" and

"recombinant host cell" are used interchangeably herein. It is understood that such terms refer not only to the particular subject cell but also to the progeny or potential progeny of such a cell. Because certain modifications may occur in succeeding generations due to either mutation or environmental influences, such progeny may not, in fact, be identical to the parent cell, but are still included within the scope of the term as used herein.

A host cell can be any prokaryotic or eukaryotic cell. For example, NOVX protein can be expressed in bacterial cells such as E. coli, insect cells, yeast or mammalian cells (such as Chinese hamster ovary cells (CHO) or COS cells). Other suitable host cells are known to those skilled in the art. Vector DNA can be introduced into prokaryotic or eukaryotic cells via conventional transformation or transfection techniques. As used herein, the terms "transformation" and "transfection" are intended to refer to a variety of art-recognized techniques for introducing foreign nucleic acid (e.g., DNA) into a host cell, including calcium phosphate or calcium chloride co-precipitation, DEAE-dextran-mediated transfection, lipofection, or electroporation. Suitable methods for transforming or transfecting host cells can be found in Sambrook, et al. (MOLECULAR CLONING: A LABORATORY MANUAL. 2nd ed., Cold Spring Harbor Laboratory, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1989), and other laboratory manuals. For stable transfection of mammalian cells, it is known that, depending upon the expression vector and transfection technique used, only a small fraction of cells may integrate the foreign DNA into their genome. In order to identify and select these integrants, a gene that encodes a selectable marker (e.g., resistance to antibiotics) is generally introduced into the host cells along with the gene of interest. Various selectable markers include those that confer resistance to drugs, such as G418, hygromycin and methotrexate. Nucleic acid encoding a selectable marker can be introduced into a host cell on the same vector as that encoding NOVX or can be introduced on a separate vector. Cells stably transfected with the introduced nucleic acid can be identified by drug selection (e.g., cells that have incoφorated the selectable marker gene will survive, while the other cells die). A host cell ofthe invention, such as a prokaryotic or eukaryotic host cell in culture, can be used to produce (i.e., express) NOVX protein. Accordingly, the invention further provides methods for producing NOVX protein using the host cells ofthe invention. In one embodiment, the method comprises culturing the host cell of invention (into which a recombinant expression vector encoding NOVX protein has been introduced) in a suitable medium such that NOVX protein is produced. In another embodiment, the method further comprises isolating NOVX protein from the medium or the host cell.

Transgenic NOVX Animals

The host cells ofthe invention can also be used to produce non-human transgenic animals. For example, in one embodiment, a host cell ofthe invention is a fertilized oocyte or an embryonic stem cell into which NOVX protein-coding sequences have been introduced. Such host cells can then be used to create non-human transgenic animals in which exogenous NOVX sequences have been introduced into their genome or homologous recombinant animals in which endogenous NOVX sequences have been altered. Such animals are useful for studying the function and/or activity of NOVX protein and for identifying and/or evaluating modulators of NOVX protein activity. As used herein, a "transgenic animal" is a non-human animal, preferably a mammal, more preferably a rodent such as a rat or mouse, in which one or more ofthe cells ofthe animal includes a transgene. Other examples of transgenic animals include non-human primates, sheep, dogs, cows, goats, chickens, amphibians, etc. A transgene is exogenous DNA that is integrated into the genome of a cell from which a transgenic animal develops and that remains in the genome ofthe mature animal, thereby directing the expression of an encoded gene product in one or more cell types or tissues ofthe transgenic animal. As used herein, a "homologous recombinant animal" is a non-human animal, preferably a mammal, more preferably a mouse, in which an endogenous NOVX gene has been altered by homologous recombination between the endogenous gene and an exogenous DNA molecule introduced into a cell ofthe animal, e.g., an embryonic cell ofthe animal, prior to development ofthe animal.

A transgenic animal ofthe invention can be created by introducing NOVX-encoding nucleic acid into the male pronuclei of a fertilized oocyte (e.g. , by microinjection, retroviral infection) and allowing the oocyte to develop in a pseudopregnant female foster animal. The human NOVX cDNA sequences SEQ ID NO:2n-l, wherein n is an integer between 1 and 33, can be introduced as a transgene into the genome of a non-human animal. Alternatively, a non-human homologue ofthe human NOVX gene, such as a mouse NOVX gene, can be isolated based on hybridization to the human NOVX cDNA (described further supra) and used as a transgene. Intronic sequences and polyadenylation signals can also be included in the transgene to increase the efficiency of expression ofthe transgene. A tissue-specific regulatory sequence(s) can be operably-linked to the NOVX transgene to direct expression of NOVX protein to particular cells. Methods for generating transgenic animals via embryo manipulation and microinjection, particularly animals such as mice, have become conventional in the art and are described, for example, in U.S. Patent Nos. 4,736,866; 4,870,009; and 4,873,191; and Hogan, 1986. In: MANIPULATING THE MOUSE EMBRYO, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. Similar methods are used for production of other transgenic animals. A transgenic founder animal can be identified based upon the presence of the NOVX transgene in its genome and/or expression of NOVX mRNA in tissues or cells ofthe animals. A transgenic founder animal can then be used to breed additional animals carrying the transgene. Moreover, transgenic animals carrying a transgene-encoding NOVX protein can further be bred to other transgenic animals carrying other transgenes. To create a homologous recombinant animal, a vector is prepared which contains at least a portion of an NOVX gene into which a deletion, addition or substitution has been introduced to thereby alter, e.g., functionally disrupt, the NOVX gene. The NOVX gene can be a human gene (e.g., the cDNA of SEQ ID NO:2n-l, wherein n is an integer between 1 and 33), but more preferably, is a non-human homologue of a human NOVX gene. For example, a mouse homologue of human NOVX gene of SEQ ID NO:2n-l, wherein n is an integer between 1 and 33, can be used to construct a homologous recombination vector suitable for altering an endogenous NOVX gene in the mouse genome. In one embodiment, the vector is designed such that, upon homologous recombination, the endogenous NOVX gene is functionally disrupted (i.e., no longer encodes a functional protein; also referred to as a "knock out" vector).

Alternatively, the vector can be designed such that, upon homologous recombination, the endogenous NOVX gene is mutated or otherwise altered but still encodes functional protein (e.g., the upstream regulatory region can be altered to thereby alter the expression of the endogenous NOVX protein). In the homologous recombination vector, the altered portion ofthe NOVX gene is flanked at its 5'- and 3 '-termini by additional nucleic acid ofthe NOVX gene to allow for homologous recombination to occur between the exogenous NOVX gene carried by the vector and an endogenous NOVX gene in an embryonic stem cell. The additional flanking NOVX nucleic acid is of sufficient length for successful homologous recombination with the endogenous gene. Typically, several kilobases of flanking DNA

(both at the 5'- and 3'-termini) are included in the vector. See, e.g., Thomas, et al, 1987. Cell 51 : 503 for a description of homologous recombination vectors. The vector is ten introduced into an embryonic stem cell line (e.g., by electroporation) and cells in which the introduced NOVX gene has homologously-recombined with the endogenous NOVX gene are selected. See, e.g., Li, et al, 1992. Cell 69: 915.

The selected cells are then injected into a blastocyst of an animal (e.g., a mouse) to form aggregation chimeras. See, e.g., Bradley, 1987. In: TERATOCARCINOMAS AND EMBRYONIC STEM CELLS: A PRACTICAL APPROACH, Robertson, ed. IRL, Oxford, pp. 113-152. A chimeric embryo can then be implanted into a suitable pseudopregnant female foster animal and the embryo brought to term. Progeny harboring the homologously- recombined DNA in their germ cells can be used to breed animals in which all cells ofthe animal contain the homologously-recombined DNA by germline transmission ofthe transgene. Methods for constructing homologous recombination vectors and homologous recombinant animals are described further in Bradley, 1991. Curr. Opin. Biotechnol. 2: 823-829; PCT International Publication Nos.: WO 90/11354; WO 91/01140; WO 92/0968; and WO 93/04169.

In another embodiment, transgenic non-humans animals can be produced that contain selected systems that allow for regulated expression ofthe transgene. One example of such a system is the cre/loxP recombinase system of bacteriophage PI . For a description of the cre/loxP recombinase system, See, e.g., Lakso, et al, 1992. Proc. Natl Acad. Sci. USA 89: 6232-6236. Another example of a recombinase system is the FLP recombinase system of Saccharomyces cerevisiae. See, O'Gorman, et al, 1991. Science 251:1351-1355. If a cre/loxP recombinase system is used to regulate expression ofthe transgene, animals containing transgenes encoding both the Cre recombinase and a selected protein are required. Such animals can be provided through the construction of "double" transgenic animals, e.g., by mating two transgenic animals, one containing a transgene encoding a selected protein and the other containing a transgene encoding a recombinase.

Clones ofthe non-human transgenic animals described herein can also be produced according to the methods described in Wilmut, et al, 1997. Nature 385: 810-813. In brief, a cell (e.g., a somatic cell) from the transgenic animal can be isolated and induced to exit the growth cycle and enter Go phase. The quiescent cell can then be fused, e.g., through the use of electrical pulses, to an enucleated oocyte from an animal ofthe same species from which the quiescent cell is isolated. The reconstructed oocyte is then cultured such that it develops to morula or blastocyte and then transferred to pseudopregnant female foster animal. The offspring borne of this female foster animal will be a clone ofthe animal from which the cell (e.g., the somatic cell) is isolated.

Pharmaceutical Compositions

The NOVX nucleic acid molecules, NOVX proteins, and anti-NOVX antibodies (also referred to herein as "active compounds") ofthe invention, and derivatives, fragments, analogs and homologs thereof, can be incoφorated into pharmaceutical compositions suitable for administration. Such compositions typically comprise the nucleic acid molecule, protein, or antibody and a pharmaceutically acceptable carrier. As used herein, "pharmaceutically acceptable carrier" is intended to include any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absoφtion delaying agents, and the like, compatible with pharmaceutical administration. Suitable carriers are described in the most recent edition of Remington's Pharmaceutical Sciences, a standard reference text in the field, which is incoφorated herein by reference. Preferred examples of such carriers or diluents include, but are not limited to, water, saline, finger's solutions, dextrose solution, and 5% human serum albumin. Liposomes and non-aqueous vehicles such as fixed oils may also be used. The use of such media and agents for pharmaceutically active substances is well known in the art. Except insofar as any conventional media or agent is incompatible with the active compound, use thereof in the compositions is contemplated. Supplementary active compounds can also be incoφorated into the compositions.

A pharmaceutical composition ofthe invention is formulated to be compatible with its intended route of administration. Examples of routes of admimstration include parenteral, e.g., intravenous, intradermal, subcutaneous, oral (e.g., inhalation), transdermal (t.e., topical), transmucosal, and rectal administration. Solutions or suspensions used for parenteral, intradermal, or subcutaneous application can include the following components: a sterile diluent such as water for injection, saline solution, fixed oils, polyethylene glycols, glycerine, propylene glycol or other synthetic solvents; antibacterial agents such as benzyl alcohol or methyl parabens; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as ethylenediaminetetiaacetic acid (EDTA); buffers such as acetates, citrates or phosphates, and agents for the adjustment of tonicity such as sodium chloride or dextrose. The pH can be adjusted with acids or bases, such as hydrochloric acid or sodium hydroxide. The parenteral preparation can be enclosed in ampoules, disposable syringes or multiple dose vials made of glass or plastic. Pharmaceutical compositions suitable for injectable use include sterile aqueous solutions (where water soluble) or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersion. For intravenous administration, suitable carriers include physiological saline, bacteriostatic water, Cremophor EL (BASF, Parsippany, N.J.) or phosphate buffered saline (PBS). In all cases, the composition must be sterile and should be fluid to the extent that easy syringeability exists. It must be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms such as bacteria and fungi. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), and suitable mixtures thereof. The proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance ofthe required particle size in the case of dispersion and by the use of surfactants. Prevention ofthe action of microorganisms can be achieved by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, ascorbic acid, thimerosal, and the like. In many cases, it will be preferable to include isotonic agents, for example, sugars, polyalcohols such as manitol, sorbitol, sodium chloride in the composition. Prolonged absoφtion ofthe injectable compositions can be brought about by including in the composition an agent which delays absoφtion, for example, aluminum monostearate and gelatin. Sterile injectable solutions can be prepared by incoφorating the active compound

(e.g. , an NOVX protein or anti-NOVX antibody) in the required amount in an appropriate solvent with one or a combination of ingredients enumerated above, as required, followed by filtered sterilization. Generally, dispersions are prepared by incoφorating the active compound into a sterile vehicle that contains a basic dispersion medium and the required other ingredients from those enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, methods of preparation are vacuum drying and freeze-drying that yields a powder ofthe active ingredient plus any additional desired ingredient from a previously sterile-filtered solution thereof.

Oral compositions generally include an inert diluent or an edible carrier. They can be enclosed in gelatin capsules or compressed into tablets. For the puφose of oral therapeutic administration, the active compound can be incoφorated with excipients and used in the form of tablets, troches, or capsules. Oral compositions can also be prepared using a fluid carrier for use as a mouthwash, wherein the compound in the fluid carrier is applied orally and swished and expectorated or swallowed. Pharmaceutically compatible binding agents, and/or adjuvant materials can be included as part ofthe composition. The tablets, pills, capsules, troches and the like can contain any ofthe following ingredients, or compounds of a similar nature: a binder such as microcrystalline cellulose, gum tragacanth or gelatin; an excipient such as starch or lactose, a disintegrating agent such as alginic acid, Primogel, or corn starch; a lubricant such as magnesium stearate or Sterotes; a glidant such as colloidal silicon dioxide; a sweetening agent such as sucrose or saccharin; or a flavoring agent such as peppermint, methyl salicylate, or orange flavoring.

For administration by inhalation, the compounds are delivered in the form of an aerosol spray from pressured container or dispenser which contains a suitable propellant, e.g., a gas such as carbon dioxide, or a nebulizer. Systemic administration can also be by transmucosal or transdermal means. For transmucosal or transdermal administration, penetrants appropriate to the barrier to be permeated are used in the formulation. Such penetrants are generally known in the art, and include, for example, for transmucosal admimstration, detergents, bile salts, and fusidic acid derivatives. Transmucosal administration can be accomplished through the use of nasal sprays or suppositories. For transdermal administration, the active compounds are formulated into ointments, salves, gels, or creams as generally known in the art.

The compounds can also be prepared in the form of suppositories (e.g., with conventional suppository bases such as cocoa butter and other glycerides) or retention enemas for rectal delivery.

In one embodiment, the active compounds are prepared with carriers that will protect the compound against rapid elimination from the body, such as a controlled release formulation, including implants and microencapsulated delivery systems. Biodegradable, biocompatible polymers can be used, such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid. Methods for preparation of such formulations will be apparent to those skilled in the art. The materials can also be obtained commercially from Alza Coφoration and Nova Pharmaceuticals, Inc. Liposomal suspensions (including liposomes targeted to infected cells with monoclonal antibodies to viral antigens) can also be used as pharmaceutically acceptable carriers. These can be prepared according to methods known to those skilled in the art, for example, as described in U.S. Patent No. 4,522,811.

It is especially advantageous to formulate oral or parenteral compositions in dosage unit form for ease of administration and uniformity of dosage. Dosage unit form as used herein refers to physically discrete units suited as unitary dosages for the subject to be treated; each unit containing a predetermined quantity of active compound calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier. The specification for the dosage unit forms ofthe invention are dictated by and directly dependent on the unique characteristics ofthe active compound and the particular therapeutic effect to be achieved, and the limitations inherent in the art of compounding such an active compound for the treatment of individuals.

The nucleic acid molecules ofthe invention can be inserted into vectors and used as gene therapy vectors. Gene therapy vectors can be delivered to a subject by, for example, intravenous injection, local administration (see, e.g., U.S. Patent No. 5,328,470) or by stereotactic injection (see, e.g., Chen, et al, 1994. Proc. Natl. Acad. Sci. USA 91: 3054-3057). The pharmaceutical preparation ofthe gene therapy vector can include the gene therapy vector in an acceptable diluent, or can comprise a slow release matrix in which the gene delivery vehicle is imbedded. Alternatively, where the complete gene delivery vector can be produced intact from recombinant cells, e.g., retroviral vectors, the pharmaceutical preparation can include one or more cells that produce the gene delivery system. The pharmaceutical compositions can be included in a container, pack, or dispenser together with instructions for administration.

Screening and Detection Methods

The isolated nucleic acid molecules ofthe invention can be used to express NOVX protein (e.g., via a recombinant expression vector in a host cell in gene therapy applications), to detect NOVX mRNA (e.g., in a biological sample) or a genetic lesion in an NOVX gene, and to modulate NOVX activity, as described further, below. In addition, the NOVX proteins can be used to screen drugs or compounds that modulate the NOVX protein activity or expression as well as to treat disorders characterized by insufficient or excessive production of NOVX protein or production of NOVX protein forms that have decreased or aberrant activity compared to NOVX wild-type protein (e.g.; diabetes (regulates insulin release); obesity (binds and transport lipids); metabolic disturbances associated with obesity, the metabolic syndrome X as well as anorexia and wasting disorders associated with chronic diseases and various cancers, and infectious disease(possesses anti-microbial activity) and the various dyshpidemias. In addition, the anti-NOVX antibodies ofthe invention can be used to detect and isolate NOVX proteins and modulate NOVX activity. In yet a further aspect, the invention can be used in methods to influence appetite, absoφtion of nutrients and the disposition of metabolic substrates in both a positive and negative fashion.

The invention further pertains to novel agents identified by the screening assays described herein and uses thereof for treatments as described, supra. Screening Assays

The invention provides a method (also referred to herein as a "screening assay") for identifying modulators, i.e., candidate or test compounds or agents (e.g., peptides, peptidomimetics, small molecules or other drugs) that bind to NOVX proteins or have a stimulatory or inhibitory effect on, e.g., NOVX protein expression or NOVX protein activity. The invention also includes compounds identified in the screening assays described herein.

In one embodiment, the invention provides assays for screening candidate or test compounds which bind to or modulate the activity ofthe membrane-bound form of an NOVX protein or polypeptide or biologically-active portion thereof. The test compounds ofthe invention can be obtained using any o the numerous approaches in combinatorial library methods known in the art, including: biological libraries; spatially addressable parallel solid phase or solution phase libraries; synthetic library methods requiring deconvolution; the "one-bead one-compound" library method; and synthetic library methods using affinity chromatography selection. The biological library approach is limited to peptide libraries, while the other four approaches are applicable to peptide, non-peptide oligomer or small molecule libraries of compounds. See, e.g., Lam, 1997. Anticancer Drug Design 12: 145. A "small molecule" as used herein, is meant to refer to a composition that has a molecular weight of less than about 5 kD and most preferably less than about 4 kD. Small molecules can be, e.g., nucleic acids, peptides, polypeptides, peptidomimetics, carbohydrates, lipids or other organic or inorganic molecules. Libraries of chemical and/or biological mixtures, such as fungal, bacterial, or algal extracts, are known in the art and can be screened with any ofthe assays ofthe invention. Examples of methods for the synthesis of molecular libraries can be found in the art, for example in: DeWitt, et al, 1993. Proc. Natl. Acad. Sci. U.S.A. 90: 6909; Erb, et al, 1994. Proc. Natl. Acad. Sci. U.S.A. 91: 11422; Zuckermann, et al, 1994. J. Med. Chem. 37:2678; Cho, et al, 1993. Science 261: 1303; Carrell, et al, 1994. Angew. Chem. Int. Ed. Engl. 33:2059; Carell, et al, 1994. Angew. Chem. Int. Ed. Engl. 33:2061; and Gallop, et al, 1994. J. Med. Chem. 37: 1233.

Libraries of compounds may be presented in solution (e.g., Houghten, 1992. Biotechniques 13: 412-421), or on beads (Lam, 1991. Nature 354: 82-84), on chips (Fodor, 1993. Nature 364: 555-556), bacteria (Ladner, U.S. Patent No. 5,223,409), spores (Ladner, U.S. Patent 5,233,409), plasmids (Cull, et al, 1992. Proc. Nat Acad. Sci. USA 89: 1865-1869) or on phage (Scott and Smith, 1990. Science 249: 386-390; Devlin, 1990. Science 249: 404-406; Cwirla, etal., 1990. Proc. Natl. Acad. Sci. U.S.A. 87: 6378-6382; Felici, 1991. J. Mol. Biol. 222: 301-310; Ladner, U.S. Patent No. 5,233,409.).

In one embodiment, an assay is a cell-based assay in which a cell which expresses a membrane-bound form of NOVX protein, or a biologically-active portion thereof, on the cell surface is contacted with a test compound and the ability ofthe test compound to bind to an NOVX protein determined. The cell, for example, can of mammalian origin or a yeast cell. Determining the ability ofthe test compound to bind to the NOVX protein can be accomplished, for example, by coupling the test compound with a radioisotope or enzymatic label such that binding ofthe test compound to the NOVX protein or biologically-active portion thereof can be determined by detecting the labeled compound in a complex. For example, test compounds can be labeled with ¹²⁵1, ³⁵S, ¹⁴C, or ³H, either directly or indirectly, and the radioisotope detected by direct counting of radioemission or by scintillation counting. Alternatively, test compounds can be enzymatically-labeled with, for example, horseradish peroxidase, alkaline phosphatase, or luciferase, and the enzymatic label detected by determination of conversion of an appropriate substrate to product. In one embodiment, the assay comprises contacting a cell which expresses a membrane-bound form of NOVX protein, or a biologically-active portion thereof, on the cell surface with a known compound which binds NOVX to form an assay mixture, contacting the assay mixture with a test compound, and determining the ability of the test compound to interact with an NOVX protein, wherein determining the ability ofthe test compound to interact with an NOVX protein comprises determining the ability ofthe test compound to preferentially bind to NOVX protein or a biologically-active portion thereof as compared to the known compound. In another embodiment, an assay is a cell-based assay comprising contacting a cell expressing a membrane-bound form of NOVX protein, or a biologically-active portion thereof, on the cell surface with a test compound and determining the ability ofthe test compound to modulate (e.g., stimulate or inhibit) the activity ofthe NOVX protein or biologically-active portion thereof. Determining the ability ofthe test compound to modulate the activity of NOVX or a biologically-active portion thereof can be accomplished, for example, by determining the ability of the NOVX protein to bind to or interact with an NOVX target molecule. As used herein, a "target molecule" is a molecule with which an NOVX protein binds or interacts in nature, for example, a molecule on the surface of a cell which expresses an NOVX interacting protein, a molecule on the surface of a second cell, a molecule in the extracellular milieu, a molecule associated with the internal surface of a cell membrane or a cytoplasmic molecule. An NOVX target molecule can be a non-NOVX molecule or an NOVX protein or polypeptide ofthe invention. In one embodiment, an NOVX target molecule is a component of a signal transduction pathway that facilitates transduction of an extracellular signal (e.g. a signal generated by binding of a compound to a membrane-bound NOVX molecule) through the cell membrane and into the cell. The target, for example, can be a second intercellular protein that has catalytic activity or a protein that facilitates the association of downstream signaling molecules with NOVX.

Determining the ability ofthe NOVX protein to bind to or interact with an NOVX target molecule can be accomplished by one ofthe methods described above for determining direct binding. In one embodiment, determining the ability ofthe NOVX protein to bind to or interact with an NOVX target molecule can be accomplished by determining the activity of the target molecule. For example, the activity ofthe target molecule can be determined by detecting induction of a cellular second messenger ofthe target (i.e. intracellular Ca²⁺, diacylglycerol, IP₃, etc.), detecting catalytic/enzymatic activity ofthe target an appropriate substrate, detecting the induction of a reporter gene (comprising an NOVX-responsive regulatory element operatively linked to a nucleic acid encoding a detectable marker, e.g., luciferase), or detecting a cellular response, for example, cell survival, cellular differentiation, or cell proliferation.

In yet another embodiment, an assay ofthe invention is a cell-free assay comprising contacting an NOVX protein or biologically-active portion thereof with a test compound and determining the ability ofthe test compound to bind to the NOVX protein or biologically- active portion thereof. Binding ofthe test compound to the NOVX protein can be determined either directly or indirectly as described above. In one such embodiment, the assay comprises contacting the NOVX protein or biologically-active portion thereof with a known compound which binds NOVX to form an assay mixture, contacting the assay mixture with a test compound, and determining the ability ofthe test compound to interact with an NOVX protein, wherein determining the ability ofthe test compound to interact with an NOVX protein comprises determining the ability ofthe test compound to preferentially bind to NOVX or biologically-active portion thereof as compared to the known compound. In still another embodiment, an assay is a cell-free assay comprising contacting

NOVX protein or biologically-active portion thereof with a test compound and determining the ability ofthe test compound to modulate (e.g. stimulate or inhibit) the activity ofthe NOVX protein or biologically-active portion thereof. Determining the ability ofthe test compound to modulate the activity of NOVX can be accomplished, for example, by determining the ability ofthe NOVX protein to bind to an NOVX target molecule by one of the methods described above for determining direct binding. In an alternative embodiment, determining the ability ofthe test compound to modulate the activity of NOVX protein can be accomplished by determining the ability ofthe NOVX protein further modulate an NOVX target molecule. For example, the catalytic/enzymatic activity ofthe target molecule on an appropriate substrate can be determined as described, supra.

In yet another embodiment, the cell-free assay comprises contacting the NOVX protein or biologically-active portion thereof with a known compound which binds NOVX protein to form an assay mixture, contacting the assay mixture with a test compound, and determining the ability ofthe test compound to interact with an NOVX protein, wherein determining the ability ofthe test compound to interact with an NOVX protein comprises determining the ability ofthe NOVX protein to preferentially bind to or modulate the activity of an NOVX target molecule.

The cell-free assays ofthe invention are amenable to use of both the soluble form or the membrane-bound form of NOVX protein. In the case of cell-free assays comprising the membrane-bound form of NOVX protein, it may be desirable to utilize a solubilizing agent such that the membrane-bound form of NOVX protein is maintained in solution. Examples of such solubilizing agents include non-ionic detergents such as n-octylglucoside, n-dodecylglucoside, n-dodecylmaltoside, octanoyl-N-methylglucamide, Triton^® X-l 14, Thesit^®, decanoyl-N-methylglucamide, Triton^® X-100, Isotridecypoly(ethylene glycol ether)_n, N-dodecyl~N,N-dimethyl-3-ammonio-l -propane sulfonate, 3-(3-cholamidopropyl) dimethylamminiol-1 -propane sulfonate (CHAPS), or

3 -(3 -cholamidopropyl)dimethylamminiol-2-hydroxy-l -propane sulfonate (CHAPSO).

In more than one embodiment o the above assay methods ofthe invention, it may be desirable to immobilize either NOVX protein or its target molecule to facilitate separation of complexed from uncomplexed forms of one or both ofthe proteins, as well as to accommodate automation ofthe assay. Binding of a test compound to NOVX protein, or interaction of NOVX protein with a target molecule in the presence and absence of a candidate compound, can be accomplished in any vessel suitable for containing the reactants. Examples of such vessels include microtiter plates, test tubes, and micro-centrifuge tubes. In one embodiment, a fusion protein can be provided that adds a domain that allows one or both ofthe proteins to be bound to a matrix. For example, GST-NO VX fusion proteins or GST- target fusion proteins can be adsorbed onto glutathione sepharose beads (Sigma Chemical, St. Louis, MO) or glutathione derivatized microtiter plates, that are then combined with the test compound or the test compound and either the non-adsorbed target protein or NOVX protein, and the mixture is incubated under conditions conducive to complex formation (e.g., at physiological conditions for salt and pH). Following incubation, the beads or microtiter plate wells are washed to remove any unbound components, the matrix immobilized in the case of beads, complex determined either directly or indirectly, for example, as described, supra. Alternatively, the complexes can be dissociated from the matrix, and the level of NOVX protein binding or activity determined using standard techniques.

Other techniques for immobilizing proteins on matrices can also be used in the screening assays ofthe invention. For example, either the NOVX protein or its target molecule can be immobilized utilizing conjugation of biotin and streptavidin. Biotinylated NOVX protein or target molecules can be prepared from biotin-NHS

(N-hydroxy-succinimide) using techniques well-known within the art (e.g., biotinylation kit, Pierce Chemicals, Rockford, 111.), and immobilized in the wells of streptavidin-coated 96 well plates (Pierce Chemical). Alternatively, antibodies reactive with NOVX protein or target molecules, but which do not interfere with binding ofthe NOVX protein to its target molecule, can be derivatized to the wells ofthe plate, and unbound target or NOVX protein trapped in the wells by antibody conjugation. Methods for detecting such complexes, in addition to those described above for the GST-immobilized complexes, include immunodetection of complexes using antibodies reactive with the NOVX protein or target molecule, as well as enzyme-linked assays that rely on detecting an enzymatic activity associated with the NOVX protein or target molecule.

In another embodiment, modulators of NOVX protein expression are identified in a method wherein a cell is contacted with a candidate compound and the expression of NOVX mRNA or protein in the cell is determined. The level of expression of NOVX mRNA or protein in the presence ofthe candidate compound is compared to the level of expression of NOVX mRNA or protein in the absence ofthe candidate compound. The candidate compound can then be identified as a modulator of NOVX mRNA or protein expression based upon this comparison. For example, when expression of NOVX mRNA or protein is greater (i.e., statistically significantly greater) in the presence ofthe candidate compound than in its absence, the candidate compound is identified as a stimulator of NOVX mRNA or protein expression. Alternatively, when expression of NOVX mRNA or protein is less (statistically significantly less) in the presence ofthe candidate compound than in its absence, the candidate compound is identified as an inhibitor of NOVX mRNA or protein expression. The level of NOVX mRNA or protein expression in the cells can be determined by methods described herein for detecting NOVX mRNA or protein.

In yet another aspect ofthe invention, the NOVX proteins can be used as "bait proteins" in a two-hybrid assay or three hybrid assay (see, e.g., U.S. Patent No. 5,283,317; Zervos, et al, 1993. Cell 72:223-232; Madura, et al, 1993. J Biol. Chem. 268: 12046-12054; Barrel, et al, 1993. Biotechniques 14: 920-924; Iwabuchi, et al, 1993. Oncogene 8: 1693-1696; and Brent WO 94/10300), to identify other proteins that bind to or interact with NOVX ("NOVX-binding proteins" or "NOVX-bp") and modulate NOVX activity. Such NOVX-binding proteins are also likely to be involved in the propagation of signals by the NOVX proteins as, for example, upstream or downstream elements ofthe NOVX pathway. The two-hybrid system is based on the modular nature of most transcription factors, which consist of separable DNA-binding and activation domains. Briefly, the assay utilizes two different DNA constructs. In one construct, the gene that codes for NOVX is fused to a gene encoding the DNA binding domain of a known transcription factor (e.g., GAL-4). In the other construct, a DNA sequence, from a library of DNA sequences, that encodes an unidentified protein ("prey" or "sample") is fused to a gene that codes for the activation domain ofthe known transcription factor. If the "bait" and the "prey" proteins are able to interact, in vivo, forming an NOVX-dependent complex, the DNA-binding and activation domains ofthe transcription factor are brought into close proximity. This proximity allows transcription of a reporter gene (e.g., LacZ) that is operably linked to a transcriptional regulatory site responsive to the transcription factor. Expression ofthe reporter gene can be detected and cell colonies containing the functional transcription factor can be isolated and used to obtain the cloned gene that encodes the protein which interacts with NOVX.

The invention further pertains to novel agents identified by the aforementioned screening assays and uses thereof for treatments as described herein. Detection Assays

Portions or fragments ofthe cDNA sequences identified herein (and the corresponding complete gene sequences) can be used in numerous ways as polynucleotide reagents. By way of example, and not of limitation, these sequences can be used to: (i) map their respective genes on a chromosome; and, thus, locate gene regions associated with genetic disease; (ii) identify an individual from a minute biological sample (tissue typing); and (Hi) aid in forensic identification of a biological sample. Some of these applications are described in the subsections, below.

Chromosome Mapping

Once the sequence (or a portion ofthe sequence) of a gene has been isolated, this sequence can be used to map the location ofthe gene on a chromosome. This process is called chromosome mapping. Accordingly, portions or fragments ofthe NOVX sequences, SEQ ID NO:2n-l, wherein n is an integer between 1 and 33, or fragments or derivatives thereof, can be used to map the location ofthe NOVX genes, respectively, on a chromosome. The mapping ofthe NOVX sequences to chromosomes is an important first step in correlating these sequences with genes associated with disease.

Briefly, NOVX genes can be mapped to chromosomes by preparing PCR primers (preferably 15-25 bp in length) from the NOVX sequences. Computer analysis ofthe NOVX, sequences can be used to rapidly select primers that do not span more than one exon in the genomic DNA, thus complicating the amplification process. These primers can then be used for PCR screening of somatic cell hybrids containing individual human chromosomes. Only those hybrids containing the human gene corresponding to the NOVX sequences will yield an amplified fragment. Somatic cell hybrids are prepared by fusing somatic cells from different mammals (e.g., human and mouse cells). As hybrids of human and mouse cells grow and divide, they gradually lose human chromosomes in random order, but retain the mouse chromosomes. By using media in which mouse cells cannot grow, because they lack a particular enzyme, but in which human cells can, the one human chromosome that contains the gene encoding the needed enzyme will be retained. By using various media, panels of hybrid cell lines can be established. Each cell line in a panel contains either a single human chromosome or a small number of human chromosomes, and a full set of mouse chromosomes, allowing easy mapping of individual genes to specific human chromosomes. See, e.g., D'Eustachio, et al, 1983. Science 220: 919-924. Somatic cell hybrids containing only fragments of human chromosomes can also be produced by using human chromosomes with translocations and deletions.

PCR mapping of somatic cell hybrids is a rapid procedure for assigning a particular sequence to a particular chromosome. Three or more sequences can be assigned per day using a single thermal cycler. Using the NOVX sequences to design oligonucleotide primers, sub-localization can be achieved with panels of fragments from specific chromosomes.

Fluorescence in situ hybridization (FISH) of a DNA sequence to a metaphase chromosomal spread can further be used to provide a precise chromosomal location in one step. Chromosome spreads can be made using cells whose division has been blocked in metaphase by a chemical like colcemid that disrupts the mitotic spindle. The chromosomes can be treated briefly with trypsin, and then stained with Giemsa. A pattern of light and dark bands develops on each chromosome, so that the chromosomes can be identified individually. The FISH technique can be used with a DNA sequence as short as 500 or 600 bases. However, clones larger than 1,000 bases have a higher likelihood of binding to a unique chromosomal location with sufficient signal intensity for simple detection. Preferably 1,000 bases, and more preferably 2,000 bases, will suffice to get good results at a reasonable amount of time. For a review of this technique, see, Verma, et al, HUMAN CHROMOSOMES: A MANUAL OF BASIC TECHNIQUES (Pergamon Press, New York 1988).

Reagents for chromosome mapping can be used individually to mark a single chromosome or a single site on that chromosome, or panels of reagents can be used for marking multiple sites and/or multiple chromosomes. Reagents corresponding to noncoding regions ofthe genes actually are preferred for mapping puφoses. Coding sequences are more likely to be conserved within gene families, thus increasing the chance of cross hybridizations during chromosomal mapping. Once a sequence has been mapped to a precise chromosomal location, the physical position ofthe sequence on the chromosome can be correlated with genetic map data. Such data are found, e.g., in McKusick, MENDELIAN INHERITANCE IN MAN, available on-line through Johns Hopkins University Welch Medical Library). The relationship between genes and disease, mapped to the same chromosomal region, can then be identified through linkage analysis (co-inheritance of physically adjacent genes), described in, e.g., Egeland, et al., 1987. Nature, 325: 783-787.

Moreover, differences in the DNA sequences between individuals affected and unaffected with a disease associated with the NOVX gene, can be determined. If a mutation is observed in some or all of the affected individuals but not in any unaffected individuals, then the mutation is likely to be the causative agent ofthe particular disease. Comparison of affected and unaffected individuals generally involves first looking for structural alterations in the chromosomes, such as deletions or translocations that are visible from chromosome spreads or detectable using PCR based on that DNA sequence. Ultimately, complete sequencing of genes from several individuals can be performed to confirm the presence of a mutation and to distinguish mutations from polymoφhisms.

Tissue Typing

The NOVX sequences ofthe invention can also be used to identify individuals from minute biological samples. In this technique, an individual's genomic DNA is digested with one or more restriction enzymes, and probed on a Southern blot to yield unique bands for identification. The sequences ofthe invention are useful as additional DNA markers for RFLP ("restriction fragment length polymorphisms," described in U.S. Patent No. 5,272,057).

Furthermore, the sequences ofthe invention can be used to provide an alternative technique that determines the actual base-by-base DNA sequence of selected portions of an individual's genome. Thus, the NOVX sequences described herein can be used to prepare two PCR primers from the 5'- and 3 '-termini ofthe sequences. These primers can then be used to amplify an individual's DNA and subsequently sequence it.

Panels of corresponding DNA sequences from individuals, prepared in this manner, can provide unique individual identifications, as each individual will have a unique set of such DNA sequences due to allelic differences. The sequences ofthe invention can be used to obtain such identification sequences from individuals and from tissue. The NOVX sequences ofthe invention uniquely represent portions ofthe human genome. Allelic variation occurs to some degree in the coding regions of these sequences, and to a greater degree in the noncoding regions. It is estimated that allelic variation between individual humans occurs with a frequency of about once per each 500 bases. Much ofthe allelic variation is due to single nucleotide polymoφhisms (SNPs), which include restriction fragment length polymoφhisms (RFLPs).

Each ofthe sequences described herein can, to some degree, be used as a standard against which DNA from an individual can be compared for identification puφoses. Because greater numbers of polymoφhisms occur in the noncoding regions, fewer sequences are necessary to differentiate individuals. The noncoding sequences can comfortably provide positive individual identification with a panel of perhaps 10 to 1,000 primers that each yield a noncoding amplified sequence of 100 bases. If predicted coding sequences, such as those in SEQ ID NO:2n-l, wherein n is an integer between 1 and 33, are used, a more appropriate number of primers for positive individual identification would be 500-2,000.

Predictive Medicine The invention also pertains to the field of predictive medicine in which diagnostic assays, prognostic assays, pharmacogenomics, and monitoring clinical trials are used for prognostic (predictive) puφoses to thereby treat an individual prophylactically. Accordingly, one aspect ofthe invention relates to diagnostic assays for determining NOVX protein and or nucleic acid expression as well as NOVX activity, in the context of a biological sample (e.g., blood, serum, cells, tissue) to thereby determine whether an individual is afflicted with a disease or disorder, or is at risk of developing a disorder, associated with aberrant NOVX expression or activity. The disorders include metabolic disorders, diabetes, obesity, infectious disease, anorexia, cancer-associated cachexia, cancer, neurodegenerative disorders, Alzheimer's Disease, Parkinson's Disorder, immune disorders, and hematopoietic disorders, and the various dyshpidemias, metabolic disturbances associated with obesity, the metabolic syndrome X and wasting disorders associated with chronic diseases and various cancers. The invention also provides for prognostic (or predictive) assays for determining whether an individual is at risk of developing a disorder associated with NOVX protein, nucleic acid expression or activity. For example, mutations in an NOVX gene can be assayed in a biological sample. Such assays can be used for prognostic or predictive propose to thereby prophylactically treat an individual prior to the onset of a disorder characterized by or associated with NOVX protein, nucleic acid expression, or biological activity. Another aspect ofthe invention provides methods for determining NOVX protein, nucleic acid expression or activity in an individual to thereby select appropriate therapeutic or prophylactic agents for that individual (referred to herein as "pharmacogenomics"). Pharmacogenomics allows for the selection of agents (e.g., drugs) for therapeutic or prophylactic treatment of an individual based on the genotype of the individual (e.g. , the genotype ofthe individual examined to determine the ability ofthe individual to respond to a particular agent.)

Yet another aspect ofthe invention pertains to monitoring the influence of agents (e.g., drugs, compounds) on the expression or activity of NOVX in clinical trials. These and other agents are described in further detail in the following sections.

Diagnostic Assays

An exemplary method for detecting the presence or absence of NOVX in a biological sample involves obtaining a biological sample from a test subject and contacting the biological sample with a compound or an agent capable of detecting NOVX protein or nucleic acid (e.g. , mRNA, genomic DNA) that encodes NOVX protein such that the presence of NOVX is detected in the biological sample. An agent for detecting NOVX mRNA or genomic DNA is a labeled nucleic acid probe capable of hybridizing to NOVX mRNA or genomic DNA. The nucleic acid probe can be, for example, a full-length NOVX nucleic acid, such as the nucleic acid of SEQ ID NO:2n-l, wherein n is an integer between 1 and 33, or a portion thereof, such as an oligonucleotide of at least 15, 30, 50, 100, 250 or 500 nucleotides in length and sufficient to specifically hybridize under stringent conditions to NOVX mRNA or genomic DNA. Other suitable probes for use in the diagnostic assays of the invention are described herein.

An agent for detecting NOVX protein is an antibody capable of binding to NOVX protein, preferably an antibody with a detectable label. Antibodies can be polyclonal, or more preferably, monoclonal. An intact antibody, or a fragment thereof (e.g., Fab or F(ab')₂) can be used. The term "labeled",, with regard to the probe or antibody, is intended to encompass direct labeling ofthe probe or antibody by coupling (i.e., physically linking) a detectable substance to the probe or antibody, as well as indirect labeling ofthe probe or antibody by reactivity with another reagent that is directly labeled. Examples of indirect labeling include detection of a primary antibody using a fluorescently-labeled secondary antibody and end-labeling of a DNA probe with biotin such that it can be detected with fluorescently-labeled streptavidin. The term "biological sample" is intended to include tissues, cells and biological fluids isolated from a subject, as well as tissues, cells and fluids present within a subject. That is, the detection method ofthe invention can be used to detect NOVX mRNA, protein, or genomic DNA in a biological sample in vitro as well as in vivo. For example, in vitro techniques for detection of NOVX mRNA include Northern hybridizations and in situ hybridizations. In vitro techniques for detection of NOVX protein include enzyme linked immunosorbent assays (ELISAs), Western blots, immunoprecipitations, and immunofluorescence. In vitro techniques for detection of NOVX genomic DNA include Southern hybridizations. Furthermore, in vivo techniques for detection of NOVX protein include introducing into a subject a labeled anti-NOVX antibody. For example, the antibody can be labeled with a radioactive marker whose presence and location in a subject can be detected by standard imaging techniques.

In one embodiment, the biological sample contains protein molecules from the test subject. Alternatively, the biological sample can contain mRNA molecules from the test subject or genomic DNA molecules from the test subject. A preferred biological sample is a peripheral blood leukocyte sample isolated by conventional means from a subject.

In another embodiment, the methods further involve obtaining a control biological sample from a control subject, contacting the control sample with a compound or agent capable of detecting NOVX protein, mRNA, or genomic DNA, such that the presence of NOVX protein, mRNA or genomic DNA is detected in the biological sample, and comparing the presence of NOVX protein, mRNA or genomic DNA in the control sample with the presence of NOVX protein, mRNA or genomic DNA in the test sample.

The invention also encompasses kits for detecting the presence of NOVX in a biological sample. For example, the kit can comprise: a labeled compound or agent capable of detecting NOVX protein or mRNA in a biological sample; means for determining the amount of NOVX in the sample; and means for comparing the amount of NOVX in the sample with a standard. The compound or agent can be packaged in a suitable container. The kit can further comprise instructions for using the kit to detect NOVX protein or nucleic acid.

Prognostic Assays The diagnostic methods described herein can furthermore be utilized to identify subjects having or at risk of developing a disease or disorder associated with aberrant NOVX expression or activity. For example, the assays described herein, such as the preceding diagnostic assays or the following assays, can be utilized to identify a subject having or at risk of developing a disorder associated with NOVX protein, nucleic acid expression or activity. Alternatively, the prognostic assays can be utilized to identify a subject having or at risk for developing a disease or disorder. Thus, the invention provides a method for identifying a disease or disorder associated with aberrant NOVX expression or activity in which a test sample is obtained from a subject and NOVX protein or nucleic acid (e.g., mRNA, genomic DNA) is detected, wherein the presence of NOVX protein or nucleic acid is diagnostic for a subject having or at risk of developing a disease or disorder associated with aberrant NOVX expression or activity. As used herein, a "test sample" refers to a biological sample obtained from a subject of interest. For example, a test sample can be a biological fluid (e.g., serum), cell sample, or tissue.

Furthermore, the prognostic assays described herein can be used to determine whether a subject can be administered an agent (e.g., an agonist, antagonist, peptidomimetic, protein, peptide, nucleic acid, small molecule, or other drug candidate) to treat a disease or disorder associated with aberrant NOVX expression or activity. For example, such methods can be used to determine whether a subject can be effectively treated with an agent for a disorder. Thus, the invention provides methods for determining whether a subject can be effectively treated with an agent for a disorder associated with aberrant NOVX expression or activity in which a test sample is obtained and NOVX protein or nucleic acid is detected (e.g., wherein the presence of NOVX protein or nucleic acid is diagnostic for a subject that can be administered the agent to treat a disorder associated with aberrant NOVX expression or activity).

The methods ofthe invention can also be used to detect genetic lesions in an NOVX gene, thereby determining if a subject with the lesioned gene is at risk for a disorder characterized by aberrant cell proliferation and/or differentiation. In various embodiments, the methods include detecting, in a sample of cells from the subject, the presence or absence of a genetic lesion characterized by at least one of an alteration affecting the integrity of a gene encoding an NOVX-protein, or the misexpression ofthe NOVX gene. For example, such genetic lesions can be detected by ascertaining the existence of at least one of: (i) a deletion of one or more nucleotides from an NOVX gene; (ii) an addition of one or more nucleotides to an NOVX gene; (Hi) a substitution of one or more nucleotides of an NOVX gene, (iv) a chromosomal rearrangement of an NOVX gene; (v) an alteration in the level of a messenger RNA transcript of an NOVX gene, (vi) aberrant modification of an NOVX gene, such as ofthe methylation pattern ofthe genomic DNA, (vii) the presence of a non-wild-type splicing pattern of a messenger RNA transcript of an NOVX gene, (viii) a non- wild-type level of an NOVX protein, (tx) allelic loss of an NOVX gene, and (x) inappropriate post-translational modification of an NOVX protein. As described herein, there are a large number of assay techniques known in the art which can be used for detecting lesions in an NOVX gene. A preferred biological sample is a peripheral blood leukocyte sample isolated by conventional means from a subject. However, any biological sample containing nucleated cells may be used, including, for example, buccal mucosal cells.

In certain embodiments, detection ofthe lesion involves the use of a probe/primer in a polymerase chain reaction (PCR) (.see, e.g., U.S. Patent Nos. 4,683,195 and 4,683,202), such as anchor PCR or RACE PCR, or, alternatively, in a ligation chain reaction (LCR) (see, e.g., Landegran, et al, 1988. Science 241 : 1077-1080; and Nakazawa, et al, 1994. Proc. Natl.

Acad. Sci. USA 91 : 360-364), the latter of which can be particularly useful for detecting point mutations in the NONX-gene (see, Abravaya, et al, 1995. Nucl. Acids Res. 23: 675-682). This method can include the steps of collecting a sample of cells from a patient, isolating nucleic acid (e.g., genomic, mRΝA or both) from the cells ofthe sample, contacting the nucleic acid sample with one or more primers that specifically hybridize to an ΝONX gene under conditions such that hybridization and amplification ofthe ΝOVX gene (if present) occurs, and detecting the presence or absence of an amplification product, or detecting the size ofthe amplification product and comparing the length to a control sample. It is anticipated that PCR and or LCR may be desirable to use as a preliminary amplification step in conjunction with any of the techniques used for detecting mutations described herein.

Alternative amplification methods include: self sustained sequence replication (see, Guatelli, et al, 1990. Proc. Natl. Acad. Sci. USA 87: 1874-1878), transcriptional amplification system (see, Kwoh, et al, 1989. Proc. Natl. Acad. Sci. USA 86: 1173-1177); Qβ Replicase (see, Lizardi, et al, 1988. BioTechnology 6: 1197), or any other nucleic acid amplification method, followed by the detection ofthe amplified molecules using techniques well known to those of skill in the art. These detection schemes are especially useful for the detection of nucleic acid molecules if such molecules are present in very low numbers.

In an alternative embodiment, mutations in an ΝOVX gene from a sample cell can be identified by alterations in restriction enzyme cleavage patterns. For example, sample and control DΝA is isolated, amplified (optionally), digested with one or more restriction endonucleases, and fragment length sizes are determined by gel electrophoresis and compared. Differences in fragment length sizes between sample and control DΝA indicates mutations in the sample DΝA. Moreover, the use of sequence specific ribozymes (see, e.g., U.S. Patent No. 5,493,531) can be used to score for the presence of specific mutations by development or loss of a ribozyme cleavage site.

In other embodiments, genetic mutations in NOVX can be identified by hybridizing a sample and control nucleic acids, e.g., DNA or RNA, to high-density arrays containing hundreds or thousands of oligonucleotides probes. See, e.g., Cronin, et al, 1996. Human Mutation 7:244-255; Kozal, et al, 1996. Nat. Med. 2: 753-759. For example, genetic mutations in NOVX can be identified in two dimensional arrays containing light-generated DNA probes as described in Cronin, et al, supra. Briefly, a first hybridization array of probes can be used to scan through long stretches of DNA in a sample and control to identify base changes between the sequences by making linear arrays of sequential overlapping probes. This step allows the identification of point mutations. This is followed by a second hybridization array that allows the characterization of specific mutations by using smaller, specialized probe arrays complementary to all variants or mutations detected. Each mutation array is composed of parallel probe sets, one complementary to the wild-type gene and the other complementary to the mutant gene.

In yet another embodiment, any of a variety of sequencing reactions known in the art can be used to directly sequence the NOVX gene and detect mutations by comparing the sequence ofthe sample NOVX with the corresponding wild-type (control) sequence. Examples of sequencing reactions include those based on techniques developed by Maxim and Gilbert, 1977. Proc. Natl. Acad. Sci. USA 74: 560 or Sanger, 1977. Proc. Natl. Acad. Sci. USA 74: 5463. It is also contemplated that any of a variety of automated sequencing procedures can be utilized when performing the diagnostic assays (see, e.g., Naeve, et al, 1995. Biotechniques 19: 448), including sequencing by mass spectromeτry (see, e.g., PCT International Publication No.' WO 94/16101; Cohen, et al, 1996. Adv. Chromatography 36: 127-162; and Griffin, et al, 1993. Appl. Biochem. Biotechnol. 38: 147-159).

Other methods for detecting mutations in the NOVX gene include methods in which protection from cleavage agents is used to detect mismatched bases in RNA/RNA or RNA DNA heteroduplexes. See, e.g., Myers, et al, 1985. Science 230: 1242. In general, the art technique of "mismatch cleavage" starts by providing heteroduplexes of formed by hybridizing (labeled) RNA or DNA containing the wild-type NOVX_. sequence with potentially mutant RNA or DNA obtained from a tissue sample. The double-stranded duplexes are treated with an agent that cleaves single-stranded regions ofthe duplex such as which will exist due to basepair mismatches between the control and sample strands. For instance, RNA/DNA duplexes can be treated with RNase and DNA/DNA hybrids treated with Si nuclease to enzymatically digesting the mismatched regions. In other embodiments, either DNA/DNA or RNA/DNA duplexes can be treated with hydroxylamine or osmium tetroxide and with piperidine in order to digest mismatched regions. After digestion ofthe mismatched regions, the resulting material is then separated by size on denaturing polyacrylamide gels to determine the site of mutation. See, e.g., Cotton, et al, 1988. Proc. Natl. Acad. Sci. USA 85: 4397; Saleeba, et al, 1992. Methods Enzymol. 217:286-295. In an embodiment, the control DNA or RNA can be labeled for detection.

In still another embodiment, the mismatch cleavage reaction employs one or more proteins that recognize mismatched base pairs in double-stranded DNA (so called "DNA mismatch repair" enzymes) in defined systems for detecting and mapping point mutations in NOVX cDNAs obtained from samples of cells. For example, the mufY enzyme of E. coli cleaves A at G/A mismatches and the thymidine DNA glycosylase from HeLa cells cleaves T at G/T mismatches. See, e.g., Hsu, et al, 1994. Carcinogenesis 15: 1657-1662. According to an exemplary embodiment, a probe based on an NOVX sequence, e.g., a wild-type NOVX sequence, is hybridized to a cDNA or other DNA product from a test cell(s). The duplex is treated with a DNA mismatch repair enzyme, and the cleavage products, if any, can be detected from electrophoresis protocols or the like. See, e.g., U.S. Patent No. 5,459,039.

In other embodiments, alterations in electrophoretic mobility will be used to identify mutations in NOVX genes. For example, single strand conformation polymoφhism (SSCP) may be used to detect differences in electrophoretic mobility between mutant and wild type nucleic acids. See, e.g., Orita, et al, 1989. Proc. Natl. Acad. Sci. USA: 86:2766; Cotton, 1993. Mutat. Res. 285: 125-144; Hayashi, 1992. Genet. Anal. Tech. Appl. 9: 73-79. Single-stranded DNA fragments of sample and control NOVX nucleic acids will be denatured and allowed to renature. The secondary structure of single-stranded nucleic acids varies according to sequence, the resulting alteration in electrophoretic mobility enables the detection of even a single base change. The DNA fragments may be labeled or detected with labeled probes. The sensitivity of the assay may be enhanced by using RNA (rather than DNA), in which the secondary structure is more sensitive to a change in sequence. In one embodiment, the subject method utilizes heteroduplex analysis to separate double stranded heteroduplex molecules on the basis of changes in electrophoretic mobility. See, e.g., Keen, et al, 1991. Trends Genet. 7: 5.

In yet another embodiment, the movement of mutant or wild-type fragments in polyacrylamide gels containing a gradient of denaturant is assayed using denaturing gradient gel electrophoresis (DGGΕ). See, e.g., Myers, et al, 1985. Nature 313: 495. When DGGΕ is used as the method of analysis, DNA will be modified to insure that it does not completely denature, for example by adding a GC clamp of approximately 40 bp of high-melting GC-rich DNA by PCR. In a further embodiment, a temperature gradient is used in place of a denaturing gradient to identify differences in the mobility of control and sample DNA. See, e.g., Rosenbaum and Reissner, 1987. Biophys. Chem. 265: 12753.

Examples of other techniques for detecting point mutations include, but are not limited to, selective oligonucleotide hybridization, selective amplification, or selective primer extension. For example, oligonucleotide primers may be prepared in which the known mutation is placed centrally and then hybridized to target DNA under conditions that permit hybridization only if a perfect match is found. See, e.g., Saiki, et α/., 1986. Nature 324: 163; Saiki, et al, 1989. Proc. Natl Acad. Sci. USA 86: 6230. Such allele specific oligonucleotides are hybridized to PCR amplified target DNA or a number of different mutations when the oligonucleotides are attached to the hybridizing membrane and hybridized with labeled target DNA. Alternatively, allele specific amplification technology that depends on selective PCR amplification may be used in conjunction with the instant invention. Oligonucleotides used as primers for specific amplification may carry the mutation of interest in the center ofthe molecule (so that amplification depends on differential hybridization; see, e.g., Gibbs, et al, 1989. Nucl. Acids Res. 17:2437-2448) or at the extreme 3'-terminus of one primer where, under appropriate conditions, mismatch can prevent, or reduce polymerase extension (see, e.g., Prossner, 1993. Tibtech. 11:238). In addition it may be desirable to introduce a novel restriction site in the region ofthe mutation to create cleavage-based detection. See, e.g., Gasparini, et al, 1992. Mol. Cell Probes 6: 1. It is anticipated that in certain embodiments amplification may also be performed using Taq ligase for amplification. See, e.g., Barany, 1991. Proc. Natl. Acad. Sci. USA 88: 189. In such cases, ligation will occur only if there is a perfect match at the 3'-terminus ofthe 5' sequence, making it possible to detect the presence of a known mutation at a specific site by looking for the presence or absence of amplification.

The methods described herein may be performed, for example, by utilizing pre-packaged diagnostic kits comprising at least one probe nucleic acid or antibody reagent described herein, which may be conveniently used, e.g., in clinical settings to diagnose patients exhibiting symptoms or family history of a disease or illness involving an NOVX gene.

Furthermore, any cell type or tissue, preferably peripheral blood leukocytes, in which NOVX is expressed may be utilized in the prognostic assays described herein. However, any biological sample containing nucleated cells may be used, including, for example, buccal mucosal cells.

Pharmacogenomics

Agents, or modulators that have a stimulatory or inhibitory effect on NOVX activity (e.g. , NOVX gene expression), as identified by a screening assay described herein can be administered to individuals to treat (prophylactically or therapeutically) disorders (The disorders include metabolic disorders, diabetes, obesity, infectious disease, anorexia, cancer- associated cachexia, cancer, neurodegenerative disorders, Alzheimer's Disease, Parkinson's Disorder, immune disorders, and hematopoietic disorders, and the various dyshpidemias, metabolic disturbances associated with obesity, the metabolic syndrome X and wasting disorders associated with chronic diseases and various cancers.) In conjunction with such treatment, the pharmacogenomics (i.e., the study ofthe relationship between an individual's genotype and that individual's response to a foreign compound or drug) ofthe individual may be considered. Differences in metabolism of therapeutics can lead to severe toxicity or therapeutic failure by altering the relation between dose and blood concentration ofthe pharmacologically active drug. Thus, the pharmacogenomics ofthe individual permits the selection of effective agents (e.g., drugs) for prophylactic or therapeutic treatments based on a consideration ofthe individual's genotype. Such pharmacogenomics can further be used to determine appropriate dosages and therapeutic regimens. Accordingly, the activity of NOVX protein, expression of NOVX nucleic acid, or mutation content of NOVX genes in an individual can be determined to thereby select appropriate agent(s) for therapeutic or prophylactic treatment ofthe individual.

Pharmacogenomics deals with clinically significant hereditary variations in the response to drugs due to altered drug disposition and abnormal action in affected persons. See e.g., Eichelbaum, 1996. Clin. Exp. Pharmacol Physiol, 23: 983-985; Linder, 1997. Clin. Chem., 43:254-266. In general, two types of pharmacogenetic conditions can be differentiated. Genetic conditions transmitted as a single factor altering the way drugs act on the body (altered drug action) or genetic conditions transmitted as single factors altering the way the body acts on drugs (altered drug metabolism). These pharmacogenetic conditions can occur either as rare defects or as polymoφhisms. For example, glucose-6-phosphate dehydrogenase (G6PD) deficiency is a common inherited enzymopathy in which the main clinical complication is hemolysis after ingestion of oxidant drugs (anti-malarials, sulfonamides, analgesics, nitrofurans) and consumption of fava beans. As an illustrative embodiment, the activity of drag metabolizing enzymes is a major determinant of both the intensity and duration of drug action. The discovery of genetic polymoφhisms of drag metabolizing enzymes (e.g., N-acetyltransferase 2 (NAT 2) and cytochrome P450 enzymes CYP2D6 and CYP2C19) has provided an explanation as to why some patients do not obtain the expected drug effects or show exaggerated drag response and serious toxicity after taking the standard and safe dose of a drug. These polymoφhisms are expressed in two phenotypes in the population, the extensive metabolizer (EM) and poor metabolizer (PM). The prevalence of PM is different among different populations. For example, the gene coding for CYP2D6 is highly polymoφhic and several mutations have been identified in PM, which all lead to the absence of functional CYP2D6. Poor metabolizers of CYP2D6 and CYP2C19 quite frequently experience exaggerated drag response and side effects when they receive standard doses. If a metabolite is the active therapeutic moiety, PM show no therapeutic response, as demonstrated for the analgesic effect of codeine mediated by its CYP2D6-formed metabolite moφhine. At the other extreme are the so called ultra-rapid metabolizers who do not respond to standard doses. Recently, the molecular basis of ultra-rapid metabolism has been identified to be due to CYP2D6 gene amplification.

Thus, the activity of NOVX protein, expression of NOVX nucleic acid, or mutation content of NOVX genes in an individual can be determined to thereby select appropriate agent(s) for therapeutic or prophylactic treatment of the individual. In addition, pharmacogenetic studies can be used to apply genotyping of polymoφhic alleles encoding drug-metabolizing enzymes to the identification of an individual's drag responsiveness phenotype. This knowledge, when applied to dosing or drug selection, can avoid adverse reactions or therapeutic failure and thus enhance therapeutic or prophylactic efficiency when treating a subject with an NOVX modulator, such as a modulator identified by one of the exemplary screening assays described herein.

Monitoring of Effects During Clinical Trials

Monitoring the influence of agents (e.g., drags, compounds) on the expression or activity of NOVX (e.g., the ability to modulate aberrant cell proliferation and/or differentiation) can be applied not only in basic drag screening, but also in clinical trials. For example, the effectiveness of an agent determined by a screening assay as described herein to increase NOVX gene expression, protein levels, or upregulate NOVX activity, can be monitored in clinical trails of subjects exhibiting decreased NOVX gene expression, protein levels, or downregulated NOVX activity. Alternatively, the effectiveness of an agent determined by a screening assay to decrease NOVX gene expression, protein levels, or downregulate NOVX activity, can be monitored in clinical trails of subjects exhibiting increased NOVX gene expression, protein levels, or upregulated NOVX activity. In such clinical trials, the expression or activity of NOVX and, preferably, other genes that have been implicated in, for example, a cellular proliferation or immune disorder can be used as a "read out" or markers ofthe immune responsiveness of a particular cell.

By way of example, and not of limitation, genes, including NOVX, that are modulated in cells by treatment with an agent (e.g., compound, drug or small molecule) that modulates NOVX activity (e.g., identified in a screening assay as described herein) can be identified. Thus, to study the effect of agents on cellular proliferation disorders, for example, in a clinical trial, cells can be isolated and RNA prepared and analyzed for the levels of expression of NOVX and other genes implicated in the disorder. The levels of gene expression (i.e., a gene expression pattern) can be quantified by Northern blot analysis or RT-PCR, as described herein, or alternatively by measuring the amount of protein produced, by one ofthe methods as described herein, or by measuring the levels of activity of NOVX or other genes. In this manner, the gene expression pattern can serve as a marker, indicative of the physiological response ofthe cells to the agent. Accordingly, this response state may be determined before, and at various points during, treatment ofthe individual with the agent. In one embodiment, the invention provides a method for monitoring the effectiveness of treatment of a subject with an agent (e.g., an agonist, antagonist, protein, peptide, peptidomimetic, nucleic acid, small molecule, or other drag candidate identified by the screening assays described herein) comprising the steps of (i) obtaining a pre-administration sample from a subject prior to administration ofthe agent; (ii) detecting the level of expression of an NOVX protein, mRNA, or genomic DNA in the preadministration sample; (Hi) obtaining one or more post-administration samples from the subject; (z^'v) detecting the level of expression or activity ofthe NOVX protein, mRNA, or genomic DNA in the post-administration samples; (v) comparing the level of expression or activity ofthe NOVX protein, mRNA, or genomic DNA in the pre-administration sample with the NOVX protein, mRNA, or genomic DNA in the post administration sample or samples; and (vz) altering the administration ofthe agent to the subject accordingly. For example, increased administration ofthe agent may be desirable to increase the expression or activity of NOVX to higher levels than detected, i.e., to increase the effectiveness ofthe agent. Alternatively, decreased administration ofthe agent may be desirable to decrease expression or activity of NOVX to lower levels than detected, i.e., to decrease the effectiveness ofthe agent.

Methods of Treatment

The invention provides for both prophylactic and therapeutic methods of treating a subject at risk of (or susceptible to) a disorder or having a disorder associated with aberrant NOVX expression or activity. The disorders include cardiomyopathy, atherosclerosis, hypertension, congenital heart defects, aortic stenosis, atrial septal defect (ASD), atrioventricular (A-V) canal defect, ductus arteriosus, pulmonary stenosis, subaortic stenosis, ventricular septal defect (VSD), valve diseases, tuberous sclerosis, scleroderma, obesity, transplantation, adrenoleukodystrophy, congenital adrenal hypeφlasia, prostate cancer, neoplasm; adenocarcinoma, lymphoma, uterus cancer, fertility, hemophilia, hypercoagulation, idiopathic thrombocytopenic puφura, immunodeficiencies, graft versus host disease, AIDS, bronchial asthma, Crohn's disease; multiple sclerosis, treatment of Albright Hereditary Ostoeodystrophy, and other diseases, disorders and conditions ofthe like. These methods of treatment will be discussed more fully, below.

Disease and Disorders

Diseases and disorders that are characterized by increased (relative to a subject not suffering from the disease or disorder) levels or biological activity may be treated with Therapeutics that antagonize (i.e., reduce or inhibit) activity. Therapeutics that antagonize activity may be administered in a therapeutic or prophylactic manner. Therapeutics that may be utilized include, but are not limited to: (i) an aforementioned peptide, or analogs, derivatives, fragments or homologs thereof; (ii) antibodies to an aforementioned peptide; (Hi) nucleic acids encoding an aforementioned peptide; (iv) administration of antisense nucleic acid and nucleic acids that are "dysfunctional" (i.e., due to a heterologous insertion within the coding sequences of coding sequences to an aforementioned peptide) that are utilized to

"knockout" endogenous function of an aforementioned peptide by homologous recombination (see, e.g., Capecchi, 1989. Science 244: 1288-1292); or (v) modulators ( i.e., inhibitors, agonists and antagonists, including additional peptide mimetic ofthe invention or antibodies specific to a peptide ofthe invention) that alter the interaction between an aforementioned peptide and its binding partner.

Diseases and disorders that are characterized by decreased (relative to a subject not suffering from the disease or disorder) levels or biological activity may be treated with Therapeutics that increase (i.e., are agonists to) activity. Therapeutics that upregulate activity may be administered in a therapeutic or prophylactic manner. Therapeutics that may be utilized include, but are not limited to, an aforementioned peptide, or analogs, derivatives, fragments or homologs thereof; or an agonist that increases bioavailability.

Increased or decreased levels can be readily detected by quantifying peptide and/or RNA, by obtaining a patient tissue sample (e.g., from biopsy tissue) and assaying it in vitro for RNA or peptide levels, structure and/or activity ofthe expressed peptides (or mRNAs of an aforementioned peptide). Methods that are well-known within the art include, but are not limited to, immunoassays (e.g., by Western blot analysis, immunoprecipitation followed by sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis, immunocytochemistry, etc.) and/or hybridization assays to detect expression of mRNAs (e.g., Northern assays, dot blots, in situ hybridization, and the like).

Prophylactic Methods

In one aspect, the invention provides a method for preventing, in a subject, a disease or condition associated with an aberrant NOVX expression or activity, by administering to the subject an agent that modulates NOVX expression or at least one NOVX activity.

Subjects at risk for a disease that is caused or contributed to by aberrant NOVX expression or activity can be identified by, for example, any or a combination of diagnostic or prognostic assays as described herein. Administration of a prophylactic agent can occur prior to the manifestation of symptoms characteristic ofthe NOVX abenancy, such that a disease or disorder is prevented or, alternatively, delayed in its progression. Depending upon the type of NOVX aberrancy, for example, an NOVX agonist or NOVX antagonist agent can be used for treating the subject. The appropriate agent can be determined based on screening assays described herein. The prophylactic methods ofthe invention are further discussed in the following subsections. Therapeutic Methods

Another aspect ofthe invention pertains to methods of modulating NOVX expression or activity for therapeutic puφoses. The modulatory method ofthe invention involves contacting a cell with an agent that modulates one or more ofthe activities of NOVX protein activity associated with the cell. An agent that modulates NOVX protein activity can be an agent as described herein, such as a nucleic acid or a protein, a naturally-occurring cognate ligand of an NOVX protein, a peptide, an NOVX peptidomimetic, or other small molecule. In one embodiment, the agent stimulates one or more NOVX protein activity. Examples of such stimulatory agents include active NOVX protein and a nucleic acid molecule encoding NOVX that has been introduced into the cell. In another embodiment, the agent inhibits one or more NOVX protein activity. Examples of such inhibitory agents include antisense NOVX nucleic acid molecules and anti-NOVX antibodies. These modulatory methods can be performed in vitro (e.g., by culturing the cell with the agent) or, alternatively, in vivo (e.g., by administering the agent to a subject). As such, the invention provides methods of treating an individual afflicted with a disease or disorder characterized by aberrant expression or activity of an NOVX protein or nucleic acid molecule. In one embodiment, the method involves administering an agent (e.g., an agent identified by a screening assay described herein), or combination of agents that modulates (e.g., up-regulates or down-regulates) NOVX expression or activity. In another embodiment, the method involves administering an NOVX protein or nucleic acid molecule as therapy to compensate for reduced or abenant NOVX expression or activity.

Stimulation of NOVX activity is desirable in situations in which NOVX is abnormally downregulated and/or in which increased NOVX activity is likely to have a beneficial effect. One example of such a situation is where a subject has a disorder characterized by aberrant cell proliferation and/or differentiation (e.g., cancer or immune associated disorders). Another example of such a situation is where the subject has a gestational disease (e.g., preclampsia).

Determination ofthe Biological Effect ofthe Therapeutic In various embodiments ofthe invention, suitable in vitro or in vivo assays are performed to determine the effect of a specific Therapeutic and whether its administration is indicated for treatment ofthe affected tissue.

In various specific embodiments, in vitro assays may be performed with representative cells ofthe type(s) involved in the patient's disorder, to determine if a given Therapeutic exerts the desired effect upon the cell type(s). Compounds for use in therapy may be tested in suitable animal model systems including, but not limited to rats, mice, chicken, cows, monkeys, rabbits, and the like, prior to testing in human subjects. Similarly, for in vivo testing, any ofthe animal model system known in the art may be used prior to administration to human subjects. Prophylactic and Therapeutic Uses of the Compositions of the Invention

The NOVX nucleic acids and proteins ofthe invention are useful in potential prophylactic and therapeutic applications implicated in a variety of disorders including, but not limited to: metabolic disorders, diabetes, obesity, infectious disease, anorexia, cancer- associated cancer, neurodegenerative disorders, Alzheimer's Disease, Parkinson's Disorder, immune disorders, hematopoietic disorders, and the various dyshpidemias, metabolic disturbances associated with obesity, the metabolic syndrome X and wasting disorders associated with chronic diseases and various cancers. As an example, a cDNA encoding the NOVX protein of the invention may be useful in gene therapy, and the protein may be useful when administered to a subject in need thereof. By way of non-limiting example, the compositions ofthe invention will have efficacy for treatment of patients suffering from: metabolic disorders, diabetes, obesity, infectious disease, anorexia, cancer-associated cachexia, cancer, neurodegenerative disorders, Alzheimer's Disease, Parkinson's Disorder, immune disorders, hematopoietic disorders, and the various dyshpidemias.

Both the novel nucleic acid encoding the NOVX protein, and the NOVX protein of the invention, or fragments thereof, may also be useful in diagnostic applications, wherein the presence or amount ofthe nucleic acid or the protein are to be assessed. A further use could be as an anti-bacterial molecule (i.e., some peptides have been found to possess anti-bacterial properties). These materials are further useful in the generation of antibodies, which immunospecifically-bind to the novel substances ofthe invention for use in therapeutic or diagnostic methods.

The invention will be further described in the following examples, which do not limit the scope of the invention described in the claims.

EXAMPLES Example A. NOVX Clone Information

Example Al.

The NOVl clone was analyzed, and the nucleotide and predicted polypeptide sequences are shown in Table 1A.

Table 1A. NOVl Sequence Analysis

SEQ IDNO:! 1046 bp

NOVla, CCATGCATGTAAAGATAAAAGCCCCAAACACTATCAGCTGTTCATTCAGCTCGTGGAA CG57883-01 ATTCTAATTCCGTGTTCATTTTTTTTTCTACAGACATTTGCCATGGCTGAGCACTTCA DNA Sequence AACAGATCATTAGATGTCCTGTCTGTCTAAAAGATCTTGAAGAAGCCGTGCAACTGAA ATGTGGATATGCCTGCTGCCTCCAGTGCCTCAATTCACTCCAGAGGGAGCCCAATGGG GAAGGTTTACTGTGCCGTTTCTGCTCTGTGGTCTCTCAGAAGGATGACATCAAGCCCA AGTACAAGCTGAGGGCGCTGGTTTCCATCATCAAGGAACTAGAGCCCAAGCTGAAATC TGTTCTAACAATGAACCCAAGGATGAGGAAGTTTCAAGTGGATATGACGTTCGATGTG GACACAGCCAACAACTATCTCATCATTTCTGAAGACCTGAGGAGTTTCCGAAGTGGGG

Sequence comparison ofthe above protein sequences yields the following sequence relationships shown in Table IB.

Table IB. Comparison of NOVla against NOVlb.

I NOVla Residues/

Protein Sequence Identities/ Similarities for the Matched Region Match Residues

NOVlb 1..287 279/287 (97%) 1..287 282/287 (98%) Further analysis of the NOVla protein yielded the following properties shown in Table IC.

Table IC. Protein Sequence Properties NOVla

PSort 0.6500 probability located in cytoplasm; 0.1000 probability located in mitochondrial analysis: matrix space; 0.1000 probability located in lysosome (lumen); 0.0000 probability located in endoplasmic reticulum (membrane)

SignalP No Known Signal Sequence Predicted analysis:

A search ofthe NOVla protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table ID.

In a BLAST search of public sequence databases, the NOVla protein was found to have homology to the proteins shown in the BLASTP data in Table IE.

PFam analysis predicts that the NOVla protein contains the domains shown in the Table IF.

Example A2.

The NOV2 clone was analyzed, and the nucleotide and predicted polypeptide sequences are shown in Table 2A.

Table 2A. NOV2 Sequence Analysis

SEQ IDNO:5 1010 bp

NOV2, ATGTCTTCATGCAGGTAAAGATGAAAGTGTCCCAAGAACTATCAGCCATTCCACTCAC CG57881- GTAAAAGCTAATACCATGCCTATTTACTCCCAGACAGTGGCCATGGCTGAACACTTTA 01 DNA AACAAGCAAGCAGTTGTCCTATCTGCCTGGATTATCTTGAAAACCCCACGCACCTGAA Sequence ATGTGGATACATCTGTTGCCTCCGATGCATGAACTCACTGCGAAAGGGGCCCGATGGG AAGGGGGTGCTGTGCCCTTTCTGCCCTGTGGTCTCTCAGAAAAATGACATCAGGCCCG CTGCCCAGCTGGGGGCGCTGGTGTCCAAGATCAAGGAACTAGAGCCCAAGGTGAGAGC TGTTCTGCAGATGAATCCAAGGATGAGAAAGTTCCAAGTGGATATGACCTTGGATGTG

Further analysis ofthe NOV2 protein yielded the following properties shown in Table

2B.

Table 2B. Protein Sequence Properties NOV2

PSort 0.6500 probability located in cytoplasm; 0.1000 probability located in mitochondrial analysis: matrix space; 0.1000 probability located in lysosome (lumen); 0.0000 probability located in endoplasmic reticulum (membrane)

SignalP No Known Signal Sequence Predicted analysis:

A search ofthe NOV2 protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 2C.

In a BLAST search of public sequence databases, the NOV2 protein was found to have homology to the proteins shown in the BLASTP data in Table 2D.

PFam analysis predicts that the NOV2 protein contains the domains shown in the Table 2E.

Example A3.

The NOV3 clone was analyzed, and the nucleotide and predicted polypeptide sequences are shown in Table 3A. Table 3A. NOV3 Sequence Analysis

SEQ ID NO:7 1770 bp

NOV3, GGAGCGGCCGCCCAGGTGCGGTCGCGTTAGTTCGGCCCAATGGCGGCACCGCTGCTTC CG58596-01 ACACGCGTTTGCCGGGAGATGCGGCCGCTTCGTCCTCTGCAGTTAAGAAGCTGGGCGC DNA GTCGAGGACTGGGATTTCAAATATGCGTGCATTAGAGAATGACTTTTTCAATTCTCCC Sequence CCAAGAAAAACTGTTCGGTTTGGTGGAACTGTGACAGAAGTCTTGCTGAAGTACAAAA AGGGTGAAACAAATGACTTTGAGTTGTTGAAGAACCAGCTGTTAGATCCAGACATAAA GGATGACCAGATCATCAACTGGCTGCTAGAATTCCGTTCTTCTATCATGTACTTGACA AAAGACTTTGAGCAACTTATCAGTATTATATTAAGATTGCCTTGGTTGAATAGAAGTC AAACAGTAGTGGAAGAGTATTTGGCTTTTCTTGGTAATCTTGTATCAGCACAGACTGT TTTCCTCAGACCGTGTCTCAGCATGATTGCTTCCCATTTTGTGCCTCCCCGAGTGATC ATTAAGGAAGGCGATGTAGATGTTTCAGATTCTGATGATGAAGATGATAATCTTCCTG CAAATTTTGACACATGTCACAGAGCCTTGCAAATAATAGCAAGATATGTACCATCGAC ACCGTGGTTTCTCATGCCAATACTGGTGGAAAAATTTCCATTTGTTCGAAAATCAGAG AGAACACTGGAATGTTACGTTCATAACTTACTAAGGATTAGTGTATATTTTCCAACCT TGAGGCATGAAATTCTGGAGCTTATTATTGAAAAACTACTCAAGCTGGATGTGAATGC ATCCCGGCAGGGTATTGAAGATGCTGAAGAAACAGCAAATCAAACTTGTGGTGGGACA GATTCCACGGAAGGATTGTTTAATATGGGATTCGCAGAGGCATTTTTGGAACATCTTT GGAAAAACTTGCAGGATCCAAGTAATCCTGCCATCATCAGGCAGGCTGCTGGAAATTA TATTGGAAGCTTTTTGGCAAGAGCTAAATTTATTTCTCTTATTACTGTAAAACCATGC CTAGATCTTTTGGTTAACTGGCTGCACATATACCTTAATAACCAGGATTCGGGAACAA AGGCATTCTGCGATGTTGCTCTCCATGGACCATTTTACTCAGCCTGCCAAGCTGTGTT CTACACCTTTGTTTTTAGACACAAGCAGCTTTTGAGCGGAAACCTGAAAGAAGGTTTG CAGTATCCTCAGAGTCTGAATTTTGAGCGGATAGTGATGAGCCAGCTAAATCCCCTGA AGATTTGCCTGCCCTCAGTGGTTAACTTTTTTGCTGCAATCACAAATAAGTACCAGCT CGTCTTCTGCTACACCATCATTGAGAGGAACAATCGCCAGATGCTGCCAGTCATTAGG AGTACCGCTGGAGGAGACTCAGTGCAGATCTGCACAAACCCGCTGGACACCTTCTTCC CCTTTGATCCCTGTGTGCTGAAGAGGTCAAAGAAATTCATTGATCCTATTTATCAGGT GTGGGAAGACATGAGTGCTGAAGAGCTACAGGAGTTCAAGAAACCCATGAAAAAGGAC ATAGTGGAAGATGAAGATGATGACTTTCTGAAAGGCGAAGTGCCCCAGAATGATACCG TGATTGGGATCACACCAAGCTCCTTTGACACGCATTTCCGAAGTCCTTCAAGTAGTGT GGGCTCCCCACCCGTGTTGTACATGCAACCCAGTCCCCTCTGACGGCAGAAATTTGTG ACTGAGATGTGACATTTGGGATTCCCCATC

ORF Start: ATG at 40 ORF Stop: TGA at 1723

SEQ ID NO:8 561 aa MW at 63593.3kD

NOV3, MAAP HTRLPGDAAASSSAVKK GASRTGISNMRA ENDFFNSPPRKTVRFGGTVTE CG58596-01 VLLKYKKGETNDFELLKNQLLDPDIKDDQIINW LEFRSSIMYLTKDFEQLISIILRL Protein P LNRSQTWEEY AFLGN VSAQTVFLRPCLSMIASHFVPPRVIIKEGDVDVSDSDD Sequence EDDNLPANFDTCHRALQIIARYVPSTPWFLMPILVEKFPFVRKSERTLECYVHNL RI SVYFPTLRHEILE IIEKLLKLDV ASRQGIEDAEETANQTCGGTDSTEG FNMGFAE AFLEHLWKNLQDPSNPAIIRQAAGNYIGSFLARAKFIS ITVKPCLDLLVN HIYLN NQDSGTKAFCDVALHGPFYSACQAVFYTFVFRHKQLLSGNLKEGLQYPQSLNFERIVM SQLNPLKICLPSWNFFAAITNKYQLVFCYTIIERNNRQ PVIRSTAGGDSVQICTN PLDTFFPFDPCV KRSKKFIDPIYQVWEDMSAEE QEFKKPMKKDIVEDEDDDFLKGE VPQNDTVIGITPSSFDTHFRSPSSSVGSPPVLYMQPSPL

Further analysis ofthe NOV3 protein yielded the following properties shown in Table

3B.

Table 3B. Protein Sequence Properties NOV3

PSort 0.3600 probability located in mitochondrial matrix space; 0.1485 probability located in analysis: microbody (peroxisome); 0.1000 probability located in lysosome (lumen); 0.0000 probability located in endoplasmic reticulum (membrane) SignalP No Known Signal Sequence Predicted analysis:

A search ofthe NOV3 protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 3C.

In a BLAST search of public sequence databases, the NOV3 protein was found to have homology to the proteins shown in the BLASTP data in Table 3D.

PFam analysis predicts that the NOV3 protein contains the domains shown in the Table 3E.

Table 3E. Domain Analysis of NOV3

Identities/ Similarities

Pfam Domain NOV3 Match Region Expect Value for the Matched Region

No Significant Matches Found To Known Sequences

Example A4.

The NOV4 clone was analyzed, and the nucleotide and predicted polypeptide sequences are shown in Table 4A.

Table 4A. NOV4 Sequence Analysis

SEQ ID NO:9 1136 bp

NOV4, GAACATTTTGTTAAATGAGCCGAGGGTGTGGAAAATATGACTTTTATATTGGTCTGGG CG57407-01 ATTAGCTATGAGCTCCAGCATTTTCACTGGAGGTAGTTTCATTTGGGGAGGGAAAAAA DNA AAAGGCTTTCTGCGACTTGCCAGAAAAGGTCCTCTGAGAGCAGGTCAAGGTGGCCACG Sequence CATATCCTAAAGAATGGTTGTGGTGGGCTAGACTGCTGTCAATGGCAGCTGGCGAGGT GGCCAACTCAGCTGCATATGGGTTTGCACCGGCCACACTGGTGACTCCACTAGGAACT CTCAGCGTCCTAGTAAGTGCTATTCTTTCTTCATACTTTCTAAATGAAAGATTTAATC CTTACGGGAAATTTGGATGTTTGCTAAGTATTTTAGGATCTACAGTTACGATCACTCA TCCTCCAAAAGAAGAGGATATTGAGACTCTAAATAAAATATCTCACAAGCTAGGTGAT CCAGGTTTTGTGGACTTTGCAACACTTGTGGTCATTATGGCCATGATATTAATCTTCC TGGTGGGTCCCCACCAGGGACAACGATCTTGTGTATGTAGCAATTTGCACATTTGTGT ATGTAACAAAATTGGCACATTTTCAGTCTCCTGGGTTAAGAGCTTAGGCAGTGCTATC AGAGAGCTGTTTGCTGGAAAGCCTGCACTGCCACATCCCCTGGCCTATGTTCTGCTGC TAAGCCTCATTGTCTGTGTGAACACACAGATTAATTACCTAAATTGCGCCCTGGATAT ATTTAACACTTCCATCATGACTCCAATACATTACATATTCTTTACGACATCAGTTTTT AAACTTGTTCAGCTATTATTTTTAAGGAGTGGCAAGATATGCCCATTGATGATGTCAC TGGTACTTTTGACTGGCTTTACAATAATCGTGGGGATATTCTTGTTGCATGCTTTTAA GAGTGTCAGCTTTAGTCTAGCAAGTCTGCCTGTGTCTCTTCGAAAAGACAAAAAAGCA ATGAATGGCAATCTCTCTAATATGTACGAAGTTCTTAATAATAATGAAGAAAGCAAAA GCTTAATCTGTGGAATCAAACTACACACTGGTGAAAATATCTCCCGAAGAAATGGAAT TCTGACAGCTTTTTAAGAAAGATATAATTAAAAG

ORF Start: ATG at 15 ORF Stop: TAA at 1116

SEQ ID NO: 10 367 aa MW at 40060.8kD

NOV4, MSRGCGKYDFYIGLG AMSSSIFTGGSFI GGKKKGF RLARKGP RAGQGGHAYPKE CG57407-01 L ARLLSMAAGEVANSAAYGFAPATLVTPLGT SVLVSAILSSYFLNERFNPYGKF Protein GC LSI GSTVTITHPPKEEDIETLN ISHK GDPGFVDFATLWIMAMI IFLVGPH Sequence QGQRSCVCSN HICVCNKIGTFSVSWVKSLGSAIRELFAGKPALPHPLAYVLLLS IV CVNTQINYLNCALDIFNTSIMTPIHYIFFTTSVFKLVQL FLRSGKICPLMMSLVLLT GFTIIVGIFLLHAFKSVSFS ASLPVSLRKDKKAMNGN SNMYEVLN NEESKSLICG IK HTGENISRRNGI TAF

Further analysis ofthe NOV4 protein yielded the following properties shown in Table

4B.

Table 4B. Protein Sequence Properties NOV4

PSort 0.6850 probability located in endoplasmic reticulum (membrane); 0.6400 probability analysis: located in plasma membrane; 0.4600 probability located in Golgi body; 0.1000 probability located in endoplasmic reticulum (lumen)

SignalP Likely cleavage site between residues 27 and 28 analysis:

A search ofthe NOV4 protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 4C.

In a BLAST search of public sequence databases, the NOV4 protein was found to have homology to the proteins shown in the BLASTP data in Table 4D.

PFam analysis predicts that the NOV4 protein contains the domains shown in the Table 4E.

Example A5.

The NOV5 clone was analyzed, and the nucleotide and predicted polypeptide sequences are shown in Table 5A.

AGCAGGTCCTGGAAGCCAACCCCATCCTGGAGGCCTTTGGAAATGCCAAGACAATCCG CAACGACAACTCAAGCCGCTTTGGGAACGGGGTGATCGAGGGCGCGCGCATCGAGCAA TTTCTCCTGGAGAAGTCCCGGGTCTGCCGGCAGGCTCCCGAGGAGCGGAACTACCATA TCTTCTACTGCATGCTCATGGGGGTGAGTGCTGAGGACAAGCAGCTGCTGAGCCTGGG CACGCCCTCCGAGTACCACTACCTGACCATGGGGAACTGCACTTCCTGTGAGGGGCTC AACGACGCCAAGGACTACGCCCACATCCGCTCGGCCATGAAGATCCTCCAGTTCTCCG ACTCCGAGAGCTGGGACGTCATCAAGCTGCTGGCTGCCATTCTCCACCTGGGGAATGT GGGGTTCATGGCTTCGGTCTTCGAGAACCTGGACGCCTCAGACGTGATGGAGACGCCC GCCTTTCCCACCGTGATGAAGTTACTGGAGGTGCAGCACCAGGAGCTCCGGGACTGTC TGATCAAGCACACCATCCTCATCCGAGGGGAATTTGTCACCAGGTCCCTGAACATTGC CCAGGCTGCTGACCGGAGGGACGCCTTTGTCAAGGGCATCTATGGGCACCTCTTCCTG TGGATTGTCAAGAAGATCAATGCCGCCATCTTCACACCACCAGCCCAGGACCCCAAAA ATGTGCGGAGGGCCATCGGCCTCCTGGACATATTTGGCTTTGAAAATTTCGAGAACAA TAGCTTCGAGCAGCTCTGCATCAACTTCGCCAACGAGCACCTGCAGCAGTTCTTTGTG CAGCACGTGTTCACCATGGAGCAAGAGGAGTACCGCTCGGAGAACATCTCCTGGGACT ATATCCACTACACCGACAATCGGCCCACCCTGGACCTGCTGGCCCTCAAGCCCATGAG CATCATCTCCCTCCTGGACGAAGAAAGCCGCTTCCCGCAGGGGACAGATCTCACCATG CTGCAAAAGCTGAACAGCGTCCATGCCAACAACAAGGCCTTCCTACAGCCCAAGAACA TCCACGATGCCAGATTTGGCATTGCCCATTTTGCCGGCGAGGTGTACTACCAAGCAGA AGGCTTCCTGGAGAAGAACCGAGACGTGCTGAGCACAGATATCCTCACCCTGGTTTAC TCCTCCAAAAACAAGTTTCTGAGGGAGATATTCAACTTGGAGTTAGCAGAGACCAAGC TGGGCGCAGACTCAAATAAACGGCCCTCCACCTTAGGAAGCCAGTTCAAACAGTCTCT GGACCAGCTGATGAAAATCCTGACCAACTGCCAGCCTTACTTCATCCGCTGCATCAAA CCTAATGAGTACAAGAAGCCGCTGCTGTTCGACCGGGAGCTGTGCCTGCGGCAGCTGC GATACTCGGGCATGATGGAGACCGTGCACATCCGCAAGTCGGGCTTCCCCATCCGCTA CACGTTCGAGGAGTTCTCGCAGAGGTTCGGCGTGTTGCTGCCCAACGCCATGCGGATG CAGCTGCAAGGCAACGTCCGCCAGATGACCCTGGGCATCACTGACGTGTGGCTGCGGA CAGACAAAGACTGGAAAGCGGGGAAGACAAAAATTTTCCTGAGGGATCATCAGGACAC TCTGCTGGAGGTACAGAGAAGCCAGGTGCTAGACAGAGCGGCGCTCAGCATCCAGAAA GTCCTTCGGGGCTACAGATACAGGAAGGAGTTCCTGAGGCAGAGGCGGGCAGCTGTGA CCCTGCAGGCCTGGTGGAGAGGCTACTGCAACAGGAGGAATTTCAAGCTGATCCTCGT GGGCTTTGAGCGCCTGCAGGCTATTGCCCGGAGCCAGCCGCTGGCGAGGCAGTACCAG GCCATGCGGCAGAGGACAGTCCAGCTGCAGGCCCTGTGCAGGGGATACCTGGTGCGCC AGCAAGTCCAGGCCAAGAGGAGGGCAGTGGTGGTCATTCAGGCCCATGGCCAGGGCAT GGCTGCCCGGCGCAACTTCCAGCAAAGGAAGGCCAATGCGCCGCTGGTCATCCCGGCC GAGGGGCAGAAAAGCCAAGGCGCTCTCCCTGCCAAGAAGCGCAGAAAAGAAAAAGAAA GAAAGAGAAAAGAAGAAAGGAAGGGAAGGAGGGAAAAAGAAAAGAAAAAAAGAAAAAA GAAAAGAAAAAAGGAAAATAAAGAAAAGCAAGCAAACAAGCAAAGCATAGTGAGAGAG AGAGGAAGGTCCATCTACGACACCGTCACTGACACGGAGATGGTGGAGAAGGTGTTCG GCTTCCTCCCTGCCATGATTGGGGGCCAGGAGGGCCAGGCCTCGCCGCACTTTGAGGA TCTGGAATCGAAGACCCAGAAGCTGCTTGAGGTTGACCTGGACACAGTCCCCATGGCG GAGGAGCCTGAGGAGGATGTGGATGGCCTGGCCGAGTACACCTTCCCCAAGTTTGCTG TGACTTACTTCCAGAAATCAGCCAGCCACACACACATCCGGCGGCCCCTCCGATACCC GTTGCTTTACCACGAAGATGACACTGACTGCTTGGCCGCCCTGGTCATATGGAACGTC ATCCTGAGGTTCATGGGTGATCTCCCAGAGCCAGTGCTGTATGCCAGGAGCAGCCAGC AGGGCAGCTCAGTGATGCGGCAGATCCATGACACGCTGGGCAGGGAGCACGGTGCCCA GGTTCCACAGCACAGTAGATCTGCACAGGCAAGTGGGGGGCAGCAGCGGGCAGAGGAG GGCGACACCTACCAGAGATCTGGCTGCAAGGACAAGGGGACCAAGGATATCTCCTCCA TGAAGCTGAAGCGGTCCTCCCGGATCACAGGCCAGGTGGCCAGCCAGCTGAACATTGG AGAGGAGGCATTGGAGCCTGATGGCCTTGGTGCAGACCGGCCCATGTCCAACCTGGAG AAGGTGCACTTCATCGTGGGCTACGCCATCCTGCGGCCCAGCCTCAGGGATGAGATTT ACTGCCAGATCTGCAAGCAGCTCTCGGAGAACTTCAAAACAAGCAGCCTGGCCCGGGG CTGGATCCTGCTCAGCCTCTGCCTCGGCTGCTTCCCACCCTCAGAGAGGTTCATGAAG TATCTACTGAACTTCATCGGCCAAGGGCCGGCGACCTACGGCCCCTTCTGTGCCGAGC GCCTGAGACGCACCTATGCCAATGGGGTGCGTGCGGAGCCCCCCACCTGGCTGGAGCT GCAGGCTGTCAAGTCCAAGAAGCACATCCCCATCCAAGTCATCTTGGCCACTGGAGAG AGCCTAACCGTCCCCGTGGACTCAGCCTCCACATCTCGGGAAATGTGCATGCACATCG CTCACAAGCAGGGCCTCAGCGACCACCTGGGCTTCTCCCTCCAGGTCGCCGTGTACGA CAAGTTCTGGTCCCTGGGCAGCGGGCGCGACCACATGATGGATGCCATCGCCCGGTGT GAGCAGATGGCCCAGGAGAGGGGCGAGAGCCAGCGCCAGTCACCCTGGCGCATCTACT TCCGGAAGGAATTCTTCACCCCCTGGCACGACTCCCGGGAGGACCCTGTCAGCACCGA GCTTATTTACCGCCAAGTCCTCCGAGGAGTCTGGTCTGGCGAGTACAGCTTCGAGAAG GAGGAAGAGCTGGTTGAGCTGCTGGCCCGGCACTGCTACGTGCAGCTCGGCGCCTCAG CAGAGAGCAAGGCTGTCCAGGAGCTGCTGCCCAGCTGCATCCCCCACAAGCTGTACAG GACCAAGCCCCCAGACAGGTGGGCGAGCCTCGTCACTGCCGCCTGCGCCAAGGCCCCA TACACTCAGAAGCAAGTCACACCACTGGCCGTGCGAGAGCAGGTGGTGGACGCCGCCC GCCTGCAGTGGCCGCTGCTCTTCTCCCGGCTCTTCGAAGTCATCACACTCTCAGGCCC CCGCCTGCCCAAGACGCAGCTGATCTTGGCTGTTAACTGGAAGGGGCTTTGCTTCCTG GACCAGCAGGAGAAGATGCTGCTGGAACTCTCTTTCCCAGAGGTCATGGGTCTGGCCA CCAACAGGGAGGCCCAGGGCGGGCAGAGGCTGCTGCTCTCCACGATGCATGAGGAGTA CGAGTTTGTGTCACCCAGCAGTGTGGCCATCGCTGAGCTGGTGGCCCTGTTCCTGGAG GGCCTGAAGGAGAGGTCCATTTTCGCCATGGCCCTGCAGGACAGGAAGGCCACAGATG ACACCACCCTCCTGGCCTTCAAGAAGGGGGACCTGTTGGTCCTCACAAAGAAGCAGGG GCTGCTGGCCTCTGAGAACTGGACCCTCGGCCAGAACGACAGGACAGGCAAGACGGGG CTGGTGCCCATGGCCTGCCTCTACACCATCCCCACGGTCACTAAGCCCTCGGCACAGC TGCTGAGCTTGCTTGCCATGTCACCAGAGAAGAGGAAGCTGGCGGCTCAGGAGGGGCA GTTCACAGAGCCACGTCCTGAGGAGCCACCCAAGGAAAAGCTGCACACCCTGGAGGAG TTCTCCTATGAGTTCTTCAGGGCTCCAGAGAAGGACATGGTGAGCATGGCCGTGCTGC CCCTGGCCCGTGCCCGTGGCCACCTGTGGGCCTATTCCTGCGAGCCGCTGCGACAGCC GCTGCTCAAGCGAGTCCACGCCAACGCCGGGGTCGGGGTCAGCGTCTATCCCCAGTCT GTGTCAGCGTGGGCTGCCCCAGCACTGTGCTCCTTGACAGCCACACCCATCCTCCGGT ACATGGGCGACTACCCTTCTCGGCAGGCCTGGCCCACCCTGGAGCTCACCGACCAGAT CTTCACACTGGCCCTGCAGCACCCGGCCCTCCAGGACGAGGTCTACTGCCAGATCCTG AAGCAGCTGACGCACAACTCCAACAGGCACAGCGAAGAGCGGGGCTGGCAGCTGCTGT GGCTGTGCACGGGCCTCTTCCCGCCCAGCAAGGGGCTGCTGCCCCATGCCCAGAAGTT TATAGACACTCGGAGGGGGAAGCTGCTGGCCCCCGACTGCAGCCGCCGAATCCAGAAG GTCCTGAGGACGGGGCCCCGGAAGCAGCCCCCGCACCAGGTGGAGGTGGAGGCCGCAG AGCAGAACGTCTCCCGCATCTGCCACAAGATCTACTTCCCCAATGACACCAGTGAGAT GCTGGAGGTGGTTGCCAACACACGGGTGCGGGATGTGTGTGACAGCATTGCCACCAGG CTGCAGCTGGCCTCCTGGGAGGGCTGCAGCCTCTTCATCAAGATTTCAGACAAGGTCA TCAGCCAGAAGGAGGGAGACTTCTTCTTTGATTCCTTGAGGGAGGTGTCTGACTGGGT GAAGAAGAACAAGCCCCAGAAAGAAGGTGCCCTGGGGGCCCCCGTGACGCTCCCCTAC CAGGTGTACTTCATGCGGAAATTGTGGCTCAACATATCTCCAGGGAAGGATGTGAATG CAGACACCATACTCCATTACCACCAGGAGCTGCCCAAGTACCTGCGCGGATTCCACAA GTGTTCGCGGGAGGATGCCATCCACCTGGCGGGCCTCATCTACAAGGCCCAGTTCAAC AACGACCGGTCCCAGCTGGCTAGTGTCCCCAAGATCCTGAGGGAACTGGTGCCTGAGA ACCTCACACGCCTGATGTCCTCGGAGGAGTGGAAAAAGAGCATCCTTCTAGCCTATGA CAAGCATAAGGACAAGACAGTGGAGGAGGCCAAGGTGGCCTTCCTGAAGTGGATCTGC CGGTGGCCCACCTTCGGATCCGCCTTCTTCGAGGTGAAGCAAACCTCGGAGCCTTCCT ACCCGGACGTCATCCTCATCGCCATCAACCGACATGGGGTTCTGCTCATCCACCCCAA GACCAAGGACCTGCTCACCACCTATCCCTTCACCAAGATCTCCAGCTGGAGCAGCGGC AGCACCTACTTCCACATGGCGCTGGGGAGCCTGGGCCGTGGCAGCCGCCTGCTGTGCG AGACCTCCCTGGGCTATAAGATGGATGACCTGCTGACCTCATATGTGCAGCAGCTCCT GAGTGCCATGAACAAGCAGCGGGGCTCCAAGGCCCCAGCCCTGGCCAGCACCTAG

ORF Start: ATG at 1 ORF Stop: TAG at 6607

SEQ ID NO: 12 2202 aa MWat 251200.3kD

NOV5, MLAEPNYRLLKRTGDHVWLEPPSTHKTGVAIGGIIKEAKPGKVLVEDDEGKEHWIRAE CG57770-01 DFGVLSPMHPNSVQGVDDMIRLGDLNEAG VHN LIRYQQHKIYTYTGSILVAVNPFQ Protein VLP YT EQVQLYYSRHMGELPPHVFAIANCYFSMKR KRDQCCIISGESGAGKTET Sequence TK I QFLATISGQHSWIEQQVLEANPILEAFGNAKTIRNDNSSRFGNGVIEGARIEQ FLLEKSRVCRQAPEERNYHIFYCM MGVSAEDKQ LSLGTPSEYHYLTMGNCTSCEGL NDAKDYAHIRSAMKILQFSDSES DVIK LAAILHLGNVGFMASVFENLDASDVMETP AFPTVMKLLEVQHQELRDC IKHTILIRGEFVTRSLNIAQAADRRDAFVKGIYGHLFL WIVKKINAAIFTPPAQDP VRRAIG DIFGFENFENNSFEQLCINFAEHLQQFFV QHVFTMEQEEYRSENIS DYIHYTDNRPTLDLLALKPMSIISLLDEESRFPQGTDLTM LQKLNSVHANNKAF QPK IHDARFGIAHFAGEVYYQAEGFLEKNRDVLSTDI TLVY SSK KFLREIFNLELAETKLGADSNKRPSTLGSQFKQS DQLMKILTNCQPYFIRCIK PJMEYKKP FDRELCLRQ RYSGMMETVHIRKSGFPIRYTFEEFSQRFGV LPNAMR QLQGNVRQMTLGITDVWLRTDKDWKAGKTKIFLRDHQDT EVQRSQVLDRAALSIQK V RGYRYRKEFLRQRRAAVT QA RGYCNRRNFKLILVGFER QAIARSQP ARQYQ AMRQRTVQLQALCRGYLVRQQVQAKRRAVWIQAHGQGMAARRNFQQRKANAPLVIPA EGQKSQGALPAKKRRKEKERKRKEERKGRREKEKKKRKKKRKKENKEKQANKQSIVRE RGRSIYDTVTDTEMVEKVFGF PAMIGGQEGQASPHFEDLESKTQKLLEVD DTVPMA ΞEPEEDVDG AEYTFPKFAVTYFQKSASHTHIRRPLRYP LYHEDDTDC AALVIWNV ILRFMGDLPEPVLYARSSQQGSSVRQIHDTLGREHGAQVPQHSRSAQASGGQQRAEE GDTYQRSGCKDKGTKDISSMKLKRSSRITGQVASQ NIGEEALEPDGLGADRPMSNLE KVHFIVGYAI RPSLRDEIYCQICKQLSENFKTSS ARGWILLSLCLGCFPPSERFMK Y LNFIGQGPATYGPFCAERLRRTYANGVRAEPPT LE QAVKSKKHIPIQVI ATGE S TVPVDSASTSREMCMHIAHKQGLSDHLGFSLQVAVYDKFWSLGSGRDHMMDAIARC EQMAQERGESQRQSPWRIYFRKEFFTP HDSREDPVSTELIYRQVLRGV SGEYSFEK EEELVEL ARHCYVQLGASAESKAVQE PSCIPHK YRTKPPDR ASLVTAACAKAP YTQKQVTPLAVREQWDAARLQ PLLFSRLFEVITLSGPRLPKTQ I AVNWKG CFL DQQEKMLLELSFPEVMGLATNREAQGGQRL LSTMHEEYEFVSPSSVAIAELVA FLE G KERSIFAMALQDRKATDDTTLLAFKKGD LV TKKQG ASENWTLGQNDRTGKTG LVPMAC YTIPTVTKPSAQ LSLLAMSPEKRKLAAQEGQFTEPRPEEPP EKLHTLEE FSYΞFFRAPEKD VSMAV PLARARGHLWAYSCEPLRQPL KRVHANAGVGVSVYPQS VSAAAPALCS TATPI RYMGDYPSRQA PTLELTDQIFTLALQHPALQDEVYCQI KQLTHNSNRHSEERGWQLL LCTGLFPPSKGL PHAQKFIDTRRGKLLAPDCSRRIQK V RTGPRKQPPHQVEVEAAEQNVSRICHKIYFPNDTSEMLEWANTRVRDVCDSIATR LQLAS EGCS FIKISDKVISQKEGDFFFDSLREVSDWVKKNKPQKEGALGAPVT PY QVYFMRKL NISPGKDVNADTILHYHQELPKY RGFHKCSREDAIH AGLIYKAQFN NDRSQDASVPKI RELVPENLTRLMSSEE KKSILLAYDKHKDKTVEEAKVAFLK IC R PTFGSAFFEVKQTSEPSYPDVILIAINRHGVLLIHPKTKDLLTTYPFTKISS SSG STYFHMALGS GRGSR LCETSLGYKMDD LTSYVQQLLSAMNKQRGSKAPALAST

Further analysis ofthe NOV5 protein yielded the following properties shown in Table

5B.

Table 5B. Protein Sequence Properties NOV5

PSort 0.9100 probability located in nucleus; 0.8500 probability located in endoplasmic analysis: reticulum (membrane); 0.4400 probability located in plasma membrane; 0.3811 probability located in microbody (peroxisome)

SignalP No Known Signal Sequence Predicted analysis:

A search ofthe NOV5 protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 5C.

In a BLAST search of public sequence databases, the NOV5 protein was found to have homology to the proteins shown in the BLASTP data in Table 5D.

PFam analysis predicts that the NOV5 protein contains the domains shown in the Table 5E.

Example A6.

The NOV6 clone was analyzed, and the nucleotide and predicted polypeptide sequences are shown in Table 6A.

SEQ LD NO:15 1649 bp

NOV6b, CTCTGACTCTGCCCTTCATACAAGTGTGATGAACCCCAGTCCCCAGGATACCTACCCA CG59233-02 GGCCCCACACCTCCCAGCATCCTGCCCAGCCGACGTGGGGGTGGTATTCTGGATGGTG DNA AAATGGACCCCAAAGTACCTGCTATTGAGGAGAACTTGCTAGATGACAAGCATTTGCT Sequence GAAGCCATGGGATGCTAAGAAGCTATCCTCATCCTCTTCCCGACCTCGGTCCTGTGAA GTCCCTGGAATTAGCATCTTTCCATCTCCTGACCAGCCTGCCAATGTGCCTGTCCTCC CACCTGCCATGAACACGGGGGGCTCCCTACCTGACCTCACCAACCTGCACTTTCCCCC ACCACTGCCCACCCCCCTGGACCCTGAAGAGACAGCCTACCCTAGCCTGAGTGGGGGC AACAGTACCTCCAATTTGACCCACACCATGACTCACCTGGGCATCAGCAGGGGCATGG GCCTGGGCCCAGGCTATGATGCACCAGGTCTTCATTCACCTCTCAGCCACCCATCCCT GCAGTCCTCCCTAAGCAATCCCAACCTCCAGGCTTCCCTGAGCAGTCCTCAGCCCCAG CTTCAGGGCTCCCACAGCCACCCCTCTCTGCCTGCCTCCTCCTTGGCCCGCCATGTAC TGCCCACCACCTCCCTGGGCCACCCCTCACTCAGTGCTCCGGCTCTCTCCTCCTCCTC TTCCTCCTCCTCCACTTCATCTCCTGTTTTGGGCGCCCCCTCTTACCCTGCTTCTACC CCTGGGGCCTCCCCCCACCACCGCCGTGTGCCCCTCAGCCCCCTGAGTTTGCTCGCGG GCCCAGCCGACGCCAGAAGGTCCCAACAGCAGCTGCCCAAACAGTTTTCGCCAACAAT GTCACCCACCTTGTCTTCCATCACTCAGGGCGTCCCCCTGGATACCAGTAAACTGTCC ACTGACCAGCGGTTACCCCCATACCCATACAGCTCCCCAAGTCTGGTTCTGCCTACCC AGCCCCACACCCCAAAGTCTCTACAGCAGCCAGGGCTGCCCTCTCAGTCTTGTTCAGT GCAGTCCTCAGGTGGGCAGCCCCCAGGCAGGCAGTCTCATTATGGGACACCGTACCCA CCTGGGCCCAGTGGGCATGGGCAACAGTCTTACCACCGGCCAATGAGTGACTTCAACC TGGGGAATCTGGAGCAGTTCAGCATGGAGAGCCCATCAGCCAGCCTGGTGCTGGATCC CCCTGGCTTTTCTGAAGGGCCTGGATTTTTAGGGGGTGAGGGGCCAATGGGTGGCCCC CAGGATCCCCACACCTTCAACCACCAGAACTTGACCCACTGTTCCCGCCATGGCTCAG GGCCTAACATCATCCTCACAGGTGACTCCTCTCCAGGTTTCTCTAAGGAGATTGCAGC AGCCCTGGCCGGAGTGCCTGGCTTTGAGGTGTCAGCAGCTGGATTGGAGCTAGGGCTT GGGCTAGAAGATGAGCTGCGCATGGAGCCACTGGGCCTGGAAGGGCTAAACATGCTGA GTGACCCCTGTGCCCTGCTGCCTGATCCTGCTGTGGAGGAGTCATTCCGCAGTGACCG GCTCCAATGAGGGCACCTCATCACCATCCCTCTTCTTGGCCCCATCCCCCACCACCAT TCCTTTCCTCCCTTCCCCCTGGCAG

ORF Start: ATG at 29 ORF Stop: TGA at 1574

SEQ ID NO: 16 515 aa MW at 53598. lkD

NOV6b, MNPSPQDTYPGPTPPSILPSRRGGGILDGE DPKVPAIEENLLDDKH LKPWDAKKLS CG59233-02 SSSSRPRSCEVPGISIFPSPDQPANVPVLPPAMNTGGS PDLTNLHFPPPLPTPLDPE Protein ETAYPSLSGGNSTSNLTHTMTHLGISRGMG GPGYDAPGLHSPLSHPS QSSLSNPNL Sequence QASLSSPQPQLQGSHSHPSLPASSLARHVLPTTSLGHPS SAPALSSSSSSSSTSSPV GAPSYPASTPGASPHHRRVPLSP SL AGPADARRSQQQ PKQFSPTMSPTLSSITQ GVPLDTSKLSTDQRLPPYPYSSPSLVLPTQPHTPKS QQPG PSQSCSVQSSGGQPPG RQSHYGTPYPPGPSGHGQQSYHRPMSDFNLGN EQFSMESPSASLVLDPPGFSEGPGF LGGEGPMGGPQDPHTFNHQNLTHCSRHGSGPNII TGDSSPGFSKEIAAALAGVPGFE VSAAGLELGLGLEDELRMEPLGLEG NMLSDPCALLPDPAVEESFRSDRLQ

Sequence comparison ofthe above protein sequences yields the following sequence relationships shown in Table 6B.

Further analysis ofthe NOV6a protein yielded the following properties shown in Table 6C.

Table 6C. Protein Sequence Properties NOV6a

PSort 0.6500 probability located in cytoplasm; 0.1000 probability located in mitochondrial analysis: matrix space; 0.1000 probability located in lysosome (lumen); 0.0000 probability located in endoplasmic reticulum (membrane)

SignalP No Known Signal Sequence Predicted analysis:

A search ofthe NOV6a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 6D.

In a BLAST search of public sequence databases, the NOV6a protein was found to have homology to the proteins shown in the BLASTP data in Table 6E.

PFam analysis predicts that the NOV6a protein contains the domains shown in the Table 6F.

Table 6F. Domain Analysis of NOV6a

Identities/ Similarities

Pfam Domain NOV6a Match Region Expect Value for the Matched Region

No Significant Matches Found To Known Sequences

Example A7.

The NOV7 clone was analyzed, and the nucleotide and predicted polypeptide sequences are shown in Table 7A.

Table 7A. NOV7 Sequence Analysis

SEQ LO NO:17 2272 bp

NOV7, CCGCCGCCGCCGCCGCCGNCGCTACTGCTGNGGGGGNCGCGGGGGCGCGCANTGGGGC CG58649-01 GCGGCTCGGAGGGGAGGNTAGGGGGGNGTGCCAGGNCCGAAGCCGAGGCGGGGCCGGG DNA ATGCGGCGCTGAGGCCCAGCATGGCCGGCCCGGGCCCCACCTTCCCGCTGCACCGGCT Sequence CGTCTGGGCGAACCGGCATCGCGAACTGGAGGCCGCACTGCACAGCCACCAGCACGAC ATTGAACAGGAGGACCCCCGCGGGCGGACCCCACTGGAGCTGGCCGTGTCTCTGGGAA ACCTGGAGTCTGTGAGAGTGCTCCTTCGACACAATGCCAACGTGGGCAAAGAGAACCG CCAGGGCTGGGCAGTCCTGCAGGAGGCAGTCAGCACTGGAGACCCCGAGATGGTGCAG CTGGTGCTCCAGTATCGGGACTACCAGAGGGCCACGCAGAGGCTGGCGGGCATTCCGG AACTGCTCAACAAACTTCGCCAGGCCCCCGATTTCTACGTTGAGATGAAGTGGGAGTT CACCAGCTGGGTGCCCCTTGTGTCTAAGATGTGCCCAAGCGATGTGTACCGCGTGTGG AAGCGGGGTGAGAGCCTGCGAGTAGACACCAGTCTCCTGGGCTTCGAGCACATGACCT GGCAGCGGGGCCGGAGGAGCTTCATCTTCAAGGGCCAGGAGGCAGGAGCCCTGGTGAT GGAAGTGGACCATGACCGGCAGGTGGTGCATGTGGAGACACTGGGGCTCACTCTGCAG GAGCCCGAAACACTGCTGGCCGCCATGCGGCCCAGCGAGGAGCATGTGGCCAGTCGCC TCACCTCTCCTATCGTCTCCACCCACCTGGACACTCGTAATGTGGCCTTTGAGAGGAA CAAATGTGGTATCTGGGGCTGGCGGTCTGAGAAGATGGAAACTGTTAGCGGCTACGAG GCCAAGGTGTACAGTGCCACCAACGTGGAGCTGGTGACACGCACACGCACGGAGCACC TCTCTGATCAGGACAAGTCGAGGAGCAAAGCGGGGAAGACTCCATTCCAGTCCTTCCT GGGGATGGCGCAGCAGCATTCCTCCCACACCGGGGCCCCCGTGCAGCAGGCAGCCAGC CCCACCAACCCCACAGCCATCTCCCCTGAGGAGTACTTCGACCCCAACTTCAGCCTGG AGTCACGGAACATTGGCCGCCCCATCGAGATGTCCAGCAAAGTACAGAGGTTCAAGGC AACACTGTGGCTGAGTGAAGAGCACCCGCTCTCCCTGGGTGACCAGGTGACCCCCATC ATCGACCTAATGGCCATCAGCAACGCTCACTTTGCCAAGCTGCGCGACTTCATCACTC TGCGCCTTCCACCTGGCTTCCCCGTCAAAATTGAGATTCCCCTTTTCCACGTGCTCAA TGCCCGCATCACCTTCAGCAACCTGTGTGGCTGTGATGAGCCCCTGAGCTCCGTGTGG GTGCCGGCCCCCAGCTCTGCTGTCGCCGCATCAGGGAACCCTTTCCCGTGCGAGGTGG ACCCCACCGTGTTTGAAGTGCCCAACGGGTACAGCGTGCTGGGCATGGAGCGCAACGA GCCCCTCCGGGACGAGGACGATGACCTCCTGCAGTTCGCCATCCAGCAGAGCCTGCTT GAAGCGGGCACTGAGGCGGAGCAGGTGACCGTCTGGGAAGCCCTGACCAACACCCGGC CCGGTGCCCGCCCTCCTCCCCAGGCCACGGTTTATGAGGAACAGCTTCAGCTGGAGCG GGCCCTCCAGGAAAGCCTGCAGCTGTCCACAGAGCCCAGGGGCCCAGGATCCCCTCCC AGGACACCCCCAGCCCCCGGTCCACCCAGCTTTGAAGAGCAGCTGCGCCTGGCCCTGG AGTTGTCTTCACGGGAGCAGGAGGAGCGGGAGCGGCGCGGGCAGCAGGAGGAGGAGGA CTTACAGCGGATCCTGCAGCTGTCACTCACTGAGCACTGAGCCATAGCCCCGGGAGGG CTGGCCAGGCCACTCCCTGCCCGCTTTTGTAATTTATTTATTTATAAACTCTCTGCTG

CTGAGCTTGGGGCCTGGAGCCCCAGGAATGAGCAGGCAGGGGAGACTGAGATGGAAAT

AAAGAGACTGTCGCAGCAAAAAAAAAAAAAAAAAAAAAACTCGAGGGGGGGCCCGGTA

CCCAATTCGCCCTATAGTGAGTCGTATTACAATTCACTGGCCGTCGTTTTACAACGTC

GTGACTGGGAAAACCCTGGCGTTACCCAACTTAATCGCCTTGCAGCACATCCCCCTTT

CGCCAGCTGG

ORF Start: ATG at 137 ORF Stop: TGA at 1952

SEQ LD NO:18 605 aa MW at 68171.1 D

NOV7, MAGPGPTFP HRLVANRHRELEAALHSHQHDIEQEDPRGRTPLELAVSLGNLESVRV CG58649-01 RHNANVGKENRQG AVLQEAVSTGDPEMVQLVLQYRDYQRATQRLAGIPEL NKLR Protein QAPDFYVEMK EFTSWVPLVSKMCPSDVYRVWKRGESLRVDTSL GFEHMTWQRGRRS Sequence FIFKGQEAGALV EVDHDRQWHVET G TLQEPETLLAA RPSEEHVASR TSPI S THLDTRNVAFERNKCGI G RSEKMETVSGYEAKVYSATNVELVTRTRTEHLSDQDKS RSKAGKTPFQSFLGMAQQHSSHTGAPVQQAASPTNPTAISPEEYFDPNFSLESRNIGR PIEMSSKVQRFKAT WLSEEHPLS GDQVTPIID AISNAHFAKLRDFIT RLPPGF PVKIEIPLFHV NARITFSNLCGCDEPLSSV VPAPSSAVAASGNPFPCEVDPTVFEV PNGYSV GMERNEPLRDEDDDLLQFAIQQSLLEAGTEAEQVTVWEALTNTRPGARPPP QATVYEEQLQLERALQESLQLSTEPRGPGSPPRTPPAPGPPSFEEQ RLALELSSREQ EERERRGQQEEEDLQRILQLSLTEH

Further analysis ofthe NOV7 protein yielded the following properties shown in Table

7B.

Table 7B. Protein Sequence Properties NOV7

PSort 0.4500 probability located in cytoplasm; 0.3000 probability located in microbody analysis: (peroxisome); 0.1000 probability located in mitochondrial matrix space; 0.1000 probability located in lysosome (lumen)

Signal? No Known Signal Sequence Predicted analysis: A search ofthe NOV7 protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 7C.

In a BLAST search of public sequence databases, the NOV7 protein was found to have homology to the proteins shown in the BLASTP data in Table 7D.

PFam analysis predicts that the NOV7 protein contains the domains shown in the Table 7E.

Example A8.

The NOV8 clone was analyzed, and the nucleotide and predicted polypeptide sequences are shown in Table 8 A.

Table 8A. NOV8 Sequence Analysis

SEQ D NO:19 3653 bp

NOV8, GTGTGGGGCGCACGGTCCCGGGATACTGGGGACGGCGGGGTGGGAGGGCGCCGTCCTG CG58645-01 GGGCCGCGGCGGCCTGGGCGGGGGAGATGGCGGCGCGATGGAGCAGCGAAAACGTGGT DNA Sequence TGTAGAGTTCCGTGACTCCCAGGCAACTGCGATGTCTGTGGACTGTCTTGGGCAGCAT GCAGTGCTTTCTGGCCGCAGATTCTTATACATCGTCAATCTAGATGCCCCTTTCGAAG GTCACCGAAAGATCTCTCGCCAGAGCAAATGGGACATTGGAGCTGTGCAGTGGAATCC TCATGACAGCTTTGCACACTATTTTGCGGCTTCGAGTAACCAACGAGTAGACCTTTAC AAGTGGAAAGACGGCAGTGGGGAAGTTGGCACAACCTTACAAGGCCACACTCGTGTCA TCAGCGACTTGGACTGGGCGGTGTTTGAGCCTGACCTCCTGGTTACCAGCTCTGTGGA CACCTACATCTACATTTGGGATATCAAAGACACAAGGAAACCTACTGTTGCACTGTCT GCTGTTGCGGGTGCCTCCCAGGTCAAATGGAATAAAAAAAATGCTAACTGCCTTGCCA CCAGCCATGACGGCGATGTGCGGATATGGGATAAGAGGAAACCCAGTACAGCAGTGGA ATATCTAGCCGCCCACCTCTCCAAAATCCATGGCCTGGACTGGCACCCAGACAGCGAG CACATTCTTGCTACCTCCAGTCAAGACAATTCTGTGAAGTTCTGGGATTACCGCCAGC CTCGGAAATACCTCAATATTCTTCCTTGCCAGGTGCCTGTCTGGAAGGCCAGATACAC ACCTTTCAGCAATGGATTGGTGACTGTGATGGTTCCCCAGCTGCGGAGGGAAAACAGC CTTCTCCTGTGGAATGTCTTTGACTTGAACACCCCAGTCCACACCTTCGTGGGGCATG ATGATGTGGTCCTGGAGTTCCAGTGGAGGAAGCAGAAGGAAGGGTCCAAGGACTATCA ACTGGTGACGTGGTCCCGGGATCAGACCTTGAGAATGTGGCGGGTGGATTCCCAGATG CAGAGGCTTTGTGCAAATGACATATTAGATGGTGTTGATGAGTTCATTGAGAGTATTT CCCTTCTGCCGGAACCTGAGAAGACCCTGCACACTGAAGATACAGATCACCAGCACAC TGCAAGCCATGGGGAGGAAGAAGCCCTAAAAGAAGATCCCCCTAGAAATCTCCTGGAA GAGAGGAAATCAGATCAACTGGGGCTGCCTCAGACCTTGCAGCAGGAATTCTCCCTGA TCAATGTGCAAATCCGGAATGTCAATGTGGAGATGGATGCGGCAGACAGGAGCTGCAC AGTGTCTGTGCACTGCAGCAACCATCGTGTCAAGATGCTGGTGAAGTTCCCTGCACAG TACCCAAACAACGCCGCCCCTTCCTTCCAGTTTATTAACCCCACAACCATCACATCCA CCATGAAAGCTAAGCTGCTGAAGATCCTGGAGTACACAGCCCTGCAGAAAGTGAAGCG TGGCCAGAGCTGCCTGGAGCCCTGCCTGCGCCAGCTCGTCTCCTGCCTTGAGTCCTTT GTGAACCAGGAAGACAGCGCTTCCAGCAACCCGTTTGCACTCCCCAACTCTGTCACTC CCCCCTTACCGACGTTTGCGCGGGTGACCACGGCTTACGGGTCGTACCAGGACGCCAA CATTCCCTTTCCTAGGACTTCTGGGGCCAGGTTCTGCGGAGCAGGTTACCTGGTATAT TTCACAAGGCCCATGACAATGCATCGGGCGGTGTCTCCCACAGAGCCTACTCCGAGAT CTCTCTCAGCCTTGTCTGCTTATCACACTGGCTTGATCGCGCCCATGAAGATCCGCAC AGAGGCCCCTGGGAACCTTCGTTTATACAGTGGGAGCCCCACTCGCAGCGAGAAAGAG CAGGTCTCCATCAGCTCCTTCTACTACAAGGAGCGGAAATCAAGACGATGGAAAAGTA AGCGTGAGGGATCAGACTCTGGCAATCGACAGATCAAGGCTGCTGGGAAAGTCATCAT CCAGGATATTGCTTGCCTCCTGCCTGTTCACAAATCGCTGGGAGAGCTGTACATATTG AATGTGAATGATATTCAGGAAACATGTCAGAAGAATGCCGCCTCTGCCTTGCTCGTTG GAAGAAAGGATCTTGTCCAGGTTTGGTCGCTGGCTACGGTAGCTACAGATCTTTGCCT TGGTCCGAAATCTGACCCAGATTTGGAAACACCCTGGGCTCGACATCCATTTGGGCGG CAGCTGCTGGAGTCCCTGTTGGCTCACTATTGCCGGCTCCGGGATGTTCAGACACTGG CGATGCTCTGTAGCGTGTTTGAAGCCCAGTCTCGGCCTCAGGGGCTACCAAACCCCTT TGGGCCTTTTCCTAACCGTTCTTCTAATCTTGTGGTGTCCCATAGTCGATATCCTAGC TTTACCTCTTCTGGTTCCTGCTCCAGTATGTCAGACCCAGGGCTCAACACTGGCGGCT GGAACATAGCGGGAAGAGAGGCAGAGCACTTGTCCTCCCCTTGGGGAGAATCCTCACC AGAAGAGCTCCGCTTTGGGAGTCTGACCTACAGTGATCCCCGTGAGCGAGAACGTGAC CAGCATGATAAAAATAAAAGGCTCCTGGACCCCGCCAATACCCAGCAATTTGATGACT TTAAGAAATGCTATGGGGAAATCCTCTACCGTTGGGGTCTGAGAGAGAAGCGAGCTGA AGTGTTGAAGTTTGTCTCCTGTCCTCCTGACCCTCACAAAGGGATCGAGTTCGGCGTG TACTGCAGCCACTGCCGGAGTGAGGTCCGTGGCACGCAGTGTGCCATCTGCAAAGGCT TCACGTTCCAGTGTGCCATCTGTCACGTGGCTGTGCGGGGATCGTCCAATTTCTGCCT GACCTGTGGGCACGGTGGCCACACCAGCCACATGATGGAGTGGTTTCGGACCCAGGAG GTGTGTCCCACCGGGTGTGGGTGCCACTGCCTGCTTGAAAGCACTTTCTGAACCTACA GAAGTTGGGTATTGTCTGAAATCCCAGAGGACCCATAAGTGCCGGTGACAAGCTGTCT

GTCAGGGGAGAGGCTCCAGAACCTGGGTTCGTCCCCAGTGAGACCGGAGGATGATCCC

CCAAGGACTGCGCAGCATCAGCTCTTGGTGGGCCTCTGCCTTCTCTTCTGTTTGGCCA

CCTGGTGTGGATGTCACTGTGTGAAGATAAGGACAGAAGTGCAGAGCTGCGCTTTGTG

TGTTGTCTATGTCGGCTGAGCTACCAAGGTGGAAGTTTTCATGGAGAAAAGCACCTGG

CTCCAGGGCCAGTGTTACAGTGTTACCCTGTAAGGTGTTAGCCTTAAACCACCGAGCA

GCGTTCTCTTGATGCCAGTGCAGAGACCAGAGTCAGATGCCCGAGGACAGTGGGTAGG

AATTTCATCAACAAATGGACCTATGGCATCATGGCTTTAGAAGCTGGTACATTTACTG

AGCTGATGGACAGTGGCCTTCTAAAATATGACACTTAAATTGTAAATATGCACTGTAC

TTAAGGATTCTTAAGATGTATTTTTTTGTTATTTCTCCTCCAGCTGCTATCCCTTGGC

TAATAAAATTCTAGTAATTTGAAAAAAAAAAAAAAGAGAGAAAGTTAAAAAAAAAAA

ORF Start: ATG at 85 ORF Stop: TGA at 3007

SEQ ID NO:20 974 aa MW at 109841.1kD

NOV8, AAR SSENWVEFRDSQATAMSVDCLGQHAVLSGRRFLYIVNLDAPFEGHRKISRQS CG58645-01 KWDIGAVQ NPHDSFAHYFAASSNQRVDLYK KDGSGEVGTTLQGHTRVISDLDWAVF Protein EPDLLVTSSVDTYIYIWDIKDTRKPTVALSAVAGASQVKNKKNANCLATSHDGDVRI Sequence DKRKPSTAVEYLAAHDSKIHG D HPDSEHI ATSSQDNSVKF DYRQPRKYLNILP CQVPVWKARYTPFSNG VTVMVPQLRRENS WNVFDLNTPVHTFVGHDDVVLEFQ RKQKEGSKDYQLVT SRDQT RMWRVDSQMQRLCANDILDGVDEFIESIS PEPEKT LHTEDTDHQHTASHGEEEALKEDPPRNLLEERKSDQLGLPQT QQEFSLINVQIRNVN VEMDAADRSCTVSVHCSNHRVKM VKFPAQYPNNAAPSFQFINPTTITSTMKAKLLKI LEYTALQ VKRGQSCLEPCLRQ VSCLESFVNQEDSASSNPFA PNSVTPP PTFARV TTAYGSYQDANIPFPRTSGARFCGAGYLVYFTRP T HRAVSPTEPTPRSLSA SAYH TG IAPMKIRTEAPGNLRLYSGSPTRSEKEQVSISSFYYKERKSRR KSKREGSDSGN RQIKAAGKVIIQDIACLLPVHKSLGELYILNVNDIQETCQKNAASALLVGRKDLVQV S ATVATDLCLGPKSDPDLETP ARHPFGRQLLESLLAHYCRLRDVQTLAMLCSVFEA QSRPQGLPNPFGPFPNRSSNLWSHSRYPSFTSSGSCSSMSDPGLNTGGWNIAGREAE HLSSPWGESSPEE RFGSLTYSDPRERERDQHDKNKRL DPANTQQFDDFKKCYGEIL YR GLREKRAEVLKFVSCPPDPHKGIEFGVYCSHCRSEVRGTQCAICKGFTFQCAICH VAVRGSSNFCLTCGHGGHTSHMMEWFRTQEVCPTGCGCHCLLESTF

Further analysis ofthe NOV8 protein yielded the following properties shown in Table

8B.

Table 8B. Protein Sequence Properties NOV8

PSort 0.8800 probability located in nucleus; 0.5150 probability located in mitochondrial analysis: matrix space; 0.4251 probability located in microbody (peroxisome); 0.2422 probability located in mitochondrial inner membrane

SignalP No Known Signal Sequence Predicted analysis:

A search ofthe NOV8 protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 8C.

In a BLAST search of public sequence databases, the NOV8 protein was found to have homology to the proteins shown in the BLASTP data in Table 8D.

PFam analysis predicts that the NOV8 protein contains the domains shown in the Table 8E.

Example A9.

The NOV9 clone was analyzed, and the nucleotide and predicted polypeptide sequences are shown in Table 9A.

Table 9A. NOV9 Sequence Analysis

SEQ LD NO:21 2134 bp

NOV9, GGCGGCTGCGACGCCCTGGTGCAGAGCAGCGCCGTCAAGATGATCGACCTCAGCGCCT

CG58632-01 TCAGCCGCAAGCCCCGGACGCTCCGGCATCTGCCCCGAACCCCGAGGCCGGAGCTGAA

DNA CGTGGCCCCATATGACCCTCACTTCCCGGCCCCGGCCCGGGATGGCTTCCCCGAGCCC

Sequence AGCATGGCGCTGCCTGGGCCAGAGCCCTTGCCCACAGAGTGTGGGTTCGAGCCACCCC

ACCTGGCCCCCCTGAGTGACCCCGAGGCCCCCAGCATGGAGTCCCCGGAGCCTGTCAA

GCCGGAACAGGGCTTCGTGTGGCAGGAGGCCAGTGAGTTCGAGGCTGACACGGCGGGT

TCGACCGTGGAACGCCACAAGAAGGCCCAGCTGGATCGGCTGGACATCAACGTGCAGA

TTGACGACTCCTATCTGGTGGAGGCGGGCGACCGCCAGAAGCGCTGGCAGTGCCGCAT

GTGCGAGAAGTCCTACACGTCCAAGTACAACCTGGTGACGCACATCCTGGGCCACAAC

GGCATCAAGCCACACTCGTGCCCACACTGCAGCAAGCTCTTCAAGCAGCCCAGCCACC

TGCAGACGCACCTGCTGACGCACCAGGGCACCCGGCCCCACAAGTGCCAGGTATGCCA

CAAGGCCTTCACGCAGACCAGCCACCTCAAGCGCCACATGCTGCTGCACTCGGAGGTC

AAGCCCTACAGCTGCCACTTCTGCGGCCGCGGCTTCGCCTACCCCAGCGAGCTCAAGG

CCCACGAAGTGAAGCATGAGAGTGGCCGCTGCCATGTCTGCGTCGAGTGCGGCCTGGA CTTCTCCACCCTGACCCAGCTC_AAGCGCC_ACCTGGCCTCCC_ACCAGGGCCCC_ACCCTC

TACCAGTGCCTCGAGTGTGACAAGTCCTTCCACTACCGCAGCCAGTTGCAGAACCACA TGCTCAAGCACCAGAACGTGCGACCCTTCGTGTGCACTGAATGCGGCATGGAGTTCAG CCAGATTCACCACCTCAAGCAGCACTCCCTCACCCACAAGGGCGTGAAGGAGTTCAAG TGCGAGGTGTGTGGCCGGGAGTTCACCCTACAGGCGAACATGAAGCGGCACATGCTGA TCCACACCAGCGTCCGGCCCTACCAGTGCCACATCTGCTTCAAGACCTTTGTACAGAA GCAGACTCTCAAGACCCACATGATTGTACACTCGCCCGTGAAGCCATTCAAATGCAAG GTGTGCGGGAAGTCCTTCAACCGCATGTACAACCTGCTGGGCCACATGCACCTGCACG CCGGCAGCAAGCCCTTCAAGTGCCCCTACTGCTCCAGCAAGTTTAATCTCAAGGGCAA CCTGAGCCGGCACATGAAGGTCAAGCATGGCGTCATGGACATCGGCCTGGACAGCCAA GACCCCATGAGTGGAGCCGACAGCGCAATGGACCCTTCAGAGCACGAAGGACACCAGG ACATGGACGCACTTCGAGGAGAACGCCTACCCAGCTATGAGGAGCGGGGACAGCAGCG CAGAGGCCAGTGTCCTCACTGAACAGGCCATGAAAGAGATGGCCTACTACAACGTGCT ATAGCGCAAGCTGGGCCACCCCTAACGGGGGCCGGGGGCGAGGGCATGGGGGTGAGAC

CCATGGGCTGCAGGCTGCACCTCCTTGCAGCCGAGACACAGTTTATGGGCCCCATTGT

TCTGAGCCCTTCCCTTCCCGAAGTCATTCGCACACTAGGGACCTTTGGACCACATGGA

GACTGTACTACTGGGCCCGGCTGGTGGGCCAGCCCGGGCCAGGGCTCCAGGCAGGGAC

AGGCAAACACGCCAGGCCCAAATCTGGGTCCCCCGGGCTGCTCCGCGGAAAGTCGGGC

ACAACAATGGCGCCCACGGGTGCAGGGTCACAAGGGCCACGGACCAGAGGGCACTACC

ACGCACAAGCGAACCCTCTACCGGGCCGTCACGAGTGACAAAGAAACCCTGATCCACG

TTCTTTCCCAACAACGCCAAAGAGAAGAAACCACTACACAGAGCACCCACACTATGAT

GACCCCCCTACGGTACGACACAAGAAGATGCACTACACGGCGCCACCACACAGACGCA

CTCGTAGGAGTAGACGCACCAGATCAGTGTACACCAGTCAGCGACG

ORF Start: ATG at 40 ORF Stop: TGA at 1528

SEQ ID NO.22 496 aa MW at 56790.7kD

NOV9, MIDLSAFSRKPRTLRHLPRTPRPELNVAPYDPHFPAPARDGFPEPSMALPGPEPLPTE CG58632-01 CGFEPPHLAPLSDPEAPSMESPEPVKPEQGFV QEASEFEADTAGSTVERHKKAQLDR Protein LDINVQIDDSYLVEAGDRQKRWQCRMCEKSYTSKYN VTHILGHNGIKPHSCPHCSKL Sequence FKQPSHLQTH LTHQGTRPHKCQVCHKAFTQTSHLKRHM LHSEVKPYSCHFCGRGFA YPSELKAHEVKHESGRCHVCVECGLDFSTLTQLKRHLASHQGPTLYQCLECDKSFHYR SQ QNHM KHQNVRPFVCTECGMEFSQIHHLKQHSLTHKGVKEFKCEVCGREFT QAN MKRHMLIHTSVRPYQCHICFKTFVQKQTLKTHMIVHSPVKPFKCKVCGKSFNRMYNLL GHMHLHAGSKPFKCPYCSSKFNLKGNLSRHMKVKHGVMDIGLDSQDPMSGADSAMDPS EHEGHQDMDALRGER PSYEERGQQRRGQCPH Further analysis ofthe NOV9 protein yielded the following properties shown in Table 9B.

Table 9B. Protein Sequence Properties NOV9

PSort j 0.6005 probability located in mitochondrial matrix space; 0.4200 probability located in analysis: j nucleus; 0.3108 probability located in microbody (peroxisome); 0.3067 probability I located in mitochondrial inner membrane

SignalP No Known Signal Sequence Predicted analysis:

A search ofthe NOV9 protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 9C.

In a BLAST search of public sequence databases, the NOV9 protein was found to have homology to the proteins shown in the BLASTP data in Table 9D.

Table 9D. Public BLASTP Results for NOV9

Protein NOV9 Identities/

Accession Residues/ Similarities for Expect

Protein/Organism/Length

Match the Matched Value

Number

Residues Portion

PFam analysis predicts that the NOV9 protein contains the domains shown in the Table 9E.

Example A10.

The NOV10 clone was analyzed, and the nucleotide and predicted polypeptide sequences are shown in Table 10 A.

Table 10A. NOV10 Sequence Analysis

SEQ LD NO:23 1678 bp

NOVlOa, GGCACGAGGCCCGGAAGTCCGGGCGGGCGCCGAGCCGGGTGCGGCACGAGGCAGAGCC CG58630-01 GTAAAGGCGCGCGGGAACATGGGGCTGTATGCTGCAGCTGCAGGCGTGTTGGCCGGCG DNA TGGAGAGCCGCCAGGGCTCTATCAAGGGGTTGGTGTACTCCAGCAACTTCCAGAACGT Sequence GAAGCAGCTGTACGCGCTGGTGTGCGAAACGCAGCGCTACTCCGCCGTGCTGGATGCT GTGATCGCCAGCGCCGGCCTCCTCCGTGCGGAGAAGAAGCTGCGGCCGCACCTGGCCA AGGTGCTAGTGTATGAGTTGTTGTTGGGAAAGGGCTTTCGAGGGGGTGGGGGCCGATG GAAGGCTCTGTTGGGCCGGCACCAGGCGAGGCTCAAGGCTGAGTTGGCTCGGCTCAAG GTTCATCGGGGTGTGAGCCGGAATGAGGACCTGTTGGAAGTGGGATCCAGGCCTGGTC CAGCCTCCCAGCTGCCTCGATTTGTGCGTGTGAACACTCTCAAGACCTGCTCCGATGA TGTAGTTGATTATTTCAAGAGACAAGGTTTCTCCTATCAGGGTCGGGCTTCCAGCCTC GATGACTTACGAGCCCTCAAGGGGAAGCATTTTCTCCTGGACCCCTTGATGCCGGAGC TGCTGGTGTTTCCCGCCCAGACAGATCTGCATGAACACCCACTGTACCGGGCCGGACA CCTCATTCTGCAGGACAGGGCCAGCTGTCTCCCAGCCATGCTGCTGGACCCCCCGCCA GGCTCCCATGTCATCGATGCCTGTGCCGCCCCAGGCAATAAGACCAGTCACTTGGCTG CTCTTCTGAAGAACCAAGGGAAGATCTTTGCCTTTGACCTGGATGCCAAGCGGCTGGC ATCCATGGCCACGCTGCTGGCCCGGGCTGGCGTCTCTTGCTGTGAACTGGCTGAGGAG GACTTCCTGGCGGTCTCCCCCTCGGATCCACGCTACCATGAGGTCCACTACATCCTGC TGGATCCTTCCTGCAGTGGCTCGGGTATGCCGAGCAGACAGCTGGAGGAGCCCGGGGC AGGCACACCTAGCCCGGTGCGTCTGCATGCCCTGGCAGGGTTCCAGCAGCGAGCCCTG TGCCACGCGCTCACTTTCCCTTCCCTGCAGCGGCTCGTCTACTCCATGTGCTCCCTCT GCCAGGAGGAGAATGAAGACATGGTACAAGATGCGCTGCAGCAGAACCCGGGCGCCTT CAGGCTAGCTCCCGCCCTGCCTGCCCGGCCCCACCGAGGCCTGAGCACGTTCCCGGGT GCCGAGCACTGCCTCCGGGCTTCCCCCAAGACCACGCTTAGCGGTGGCTTCTTCGTTG CTGTAATTGAACGGGTCGAGATGCCGACCACCTCACAGGCCAAAGCATCAGCACCAGA ACGCACACCCAGCCCAGCCCCAAAGAGAAAGAAGAGAGCAAAAAGCTGCAGCCGGTGC TTGCACACCGCCTTGCACATAGCAGAGGCTCCGGGCTCACTCCTTCCTGGTGGGAAAG GAAGATGCCTGTCCTCTCCGTGGAAGACCCTGGGCCCTCACCGCAGGCAGCAGTTTGC GTTTTGAAAGGTTATTGGGTCCCTTCCTCGGGCTGTGTTCTTGCTGGTGAGCAAAAGT GTTGCCTGCAGAAATAAAATGCAGAACGTACTCTACGATAAAAAAAAAAAAAAA

ORF Start: ATG at 77 ORF Stop: TGA at 1571

SEQ LD NO:24 498 aa MW at 54070.7kD

NOVlOa, MGLYAAAAGV AGVESRQGSIKG VYSSNFQNVKQ YAVCETQRYSAV DAVIASAG CG58630-01 LRAEKKLRPH AKV VYELLLGKGFRGGGGR KALLGRHQAR KAELARLKVHRGVS Protein RNED LEVGSRPGPASQ PRFVRV T KTCSDDWDYFKRQGFSYQGRASSLDDLRAL Sequence KGKHF LDP MPELLVFPAQTDLHEHPLYRAGHLILQDRASCLPAM LDPPPGSHVID ACAAPGNKTSH AA KNQGKIFAFD DAKRLASMAT LARAGVSCCELAEEDFLAVS PSDPRYHEVHYIL DPSCSGSGMPSRQLEEPGAGTPSPVRLHALAGFQQRA CHALTF PSLQRLVYSMCS CQEENEDMVQDALQQNPGAFRLAPA PARPHRGLSTFPGAEHC R ASPKTT SGGFFVAVIERVEMPTTSQAKASAPERTPSPAPKRKKRAKSCSRCLHTALH IAEAPGS PGGKGRCLSSPWKTLGPHRRQQFAF

SEQ LD NO:25 1219 bp

NOVlOb, AAGGCGCGCGGGAACATGGGGCTGTACGCTGCGGTGGCAGGCGTGCTGGCCGGCGTGG CG58630-02 AGAGCCGCCAGGGCTCTATCAAGGGGCTGGTGTACTCCAGCAACTTCCAGCCTCGATG DNA ACTTACGAGCCCTCAAGGGGAAGCATTTTCTCCTGGACCCCTTGATGCCGGAGCTGCT Sequence GGTGTTTCCCGCCCAGACAGATCTGCATGAACACCCACTGTACCGGGCCGGACACCTC ATTCTGCAGGACAGGGCCAGCTGTCTCCCAGCCATGCTGCTGGACCCCCGCCAGGCTC CCATGTCATGGATGCCTGTGCCACCCCAGGCAATAAAGACCAGTCACTTGGCTGCTCT TCTGAAGAACCAAGGGAAGATCTTTGCCTTTGACCTGGGTGCCAGGCGGCTGGCATCC ATGGCCACGCTGCTGGCCTGGGCTGGCGTCTCCTGCTGTGAGCTGGCTGAGGAGGACT TCCTGGCGGTCTCCCCCTTAGATCCGCGCTATCGTGAGGTCCACTATGTCCTGCTGGA TCCTTCCTGCAGTGGCTCGGGTGAGATGGTATGCCGAGCAGACAGCTGGAGGAGCCCG GGGCAGGGACACCTTAGCCCGGTGCGTCTGCATGCCCTGGCAGGGTTCCAGCAGCGAG CCCTGTGCCACGCGCTCACTTTCCCTTCCCTGCAGCGGCTCGTCTACTCCATGTGCTC CCTCTGCCAGGAGGAGAATGAAGACATGGTACAAGATGCGCTGCAGCAGAACCCGGGC GCCTTCAGGCTAGCTCCCGCCCTGCCTGCCCGGCCCCACCGAGGCCTGAGCACGTTCC CGGGTGCCGAGCACTGCCTCCGGGCTTCCCCCAAGACCACGCTTAGCGGTGGCTTCTT CGTTGCTGTAATTGAACGGGTCGAGATGCCGACCACCTCACAGGCCAAAGCATCAGCA CCAGAACGCACACCCAGCCCAGCCCCAAAGAGAAAGAAGAGAGCAAAAAGCTGCAGCC GGTGCTTGCACACCGCCTTGCACATAGCAGAGGCTCCGGGCTCACTCCTTCCTGGTGG GAAAGGAAGATGCCTGTCCTCTCCGTGGAAGACCCTGGGCCCTCACCGCAGGCAGCAG TTTGCGTTTTGAAAGGTTATTGGGTCCCTTCCTCGGGCTGTGTTCTTGCTGGTGAGCA AAAGTGTTGCCTGCAGAAATAAAATGCAGAACGTACTCTACGATAAAAAAAAAAAAAA

A

ORF Start: ATG at 161 ORF Stop: TGA at 1112

SEQ ID NO:26 317 aa MW at 34690.7kD

NOVlOb, MPELLVFPAQTDLHEHP YRAGHLILQDRASC PAMLLDPRQAP S MPVPPQAIKTS CG58630-02 H AALLKNQGKIFAFD GARR ASMATL AAGVSCCE AEEDFLAVSP DPRYREVH Protein YVLLDPSCSGSGEMVCRADS RSPGQGHLSPVRLHALAGFQQRALCHALTFPSLQRLV Sequence YSMCSLCQEENEDMVQDALQQNPGAFRLAPALPARPHRG STFPGAEHCLRASPKTT SGGFFVAVIERVEMPTTSQAKASAPERTPSPAPKRKKRAKSCSRCLHTA HIAEAPGS LPGGKGRCLSSPWKT GPHRRQQFAF

Sequence comparison ofthe above protein sequences yields the following sequence relationships shown in Table 10B.

Further analysis of the NOVlOa protein yielded the following properties shown in Table IOC.

Table IOC. Protein Sequence Properties NOVlOa

PSort 0.3700 probability located in outside; 0.1900 probability located in plasma membrane; analysis: 0.1000 probability located in endoplasmic reticulum (membrane); 0.1000 probability located in endoplasmic reticulum (lumen) SignalP No Known Signal Sequence Predicted analysis:

A search ofthe NOVlOa protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 10D.

In a BLAST search of public sequence databases, the NOVlOa protein was found to have homology to the proteins shown in the BLASTP data in Table 10E.

PFam analysis predicts that the NOVlOa protein contains the domains shown in the Table 10F.

Example All.

The NOVl 1 clone was analyzed, and the nucleotide and predicted polypeptide sequences are shown in Table 11 A.

Further analysis ofthe NOVl 1 protein yielded the following properties shown in Table 1 IB. Table 11B. Protein Sequence Properties NOVll

PSort 0.4856 probability located in mitochondrial matrix space; 0.3000 probability located in analysis: microbody (peroxisome); 0.2654 probability located in lysosome (lumen); 0.1962 probability located in mitochondrial inner membrane

SignalP No Known Signal Sequence Predicted analysis:

A search ofthe NOVl 1 protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 1 IC.

In a BLAST search of public sequence databases, the NOVl 1 protein was found to have homology to the proteins shown in the BLASTP data in Table 1 ID.

PFam analysis predicts that the NOVl 1 protein contains the domains shown in the Table HE. Table HE. Domain Analysis of NOVll

Identities/ Similarities

Pfam Domain NOVll Match Region Expect Value for the Matched Region

No Significant Matches Found To Known Sequences

Example A12.

The NOVl 2 clone was analyzed, and the nucleotide and predicted polypeptide sequences are shown in Table 12 A.

Table 12A. NOV12 Sequence Analysis

SEQ ED N0.29 4380 bp

NOV12, GCGGCGGCACGGCGGCGGGTGATGGCTCCTCCGGGCTGCCCGGGTTCGTGCCCCAACT CG57703-01 TCGCCGTAGTCTGCTCCTTCTTGGAGCGCTACGGGCCGCTGCTAGACCTGCCTGAGTT DNA GCCGTCCCTGAGCTGGACGGGTGTGCAGGCGCCGCGCGGACGTCGCAACGGAGAAGTA Sequence CCAAAAGAATTGGTGGAGCTCCATTTGAAGCTGATGAGGAAAATTGGCAAATCTGTTA CTGCAGACAGATGGGAAAAATATTTGATCAAGTACCTCTGTGAGTGTCAGTTTGATGA CAATCTCAAATTCAAGAATATTATTAATGAGGAGGATGCCGATACTATGCGTCTCCAG CCAATTGGTCGAGACAAAGATGGCCTCATGTACTGGTACCAATTGGATCAAGATCACA ATGTCAGAATGTACATAGAAGAACAAGATGATCAAGATGGCTCTTCATGGAAATGCAT TGTCAGAAATCGAAACGAGTTGGCTGAGACTCTTGCACTCCTGAAAGCACAAATTGAT CCTGTACTATTGAAAAACTCTAGCCAACAAGACAACTCTTCTCGGGAAAGTCCCAGCT TAGAGGATGAGGAGACTAAAAAAGAGGAAGAAACACCTAAACAAGAGGAACAGAAAGA AAGTGAAAAGATGAAAAGTGAGGAGCAGCCTATGGATTTAGAAAACCGTTCTACAGCC AATGTTCTAGAAGAGACTACTGTGAAAAAAGAAAAAGAAGATGAAAAGGAACTTGTGA AACTGCCAGTCATAGTGAAGCTAGAAAAACCTTTGCCAGAAAATGAAGAAAAAAAGAT TATCAAAGAAGAAAGTGATTCCTTCAAGGAAAATGTCAAACCCATTAAAGTTGAGGTG AAGGAATGTAGAGCAGATCCTAAAGATACCAAAAGTAGCATGGAGAAGCCAGTGGCAC AGGAGCCTGAAAGGATCGAATTTGGTGGCAATATTAAATCTTCTCACGAAATTACTGA GAAATCTACTGAAGAAACTGAGAAACTTAAAAATGACCAGCAGGCCAAGATACCACTA AAAAAACGAGAAATTAAACTGAGTGATGATTTTGACAGTCCAGTCAAGGGACCTTTGT GTAAATCAGTTACTCCAACAAAAGAGTTTTTGAAAGATGAAATAAAACAAGAGGAAGA GACTTGTAAAAGGATCTCTACAATCACTGCTTTGGGTCATGAAGGGAAACAGCTGGTA AATGGAGAAGTTAGTGATGAAAGGGTAGCTCCAAATTTTAAGACAGAACCAATAGAGA CAAAGTTTTATGAGACAAAGGAAGAGAGCTATAGCCCCTCTAAGGACAGAAATATCAT CACGGAGGGAAATGGAACAGAGTCCTTAAATTCTGTCATAACAAGTATGAAAACAGGT GAGCTTGAGAAAGAAACAGCCCCTTTGAGGAAAGATGCAGATAGTTCAATATCAGTCT TAGAGATCCATAGTCAAAAAGCACAAATAGAGGAACCCGATCCTCCAGAAATGGAAAC TTCTCTTGATTCTTCTGAGATGGCAAAAGATCTCTCTTCAAAAACTGCTTTATCTTCC ACCGAGTCGTGTACCATGAAAGGTGAAGAGAAGTCTCCCAAAACTAAGAAGGATAAGC GCCCACCAATCCTAGAATGTCTTGAAAAGTTAGAGAAGTCCAAAAAGACTTTTCTTGA TAAGGACGCACAAAGATTGAGTCCAATACCAGAAGAAGTTCCAAAGAGTACTCTAGAG TCAGAAAAGCCTGGCTCTCCTGAGGCAGCTGAAACTTCTCCACCATCTAATATCATTG ACCACTGTGAGAAACTAGCCTCAGAAAAAGAAGTGGTAGAATGCCAGAGTACAAGTAC TGTTGGTGGCCAGTCTGTGAAAAAAGTAGACCTAGAAACCCTAAAAGAGGATTCTGAG TTCACAAAGGTAGAAATGGATAATCTGGACAATGCCCAGACCTCTGGCATAGAGGAGC CTTCTGAGACAAAGGGTTCTATGCAAAAAAGCAAATTCAAATATAAGTTGGTTCCTGA AGAAGAAACCACTGCCTCAGAAAATACAGAGATAACCTCTGAAAGGCAGAAAGAGGGC ATCAAATTAACAATCAGGATATCAAGTCGGAAAAAGAAGCCCGATTCTCCCCCCAAAG TTCTAGAACCAGAAAACAAGCAAGAGAAGACAGAAAAGGAAGAGGAGAAAACAAATGT GGGTCGTACTTTAAGAAGATCTCCAAGAATATCTAGACCCACTGCAAAAGTGGCTGAG ATCAGAGATCAGAAAGCTGATAAAAAAAGAGGGGAAGGAGAAGATGAGGTGGAAGAAG AGTCAACAGCTTTGCAAAAAACTGACAAAAAGGAAATTTTGAAAAAATCAGAGAAAGA TACAAATTCTAAAGTAAGCAAGGTAAAACCCAAAGGCAAAGTTCGATGGACTGGTTCT CGGACACGTGGCAGATGGAAATATTCCAGCAATGATGAAAGTGAAGGGTCTGGCAGTG AAAAATCATCTGCAGCTTCAGAAGAGGAGGAAGAAAAGGAAAGTGAAGAAGCCATCCT AGCAGATGATGATGAACCATGCAAAAAATGTGGCCTTCCAAACCATCCTGAGCTAATT CTTCTGTGTGACTCTTGCGATAGTGGATACCATACTGCCTGCCTTCGCCCTCCTCTGA TGATCATCCCAGATGGAGAATGGTTCTGCCCACCTTGCCAACATAAACTGCTCTGTGA AAAATTAGAGGAACAGTTGCAGGATTTGGATGTTGCCTTAAAGAAGAAAGAGCGTGCC GAACGAAGAAAAGAACGCTTGGTGTATGTTGGTATCAGTATTGAAAACATCATTCCTC CACAAGAGCCAGACTTTTCTGAAGATCAAGAAGAAAAGAAAAAAGATTCAAAAAAATC CAAAGCAAACTTGCTTGAAAGGAGGTCAACAAGAACAAGGAAATGTATAAGCTACAGA TTTGATGAGTTTGATGAAGCAATTGATGAAGCTATTGAAGATGACATCAAAGAAGCCG ATGGAGGAGGAGTTGGCCGAGGAAAAGATATCTCCACCATCACAGGTCATCGTGGGAA AGACATCTCTACTATTTTGGATGAAGAAAGAAAAGAAAATAAACGACCCCAGAGGGCA GCTGCTGCTCGAAGGAAGAAACGCCGGCGATTAAATGATCTGGACAGTGATAGCAACC TGGATGAAGAAGAGAGCGAGGATGAATTCAAGATCAGTGATGGATCTCAAGATGAGTT TGTTGTGTCTGATGAAAACCCAGATGAAAGTGAAGAAGATCCGCCATCTAATGATGAC AGTGACACTGACTTTTGTAGCCGTAGACTGAGGCGACACCCCTCTCGGCCAATGAGGC AGAGCAGGCGTTTGCGAAGAAAGACCCCAAAGAAAAAATATTCCGATGATGATGAAGA GGAGGAATCTGAGGAGAATAGTAGAGACTCTGAAAGTGACTTCAGTGATGATTTTAGT GATGATTTTGTAGAAACTCGGCGAAGGCGGTCAAGGAGAAATCAGAAAAGACAAATTA ACTACAAAGAAGACTCAGAAAGTGACGGTTCCCAGAAGAGTTTGCGACGTGGTAAAGA AATAAGGCGAGTACACAAGCGAAGACTTTCCAGCTCAGAGAGTGAAGAGAGCTATTTG TCCAAGAACTCTGAAGATGATGAGCTAGCTAAAGAATCAAAGCGGTCAGTTCGAAAGC GGGGCCGAAGCACAGACGAGTATTCAGAAGCAGATGAGGAGGAGGAGGAAGAGGAAGG CAAACCATCCCGCAAACGGCTACACCGGATTGAGACGGATGAGGAGGAGAGTTGTGAC AATGCTCATGGAGATGCAAATCAGCCTGCCCGTGACAGCCAGCCTAGGGTCCTGCCCT CAGAACAAGAGAGCACCAAGAAGCCCTACCGGATAGAAAGTGATGAGGAAGAGGACTT TGAAAATGTAGGCAAAGTGGGGAGCCCATTGGACTATAGCTTAGTGGACTTACCTTCA ACCAATGGACAGAGCCCTGGCAAAGCCATTGAGAACTTGATTGGCAAGCCTACTGAGA AGTCTCAGACCCCCAAGGACAACAGCACAGCCAGTGCAAGCCTAGCCTCCAATGGGAC AAGTGGTGGGCAGGAGGCAGGAGCACCAGAAGAGGAGGAAGATGAGCTTTTGAGAGTG ACTGACCTTGTTGATTATGTCTGTAACAGTGAACAGTTATAAGACTTTTTTTCCATTT TTGTGCTAATTTATTCCACGGTAGCTCTCACACCAGCGGGCCAGTTATTAAAAGCTGT

TTAATTTTTCCTAGAAAACTCCACTACAGAATGACTTTTAGAAGAAAAATTTCAACAA

ATCCTGAAGTCTTTCTGTGAAGTGACCAGT

ORF Start: ATG at 22 ORF Stop: TAA at 4216

SEQ ID NO30 1398 aa MW at 159105.0kD

NOV12, MAPPGCPGSCPNFAWCSFLERYGP LDLPELPSLSWTGVQAPRGRRNGEVPKELVEL CG57703-01 HLKLMRKIGKSVTADRWEKYLIKYLCECQFDDNLKFKNIINEEDADTMRLQPIGRDKD Protein GLMY YQLDQDHNVRMYIEEQDDQDGSSWKCIVRNRNELAETLALLKAQIDPVLLKNS Sequence SQQDNSSRESPSLEDEETKKEEETPKQEEQKESEKMKSEEQPMDLENRSTANVLEETT VKKE EDEKELVK PVIVKLEKP PENEEKKIIKEESDSFKENVKPIKVEVKECRADP KDTKSS EKPVAQEPERIEFGGNIKSSHEITEKSTEETEKLKNDQQAKIPLKKREIK SDDFDSPVKGPLCKSVTPTKEF KDEIKQEEETCKRISTITALGHEGKQLVNGEVSDE RVAPNFKTEPIETKFYETKEESYSPSKDRNIITEGNGTES NSVITSMKTGE EKETA P RKDADSSISVLEIHSQKAQIEEPDPPEMETSLDSSEMAKD SSKTA SSTESCTMK GEEKSPKTKKDKRPPILECLEKLEKSKKTF DKDAQRLSPIPEEVPKSTLESEKPGSP EAAETSPPSNIIDHCEKLASEKEWECQSTSTVGGQSVKKVDLETLKEDSEFT VEMD NLDNAQTSGIEEPSETKGSMQKSKFKYKLVPEEETTASENTEITSERQKEGIKLTIRI SSR KKPDSPPKVLEPENKQEKTEKEEEKTNVGRTLRRSPRISRPTAKVAEIRDQKAD KKRGEGEDEVEEESTA QKTDKKEILKKSEKDTNSKVSKVKPKGKVR TGSRTRGRWK YSSNDESEGSGSEKSSAASEEEEEKESEEAILADDDEPCKKCGLPNHPELI LCDSCD SGYHTACLRPPLMIIPDGE FCPPCQHKL CEKLEEQ QDLDVALKKKERAERRKERL VYVGISIENIIPPQEPDFSEDQEEKKKDSKKSKANLLERRSTRTRKCISYRFDΞFDEA IDEAIEDDIKEADGGGVGRGKDISTITGHRGKDISTILDEERKENKRPQRAAAARRKK RRRLNDLDSDSNLDEEESEDEFKISDGSQDEFWSDENPDESEEDPPSNDDSDTDFCS RRLRRHPSRPMRQSRRLRRKTP KKYSDDDEEEESEENSRDSESDFSDDFSDDFVETR RRRSRRNQKRQINYKEDSESDGSQKSLRRGKEIRRVHKRRLSSSESEESY SKNSEDD ELAKESKRSVRKRGRSTDEYSEADEEEEEEEGKPSRKRLHRIETDEEESCDNAHGDA QPARDSQPRVLPSEQESTKKPYRIESDEEEDFENVGKVGSPLDYSLVDLPSTNGQSPG KAIEN IGKPTEKSQTPKDNSTASASLASNGTSGGQEAGAPEEEEDE LRVTDLVDYV

A search ofthe NOVl 2 protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 12C.

In a BLAST search of public sequence databases, the NOV12 protein was found to have homology to the proteins shown in the BLASTP data in Table 12D.

PFam analysis predicts that the NOV 12 protein contains the domains shown in the Table 12E.

Example A13.

The NOV 13 clone was analyzed, and the nucleotide and predicted polypeptide sequences are shown in Table 13 A.

CTTCGGGACGGTTTTGGCGGGCAGGATGGTGGTGAGCTGCGGCCGCTGCAGAGTGAAG GCGCTGCAGCGCTGGTCACCAAGGGGTGCCAGCGATTGGCAGCCCAGGGCGCACGGCC TGAGGCCCCCAAACGGAAATGGGCCGAGGATGGTGGGGATGCCCCTTCACCCAGCAAA CGGCCCTGGGCCAGGCAAGAGAACCAGGAGGCAGAGCGGGAGGGTGGCATGAGCTGCA GCTGCAGCAGTGGCAGTGGTGAGGCCAGTGCTGGGCTGATGGAGGAGGCGCTGCCCTC TGCGCCCGAGCGCCTGGCCCTGGACTATATCGTGCCCTGCATGCGGTACTACGGCATC TGCGTCAAGGACAGCTTCCTGGGGGCAGCACTGGGCGGTCGCGTGCTGGCCGAGGTGG AGGCCCTCAAACGGGGTGGGCGCCTGCGAGACGGGCAGCTAGTGAGCCAGAGGGCGAT CCCGCCGCGCAGCATCCGTGGGGACCAGATTGCCTGGGTGGAAGGCCATGAACCAGGC TGTCGAAGCATTGGTGCCCTCATGGCCCATGTGGACGCCGTCATCCGCCACTGCGCAG GGCGGCTGGGCAGCTATGTCATCAATGGGCGCTGCATCACCTGTATCTATTACCTGAA TCAGAACTGGGACGTTAAGGTGCATGGCGGCCTGCTGCAGATCTTCCCTGAGGGCCGG CCCGTGGTAGCCAACATCGAGCCACTCTTTGACCGGTTGCTCATTTTCTGGTCTGACC GGCGGAACCCCCACGAGGTGAAGCCAGCCTATGCCACCAGGTACGCCATCACTGTCTG GTATTTTGATGCCAAGGAGCGGGCAGCAGCCAAAGACAAGTATCAGCTAGCATCAGGA CAGAAAGGTGTCCAAGTACCTGTATCACAGCCGCCTACGCCCACCTAGTGGCCAGTCC CAGAGCCGCATGGCAGACAGCTTAAATGACTTCAGGAGAGCCCTGGGCCTGTGCTGGC

TGCTCCTTCCCTGCCACCGCTGCTGCTTCTGACTTTGCCTCTGTCCTGCCTGGTGTGG

AGGGCTCTGTCTGTTGCTGAGGACCAAGGAGGAGAAGAGACCTTTGCTGCCCCATCAT

GGGGGCTGGGTTGTCACCTGGACAGGGGGCAGCCGTGGAGGCCACCGTTACCAACTGA

AGCTGGGGGCCTGGGTCCTACCCTGTCTGGTCATGACCCCATTAGGTATGGAGAGCTG

GCGAGCGAGGCATTGTTCACTTCCCACCAGGATGCAGGACTTGGGGTTGAACGTGAGT

CATGGGCCTCTTGCTGGGAATGGGGTGGGCAGGAGTACCCCCAAGTTCTTCTCATCCT

CCCACCTGGAATGTTGACTGAATTCCCCAAACCTTGGGC

ORF Start: ATG at 12 ORF Stop: TAG at 1206

SEQ LD N032 398 aa MW at 42435.6kD

NOV13a, MGPGGALHARGMTTATAAMDSPCQPQP SQALPQ PGSSSEPLEPEPGRARMGVESY CG58651-01 PCPLLPSYHCPGVPSEASAGSGTPRATATSTTASP RDGFGGQDGGELRPLQSEGAAA Protein LVTKGCQRLAAQGARPEAPKRKAEDGGDAPSPSKRP ARQENQEAEREGGMSCSCSS Sequence GSGEASAGLMEEALPSAPERLA DYIVPCMRYYGICVKDSFLGAALGGRVLAEVEALK RGGRLRDGQ VSQRAIPPRSIRGDQIAWVEGHEPGCRSIGALMAHVDAVIRHCAGRLG SYVINGRCITCIYYLNQN DVKVHGG LQIFPEGRPWANIEP FDRLLIF SDRRNP HEVKPAYATRYAITVWYFDAKERAAAKDKYQLASGQKGVQVPVSQPPTPT

SEQ ID NO:33 1589 bp

NOVl 3b, AAGTTGAAACAAGACGAGCGCCGGGGCCGGACGAAAAGCCTCGCCCCCCTGAAGGTAC CG58651-02 CCTTCCCAAGCCCTTAGGGACCGCAGAGGACTTGGGGACCAGCAAGCAACCCCCAGGG DNA CACGAGAAGAGCTCTTGCTGTCTGCCCTGCCTCACCCTGCCCCACGCCAGGCCCGGTG Sequence GCCCCCAGCTGCATCAAGTGGAGGCGGAGGAGGAGGCGGAGGAGGGTGGCACCATGGG

CCCGGGCGGTGCCCTCCATGCCCGGGGGATGAAGACACTGCTGCCATGGACAGCCCGT

GCCAGCCGCAGCCCCTAAGTCAGGCTCTCCCTCAGTTACCAGGGTCTTCGTCAGAGCC CTTGGAGCCTGAGCCTGGCCGGGCCAGGATGGGAGTGGAGAGTTACCTGCCCTGTCCC CTGCTCCCCTCCTACCACTGTCCAGGAGTGCCTAGTGAGGCCTCGGCAGGGAGTGGGA CCCCCAGAGCCACAGCCACCTCTACCACTGCCAGCCCTCTTCGGGACGGTTTTGGCGG GCAGGATGGTGGTGAGCTGCGGCCGCTGCAGAGTGAAGGCGCTGCAGCGCTGGTCACC AAGGGGTGCCAGCGATTGGCAGCCCAGGGCGCACGGCCTGAGGCCCCCAAACGGAAAT GGGCCGAGGATGGTGGGGATGCCCCTTCACCCAGCAAACGGCCCTGGGCCAGGCAAGA GAACCAGGAGGCAGAGCGGGAGGGTGGCATGAGCTGCAGCTGCAGCAGTGGCAGTGGT GAGGCCAGTGCTGGGCTGATGGAGGAGGCGCTGCCCTCTGCGCCCGAGCGCCTGGCCC TGGACTATATCGTGCCCTGCATGCGGTACTACGGCATCTGCGTCAAGGACAGCTTCCT GGGGGCAGCACTGGGCGGTCGCGTGCTGGCCGAGGTGGAGGCCCTCAAACGGGGTGGG CGCCTGCGAGACGGGCAGCTAGTGAGCCAGAGGGCGATCCCGCCGCGCAGCATCCGTG GGGACCAGATTGCCTGGGTGGAAGGCCATGAACCAGGCTGTCGAAGCATTGGTGCCCT CATGGCCCATGTGGACGCCGTCATCCGCCACTGCGCAGGGCGGCTGGGCAGCTATGTC ATCAACGGGCGCACCAAGGCCATGGTGGCGTGTTACCCAGGCAACGGGCTCGGGTACG TAAGGCACGTTGACAATCCCCACGGCGATGGGCGCTGCATCACCTGTATCTATTACCT GAATCAGAACTGGGACGTTAAGGTAGTGCATGGCGGCCTGCTGCAGATCTTCCCTGAG GGCCGGCCCGTGGTAGCCAACATCGAGCCACTCTTTGACCGGTTGCTCATTTTCTGGT CTGACCGGCGGAACCCCCACGAGGTGAAGCCAGCCTATGCCACCAGGTATGCCATCAC TGTCTGGTATTTTGATGCCAAGGAGCGGGCAGCAGCCAAAGACAAGTATCAGCTAGGT

Sequence comparison ofthe above protein sequences yields the following sequence relationships shown in Table 13B.

Further analysis ofthe NOVl 3 a protein yielded the following properties shown in Table 13C.

Table 13C. Protein Sequence Properties NOV13a

PSort 03000 probability located in nucleus; 0.1893 probability located in lysosome (lumen); analysis: 0.1000 probability located in mitochondrial matrix space; 0.0000 probability located in endoplasmic reticulum (membrane)

SignalP No Known Signal Sequence Predicted analysis:

A search ofthe NOVl 3a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 13D.

Table 13D. Geneseq Results for NO 13a

NOV13a Identities/

Geneseq Protein/Organism/Length [Patent #, Residues/ Similarities for Expect Identifier Date] Match the Matched Value

Residues Region

In a BLAST search of public sequence databases, the NOVl 3 a protein was found to have homology to the proteins shown in the BLASTP data in Table 13E.

PFam analysis predicts that the NOVl 3a protein contains the domains shown in the Table 13F.

Table 13F. Domain Analysis of NOV13a

Identities/ Similarities

Pfam Domain NOV13a Match Region Expect Value for the Matched Region

No Significant Matches Found To Known Sequences Example A14.

The NOVl 4 clone was analyzed, and the nucleotide and predicted polypeptide sequences are shown in Table 14 A.

Further analysis of the NOVl 4 protein yielded the following properties shown in Table 14B.

Table 14B. Protein Sequence Properties NOV14

PSort 0.7000 probability located in nucleus; 0.1000 probability located in mitochondrial analysis: matrix space; 0.1000 probability located in lysosome (lumen); 0.0000 probability located in endoplasmic reticulum (membrane)

SignalP No Known Signal Sequence Predicted analysis:

A search ofthe NOVl 4 protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 14C.

Table 14C. Geneseq Results for NOV14

NOV14

Identities/

Geneseq Protein/Organism/Length [Patent #, Residues/ Expect Similarities for the Identifier Date] Match Value Matched Region

Residues

In a BLAST search of public sequence databases, the NOV14 protein was found to have homology to the proteins shown in the BLASTP data in Table 14D.

PFam analysis predicts that the NOV14 protein contains the domains shown in the Table 14E.

Example A15.

The NOVl 5 clone was analyzed, and the nucleotide and predicted polypeptide sequences are shown in Table 15A.

Table 15A. NOV15 Sequence Analysis

SEQ LD NO:37 1409 bp

NOVl 5, GGCACAGCCCAGGAACGTTGCTTTGGAGAATCCTGCAGATAAGGCTTTTCCAAAAAGC

CG59536-01 GCGAGCATCTTGTTGTATTCAGATACCCTATCGTCGTCAGTCATGGCTAGCATCACTG

DNA CGTGTGTGGGTAACAGCAGGCAGCAGAATGCACCTTTGCCGCCTTGGGCCCATTCCAT

Sequence GTTGAGGTCTCTGGGGAGGAGTCTCTGTCCTTTAGTGGTCAAAATGGCAGAGAGAAAC ATGAAGTTGTTCTCAGGAAGAGTGGTGCCAGCCCAGGGGAAAGAAACCTTTGAAAACT GGCTGATCCAAGTCAATGAGGTCCTGCCAGATTGGAGTATGTCTGAGGAGGAAAAACT CAAGCGCTTGATGAAAACACTTAGGGGCCCTGCCCGGGAGGTCATGCGTTTGCTTCAG GCGGCCAACCCCAACCTAAGTGTAGCAGATTTCTTGCGGGCAATGAAATTGGTGTTTG GGGAGTCTGAAAGCAGTGTGACTGCCCATGGTAAATTTTTTAACACCCTGCAGGCACA AGGGGAGAAAGCCTCCCTTTATGTGATCCGTTTAGAGGTGCAGCTCCAGAATGCTATT CAGGCAGGCATCCTAGCTGAGAAAGATGCAAACCAGACTCGCTTGCAACAGCTTCTTT TAGGCGCTGAGCTGAATAGGGACCTGCGCTTCAGGCTTAAGCATCTTCTCAGGATGTA TGCAAATAAGCAGGAGCGGCTTCCCAATTTCCTGGAGTTAATCAAGATGATAAGGGAG GAAGAGGATTGGGATGATGCTTTTATTAAACGGAAGCGGCCGAAAAGGTCTGAGCCAA TAATGGAGAGGGCAGCCAGCCCTGTGGCATTTCAGGGCGCCCAGCCAATAGCAATCAG CAGTGCTGACTGTAACTGCAACGTGATAGAAATAGATGATACCCTTGATGACTCTGAT GAGGATGTGATCCTGGTGGTGTCTCTGTACCCTTCACTGACACCTACAGGTGCCCCTC CCTTCAGAGGAAGAGCCAGACCTCTGGATCAAGTGCTGGTTATTGATTCCCCCAACAA TTCTGGGGCTCAGTCTCTTTCTACCAGTGGTGGTTCTGGGTATAAGAATGATGGTCCT GGGAATATTCGTAGAGCCAGGAAGCGAAAATACACAACCCGCTGTTCATATTGTGGGG AGGAGGGCCACTCAAAAGAAACCTGTGACAATGAGAGCAACAAGGCCCAGGTTTTTGA GAATCTGATCATCACCCTGCAGGAGCTGACACATACAGAGGAGAGGTCAAAAGAGGTC CCTGGAGAACACAGTGATGCTTCTGAGCCACAGTAAGGATCTAGTCCAGCCCTAAATG AGTCCTTGACTGTATTCAGAGTCTGGTAATGGGAATAACAGGAGAGGGGGGTGGGTTT

CTAACTGCATGAATTAA

ORF Start: ATG at 101 ORF Stop: TAA at 1310

SEQ ID N038 403 aa MW at 45159.8kD

NOV15, ASITACVGNSRQQNAP PPWAHSM RSLGRSLCPLWKMAERNMKLFSGRWPAQGK CG59536-01 ETFENW IQVNEVLPD SMSEEEKLKRLMKTLRGPAREVMRL QAANPNLSVADF RA Protein MKLVFGESESSVTAHGKFFNTLQAQGEKASLYVIRLEVQLQNAIQAGI AEKDANQTR Sequence LQQLLLGAELNRDLRFRLKHLLRMYA KQER PNFLELIKMIREEEDWDDAFIKRKRP KRSEPIMERAASPVAFQGAQPIAISSADCNCNVIEIDDT DDSDEDVILWS YPSLT PTGAPPFRGRARPLDQVLVIDSPNNSGAQS STSGGSGYKNDGPGNIRRARKRKYTTR CSYCGEEGHSKETCDNESNKAQVFENLIITLQELTHTEERSKEVPGEHSDASEPQ

Further analysis of the NOVl 5 protein yielded the following properties shown in Table 15B. Table 15B. Protein Sequence Properties NOV15

PSort 0.7000 probability located in nucleus; 0.1000 probability located in mitochondrial analysis: matrix space; 0.1000 probability located in lysosome (lumen); 0.0000 probability located in endoplasmic reticulum (membrane)

SignalP No Known Signal Sequence Predicted analysis:

A search ofthe NOVl 5 protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several , homologous proteins shown in Table 15C.

In a BLAST search of public sequence databases, the NOVl 5 protein was found to have homology to the proteins shown in the BLASTP data in Table 15D.

PFam analysis predicts that the NOVl 5 protein contains the domains shown in the Table 15E.

Example A16.

The NOVl 6 clone was analyzed, and the nucleotide and predicted polypeptide sequences are shown in Table 16A.

Table 16A. NOVl 6 Sequence Analysis

SEQ ID N039 1746 bp

NOVl 6, ATGGTGCTGGCGCGGGTGGCGCGCCGGCCGGCGGGGTCTGCGGAGGCCCAGGCGTCCC CG59299-01 TGGCGGGGAAGGCGGCGCTTCCGGAGCGTGCCAGGAAGATGTCGAGCCCGGGCATCGA DNA CGGCGACCCCAAGCCTCCATGCTTGCCTCGAAACGGTCTGGTGAAGCTGCCGGGCCAG Sequence CCCAACGGCCTGGGTGCGGCCAGCATCACCAAGGGCACGCCAGCCACCAAGAACCGCC CCTGCCAGCCACCACCCCCACCCACCCTCCCACCACCCAGCCTGGCTGCTCCACTGTC CCGGGCTGCCCTGGCTGGGGGCCCGTGCACCCCGGCAGGTGGACCAGCCTCAGCCTTG GCACCTGGGCACCCAGCGGAGCGGCCGCCGCTGGCCACGGACGAGAAGATCCTCAATG GGCTCTTCTGGTATTTCTCGGCCTGCGAGAAGTGTGTGCTGGCCCAGGTGTGCAAGGC CTGGCGGCGCGTGCTGTACCAGCCCAAGTTCTGGGCAGGCCTCACGCCGGTGCTGCAT GCCAAGGAGCTCTACAACGTGCTGCCTGGTGGCGAGAAGGAGTTCGTGAACCTGCAGG GTTTTGCCGCCAGAGGCTTCGAGGGCTTCTGCCTGGTTGGCGTCTCCGACCTGGACAT CTGTGAGTTCATTGACAACTATGCGCTCTCCAAGAAGGGTGTCAAAGCCATGAGCCTC AAGCGCTCCACCATCACGGACGCAGGCCTCGAGGTTATGCTTGAACAGATGCAGGGCG TGGTGCGTCTGGAGCTGTCGGGCTGCAACGACTTCACCGAGGCCGGGCTGTGGTCCAG CCTGAGCGCGCGCATCACCTCGCTGAGCGTGAGTGACTGCATCAACGTGGCCGACGAC GCCATCGCGGCCATCTCGCAGCTGCTGCCCAACCTGGCGGAGCTGAGCCTGCAGGCCT ACCACGTGACGGACACGGCGCTGGCCTACTTCACGGCGCGCCAGGGCCACAGCACGCA CACGCTGCGCCTGCTCTCCTGCTGGGAGATCACCAACCACGGCGTGGTCAACGTGGTG CACAGCCTGCCCAACCTCACCGCGCTCAGCCTCTCGGGCTGCTCCAAGGTCACCGACG ACGGCGTGGAGCTCGTGGCCGAGAACCTGCGCAAGCTGCGCAGCCTTGACCTCTCGTG GTGCCCACGCATCACCGACATGGCGCTGGAGTACGTGGCCTGCGACCTGCACCGCCTA GAGGAGCTCGTGCTCGACAGGTGTGTACGCATCACGGACACTGGCCTCAGCTATCTGT CCACCATGTCGTCCCTCCGCAGCCTCTACCTGCGATGGTGCTGCCAGGTGCAAGACTT

Further analysis ofthe NOVl 6 protein yielded the following properties shown in Table 16B.

Table 16B. Protein Sequence Properties NOV16

PSort 0.6586 probability located in mitochondrial matrix space; 0.3512 probability located in analysis: mitochondrial inner membrane; 0.3512 probability located in mitochondrial intermembrane space; 0.3512 probability located in mitochondrial outer membrane

SignalP No Known Signal Sequence Predicted analysis:

A search ofthe NOVl 6 protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 16C.

In a BLAST search of public sequence databases, the NOVl 6 protein was found to have homology to the proteins shown in the BLASTP data in Table 16D.

PFam analysis predicts that the NOVl 6 protein contains the domains shown in the Table 16E.

Example A17.

The NOVl 7 clone was analyzed, and the nucleotide and predicted polypeptide sequences are shown in Table 17 A.

Sequence comparison ofthe above protein sequences yields the following sequence relationships shown in Table 17B.

Table 17B. Comparison of NOV17a against NO V17b. _β . . _e NOV17a Residues/NOV17b Identities/ Similarities for the

Protein Sequence ! ■ M,.-at,c.h ■ R„esi .d ,ues Matched Region

Further analysis ofthe NOVl 7a protein yielded the following properties shown in Table 17C.

Table 17C. Protein Sequence Properties NOV17a

PSort 0.4385 probability located in mitochondrial matrix space; 0.3000 probability located in analysis: microbody (peroxisome); 0.1789 probability located in lysosome (lumen); 0.1227 probability located in mitochondrial inner membrane

SignalP No Known Signal Sequence Predicted analysis:

A search ofthe NOV 17a protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 17D.

In a BLAST search of public sequence databases, the NOVl 7a protein was found to have homology to the proteins shown in the BLASTP data in Table 17E.

PFam analysis predicts that the NOVl 7a protein contains the domains shown in the Table 17F.

Table 17F. Domain Analysis of NOV17a

Identities/ Similarities

Pfam Domain NOV17a Match Region Expect Value for the Matched Region

No Significant Matches Found To Known Sequences

Example A18.

The NOVl 8 clone was analyzed, and the nucleotide and predicted polypeptide sequences are shown in Table 18 A.

Further analysis of the NOVl 8 protein yielded the following properties shown in Table 18B.

Table 18B. Protein Sequence Properties NOV18

PSort 0.6500 probability located in cytoplasm; 0.3053 probability located in lysosome analysis: (lumen); 0.1000 probability located in mitochondrial matrix space; 0.0000 probability located in endoplasmic reticulum (membrane)

SignalP No Known Signal Sequence Predicted analysis:

A search ofthe NOVl 8 protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 18C.

Table 18C. Geneseq Results for NOV18

NOV18

Identities/

Geneseq Protein/Organism/Length [Patent #, Residues/ Expect Similarities for the Identifier Date] Match Value Matched Region Residues

AAG93258 Human protein HP10582 - Homo 15..222 59/226 (26%) 3e-09 sapiens, 614 aa. [WO200142302-A1, 129.336 93/226 (41%) 14-JUN-2001]

In a BLAST search of public sequence databases, the NOVl 8 protein was found to have homology to the proteins shown in the BLASTP data in Table 18D.

PFam analysis predicts that the NOVl 8 protein contains the domains shown in the Table 18E.

Table 18E. Domain Analysis of NOVl 8

Identities/ Similarities

Pfam Domain NOV18 Match Region Expect Value for the Matched Region

No Significant Matches Found To Known Sequences

Example A19.

The NOVl 9 clone was analyzed, and the nucleotide and predicted polypeptide sequences are shown in Table 19A.

Further analysis ofthe NOV 19 protein yielded the following properties shown in Table 19B. Table 19B. Protein Sequence Properties NOV19

PSort 0.4558 probability located in mitochondrial matrix space; 0.2393 probability located in analysis: microbody (peroxisome); 0.1497 probability located in mitochondrial inner membrane; 0.1497 probability located in mitochondrial intermembrane space

SignalP Likely cleavage site between residues 12 and 13 analysis:

A search ofthe NOVl 9 protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 19C.

In a BLAST search of public sequence databases, the NOVl 9 protein was found to have homology to the proteins shown in the BLASTP data in Table 19D.

(BM401L17.7.1) (NOVEL PROTEIN (ISOFORM 1)) - Mus musculus (Mouse), 115 aa.

Q9H3Y7 DJ697K14.9.2 (NOVEL PROTEIN, 1..66 66/66 (100%) 2e-34 ISOFORM 2) - Homo sapiens (Human), 70 1..66 66/66 (100%) aa.

PFam analysis predicts that the NOVl 9 protein contains the domains shown in the Table 19E.

Table 19E. Domain Analysis of NOV19

Identities/ Similarities

Pfam Domain NOV19 Match Region Expect Value for the Matched Region

No Significant Matches Found To Known Sequences

Example A20.

The NOV20 clone was analyzed, and the nucleotide and predicted polypeptide sequences are shown in Table 20A.

Further analysis ofthe NOV20 protein yielded the following properties shown in Table 20B.

Table 20B. Protein Sequence Properties NO 20

PSort 0.4500 probability located in cytoplasm; 0.3000 probability located in microbody analysis: (peroxisome); 0.2185 probability located in lysosome (lumen); 0.1000 probability located in mitochondrial matrix space SignalP No Known Signal Sequence Predicted analysis:

A search ofthe NOV20 protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 20C.

In a BLAST search of public sequence databases, the NOV20 protein was found to have homology to the proteins shown in the BLASTP data in Table 20D.

PFam analysis predicts that the NOV20 protein contains the domains shown in the Table 20E.

Table 20E. Domain Analysis of NOV20

Identities/ Similarities

Pfam Domain NOV20 Match Region Expect Value for the Matched Region

No Significant Matches Found To Known Sequences

Example A21.

The NOV21 clone was analyzed, and the nucleotide and predicted polypeptide sequences are shown in Table 21 A.

Table 21A. NOV21 Sequence Analysis

SEQ ID N0.51 4413 bp

NOV21, CTCACATTCCTGGCATAGCAGCCGCCTCCGGCGCGGGCCGACCCTGGGGCTGCGCGCT

CG59636-01 GGGGCGCGAACAGCCAGAGCGTCGGCGCCACGGCCGAGAACACATCTTCGCCGCCGAG

DNA CTGAGCTGGGCCGAGCCGGAGGTTGTGGTCTGACTGCGCTGGGCACCCTCGGGCCGCA

Sequence GCGGTGCTCTGGGGCCAGGTGCCACCGGCCATTGTCCAGGCAGCTGTGTGCAAGCCAA

AGAAGCATGAGGACACTGGAAGACTCCTCGGGGACAGTCCTGCACCGCCTCATCCAGG

AGCAGCTGCGCTACGGCAACCTGACTGAGACGCGCACGCTGCTAGCCATCCAGCAGCA

GGCCCTGAGGGGTGGGGCTGGAACTGGGGGTACAGGGAGCCCCCAGGCCTCCCTGGAG

ATCCTGGCCCCAGAGGACAGTCAGGTGCTGCAGCAGGCCACCAGGCAGGAGCCCCAGG

GCCAGGAGCACCAGGGCGGTGAGAACCACCTGGCAGAGAACACCCTCTACCGGCTATG

CCCACAGCCCAGCAAGGGAGAGGAGCTGCCCACCTATGAGGAGGCCAAAGCCCACTCG

CAGTACTATGCGGCCCAGCAGGCAGGGACCCGGCCACATGCGGGGGACCGAGATCCCC

GTGGGGCCCCGGGAGGCAGTCGGAGGCAGGACGAGGCCCTGCGGGAGCTGAGGCATGG

GCACGTGCGCTCGTTGAGTGAACGGCTCCTTCAGTTGTCCCTGGAGAGGAACGGCGCC

CGGGCCCCCAGCCACATGAGCTCCTCCCACAGCTTCCCACAGCTGGCCCGCAACCAGC

AGGGCCCCCCACTGAGGGGCCCCCCTGCTGAGGGCCCAGAGTCCCGAGGACCCCCACC

TCAGTACCCTCATGTTGTACTAGCTCATGAGACCACCACTGCTGTCACTGACCCACGG

TACCGTGCCCGCGGCAGCCCGCACTTCCAGCATGCTGAAGTCAGGATCCTGCAGGCCC

AGGTGCCTCCTGTGTTCCTCCAACAGCAGCAGCAGTACCAGTACCTGCAGCAATCTCA

GGAGCACCCCCCTCCCCCACATCCAGCTGCTCTCGGCCATGGCCCCCTGAGCTCCCTC

AGTCCACCTGCTGTGGAGGGGCCAGTGAGTGCCCAGGCCTCCTCAGCCACCTCGGGCA

GTGCCCACCTGGCCCAGNATGGAGGCCGTGCTGAGGGAGAATGCCAGGCTGCAGAGAG

ACAATGAGCGGCTGCAGAGGGAGCTGGAGAGCTCTGCGGAGAAGGCTGGCCGCATTGA GAAGCTGGAAAGCGAAATCCAGCGGCTCTCTGAGGCCCATGAGAGCCTGACCAGAGCC TCCTCCAAGCGTGAGGCCCTGGAGAAGACCATGCGGAACAAGATGGACAGTGAAATGA GGAGGCTGCAAGACTTCAACCGGGATCTTAGAGAGAGATTGGAATCTGCAAATCGCCG CCTGGCAAGCAAGACACAGGAGGCCCAGGCCGGCAGTCAGGACATGGTGGCCAAGCTG CTTGCTCAGAGCTACGAACAGCAGCAGGAGCAAGAGAAGCTGGAGCGAGAGATGGCAC TGCTGCGCGGCGCCATCGAGGACCAGCGGCGGCGTGCCGAGCTGCTGGAGCAGGCTCT GGGCAATGCGCAGGGCCGGGCAGCTCGAGCCGAAGAGGAGCTGCGCAAGAAGCAGGCC TATGTGGAGAAAGTGGAGCGGCTGCAGCAGGCGCTCGGGCAGCTGCAGGCAGCCTGTG AGAAGCGGGAGCAGCTGGAGCTGCGTCTGCGGACTCGCCTGGAGCAGGAACTCAAGGC CCTGCGTGCACAGCAGAGACAGGCAGGTGCCCCAGGTGGTAGCAGTGGCAGTGGTGGG TCTCCAGAGCTCAGCGCCCTGCGACTGTCAGAACAACTGCGAGAGAAGGAGGAGCAGA TCCTGGCGCTGGAGGCCGACATGACCAAGTGGGAGCAGAAGTATTTGGAGGAACGTGC CATGAGGCAGTTTGCCATGGATGCGGCTGCCACGGCTGCTGCTCAGCGTGACACCACT CTCATCCGACATTCCCCCCAGCCCTCACCCAGCAGCAGCTTCAATGAGGGTCTGCTCA CTGGTGGCCACAGGCATCAGGAGATGGAAAGCAGGTTAAAGGTGCTCCATGCCCAGAT CCTGGAGAAGGATGCAGTGATCAAGGTCCTTCAGCAGCGCTCCAGGAGAGACCCTGGC AAGGCCATCCAGGGCTCCCTGCGGCCTGCCAAGTCGGTGCCATCTGTTTTCGCGGCTG CGGCAGCAGGAACCCAGGGCTGGCAAGGGCTCTCTTCTAGTGAGCGACAAACAGCAGA CGCCCCTGCTCGGCTGACTACAGCAGACAGAGCACCCACAGAGGAGCCAGTGGTCACA GCTCCCCCTGCTGCCCATGCCAAACACGGGAGCAGAGATGGGAGCACCCAGACTGACG GCCCCCCAGACAGCACCTCCACCTGCCTGCCACCGGAGCCTGACAGCCTTCTGGGGTG CAGCAGTAGCCAGAGAGCAGCCTCTCTGGACTCTGTAGCTACATCCAGAGTCCAGGAC TTGTCAGACATGGTGGAGATACTGATCTGAAGGAGGTGGTGCTTCAGGACTCTGAGCC ATTCTCTCCCCTCCTCTGCCCTGTGCCACTCTCAGCCATTTCAGCAGCCCCGTCAACC

GCTGCTCCGTCCCTTTCCCCAGCCAGACACTCATTCCCATTGACCATCTGGTCCCAGG

AGCTCAGGAGGAGGACCCCAGGGGAGAGGAGAGCTGTGAGAGCACCGGCACCCCCAGA

AGACTCTGCTTCTTAGCCCACATTCCTCCGGGCCTTATGGAGAATGAGGATTCAGCCT

TGACTTCTTGCCCAAGGCCTGCTACTGGGGTAGCAACTGACAGCTCAGAAAGGAGCTG

AGCTCCCTCTGCCCTGCCAGTTGTCAGTCAGGCAGGGAGGGAGTGGCTGTGTTGGTTT

GGGGAACTAATTTCCAAGGACGGCTGCCCGTGGACACCAGGTGGACTGGTTCACTAAT

CAAGTCAGCCATATTGTTCTCTGGCTAAGTTTGGTTCCAGCCAACGTCATCTGCTCTT

CAGTTCCTCACTGCCTTCTTGGGATACTAAGACTTGAATTTTTTGGGGACTATTAAGG

GTGTTAGTCTTGGAGAAGACACAGCCTCACCTTCTCACTTGCTGTGGGTGAGGGGCCA

TTTAAGTGGACTGGGAGACAGTGCGCAGTTTGTATATAATTCCCTTTCTTGTGGAACA

GAAGACTGAGGCCTGCAGGTTCCGATGTGTCTCCATGGGCTGTGCTCCCCTCTTCCTA

CTGTCAGTTTCTGAAACTTCTGACTGGCCTCCCAGTTATGCCTCCTCCTCAAGTTCCT

GGCCCGTGGATGTTAAAGCTGCTCGATTCCCAGGATCTCGGCTGCCTTTTCCTCTATC

TTGAGCCCTATAAATGCCCACGGGACCCCCACCACCAGCCTCTTGAAGTGGCTCCACA

GCTCCTGTCCCTGGAACATCCTGTCAGTTTGGTCATAAACCCTGAGCCAGATGAAATG

AGCCACCGTGAACAGACATCTGCCATGCCCCCAGGTGGGCTTCGGTGGCCCTACCCGG

TACCAGTTCTCTCTGAGAAACTGGAGATGTCTTGTTAGCATAAGTGTCTTCATTCCCA

CCTGGAGGGTTTGGGAGAGGAGCAAAGCAGTTGAAAACTAGTTAATGAGCTACAAGAG

TCAAATAGTCCTCTGAATGGAGCCCCCATCACAAAACAGTGCCCAGGAGGCTGGCTCC

TCAAGCTACCCATGCCCAGCGCCCTAAAGCAGGACCAGATGCTTTGGAATTGGGGTGA

AACACCCACATGGCAGCCTGCTAGCAGCAGTGACTTTGACTTCTGGTCTTAAAGAGTC

CCTCACTTCAGCCCCAGGAGCTATTGGTGGGTTTTAGCAGTTTTGTCTTTACCGTTTT

TAGTTCTCCTTGATTCTTTGTTTTCTTCCTTTATCGTTTTTAGGTTTGGTATGTGTTG

TTTTATTTCCATGGTTCCTCAAGTTTCCTTTTTAAACATTTGCATTTGCTGGACAATT

GCAATTTTTTTTAAAAAATTCCCCTACCCCTGTTTAAAGCTGAAAAATACATTTGGTT

CATGTGCATTGTTTACAAAGCAAAAAGAAAAAAGAGGAAAAAAAGGCAAAAAATATTG

TGAAAGAAAAAAAACAACTTAATATATTTTGGATTAATATTTGGTATTTCTTTTAAAG

TATTTTTTGTGCTGTGAACATTTTCTGCCAAAGACCATGATGTGTGTCTGTATGTTTA

AGTTATCGTAAATATTTAAAATGTAAACATGGCTGTTTTGTTATGCCACCCTGTACCA

GGATTGCTGCCGCATTCCACTGGGTATAACAGTATTTTAATTAAAAAATAATAATTAA

AAGTG

ORF Start: ATG at 1179 ORF Stop: TGA at 2580

SEQ LD NO:52 467 aa MW at 51871.4kD

NOV21, MEAVLRENARLQRDNER QRELESSAEKAGRIEKLESEIQRLSEAHESLTRASSKREA CG59636-01 EKTMRNKMDSEMRRLQDFNRD RERLESANRRLASKTQEAQAGSQDMVAKLLAQSYE Protein QQQEQEK EREMALLRGAIEDQRRRAE LEQALGNAQGRAARAEEE RKKQAYVEKVE Sequence RLQQALGQLQAACEKREQ ELRLRTRLEQE KA RAQQRQAGAPGGSSGSGGSPELSA LR SEQLREKEEQILALEADMTK EQKY EERAMRQFAMDAAATAAAQRDTTLIRHSP QPSPSSSFNEG TGGHRHQEMESR KVLHAQILEKDAVI VLQQRSRRDPGKAIQGS LRPAKSVPSVFAAAAAGTQGWQG SSSERQTADAPAR TTADRAPTEEPWTAPPAAH AKHGSRDGSTQTDGPPDSTSTC PPEPDSLLGCSSSQRAAS DSVATSRVQD SD VE ILI

Further analysis ofthe NOV21 protein yielded the following properties shown in Table 21B.

Table 21B. Protein Sequence Properties NOV21

PSort 0.4593 probability located in mitochondrial matrix space; 0.3000 probability located in analysis: microbody (peroxisome); 0.1552 probability located in mitochondrial inner membrane; 0.1552 probability located in mitochondrial intermembrane space

SignalP No Known Signal Sequence Predicted analysis:

A search ofthe NOV21 protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 21C.

In a BLAST search of public sequence databases, the NOV21 protein was found to have homology to the proteins shown in the BLASTP data in Table 21D.

PFam analysis predicts that the NOV21 protein contains the domains shown in the Table 2 IE.

Example A22.

The NOV22 clone was analyzed, and the nucleotide and predicted polypeptide sequences are shown in Table 22A.

Table 22A. NOV22 Sequence Analysis

SEQ LD NO:53 2717 bp

NOV22, TTCAGAAGGAGGAGAGACACCGGGCCCAGGGCACCCTCGCGGGCGGGCGGACCCAAGC CG59675-01 AGTGAGGGCCTGCAGCCGGCCGGCCAGGGCAGCGGCAGGCGCGGCCCGGACCTACGGG DNA AGGAAGCCCCGAGCCCTCGGCGGGCTGCGAGCGACTCCCCGGCGATGCCTCACAACTC Sequence CATCAGATCTGGCCATGGAGGGCTGAACCAGCTGGGAGGGGCCTTTGTGAATGGCAGA CCTCTGCCGGAAGTGGTCCGCCAGCGCATCGTAGACCTGGCCCACCAGGGTGTAAGGC CCTGCGACATCTCTCGCCAGCTCCGCGTCAGCCATGGCTGCGTCAGCAAGATCCTTGG CAGGAGTAGGTACTACGAGACTGGCAGCATCCGGCCTGGAGTGATAGGGGGCTCCAAG CCCAAGGTGGCCACCCCCAAGGTGGTGGAGAAGATTGGGGACTACAAACGCCAGAACC CTACCATGTTTGCCTGGGAGATCCGAGACCGGCTCCTGGCTGAGGGCGTCTGTGACAA TGACACTGTGCCCAGTGTCAGCTCCATTAATAGAATCATCCGGACCAAAGTGCAGCAA CCATTCAACCTCCCTATGGACAGCTGCGTGGCCACCAAGTCCCTGAGTCCCGGACACA CGCTGATCCCCAGCTCAGCTGTAACTCCCCCGGAGTCACCCCAGTCGGATTCCCTGGG CTCCACCTACTCCATCAATGGGCTCCTGGGCATCGCTCAGCCTGGCAGCGACAAGAGG AAAATGGATGACAGTGATCAGGATAGCTGCCGACTAAGCATTGACTCACAGAGCAGCA GCAGCGGACCCCGAAAGCACCTTCGCACGGATGCCTTCAGCCAGCACCACCTCGAGCC GCTCGAGTGCCCATTTGAGCGGCAGCACTACCCAGAGGCCTATGCCTCCCCCAGCCAC ACCAAAGGCGAGCAGGGCCTCTACCCGCTGCCCTTGCTCAACAGCACCCTGGACGACG GGAAGGCCACCCTGACCCCTTCCAACACGCCACTGGGGCGCAACCTCTCGACTCACCA GACCTACCCCGTGGTGGCAGATCCTCACTCACCCTTGGCCATAAAGCAGGAAACCCCC GAGGTGTCCAGTTCTAGCTCCACCCCTTGCTCTTTATCTAGCTCCGCCCTTTTGGATC TGCAGCAAGTCGGCTCCGGGGTCCCGCCCTTCAATGCCTTTCCCCATGCTGCCTCCGT GTACGGGCAGTTCACGGGCCAGGCCCTCCTCTCAGGGCGAGAGATGGTGGGGCCCACG CTGCCCGGATACCCACCCCACATCCCCACCAGCGGACAGGGCAGCTATGCCTCCTCTG CCATCGCAGGCATGGTGGCAGGAAGTGAATACTCTGGCAATGCCTATGGCCACACCCC CTACTCCTCCTACAGCGAGGCCTGGGGCTTCCCCAACTCCAGCTTGCTGAGTTCCCCA TATTATTACAGTTCCACATCAAGGCCGAGTGCACCGCCCACCACTGCCACGGCCTTTG ACCATCTGTAGTTGCCATGGGGACAGTGGGAGCGACTGAGCAACAGGAGGACTCAGCC TGGGACAGGCCCCAGAGAGTCACACAAAGGAATCTTTATTATTACATGAAAAATAACC

ACAAGTCCAGCATTGCGGCACACTCCCTGTGTGGTTAATTTAATGAACCATGAAAGAC

AGGATGACCTTGGACAAGGCCAAACTGTCCTCCAAGACTCCTTAATGAGGGGCAGGAG

TCCCAGGGAAAGAGAACCATGCCATGCTGAAAAAGACAAAATTGAAGAAGAAATGTAG

CCCCAGCCGGTACCCTCCAAAGGAGAGAAGAAGCAATAGCCGAGGAACTTGGGGGGAT

GGCGAATGGTTCCTGCCCGGGCCCAAGGGTGCACAGGGCACCTCCATGGCTCCATTAT

TAACACAACTCTAGCAATTATGGACCATAAGCACTTCCCTCCAGCCCACAAGTCACAG

CCTGGTGCCGAGGCTCTGCTCACCAGCCACCCAGGGAGTCACCTCCCTCAGCCTCCCG

CCTGCCCCACACGGAGGCTCTGGCTGTCCTCTTTCCTCCACTCCATTTGCTTGGCTCT

TTCTACACCTCCCTCTTGGATGGGCTGAGGGCTGGAGCGAGTCCCTCAGAAATTCCAC

CAGGCTGTCAGCTGACCTCTTTTTCCTGCTGCTGTGAAGGTATAGCACCACCCAGGTC

CTCCTGCAGTGCGGCATCCCCTTGGCAGCTGCCGTCAGCCAGGCCAGCCCCAGGGAGC

TTAAAACAGACATTCCACAGGGCCTGGGCCCCTGGGAGGTGAGGTGTGGTGTGCGGCT

TCACCCAGGGCAGAACAAGGCAGAATCGCAGGAAACCCGCTTCCCCTTCCTGACAGCT

CCTGCCAAGCCAAATGTGCTTCCTGCAGCTCACGCCCACCAGCTACTGAAGGGACCCA

AGGCACCCCCTGAAGCCAGCGATAGAGGGTCCCTCTCTGCTCCCCAGCAGCTCCTGCC

CCCAAGGCCTGACTGTATATACTGTAAATGAAACTTTGTTTGGGTCAAGCTTCCTTCT

TTCTAACCCCCAGACTTTGGCCTCTGAGTGAAATGTCTCTCTTTGCCCTGTGGGGCTT

CTCTCCTTGATGCTTCTTTCTTTTTTTAAAGACAACCTGCCATTACCACATGACTCAA

TAAACCATTGCTCTTCAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA

ORF Start: ATG at 161 ORF Stop: TAG at 1517

SEQ LD NO:54 452 aa MW at 483093kD

NOV22, MPHNSIRSGHGGLNQLGGAFVNGRP PEWRQRIVD AHQGVRPCDISRQLRVSHGCV CG59675-01 SKILGRSRYYETGSIRPGVIGGSKP VATPKWEKIGDYKRQNPTMFAWEIRDRLLAE Protein GVCDNDTVPSVSSINRIIRTKVQQPFNLP DSCVATKSLSPGHTLIPSSAVTPPESPQ Sequence SDSLGSTYSINGLLGIAQPGSDKRKMDDSDQDSCRLSIDSQSSSSGPRKH RTDAFSQ HHLEPLECPFERQHYPEAYASPSHTKGEQGLYPLPLLNST DDGKATLTPSNTPLGRN LSTHQTYPWADPHSP AIKQETPEVSSSSSTPCSLSSSALLD QQVGSGVPPFNAFP HAASVYGQFTGQA LSGREMVGPTLPGYPPHIPTSGQGSYASSAIAGMVAGSEYSGNA YGHTPYSSYSEA GFPNSSLLSSPYYYSSTSRPSAPPTTATAFDHL

Further analysis of the NOV22 protein yielded the following properties shown in Table 22B.

Table 22B. Protein Sequence Properties NOV22

PSort 0.6500 probability located in cytoplasm; 0.1000 probability located in mitochondrial analysis: matrix space; 0.1000 probability located in lysosome (lumen); 0.0000 probability located in endoplasmic reticulum (membrane)

No Known Signal Sequence Predicted analysis:

A search ofthe NOV22 protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 22C.

In a BLAST search of public sequence databases, the NOV22 protein was found to have homology to the proteins shown in the BLASTP data in Table 22D.

Q00288 Paired box protein PAX-8 - Mus 1..452 438/459 (95%) 0.0 musculus (Mouse), 457 aa. 1..457 441/459 (95%)

PFam analysis predicts that the NOV22 protein contains the domains shown in the Table 22E.

Example A23.

The NOV23 clone was analyzed, and the nucleotide and predicted polypeptide sequences are shown in Table 23 A.

Table 23A. NOV23 Sequence Analysis

SEQ LDNO:55 5447 bp

NOV23, TGCCCGTGGACTATGACCACCTATCGGGCCATCCCCAGCGATGGTGTGGACCTGGCAG CG59719-01 CCAGCTGTGGCGCCAGGGTGGGCGATGTCCTCCCTGGGCCACACACAGGGGACTACGC DNA TCCCTTGGGATTCTGGGCCCAGAATGGCAGCATGTCCCAGCCTCTTGGCGAGAGCCCG Sequence GCCACCGCCACCGCCACCGCCACCGCCACCACCCGCCCCAGCCCCACCACTCCCGCAA TGCCCAAGATGGGCGTGCGCGCAAGGGTGGCCGACTGGCCGCCCAAGGGGGAGGCCCT GAGAGAGCACAGCAACCCAAGCCCCTCCCAGGACACAGATGGCACAAAGGCCACCAAG ATGGCCCATTCCATGAGGAGCATACAGAACGGACAGCCCCCCACCAGCACCCCGGCTT CCTCAGGGTCCAAAGCCTTCCACCGACTCTCCAGGAGAAGGTCCAAAGACGTGGAGTT CCAGGACGGGTGGCCCCGGTCCCCCGGCAGGGCCTTCCTCCCCCTTCGGCACCGCAGC AGCAGCGAGATCACCCTCAGCGAGTGTGACGCGGAGGACGCGGGGGAGCCGCGGGGGG CCCGGCACACGGGGGCGCTGCCCCTCTTCCGCGAGTACGGGAGCACCTCGTCCATCGA CGTGCAGGGCATGCCCGAGCAGAGCTTCTTCGACATCCTGAACGAGTTCCGCAGCGAG CAGCCCGACGCCCGAGGGTGCCAGGCCCTCACCGAGCTCCTCCGGGCAGATCCTGGCC CACACCTCATGGGGGGCGGCGGCGGAGCCAAGGGGGACTCCCACAACGGGCAGCCCGC CAAGGACAGCCTCCTGCCACTGCAGCCCACGAAGGAGAAGGAGAAGGCCCGGAAGAAA CCTGCGCGGGGCCTCGGCGGCGGGGACACGGTGGACTCGTCCATCTTTCGGAAGCTAA GGAGCAGCAAACCCGAGGGGGAGGCTGGGCGTTCCCCGGGGGAGGCCGACGAGGGCCG GAGCCCCCCGGAAGCCAGCAGGCCGTGGGTGTGTCAGAAGAGCTTCGCCCACTTCGAC GTGCAGAGCATGCTGTTCGACCTCAACGAGGCGGCCGCCAACAGGGTGTCGGTGTCGC AGCGGCGGAACACCACCACGGGTGCTTCGGCCGCTTCCGCCGCCTCGGCCATGGCCTC CCTCACGGCCTCGCGGGCCCACAGCCTCGGAGGCCTGGACCCGGCCTTCACCAGCACA GAGGACCTAAACTGCAAGGAGAACTTGGAGCAGGACCTCGGCGATGACAACAGCAACG ACCTGCTGCTCAGCTGCCCGCACTTCCGCAATGAGATCGGGGGCGAGTGTGAGCGCAA CGTGAGCTTCTCCCGGGCTTCCGTGGGCTCCCCGAGCAGCGGCGAGGGCCACCTGGCA GAGCCCGCCCTGAGCGCCTACCGCACCAACGCCAGCATCTCGGTGTTGGAAGTTCCCA AGGAGCAGCAGCGGACGCAGAGTCGGCCCCGGCAGTACAGCATCGAGCATGTGGACCT GGGCGCCCGCTACTACCAGGATTACTTCGTGGGCAAAGAACATGCCAATTACTTCGGC GTGGATGAGAAGCTGGGGCCAGTGGCTGTGAGCATTAAGCGGGAGAAGCTGGAAGACC ACAAGGAGCACGGACCTCAGTACCAGTACAGGATCATCTTCCGGACCCGCGAGCTCAT CACCCTGCGGGGCTCCATCCTGGAAGATGCTACGCCCACAGCCACCAAGCATGGGACC GGGCGGGGCCTGCCCTTGAAGGATGCCCTGGAGTATGTCATCCCCGAGCTCAACATCC ACTGCCTGCGGCTGGCCCTCAACACCCCCAAGGTGACGGAGCAACTGCTGAAGCTCGA TGAGCAAGGGGTGAGTCAGGGGAAGCACAAGGTGGGCATCCTCTATTGCAAGGCCGGC CAGAGCTCCGAGGAGGAGATGTACAACAATGAGGAGGCCGGCCCCGCCTTTGAGGAGT TCCTCTCCCTCATCGGCGAGAAGGTCTGCCTGAAGGGCTTCACCAAGTACGCTGCCCA GCTGGACGTCAAGACCGACTCCACGGGAACCCACTCCCTCTACACGATGTACCAGGAC TACGAGATCATGTTCCATGTCTCCACCCTGCTCCCTTACACCCCCAACAACAGGCAGC AGCTGCTACGGAAGAGGCACATAGGAAATGACATCGTGACGATCATCTTCCAGGAGCC TGGCGCGCTACCGTTCACCCCCAAGAACATCCGCTCCCACTTCCAGCACGTCTTCATC ATTGTCCGAGTCCACAACCCCTGCACTGATAACGTCTGTTACAGTATGGCTGTGACCC GATCCAAAGACGCTCCTCCTTTCGGCCCCCCCATCCCCAGTGGAACCACATTCCGCAA ATCCGACGTCTTCAGAGACTTCTTGCTGGCCAAGGTGATTAACGCTGAGAACGCCGCG CACAAGTCCGACAAGTTCCACACCATGGCCACCAGGACCCGCCAGGAGTATCTCAAGG ACCTGGCCGAAAACTGTGTCTCCAACACCCCCATCGACTCCACCGGCAAATTCAACCT CATCTCCCTGACCTCCAAGAAGAAGGAAAAGACAAAAGCACGGGCTGGCGCTGAGCAG CACAGTGCAGGGGCCATCGCCTGGAGGGTGGTGGCCCAGGACTACGCCCAGGGGGTGG AAATCGACTGCATTTTGGGAATTTCCAATGAGTTTGTGGTGCTCCTGGACTTACGCAC CAAGGAGGTGGTGTTCAACTGCTACTGCGGGGATGTCATTGGCTGGACTCCAGACTCC TCCACACTCAAAATCTTCTATGGACGAGGAGACCACATCTTCCTACAGGCGACAGAGG GTTCTGTGGAGGACATAAGGGAGATAGTGCAGAGACTGAAGGTGATGACCAGTGGCTG GGAGACGGTGGACATGACGCTTCGGCGGAACGGGCTCGGGCAGCTGGGCTTCCACGTG AAGTACGACGGCACGGTGGCCGAGGTTGAGGACTATGGGTTCGCCTGGCAGGCCGGCC TCCGGCAGGGCAGCCGACTAGTGGAGATCTGCAAGGTGGCCGTGGTCACACTGACCCA CGACCAGATGATCGACCTGCTGCGCACCTCTGTCACTGTGAAGGTGGTCATCATCCCG CCTTTTGAGGACGGCACTCCCCGGAGGGGTTGGCCGGAGACCTACGACATGAATACCT CGGAGCCCAAGACGGAGCAGGAAAGCATCACTCCTGGGGGCCGGCCCCCCTACCGCAG CAATGCTCCCTGGCAGTGGAGCGGGCCCGCATCCCATAACTCTCTACCAGCCTCCAAG TGGGCCACTCCAACCACTCCCGGCCATGCCCAGTCCCTGAGCCGGCCCCTGAAGCAGA CCCCCATAGTCCCCTTCCGGGAGTCCCAGCCACTGCACAGCAAGAGGCCTGTCAGCTT CCCAGAAACCCCTTACACAGTATCACCAGCAGGGGCCGACAGAGTCCCTCCCTACCGA CAGCCTTCTGGGAGCTTCTCCACCCCCGGTTCGGCCACCTACGTGAGATACAAGCCAT CCCCAGAAAGGTACACGGCTGCCCCACACCCCCTGCTATCTCTTGATCCCCACTTCAG CCACGATGGGACGTCCAGCGGCGACTCCTCTTCCGGCGGCCTGACCAGCCAGGAGAGC ACCATGGAACGCCAGAAGCCAGAGCCTTTGTGGCATGTGCCTGCCCAGGCCAGGCTCT CAGCCATAGCCGGAAGCAGCGGGAACAAGCACCCGTCCAGGCAGGATGCAGCAGGCAA AGATTCTCCCAACAGGCATTCCAAAGGAGAACCTCAATACTCAAGTCATTCCAGCAGC AACACCCTCTCCAGCAACGCATCCAGCAGCCACAGCGACGACCGCTGGTTCGACCCCC TGGACCCCCTGGAGCCAGAGCAAGACCCCCTCTCCAAGGGTGGCTCTAGTGACAGCGG CATCGACACCACCCTCTACACCTCCAGCCCTAGCTGCATGTCCCTGGCCAAGGCTCCA CGGCCCGCCAAGCCACACAAGCCCCCTGGAAGTATGGGCCTTTGTGGCGGGGGTCGCG AGGCCGCTGGGAGGTCCCACCACGCAGACAGGCGGCGGGAGGTCTCCCCTGCCCCCGC AGTTGCCGGCCAAAGCAAGGGCTACCGACCGAAGCTGTACTCCTCCGGCTCCAGCACC CCCACGGGACTGGCGGGGGGCAGCCGAGACCCACCGAGGCAGCCCAGTGACATGGGCT CGAGGGTTGGCTACCCCGCTCAGGTTTACAAAACTGCCAGTGCAGAGACTCCTCGGCC CTCCCAGCTGGCCCAGCCCAGCCCCTTTCAGCTCTCCGCCTCCGTCCCCAAGTCCTTC TTCTCCAAGCAGCCTGTACGCAATAAGCACCCAACAGGGTGGAAGAGAACGGAGGAGC CCCCACCACGGCCACTCCCCTTCAGTGACCCAAAGAAGCAGGTGGACACGAACACCAA AAATGTCTTTGGGCAACCGAGGTTGAGGGCATCCCTCCGAGACCTCCGGTCACCACGG AAGAACTACAAATCCACCATCGAGGATGACCTGAAGAAACTCATCATCATGGACAACC TGGGGCCAGAGCAGGAGAGAGACACGGGACAGTCACCGCAGAAGGGCCTGCAGCGGAC GCTGTCGGACGAGAGCCTGTGCAGCGGGCGCCGGGAGCCCAGCTTCGCCAGCCCCGCT GGCCTAGAGCCAGGGCTGCCCAGCGACGTGCTCTTCACCAGCACCTGCGCCTTCCCGT CCAGCACGCTGCCTGCACGCCGCCAGCACCAGCACCCCCACCCGCCCGTCGGCCCCGG TGCCACCCCTGCCGCCGGCAGCGGCTTTCCCGAGAAGAAATCCACCATCTCAGCCTCG GAGCTCTCGCTGGCTGATGGGCGGGACCGCCCCCTGCGGCGCCTGGACCCTGGGCTGA TGCCCCTGCCTGACACAGCTGCTGGCCTCGAGTGGTCCAGCCTGGTGAACGCAGCCAA GGCATACGAAGTGCAAAGAGCCGTCTCACTCTTCTCTCTGAACGACCCGGCCCTGAGC CCGGACATCCCGCCTGCACACAGTCCTGTCCACAGCCACCTGAGCCTGGAGAGGGGAC CCCCGACCCCCAGGACCACCCCTACCATGAGCGAGGAGCCACCCCTGGATCTGACAGG CAAGGTGTACCAGCTGGAGGTGATGCTGAAACAGCTGCACACTGACCTGCAGAAGGAG AAGCAGGACAAGGTGGTGCTCCAGTCAGAGGTGGCCAGCCTGCGGCAGAACAACCAGC GGCTGCAGGAGGAGTCGCAGGCCGCCAGCGAGCAGCTGCGCAAGTTTGCGGAGATCTT CTGCAGGGAGAAGAAGGAGCTCTGAGGTGGGAGGCCGCCGCCCGCCTTCGCTCCTTCC CCTCAGGCCGTGGCCCTGCTGCCTCTCTCCCTCCACTCAGCTCCCAGCTGCCG

ORF Start: ATG at 13 ORF Stop: TGA at 5359 SEQLDNO:56 1782 aa MW tl94606.5kD

NOV23, MTTYRAIPSDGVDLAASCGARVGDVLPGPHTGDYAPLGFWAQWGSMSQP GESPATAT CG59719-01 ATATATTRPSPTTPAMPK GVRARVADWPPKREALREHSNPSPSQDTDGTKΆTKMAHS Protein MRSIQNGQPPTSTPASSGSKAFHR SRRRSKDVEFQDG PRSPGRAFLP RHRSSSEI Sequence TLSECDAEDAGEPRGARHTGALPLFREYGSTSSIDVQGMPEQSFFDILNEFRSEQPDA RGCQALTE LRADPGPHLMGGGGGAKGDSHNGQPAKDSLLPLQPTKEKEKARKKPARG GGGDTVDSSIFRKLRSSKPEGEAGRSPGEADEGRSPPEASRP VCQKSFAHFDVQSM FD NEAAANRVSVSQRRNTTTGASAASAASAMASLTASRAHSLGGLDPAFTSTEDLN CKENLEQDLGDDNSNDL LSCPHFRNEIGGECERNVSFSRASVGSPSSGEGHLAEPA SAYRTNASISVLEVPKEQQRTQSRPRQYSIEHVDLGARYYQDYFVGKEHANYFGVDEK LGPVAVSIKREKLEDHKEHGPQYQYRIIFRTRELIT RGSILEDATPTATKHGTGRGL P DALEYVIPELNIHCLRLALNTPKVTEQLLKLDEQGVSQGKHKVGILYCKAGQSSE EEMYN EEAGPAFEEFLSLIGEKVC KGFTKYAAQLDVKTDSTGTHSLYTMYQDYEI FHVST LPYTPNNRQQLLRKRHIGNDIVTIIFQEPGALPFTPKNIRSHFQHVFIIVRV HNPCTD VCYSMAVTRSKDAPPFGPPIPSGTTFRKSDVFRDFL AKVINAENAAHKSD KFHTMATRTRQEYLKD AENCVSNTPIDSTGKFNLISLTSKKKEKTKARAGAEQHSAG

AIA RWAQDYAQGVEIDCI GISNEFVΛΠJLDLRTKEWFNCYCGDVIG TPDSSTLK

IFYGRGDHIFLQATEGSVEDIREIVQRLKVMTSG ETVD TLRRNGLGQLGFHVKYDG TVAEVEDYGFAWQAGLRQGSRLVEIC VAWTLTHDQMID LRTSVTV WIIPPFED GTPRRGWPETYD TSEPKTEQESITPGGRPPYRSNAPWQWSGPASHNSLPASK ATP TTPGHAQSLSRPLKQTPIVPFRESQPLHSKRPVSFPΞTPYTVSPAGADRVPPYRQPSG SFSTPGSATYVRYKPSPERYTAAPHP LSLDPHFSHDGTSSGDSSSGGLTSQESTMER QKPEPL HVPAQARLSAIAGSSGNKHPSRQDAAGKDSPNRHSKGEPQYSSHSSSNTLS SNASSSHSDDR FDPLDPLEPEQDPLSKGGSSDSGIDTT YTSSPSCMSLAKAPRPAK PHKPPGSMG CGGGREAAGRSHHADRRREVSPAPAVAGQSKGYRPKLYSSGSSTPTGL AGGSRDPPRQPSDMGSRVGYPAQVYKTASAETPRPSQLAQPSPFQLSASVPKSFFSKQ PVRNKHPTGWKRTEEPPPRPLPFSDPKKQVDTNTKNVFGQPRLRASLRD RSPRKNYK STIEDDLKKLIIMDNLGPEQERDTGQSPQKGLQRTLSDESLCSGRREPSFASPAG EP GLPSDVLFTSTCAFPSSTLPARRQHQHPHPPVGPGATPAAGSGFPEKKSTISASELS ADGRDRPLRRLDPGLMPLPDTAAGLEWSSLVNAAKAYEVQRAVSLFSLNDPALSPDIP PAHSPVHSHLSLERGPPTPRTTPT SEEPPLDLTGKVYQLEVM KQLHTDLQKEKQDK WLQSEVASLRQNNQRLQEESQAASEQLRKFAEIFCREKKEL

Further analysis ofthe NOV23 protein yielded the following properties shown in Table 23B.

Table 23B. Protein Sequence Properties NOV23

PSort 0.6000 probability located in nucleus; 0.3000 probability located in microbody analysis: (peroxisome); 0.1000 probability located in mitochondrial matrix space; 0.1000 probability located in lysosome (lumen)

SignalP No Known Signal Sequence Predicted analysis:

A search ofthe NOV23 protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 23 C.

Table 23C. Geneseq Results for NOV23

NOV23

Geneseq Protein/Organism/Length [Patent #, Identities/

Residues/ Expect Identifier Date] Similarities for the

Match Value Matched Region

Residues

In a BLAST search of public sequence databases, the NOV23 protein was found to have homology to the proteins shown in the BLASTP data in Table 23D.

PFam analysis predicts that the NOV23 protein contains the domains shown in the Table 23E.

Table 23E. Domain Analysis of NOV23

Identities/ Similarities

Pfam Domain NOV23 Match Region Expect Value for the Matched Region

Example A24.

The NOV24 clone was analyzed, and the nucleotide and predicted polypeptide sequences are shown in Table 24A.

Table 24A. NOV24 Sequence Analysis

SEQ LD NO:57 4894 bp

NOV24, TTCCTCAACATCAACGAGACCTTCAAGTTAATGGAGCAGCTTGCCAACATAGCCATGA CG59777-01 GGCAACTCTTAGACAATGAGGGATTTGAACAAGATCGAGATCTTGATGCCAGGGCAAA DNA GAGTGAGAGATACCGTGCACTTTTCCGGCTGCCCAAAGATGAAAAATTAGATGGCCAC Sequence ACAGACTGCACTCTCTGGACTCCATTTAACAAAATGCACATTTTGGGGCAGATGTTTG TGTCCACAAATTACATCTGTTTTACCAGCAAGGAGGAGAACTTATGTAGCCTCATTAT CCCGCTCCGTGAGGTAACAATTGTGGAAAAGGCAGACAGCTCCAGTGTGCTCCCCAGT CCCTTATCCATCAGCACCCGAAACAGGATGACCTTCCTATTTGCCAACTTGAAAGATA GAGACTTTCTAGTGCAGAGGATCTCAGATTTCCTGCAACAGACTACTTCCAAAATATA TTCTGACAAGGAGTTTGCAGGAAGTTACAACAGTTCAGATGATGAGGTGTACTCTCGA CCCAGCAGCCTCGTCTCCTCCAGCCCCCAGAGAAGCACGAGCTCTGATGCTGATGGAG AGCGCCAGTTTAACCTAAATGGCAACAGCGTCCCCACAGCCACACAGACCCTGATGAC CATGTATCGGCGGCGGTCTCCCGAGGAGTTCAACCCGAAATTGGCCAAAGAGTTTCTG AAAGAGCAAGCCTGGAAGATTCACTTTGCTGAGTATGGGCAAGGGATCTGCATGTACC GCACAGAGAAAACGCGGGAGCTGGTGTTGAAGGGCATCCCGGAGAGCATGCGTGGGGA GCTCTGGCTGCTGCTGTCAGGTGCCATCAATGAGAAGGCCACACATCCTGGGTACTAT GAAGACCTAGTGGAGAAGTCCATGGGGAAGTATAATCTCGCCACGGAGGAGATTGAGA GGGATTTACACCGCTCCCTTCCAGAACACCCAGCTTTTCAGAATGAAATGGGCATTGC TGCACTAAGGAGAGTCTTAACAGCTTATGCTTTTCGAAATCCCAACATAGGGTATTGC CAGGCCATGAATATTGTCACTTCAGTGCTGCTGCTTTATGCCAAAGAGGAGGAAGCTT TCTGGCTGCTTGTGGCTTTGTGTGAGCGCATGCTCCCAGATTACTACAACACCAGAGT TGTGGGTGCACTGGTGGACCAAGGTGTCTTTGAGGAGCTAGCACTAGACTACGTCCCA CAGCTGTACGACTGCATGCAAGACCTGGGCGTGATTTCCACCATCTCCCTGTCTTGGT TCCTCACACTATTTCTCAGTGTGATGCCTTTTGAGAGTGCAGTTGTGGTTGTTGACTG TTTCTTCTATGAAGGAATTAAAGTGATATTCCAGTTGGCCCTAGCTGTGCTGGATGCA AATGTGGACAAACTGTTGAACTGCAAGGATGATGGGGAGGCCATGACCGTTTTGGGAA GGTATTTAGACAGTGTGACCAATAAAGACAGCACACTGCCTCCCATTCCTCACCTCCA CTCCTTGCTCAGCGATGATGTGGAACCTTACCCTGAGGTAGACATCTTTAGACTCATC AGAACTTCCTACGAGAAATTCGGAACTATCCGGGCAGATTTGATTGAACAGATGAGAT TCAAACAGAGACTGAAAGTGATCCAGACGCTGGAGGATACTACGAAACGCAACGTGGT ACGAACCATTGTGACAGAAACTTCCTTTACCATTGATGAGCTGGAAGAACTTTATGCT CTTTTCAAGGCAGAACATCTCACCAGCTGCTACTGGGGCGGGAGCAGCAACGCGCTGG ACCGGCATGACCCCAGCCTGCCCTACCTGGAACAGTATCGCATTGACTTCGAGCAGTT CAAGGGAATGTTTGCTCTTCTCTTTCCTTGGGCATGTGGAACTCACTCTGACGTTCTG GCCTCCCGCTTGTTCCAGTTATTAGATGAAAATGGAGACTCTTTGATTAACTTCCGGG AGTTTGTCTCTGGGCTAAGTGCTGCATGCCATGGGGACCTCACAGAGAAGCTCAAACT CCTGTACAAAATGCACGTCTTGCCTGAGCCATCCTCTGATCAAGATGAACCAGATTCT GCTTTTGAAGCAACTCAGTACTTCTTTGAAGATATTACCCCAGAATGTACACATGTTG TTGGATTGGATAGCAGAAGCAAACAGGGTGCAGATGATGGCTTTGTTACGGTGAGCCT AAAGCCAGACAAAGGGAAGAGAGCAAATTCCCAAGAAAATCGTAATTATTTGAGACTG TGGACTCCAGAAAATAAATCTAAGTCAAAGAATGCAAAGGATTTACCCAAATTAAATC AGGGGCAGTTCATTGAACTGTGTAAGACAATGTATAACATGTTCAGCGAAGACCCCAA TGAGCAGGAGCTGTACCATGCCACGGCAGCAGTGACCAGCCTCCTGCTGGAGATTGGG GAGGTCGGCAAGTTGTTCGTGGCCCAGCCTGCAAAGGAGGGCGGGAGCGGAGGCAGTG GGCCGTCCTGCCACCAGGGCATCCCAGGCGTGCTCTTCCCCAAGAAAGGGCCAGGCCA GCCTTACGTGGTGGAGTCTGTTGAGCCCCTGCCGGCCAGCCTGGCCCCCGACAGCGAG GAACACTCCCTTGGAGGACAAATGGAGGACATCAAGCTGGAGGACTCCTCGCCCCGGG ACAACGGGGCCTGCTCCTCCATGCTGATCTCTGACGACGACACCAAGGACGACAGCTC CATGTCCTCATACTCGGTGCTGAGTGCCGGCTCCCACGAGGAGGACAAGCTGCACTGC GAGGACATCGGAGAGGACACGGTCCTGGTGCGGAGCGGCCAGGGCACGGCGGCACTGC CCCGGAGCACCAGCCTGGACCGGGACTGGGCCATCACCTTCGAGCAGTTCCTGGCCTC CCTCTTAACTGAGCCTGCCCTGGTCAAGTACTTTGACAAGCCCGTGTGCATGATGGCC AGGATTACCAGTGCAAAAAACATCCGGATGATGGGCAAGCCCCTCACCTCGGCCAGTG ACTATGAAATCTCGGCCATGTCCGGCTGACACGGGCGCCTTCCCGGGGGAGTGGGAGG AGAGGGAGGGGAGGGATTTTTTATGTTCTTCTGTGTTGAGTTTTTTCTTTCTTTCTTT

TAAATTAAATATTTATTAGTACCTGGCTTGAAGCCTAGTGTTTTCATAATGTAATTCA

ATGAAAACTGTTGGAGAAATATTTAAACACCTCAATGTAGGTACATTACACTCTTGTT

GCGGGGAGGGGATTTACCAGAATACAGTTTATTTCGTGAATTCTAAAAAACAAAAAGA

TGAATCTGTCAGTGATATGTGTGTATTATAACTTATTAATCTTGCTGTTGAGCTGTAT

ACATGGTTTAAAAAATAGTACTGTTTAATGCTAAGTAAGGCAGCAGTCATTTGTGTAT

TCAGGCTTTTTAAATAAAATTAGAGCTGTAAGGAAAATGAAAAGCCACAAATGCAAGA

CTGTTCTTAAATGGAAGGCATAGTCAGCGAGGGTAAATCCTATACCACTTTAGGAAGT

ATTAAAAATATTTTTAAGATTTGAAATATATTTCATAGAAGTCCTCTATTCAAAATCA

TATTCCACAGATGTTCCCCTTCAAAGGGAAAACATTTGGGGTTCTAAACAGTTATGAA

AGTAAGTGATTTTTACATGATTCCAGAATAACACTTGTATTGACCAATTTAGACAGAT

ACCAGACCAATTTTGCATTTAAGAAATTGTTCTGATTATTTACGTCAACTCATTAGAA

TTCAGTGAAAAGTAACAGTCTTTTGTCACAGAGAATCTGAAAGTAGCAGCAAAGACAG

AGGGCTCATGACAGGTTTTTGCTTTTGCTTTGCTTTTGTTTTTGAAAGAGTAAAAGTA

CTGATGCTTCTGATACTGGATGTTTAGCTTCTTACTGCAAAAACATAAGTAAAACAGT

CAACTTTACCATTTCCGTATTCTCCATAGATTGAAGAAATTTATACCACATATCGCAT

ATGACCATCTTTCCATCAAATCAATGTAGAGATAATGTAAACTGAAAAAAAATCTGCA

AGATAATGTAACTGAATGTTTTAAAAACAGAACTTGTCACTTTATATAAAAGAATAGT

ATGCTCTATTTCCTGAATGGATGTGGAAATGAAAGCTAGCGCACCTGCACTTTGAATT

CTTGCTTCTTTTTTATTACTGTTATGATTTTGCTTTTTACAGATGTTGGACGATTTTT

TCTTCTGATTGTTGAATTCATAATCATGGTCTCATTTCCTTTGCTTCTTTGGAATATT

TCTTTCAACACATTCCTTTATTTTATTATACATTGTGTCCTTTTTTTAGCTATTGCTG

CTGTTGTTTTTTATTCTATTTACAGGATGATTTTTAAACTGTCAAATGAAGTAGTGTT

AACCTCAAATAGGCTAAATGTGAACAAATAAAATACAGCAAATACTCAGATACAGCTT

TTTATCTTTGTGCTTGAGTTCCTGCCTAAGGCAATAACATTATTCTTTTGACAACTTT

TGCAGGGGAAATTATATCAGGCAACCATTTTGATTAAGTAAATAAATTTTATAGGCAA

ACATATAGAGAGATATACAATTTGTAGTATATCAATGACTATATTTATAATAAGGAAT

ATAATTGTTATCAGTTATCTAACTTAAAATGCTTATCCATAATGATCAGTGATATTCA

GCTTTTTAAAATATGCTTGTTGGTTGCATGTCTGTCTTCATATCCACATTGAGGATTC

CATCTCACACCTAGTTCATTAAATAGGGCATATTAGTTTCAGATGTTTGTCGTGGTTT

GTTTAGGTTTACTACACATATTTTCCGTTTGTGGGGAGTTGTTCCTTTGTTGCATCCA

TTGTTATTAAGGCTATGTGGGC

ORF Start: ATG at 31 ORF Stop: TGA at 3043

SEQ LD N0.58 1004 aa MW at ll3034.5kD

NOV24, MEQ A IAMRQ LDNEGFEQDRD DARAKSERYRALFRLPKDEKLDGHTDCTL TPFN CG59777-01 HILGQMFVSTNYICFTSKEENLCSLIIPLREVTIVEKADSSSVLPSPLSISTRNRM Protein TFLFANLKDRDFLVQRISDFLQQTTSKIYSDKEFAGSYNSSDDEVYSRPSSLVSSSPQ Sequence RSTSSDADGERQFN NGNSVPTATQTLMTMYRRRSPEEFNPKLAKEFLKEQAWKIHFA EYGQGICMYRTEKTRELVLKGIPESMRGELW LLSGAINEKATHPGYYED VEKSMGK YNLATEEIERDLHRSLPEHPAFQNEMGIAALRRVLTAYAFRNPNIGYCQAMNIVTSVL LLYAKEEEAF LLVA CERMLPDYYNTRWGALVDQGVFEELALDYVPQLYDC QDLG VISTISLSWFLTLFLSVMPFESAWWDCFFYEGIKVIFQ ALAVLDAVDKLLNCKD DGEAMTVLGRYLDSVTNKDST PPIPHLHS LSDDVEPYPEVDIFRLIRTSYE FGTI RADLIEQMRFKQRLKVIQTLEDTTKRNWRTIVTETSFTIDELEELYA FKAEHLTSC Y GGSSNALDRHDPSLPYLEQYRIDFEQFKGMFALLFP ACGTHSDVLASRLFQL DE NGDSLINFREFVSG SAACHGD TEKLKLLYKMHVLPEPSSDQDEPDSAFEATQYFFE DITPECTHWGLDSRSKQGADDGFVTVSLKPDKGKRA SQENRNYLRLWTPENKSKSK NAKDLPKLNQGQFIELCKTMY MFSEDPNEQELYHATAAVTSLL EIGEVGKLFVAQP

A search ofthe NOV24 protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 24C.

In a BLAST search of public sequence databases, the NOV24 protein was found to have homology to the proteins shown in the BLASTP data in Table 24D.

PFam analysis predicts that the NOV24 protein contains the domains shown in the Table 24E.

Example A25.

The NOV25 clone was analyzed, and the nucleotide and predicted polypeptide sequences are shown in Table 25A.

Table 25A. NOV25 Sequence Analysis

SEQ LD NO:59 1916 bp

NOV25, AAGCAAGTCCTTGGATTTACTGGGTTTTACTTGGCTTTAATGGGAGGAGACAGTAAGA CG59658-01 AAAATACAGAACTGAGACCATTTCAGATAGAGATAAGCACTATGAGGGAAGCAAAGCT DNA GCGTGTGTGCGGCAGGGGGCGTGGGCGCGGCGGCGCGGGCGGAGCGGCGCGGCCCGGG Sequence CGCGGCGTGGGTAGAGCCGAGGCGGCGGCGGCGGCCGGCCTCTATCAGGATATCTACC CTGCAGTACACACACTCACACAGGCACAAACACACACCACCCTCTGTCTCTCACATAC ACTCACTCTCACACACACACACCCTCCCCCACACACTCTCACACACACACTTTCTCAC ACACTCACACACACACTCTCACACACACTCTCTCACCCCCACACACTCACACACACTC ACCCCACACACTCTCACACTCACTCTCACACACACTCACCCTACACTCCCGACTCCCC TCAGACCCCCCCTCCTCTGGAGGAGACCCACATCTCATGCCTGTTCCCGGAGCTGCTG GCCATGATCTTCGGCTACCTGGACGTCCGGGACAAGGGGCGCGCGGCGCAGGTGTGCA CCGCCTGGCGGGACGCCGCCTACCACAAGTCGGTGTGGCGGGGGGTGGAGGCCAAGCT GCACCTGCGCCGGGCCAACCCGTCGCTGTTCCCCAGCCTGCAGGCCCGGGGCATCCGC CGGGTGCAGATCCTGAGCCTCCGCCGCAGCCTCAGCTACGTGATCCAGGGCATGGCCA ACATCGAGAGCCTCAACCTCAGCGGCTGCTACAACCTCACCGACAACGGGCTGGGCCA CGCGTTTGTGCAGGAGATCGGCTCCCTGCGCGCTCTCAACCTGAGCCTCTGCAAGCAG ATCACTGACAGCAGCCTGGGCCGCATAGCCCAGTACCTCAAGGGCCTGGAGGTGCTGG AGCTGGGAGGTTGCAGCAACATCACCAACACTGGCCTTCTGCTCATCGCCTGGGGTCT GCAGCGCCTCAAGAGCCTTAACCTCCGCAGCTGCCGCCACCTTTCGGATGTGGGCATC GGGCACCTGGCCGGCATGACGCGCAGCGCGGCGGAGGGCTGCCTGGGCCTGGAGCAGC TCACGCTACAGGACTGCCAGAAGCTCACAGATCTTTCTCTAAAGCACATCTCCCGAGG GCTGACGGGCCTGAGGCTCCTCAACCTCAGCTTCTGTGGGGGAATCTCGGACGCTGGC CTCCTGCACCTGTCGCACATGGGCAGCCTGCGCAGCCTCAACCTGCGCTCCTGTGACA ACATCAGTGACACGGGCATCATGCATCTGGCCATGGGCAGCCTGCGCCTCTCGGGGCT GGATGTTTCGTTCTGTGACAAGGTGGGAGACCAGAGTCTGGCTTACATAGCCCAGGGG CTGGATGGCCTCAAGTCTCTCTCCCTCTGCTCCTGCCACATCAGTGATGATGGCATCA ACCGCATGGTGCGGCAGATGCACGGGCTGCGCACGCTCAACATTGGACAGTGTGTGCG CATCACGGACAAGGGCCTGGAGCTGATCGCTGAGCACCTGAGCCAACTCACCGGCATA GACCTGTACGGCTGCACCCGAATCACCAAGCGCGGCCTGGAGCGCATCACGCAGCTGC CGTGCCTCAAGGTACTCAACCTGGGACTCTGGCAGATGACGGACAGTGAGAAGGTCAG GTGAGGGCGGCAGCACCAGCTCCCCTTGTCCCGCCCTGTTCATCCTCCCATTACCACC GCCCCCACACACTCACACGCACACTTACGCACAGATCATTGCAGCGGATGAGATGGGG

CTATGACAGAAGCCTCAGGCTCGTTTCCTCCTCCCTCCTCCAGCCCCCTCCCGGCTTC

CACTTAGATCTGCAGCCCTACCCACAACCACCTAAGCCTACTACCAGCTCCTTTTACA

CG

ORF Start: ATG at 40 ORF Stop: TGA at 1684

SEQ 1DNO.60 548 aa MW at 59767.0kD

NOV25, MGGDSKKNTELRPFQIEISTMREAKLRVCGRGRGRGGAGGAARPGRGVGRAEAAAAAG CG59658-01 LYQDIYPAVHTLTQAQTHTTLCLSHTLTLTHTHPPPHTLTHTLSHTLTHTLSHTLSHP Protein HT THTHPTHSHTHSHTHSPYTPDSPQTPPPLEETHISCLFPEL AMIFGYLDVRDKG Sequence RAAQVCTAWRDAAYHKSVWRGVEAKLHLRRA PS FPSLQARGIRRVQILSLRRSLSY VIQG A IESLNLSGCYN TDNGLGHAFVQEIGS RA N SLCKQITDSS GRIAQY KGLEVLELGGCSNITNTG L IA GLQRLKS NLRSCRHLSDVGIGHLAGMTRSAAEG CLGLEQ TLQDCQKLTDLSLKHISRGLTGLRL NLSFCGGISDAGLLHLSHMGS RSL NLRSCDNISDTGIMHLAMGSLRLSG DVSFCD VGDQSLAYIAQG DGLKSLSLCSCH ISDDGINRMVRQMHGLRTLNIGQCVRITDKGLE IAEH SQ TGIDLYGCTRITKRGL ERITQ PCLKVLNLGL QMTDSEKVR

Further analysis ofthe NOV25 protein yielded the following properties shown in Table 25B.

Table 25B. Protein Sequence Properties NOV25

PSort 0.4500 probability located in cytoplasm; 03000 probability located in microbody analysis: (peroxisome); 0.2360 probability located in lysosome (lumen); 0.1000 probability located in mitochondrial matrix space

SignalP No Known Signal Sequence Predicted analysis: A search ofthe NOV25 protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 25C.

In a BLAST search of public sequence databases, the NOV25 protein was found to have homology to the proteins shown in the BLASTP data in Table 25D.

PFam analysis predicts that the NOV25 protein contains the domains shown in the Table 25E.

Example A26.

The NOV26 clone was analyzed, and the nucleotide and predicted polypeptide sequences are shown in Table 26A.

Further analysis ofthe NOV26 protein yielded the following properties shown in Table 26B.

Table 26B. Protein Sequence Properties NOV26

PSort 0.4500 probability located in cytoplasm; 0.3866 probability located in microbody analysis: (peroxisome); 0.1564 probability located in lysosome (lumen); 0.1000 probability located in mitochondrial matrix space

SignalP No Known Signal Sequence Predicted analysis:

A search ofthe NOV26 protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 26C.

Table 26C. Geneseq Results for NOV26

Geneseq Protein/Organism/Length [Patent #, NOV26 Identities/ Expect Identifier Date] Value

In a BLAST search of public sequence databases, the NOV26 protein was found to have homology to the proteins shown in the BLASTP data in Table 26D.

PFam analysis predicts that the NOV26 protein contains the domains shown in the Table 26E. Table 26E. Domain Analysis of NOV26

Identities/ Similarities

Pfam Domain NOV26 Match Region { Expect Value for the Matched Region

TBC: domain 1 of l 98.315 60/343 (17%) 2.3e-09 133/343 (39%)

Example A27.

The NOV27 clone was analyzed, and the nucleotide and predicted polypeptide sequences are shown in Table 27A.

Table 27A. NOV27 Sequence Analysis

SEQ LD NO:63 5973 bp

NOV27, TGGACGGCATCTTTTTGAACCAAATTATGCTGCAAATAGATCCCAGGCCCACAAATCA CG59903-01 ACGCATCAATAAGCACGTCAACAATGATGTGAACCTTCGCATTCAGAATTTGACCATC DNA TTGGTGAGAAACATTAAGACCTACTACCAGGAAGTTCTCCAGCAGCTGATTGTAATGA Sequence ATTTGCCCAATGTTTTGATGATTGGCAGAGACCCACTGTCTGGTGAGAGCATGGAGGA AATCAAGAAGGTGCTGCTGCTGGTGCTGGGCTGTGCTGTCCAGTGTGAGAGGAAAGAG GAGTTCATTGAAAGAATCAAACAGCTGGACATTGAGACCCAGGCTGGCATCGTGGCCC ATATCCAGGAGGTAACTCACAACCAAGAGAACGTGTTTGACCTGCAGTGGCTGGAGCT GCCCGACGTGGCTCCGGAGGAGCTGGAGGCCCTGTCGAGGAGCATGGTGCTCCACCTG CGGAGGCTCATCGACCAGCGGGACGAGTGCACCGAGCTGATCGTGGACCTCACTCAGG AACGGGACTACCTGCAGGCACAGCATCCACCCAGCCCCATCAAGTCCTCCAGCGCCGA CTCCACTCCCAGCCCCACCAGCAGCCTCTCTAGCGAAGACAAGCAGCACCTGGCCGTA GAGCTGGCCGACACCAAGGCCAGGCTGCGGCGCGTCAGGCAGGAGCTGGAGGATAAGA CAGAGCAGCTTGTGGACACCAGACATGAGGTGGACCAGCTGGTGCTGGAACTGCAGAA AGTTAAGCAGGAGAACATCCAGCTAGCGGCAGACGCCCGGTCTGCTCGTGCCTATCGA GACGAGCTGGATTCCCTGCGGGAGAAGGCGAACCGCGTGGAGAGGCTGGAGCTGGAGC TGACCCGCTGCAAGGAGAAGCTGCACGACGTGGACTTCTACAAGGCCCGCATGGAGGA GCTGAGAGAAGATAATATCATTTTAATTGAAACCAAGGCCATGCTGGAGGAACAGCTG ACTGCTGCTCGGGCCCGGGGCGATAAAGTCCATGAGCTGGAAAAGGAGAACCTGCAGC TGAAATCCAAGCTTCACGACCTGGAATTGGACCGGGACACAGATAAGAAACGAATTGA GGAGCTGCTGGAAGAAAACATGGTCCTTGAGATTGCACAGAAGCAGAGCATGAACGAA TCTGCCCACCTTGGCTGGGAGCTGGAGCAGCTGTCCAAGAACGCAGACTTGTCATCAG CCTCCAGGAAGTCGTTTGTGTTTGAGCTGAACGAATGTGCGTCCAGCCGCATCCTGAA GCTGGAGAAGGAGAATCAGAGCCTCCAGAGCACCATCCAGGGGCTGCGGGACGCGTCC CTGGTGTTGGAGGAGAGCGGCCTCAAGTGCGGGGAGCTGGAGAAGGAGAACCACCAGC TCAGCAAGAAGGTAGAAAAGTTACAAACCCAGCTGGAGAGAGAAAAGCAGAGCAACCA AGATCTGGAGACCCTCAGTGAGGAGCTGATCAGAGAGAAGGAGCAGCTGCAGAGTGAC ATGGAGACCCTGAAGGCTGACAAAGCCAGGCAGATCAAGGACCTTGAGCAGGAAAAGG ACCACCTCAACCGAGCCATGTGGTCGCTGCGGGAGAGGTCGCAGGTCAGCAGTGAGGC CCGCATGAAAGACGTGGAGAAGGAGAACAAAGCCCTCCACCAGACGGTGACGGAGGCC AATGGCAAGCTCAGCCAGTTGGAGTTTGAGAAGCGGCAGCTGCACAGGGACTTGGAGC AGGCCAAGGAGAAGGGGGAGCGGGCAGAGAAGCTGGAGAGGGAGCTACAGCGACTCCA GGAGGAGAACGGGAGGCTGGCCAGGAAGGTGACCTCCCTGGAGACAGCCACCGAGAAA GTCGAGGCCCTGGAGCATGAGAGCCAGGGCCTGCAGCTGGAGAACCGGACTCTGAGGA AGTCTCTGGACACCTTGCAGAACGTGTCCCTGCAGCTTGAGGGCCTGGAGCGTGACAA CAAGCAGCTGGACGCAGAGAACCTGGAGCTGCGCAGGCTGGTGGAGACCATGCGCTTC ACCAGCACCAAGCTGGCACAGATGGAGAGGGAGAACCAGCAGCTGGAGCGTGAGAAGG AGGAGCTGAGGAAGAACGTGGATCTGCTCAAGGCGCTGGGCAAGAAGTCAGAGCGCCT GGAGCTCAGCTACCAGAGCGTGAGCGCTGAGAACCTCCGGCTGCAGCAGAGCCTGGAG AGCAGCAGCCACAAGACGCAGACCTTGGAGAGTGAGCTGGGCGAGCTGGAGGCTGAGC GCCAGGCGCTGCGGCGGGACCTGGAGGCCCTCCGGCTGGCCAATGCACAGTTGGAGGG GGCCGAGAAGGACAGGAAGGCCCTGGAGCAGGAGGTGGCCCAGCTCGAGAAGGATAAG AAGCTGCTGGAGAAGGAGGCCAAGCGGCTGTGGCAGCAGGTGGAGCTCAAGGATGCAG TCTTGGACGATAGCACTGCCAAACTGTCCGCCGTTGAGAAGGAGAGCCGCGCGCTGGA CAAGGAGCTGGCCCGCTGCAGGGACGCAGCCGGCAAGCTGAAGGAGCTGGAGAAGGAC AACCGGGACCTCACCAAGCAAGTCACCGTGCATGCAAGGACACTGACAACTCTGAGGG AGGACCTGGTGCTCGAGAAGCTGAAGAGCCAGCAGCTCAGCAGTGAGCTGGACAAGCT GAGCCAGGAACTGGAGAAGGTCGGCCTCAACAGGGAGCTGCTGTTGCAGGAGGACGAC AGCGGCAGTGACACAAAATACAAGATTTTGGAGGGCAGAAATGAATCAGCATTAAAAA CAACACTAGCCATGAAAGAAGAAAAGATTGTGCTCTTAGAAGCACAGATGGAAGAGAA AGCGAGCCTAAATCGCCAGTTAGAGAGTGAGCTGCAGATGCTAAAGAAGGAGTGTGAG ACCCTCAGGCAGAACCAGGGAGAGGGGCAGCACTTGCAGAACTCTTTCAAGCACCCTG CGGGGAAGACAGCCGCCAGTCACCAGGGGAAGGAGGCCTGGGGGCCCGGCCATAAGGA AGCCACCATGGAGCTTCTCCGAGTGAAGGACCGGGCCATCGAGCTGGAGCGGAATAAT GCAGCTCTGCAGGCTGAGAAGCAGCTGCTAAAGGAACAGCTGCAGCACCTGGAGACCC AGAACGTGACCTTCAGCAGCCAGATCTTGACACTGCAGAAACAGAGCGCCTTCCTGCA GGAGCACAACACCACACTGCAGACCCAGACCGCCAAGCTGCAGGTGGAGAACTCCACG CTGAGTTCCCAGAGCGCAGCGCTCACCGCGCAGTACACGCTGCTGCAGAACCACCACA CGGCCAAGGAGACGGAGAACGAAAGCCTGCAGAGGCAGCAGGAGCAACTTACAGCGGC CTACGAGGCCCTGCTGCAGGACCACGAGCACCTGGGCACGCTGCACGAGCGGCAATCG GCCGAGTACGAGGCCCTCATCCGCCAGCACAGCTGCCTAAAGACACTGCATCGGAATC TGGAGCTGGAGCACAAGGAGCTCGGGGAGAGGCACGGTGACATGCTGAAGCGCAAGGC GGAGCTGGAGGAGCGGGAGAAGGTCTTGACCACTGAGCGAGAGGCGCTGCAGCAGGAG CAGAGGACAAACGCCCTCGCCATGGGCGAGAACCAGAGGCTGCGGGGCGAGCTGGACA GGGTCAATTTCCTGCACCACCAGCTGAAGGGGGAGTACGAGGAGCTGCACGCCCACAC CAAGGAGCTGAAAACCTCACTGAACAACGCGCAGCTGGAGCTCAACCGCTGGCAGGCC CGCTTCGACGAGCTGAAGGAGCAGCACCAGACCATGGACATCTCGCTGACCAAGCTGG ACAACCACTGTGAGCTGCTCTCCCGTCTCAAGGGGAACTTGGAGGAAGAAAATCATCA CCTCCTGAGCCAGATCCAGCTGTTGAGCCAGCAGAACCAGATGCTTCTGGAGCAGAAC ATGGAGAACAAGGAGCAGTACCATGAGGAGCAGAAGCAGTACATAGACAAATTAAATG CCTTACGAAGACATAAGGAAAAGCTGGAAGAAAAAATCATGGATCAATACAAGTTCTA TGATCCTCCTCCAAAGAAGAAGAACCACTGGATTGGAGCCAAAGCCTTAGTCAAACTC ATCAAACCAAAGAAAGAGGGTTCGAGGGAACGCTTAAAATCCACCGTGGACAGCCCTC CCTGGCAGCTGGAGTCCTCAGACCCCGCCTCGCCGGCGGCCTCTCAGCCGCTCAGATC ACAGGCCGAGAACCCCGACACCCCCGCACTGGGCTCCAACTGTGCAGAAGAGCGCGAC GCCCACAACGGGTCTGTGGGGAAAGGCCCTGGGGATCTAAAACCAAAGCGAGGCTCCC CACACAGAGGCAGCCTTGACCGCACAGATGCCTCCACCGATCTGGCCATGAGGTCCTG GCCCTCGGAGCTGGGCTCCCGGACTTGCTCAACTTCAGCCACCACTACAGCCCCTTCC AACTCCACCCCCATCGCCCGGCACCCAGGCCGCACCAAAGGCTATAACTCAGATGACA ACCTCTGTGAGCCATCCCTGGAGTTTGAGGTCCCCAACCACAGGCAGTACGTGTCGCG GCCAAGTAGCTTAGAGAGCAGTAGAAACACATCCAGCAACAGCTCACCTCTTAACCTA AAAGGCTCCTCCGAGCAGCTCCATGGCCGGTCTGAGAGCTTCAGCAGCGAAGACCTGA TCCCCAGCAGGGACCTGGCCACTTTGCCCCGGGAAGCCAGCACACCGGGACGCAACGC CCTCGGCCGCCACGAGTACCCCTTGCCTCGGAACGGGCCTCTCCCACAGGAGGGTGCC CAGAAGAGGGGCACAGCCCCTCCCTACGTCGGAGTGCGGCCCTGCTCGGCCTCCCCCA GCAGTGAGATGGTCACCTTGGAGGAGTTCCTGGAGGAGAGCAACCGCAGCTCCCCCAC CCATGACACTCCCAGTTGCCGGGATGACCTGCTGAGTGACTACTTCCGAAAGGCCAGC GATCCCCCAGCCATCGGAGGCCAACCAGGACCACCTGCCAAGAAAGAAGGGGCCAAGA TGCCCACCAACTTTGTGGCCCCCACCGTCAAAATGGCCGCCCCCACCTCGGAGGGGAG GCCGCTGAAGCCCGGGCAGTACGTAAAGCCAAACTTCAGACTGACTGAGGCCGAGGCC CCACCCAGCGTGGCCCCGAGACAGGCCCAGCCTCCCCAGAGCCTGTCTCTGGGCAGAC CCCGGCAGGCTCCGGTGCCCCCAGCTTCCCATGCACCTGCCAGCCGCAGTGCCTCCTT GAGCCGGGCCTTCAGCCTGGCCTCAGCTGACCTTCTCCGGGCCAGCGGGCCAGAGGCC TGCAAACAGGAGTCCCCTCAGAAGCTGGGGGCTCCTGAGGCCTTAGGGGGCAGAGAGA CAGGCAGCCACACCCTGCAAAGCCCCGCACCCCCCAGCTCCCATAGCCTGGCCCGGGA GCGGACCCCACTTGTGGGAAAGGCTGGCAGCTCCTGTCAGGGCCCAGGTCCCCGCAGC CGGCCGCTGGACACGAGGCGCTTCTCCCTGGCTCCCCCAAAGGAGGAGAGGCTGGCCC CCCTGCATCAGTCTGCCACAGCCCCCGCCATTGCCACTGCAGGTGCTGGTGCTGCTGC TGCTGGCAGTGGCAGCAACTCCCAGCTCCTGCACTTCTCACCTGCTGCAGCCCCGGCT GCCAGGACCAAGCCCAAGGCGCCCCCACGCTCAGGGGAGGTGGCCACCATCACCCCTG TCCGGGCAGGGCTCAGCCTCTCAGAGGGAGACGGGGTCCCGGGGCAGGGCTGCAGTGA GGGGCTTCCGGCCAAGAGCCCAGGTCGGTCTCCCGATTTGGCTCCCCACCTCGGCCGG GCCCTGGAGGACTGCAGTCGAGGGAGCGTCTCAAAGAGCAGTCCGGCCTCCCCGGAGC CCGGCGGGGATCCGCAGACCGTGTGGTATGAGTACGGCTGTGTGTGACTGTCTCGTG

ORF Start: ATG at27 ORF Stop: TGA at 5961

SEQLDN0.64 1978 aa MW at 222593.2kD

NOV27, M QIDPRPTNQRINKHVNNDVN RIQNLTILVRNIKTYYQEV QQLIVMNLPNVLMIG CG59903-01 RDPLSGESMEEIKKV LLVLGCAVQCER EEFIERIKQ DIETQAGIVAHIQEVTHNQ Protein ENVFDLQWLELPDVAPEELEALSRSMVLHLRRLIDQRDECTELIVDLTQERDYLQAQH Sequence PPSPIKSSSADSTPSPTSSLSSEDKQH AVE ADTKARLRRVRQELEDKTEQ VDTRH EVDQLVLELQ VKQENIQLAADARSARAYRDELDSLREKANRVERLELELTRCKEKLH DVDFYKARMEELREDNIILIETKAMLEEQLTAARARGDKVHE EKENLQLKSK HDLE LDRDTDKKRIEELLEENMVLEIAQKQSMNESAHLGWELEQLSKNADLSSASRKSFVFE LNECASSRILKLEKENQSLQSTIQGLRDASLVIIEESGLKCGELEKENHQLSKIVEKLQ TQLEREKQSNQD ETLSEELIREKEQLQSDMETLKADKARQIKDLEQEKDHLNRAMWS LRERSQVSSEARMKDVEKENKALHQTVTEANGKLSQLEFEKRQLHRDLEQAKEKGERA EKLERELQRLQEENGRLARKVTSLETATEKVEALEHESQGLQLENRTLRKSLDTLQNV SLQLEGLERDNKQLDAENLELRRLVETMRFTSTKLAQMERENQQLEREKEELRKNVDL LKALGKKSERLELSYQSVSAENLRLQQSLESSSHKTQTLESΞLGELEAERQALRRDLE ALR ANAQLEGAEKDRKALEQEVAQ EKDKKLLEKEAKRL QQVELKDAVLDDSTAKL SAVEKESRALDKELARCRDAAGKLKELEKDNRDLTKQVTVHARTLTTLREDLVLEKLK SQQLSSELDKLSQELEKVGLNRELLLQEDDSGSDTKYKILEGRNESALKTTLAMKEEK IVLLEAQMEEKASLNRQLESELQMLKKECETLRQNQGEGQHLQNSFKHPAGKTAASHQ GKEA GPGHKEATMELLRVKDRAIELER NAA QAEKQL KEQLQHLETQNVTFSSQI LTLQKQSAFLQEHNTTLQTQTAKLQVENSTLSSQSAALTAQYTLLQNHHTAKETENES LQRQQEQLTAAYEALLQDHEHLGTLHERQSAEYEALIRQHSCLKTLHRNLELEHKELG ERHGDMLKRKAELEEREKVLTTEREA QQEQRTNALAMGENQRLRGELDRV FLHHQL KGEYEE HAHTKELKTSLNNAQ E NRWQARFDELKEQHQTMDISLTKLDNHCELLSR LKGNLEEENHHLLSQIQLLSQQNQMLLΞQNMENKEQYHEΞQKQYIDKLNALRRHKEKL EEKIMDQYKFYDPPPKKK HWIGAKALVK IKPKKEGSRERLKSTVDSPP QLESSDP ASPAASQPLRSQAENPDTPALGSNCAEERDAHNGSVGKGPGDLKPKRGSPHRGSLDRT DASTDLAMRS PSELGSRTCSTSATTTAPSNSTPIARHPGRTKGYNSDDNLCEPS EF EVPNHRQYVSRPSSLESSRNTSSNSSPLNLKGSSEQLHGRSESFSSEDLIPSRDLATL PREASTPGRNALGRHEYPLPRNGPLPQEGAQKRGTAPPYVGWPCSASPSSEMVTI.EE FLEESNRSSPTHDTPSCRDDLLSDYFRKASDPPAIGGQPGPPAKKEGAKMPTNFVAPT VKMAAPTSEGRPLKPGQYVKPNFRLTEAEAPPSVAPRQAQPPQSLSLGRPRQAPVPPA SHAPASRSASLSRAFSLASADLLRASGPEACKQESPQKLGAPEALGGRETGSHTLQSP APPSSHSLARERTPLVGKAGSSCQGPGPRSRPLDTRRFSLAPPKEERLAPLHQSATAP AIATAGAGAAAAGSGSNSQLLHFSPAAAPAARTKPKAPPRSGEVATITPVRAGLSLSE GDGVPGQGCSEGLPAKSPGRSPDLAPHLGRALEDCSRGSVSKSSPASPEPGGDPQTV

YEYGCV

Further analysis ofthe NOV27 protein yielded the following properties shown in Table 27B.

Table 27B. Protein Sequence Properties NOV27

PSort 0.9600 probability located in nucleus; 0.3000 probability located in microbody analysis: (peroxisome); 0.1000 probability located in mitochondrial matrix space; 0.1000 probability located in lysosome (lumen)

SignalP No Known Signal Sequence Predicted analysis:

A search ofthe NOV27 protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 27C.

In a BLAST search of public sequence databases, the NOV27 protein was found to have homology to the proteins shown in the BLASTP data in Table 27D.

PFam analysis predicts that the NOV27 protein contains the domains shown in the Table 27E.

Example A28.

The NOV28 clone was analyzed, and the nucleotide and predicted polypeptide sequences are shown in Table 28A.

IHKNQEYKLDCCLSERDTHWSCSEDGKVFF DLVEGALALALPVGSGWQSLAYHPTE PCLLTAMGGSVQC REEAYEAEDGAG

Further analysis ofthe NOV28 protein yielded the following properties shown in Table 28B.

Table 28B. Protein Sequence Properties NOV28

PSort 0.6500 probability located in cytoplasm; 0.1000 probability located in mitochondrial analysis: matrix space; 0.1000 probability located in lysosome (lumen); 0.0000 probability located in endoplasmic reticulum (membrane)

SignalP No Known Signal Sequence Predicted analysis:

A search ofthe NOV28 protein against the Geneseq database, a proprietary database that contains sequences published in patents and patent publication, yielded several homologous proteins shown in Table 28C.

In a BLAST search of public sequence databases, the NOV28 protein was found to have homology to the proteins shown in the BLASTP data in Table 28D.

PFam analysis predicts that the NOV28 protein contains the domains shown in the Table 28E.

Example B: Sequencing Methodology and Identification of NOVX Clones

1. GeneCalling™ Technology: This is a proprietary method of performing differential gene expression profiling between two or more samples developed at CuraGen and described by Shimkets, et al., "Gene expression analysis by transcript profiling coupled to a gene database query" Nature Biotechnology 17:198-803 (1999). cDNA was derived from various human samples representing multiple tissue types, normal and diseased states, physiological states, and developmental states from different donors. Samples were obtained as whole tissue, primary cells or tissue cultured primary cells or cell lines. Cells and cell lines may have been treated with biological or chemical agents that regulate gene expression, for example, growth factors, chemokines or steroids. The cDNA thus derived was then digested with up to as many as 120 pairs of restriction enzymes and pairs of linker-adaptors specific for each pair of restriction enzymes were ligated to the appropriate end. The restriction digestion generates a mixture of unique cDNA gene fragments. Limited PCR amplification is performed with primers homologous to the linker adapter sequence where one primer is biotinylated and the other is fluorescently labeled. The doubly labeled material is isolated and the fluorescently labeled single strand is resolved by capillary gel electrophoresis. A computer algorithm compares the electropherograms from an experimental and control group for each ofthe restriction digestions. This and additional sequence-derived information is used to predict the identity of each differentially expressed gene fragment using a variety of genetic databases. The identity ofthe gene fragment is confirmed by additional, gene- specific competitive PCR or by isolation and sequencing ofthe gene fragment.

2. SeqCalling™ Technology: cDNA was derived from various human samples representing multiple tissue types, normal and diseased states, physiological states, and developmental states from different donors. Samples were obtained as whole tissue, primary cells or tissue cultured primary cells or cell lines. Cells and cell lines may have been treated with biological or chemical agents that regulate gene expression, for example, growth factors, chemokines or steroids. The cDNA thus derived was then sequenced using CuraGen's proprietary SeqCalling technology. Sequence traces were evaluated manually and edited for corrections if appropriate. cDNA sequences from all samples were assembled together, sometimes including public human sequences, using bioinformatic programs to produce a consensus sequence for each assembly. Each assembly is included in CuraGen Corporation's database. Sequences were included as components for assembly when the extent of identity with another component was at least 95% over 50 bp. Each assembly represents a gene or portion thereof and includes information on variants, such as splice forms single nucleotide polymorphisms (SNPs), insertions, deletions and other sequence variations.

3. PathCalling™ Technology:

The NOVX nucleic acid sequences are derived by laboratory screening of cDNA library by the two-hybrid approach. cDNA fragments covering either the full length ofthe DNA sequence, or part ofthe sequence, or both, are sequenced. In silico prediction was based on sequences available in CuraGen Corporation's proprietary sequence databases or in the public human sequence databases, and provided either the full length DNA sequence, or some portion thereof. The laboratory screening was performed using the methods summarized below: cDNA libraries were derived from various human samples representing multiple tissue types, normal and diseased states, physiological states, and developmental states from different donors. Samples were obtained as whole tissue, primary cells or tissue cultured primary cells or cell lines. Cells and cell lines may have been treated with biological or chemical agents that regulate gene expression, for example, growth factors, chemokines or steroids. The cDNA thus derived was then directionally cloned into the appropriate two- hybrid vector (Gal4-activation domain (Gal4-AD) fusion). Such cDNA libraries as well as commercially available cDNA libraries from Clontech (Palo Alto, CA) were then transferred from E. coli into a CuraGen Corporation proprietary yeast strain (disclosed in U. S. Patents 6,057,101 and 6,083,693, incorporated herein by reference in their entireties).

Gal4-binding domain (Gal4-BD) fusions of a CuraGen Corporation proprietary library of human sequences was used to screen multiple Gal4-AD fusion cDNA libraries resulting in the selection of yeast hybrid diploids in each of which the Gal4-AD fusion contains an individual cDNA. Each sample was amplified using the polymerase chain reaction (PCR) using non-specific primers at the cDNA insert boundaries. Such PCR product was sequenced; sequence traces were evaluated manually and edited for corrections if appropriate. cDNA sequences from all samples were assembled together, sometimes including public human sequences, using bioinformatic programs to produce a consensus sequence for each assembly. Each assembly is included in CuraGen Corporation's database. Sequences were included as components for assembly when the extent of identity with another component was at least 95% over 50 bp. Each assembly represents a gene or portion thereof and includes information on variants, such as splice forms single nucleotide polymorphisms (SNPs), insertions, deletions and other sequence variations. Physical clone: the cDNA fragment derived by the screening procedure, covering the entire open reading frame is, as a recombinant DNA, cloned into pACT2 plasmid (Clontech) used to make the cDNA library. The recombinant plasmid is inserted into the host and selected by the yeast hybrid diploid generated during the screening procedure by the mating of both CuraGen Corporation proprietary yeast strains N106' and YULH (U. S. Patents 6,057,101 and 6,083,693).

4. RACE: Techniques based on the polymerase chain reaction such as rapid amplification of cDNA ends (RACE), were used to isolate or complete the predicted sequence ofthe cDNA ofthe invention. Usually multiple clones were sequenced from one or more human samples to derive the sequences for fragments. Various human tissue samples from different donors were used for the RACE reaction. The sequences derived from these procedures were included in the SeqCalling Assembly process described in preceding paragraphs.

5. Exon Linking: The NOVX target sequences identified in the present invention were subjected to the exon linking process to confirm the sequence. PCR primers were designed by starting at the most upstream sequence available, for the forward primer, and at the most downstream sequence available for the reverse primer. Table Bl shows the sequences ofthe PCR primers used for obtaining different clones. In each case, the sequence was examined, walking inward from the respective termini toward the coding sequence, until a suitable sequence that is either unique or highly selective was encountered, or, in the case ofthe reverse primer, until the stop codon was reached. Such primers were designed based on in silico predictions for the full length cDNA, part (one or more exons) ofthe DNA or protein sequence ofthe target sequence, or by translated homology ofthe predicted exons to closely related human sequences from other species. These primers were then employed in PCR amplification based on the following pool of human cDNAs: adrenal gland, bone marrow, brain - amygdala, brain - cerebellum, brain - hippocampus, brain - substantia nigra, brain - thalamus, brain -whole, fetal brain, fetal kidney, fetal liver, fetal lung, heart, kidney, lymphoma - Raji, mammary gland, pancreas, pituitary gland, placenta, prostate, salivary gland, skeletal muscle, small intestine, spinal cord, spleen, stomach, testis, thyroid, trachea, uterus. Usually the resulting amplicons were gel purified, cloned and sequenced to high redundancy. The PCR product derived from exon linking was cloned into the pCR2.1 vector from Invitrogen. The resulting bacterial clone has an insert covering the entire open reading frame cloned into the pCR2.1 vector. The resulting sequences from all clones were assembled with themselves, with other fragments in CuraGen Corporation's database and with public ESTs. Fragments and ESTs were included as components for an assembly when the extent of their identity with another component ofthe assembly was at least 95% over 50 bp. In addition, sequence traces were evaluated manually and edited for corrections if appropriate. These procedures provide the sequence reported herein. 6. Physical Clone:

Exons were predicted by homology and the intron/exon boundaries were determined using standard genetic rules. Exons were further selected and refined by means of similarity determination using multiple BLAST (for example, tBlasfN, BlastX, and BlasfN) searches, and, in some instances, GeneScan and Grail. Expressed sequences from both public and proprietary databases were also added when available to further define and complete the gene sequence. The DNA sequence was then manually corrected for apparent inconsistencies thereby obtaining the sequences encoding the full-length protein.

The PCR product derived by exon linking, covering the entire open reading frame, was cloned into the pCR2.1 vector from Invitrogen to provide clones used for expression and screening purposes.

Example C. Quantitative expression analysis of clones in various cells and tissues

The quantitative expression of various clones was assessed using microtiter plates containing RNA samples from a variety of normal and pathology-derived cells, cell lines and tissues using real time quantitative PCR (RTQ PCR). RTQ PCR was performed on an Applied Biosystems ABI PRISM® 7700 or an ABI PRISM® 7900 HT Sequence Detection System. Various collections of samples are assembled on the plates, and referred to as Panel 1 (containing normal tissues and cancer cell lines), Panel 2 (containing samples derived from tissues from normal and cancer sources), Panel 3 (containing cancer cell lines), Panel 4 (containing cells and cell lines from normal tissues and cells related to inflammatory conditions), Panel 5D/5I (containing human tissues and cell lines with an emphasis on metabolic diseases), AI_comprehensive_panel (containing normal tissue and samples from autoimmune diseases), Panel CNSD.01 (containing central nervous system samples from normal and diseased brains) and CNS_neurodegeneration_panel (containing samples from normal and Alzheimer's diseased brains). RNA integrity from all samples is controlled for quality by visual assessment of agarose gel electropherograms using 28S and 18S ribosomal RNA staining intensity ratio as a guide (2:1 to 2.5:1 28s:18s) and the absence of low molecular weight RNAs that would be indicative of degradation products. Samples are controlled against genomic DNA contamination by RTQ PCR reactions run in the absence of reverse transcriptase using probe and primer sets designed to amplify across the span of a single exon.

First, the RNA samples were normalized to reference nucleic acids such as constitutively expressed genes (for example, β-actin and GAPDH). Normalized RNA (5 ul) was converted to cDNA and analyzed by RTQ-PCR using One Step RT-PCR Master Mix Reagents (Applied Biosystems; Catalog No. 4309169) and gene-specific primers according to the manufacturer's instructions.

In other cases, non-normalized RNA samples were converted to single strand cDNA (sscDNA) using Superscript II (Invitrogen Corporation; Catalog No. 18064-147) and random hexamers according to the manufacturer's instructions. Reactions containing up to 10 μg of total RNA were performed in a volume of 20 μl and incubated for 60 minutes at 42°C. This reaction can be scaled up to 50 μg of total RNA in a final volume of 100 μl. sscDNA samples are then normalized to reference nucleic acids as described previously, using IX TaqMan® Universal Master mix (Applied Biosystems; catalog No. 4324020), following the manufacturer's instructions.

Probes and primers were designed for each assay according to Applied Biosystems Primer Express Software package (version I for Apple Computer's Macintosh Power PC) or a similar algorithm using the target sequence as input. Default settings were used for reaction conditions and the following parameters were set before selecting primers: primer concentration = 250 nM, primer melting temperature (Tm) range = 58°-60°C, primer optimal Tm = 59°C, maximum primer difference = 2°C, probe does not have 5'G, probe Tm must be 10°C greater than primer Tm, amplicon size 75bp to lOObp. The probes and primers selected (see below) were synthesized by Synthegen (Houston, TX, USA). Probes were double purified by HPLC to remove uncoupled dye and evaluated by mass spectroscopy to verify coupling of reporter and quencher dyes to the 5' and 3' ends ofthe probe, respectively. Their final concentrations were: forward and reverse primers, 900nM each, and probe, 200nM. PCR conditions: When working with RNA samples, normalized RNA from each tissue and each cell line was spotted in each well of either a 96 well or a 384-well PCR plate (Applied Biosystems). PCR cocktails included either a single gene specific probe and primers set, or two multiplexed probe and primers sets (a set specific for the target clone and another gene-specific set multiplexed with the target probe). PCR reactions were set up using TaqMan® One-Step RT-PCR Master Mix (Applied Biosystems, Catalog No.4313803) following manufacturer's instructions. Reverse transcription was performed at 48°C for 30 minutes followed by amplification PCR cycles as follows: 95°C 10 min, then 40 cycles of 95°C for 15 seconds, 60°C for 1 minute. Results were recorded as CT values (cycle at which a given sample crosses a threshold level of fluorescence) using a log scale, with the difference in RNA concentration between a given sample and the sample with the lowest CT value being represented as 2 to the power of delta CT. The percent relative expression is then obtained by taking the reciprocal of this RNA difference and multiplying by 100.

When working with sscDNA samples, normalized sscDNA was used as described previously for RNA samples. PCR reactions containing one or two sets of probe and primers were set up as described previously, using IX TaqMan® Universal Master mix (Applied Biosystems; catalog No. 4324020), following the manufacturer's instructions. PCR amplification was performed as follows: 95°C 10 min, then 40 cycles of 95°C for 15 seconds, 60°C for 1 minute. Results were analyzed and processed as described previously.

Panels 1, 1.1, 1.2, and 1.3D

The plates for Panels 1, 1.1, 1.2 and 1.3D include 2 control wells (genomic DNA control and chemistry control) and 94 wells containing cDNA from various samples. The samples in these panels are broken into 2 classes: samples derived from cultured cell lines and samples derived from primary normal tissues. The cell lines are derived from cancers of the following types: lung cancer, breast cancer, melanoma, colon cancer, prostate cancer, CNS cancer, squamous cell carcinoma, ovarian cancer, liver cancer, renal cancer, gastric cancer and pancreatic cancer. Cell lines used in these panels are widely available through the American Type Culture Collection (ATCC), a repository for cultured cell lines, and were cultured using the conditions recommended by the ATCC. The normal tissues found on these panels are comprised of samples derived from all major organ systems from single adult individuals or fetuses. These samples are derived from the following organs: adult skeletal muscle, fetal skeletal muscle, adult heart, fetal heart, adult kidney, fetal kidney, adult liver, fetal liver, adult lung, fetal lung, various regions ofthe brain, the spleen, bone marrow, lymph node, pancreas, salivary gland, pituitary gland, adrenal gland, spinal cord, thymus, stomach, small intestine, colon, bladder, trachea, breast, ovary, uterus, placenta, prostate, testis and adipose.

In the results for Panels 1, 1.1, 1.2 and 1.3D, the following abbreviations are used: ca. = carcinoma,

* = established from metastasis, met = metastasis, s cell var = small cell variant, non-s = non-sm = non-small, squam = squamous, pi. eff = pi effusion = pleural effusion, glio = glioma, astro = astrocytoma, and neuro = neuroblastoma.

Gener al_screenin jpanel_vl .4

The plates for Panel 1.4 include 2 control wells (genomic DNA control and chemistry control) and 94 wells containing cDNA from various samples. The samples in Panel 1.4 are broken into 2 classes: samples derived from cultured cell lines and samples derived from primary normal tissues. The cell lines are derived from cancers ofthe following types: lung cancer, breast cancer, melanoma, colon cancer, prostate cancer, CNS cancer, squamous cell carcinoma, ovarian cancer, liver cancer, renal cancer, gastric cancer and pancreatic cancer. Cell lines used in Panel 1.4 are widely available through the American Type Culture Collection (ATCC), a repository for cultured cell lines, and were cultured using the conditions recommended by the ATCC. The normal tissues found on Panel 1.4 are comprised of pools of samples derived from all major organ systems from 2 to 5 different adult individuals or fetuses. These samples are derived from the following organs: adult skeletal muscle, fetal skeletal muscle, adult heart, fetal heart, adult kidney, fetal kidney, adult liver, fetal liver, adult lung, fetal lung, various regions ofthe brain, the spleen, bone marrow, lymph node, pancreas, salivary gland, pituitary gland, adrenal gland, spinal cord, thymus, stomach, small intestine, colon, bladder, trachea, breast, ovary, uterus, placenta, prostate, testis and adipose. Abbreviations are as described for Panels 1, 1.1, 1.2, and 1.3D.

Panels 2D and 2.2

The plates for Panels 2D and 2.2 generally include 2 control wells and 94 test samples composed of RNA or cDNA isolated from human tissue procured by surgeons working in close cooperation with the National Cancer Institute's Cooperative Human Tissue Network (CHTN) or the National Disease Research Initiative (NDRI). The tissues are derived from human malignancies and in cases where indicated many malignant tissues have "matched margins" obtained from noncancerous tissue just adjacent to the tumor. These are termed normal adjacent tissues and are denoted "NAT" in the results below. The tumor tissue and the "matched margins" are evaluated by two independent pathologists (the surgical pathologists and again by a pathologist at NDRI or CHTN). This analysis provides a gross histopathological assessment of tumor differentiation grade. Moreover, most samples include the original surgical pathology report that provides information regarding the clinical stage of the patient. These matched margins are taken from the tissue suπounding (i.e. immediately proximal) to the zone of surgery (designated "NAT", for normal adjacent tissue, in Table RR). In addition, RNA and cDNA samples were obtained from various human tissues derived from autopsies performed on elderly people or sudden death victims (accidents, etc.). These tissues were ascertained to be free of disease and were purchased from various commercial sources such as Clontech (Palo Alto, CA), Research Genetics, and Invitrogen.

Panel 3D

The plates of Panel 3D are comprised of 94 cDNA samples and two control samples. Specifically, 92 of these samples are derived from cultured human cancer cell lines, 2 samples of human primary cerebellar tissue and 2 controls. The human cell lines are generally obtained from ATCC (American Type Culture Collection), NCI or the German tumor cell bank and fall into the following tissue groups: Squamous cell carcinoma ofthe tongue, breast cancer, prostate cancer, melanoma, epidermoid carcinoma, sarcomas, bladder carcinomas, pancreatic cancers, kidney cancers, leukemias/lymphomas, ovarian/uterine/cervical, gastric, colon, lung and CNS cancer cell lines. In addition, there are two independent samples of cerebellum. These cells are all cultured under standard recommended conditions and RNA extracted using the standard procedures. The cell lines in panel 3D and 1.3D are ofthe most common cell lines used in the scientific literature.

Panels 4D, 4R, and 4.1D Panel 4 includes samples on a 96 well plate (2 control wells, 94 test samples) composed of RNA (Panel 4R) or cDNA (Panels 4D/4.1D) isolated from various human cell lines or tissues related to inflammatory conditions. Total RNA from control normal tissues such as colon and lung (Stratagene, La Jolla, CA) and thymus and kidney (Clontech) was employed. Total RNA from liver tissue from cirrhosis patients and kidney from lupus patients was obtained from BioChain (Biochain Institute, Inc., Hayward, CA). Intestinal tissue for RNA preparation from patients diagnosed as having Crohn's disease and ulcerative colitis was obtained from the National Disease Research Interchange (NDRI) (Philadelphia, PA).

Astrocytes, lung fibroblasts, dermal fibroblasts, coronary artery smooth muscle cells, small airway epithelium, bronchial epithelium, microvascular dermal endothelial cells, microvascular lung endothelial cells, human pulmonary aortic endothelial cells, human umbilical vein endothelial cells were all purchased from Clonetics (Walkersville, MD) and grown in the media supplied for these cell types by Clonetics. These primary cell types were activated with various cytokines or combinations of cytokines for 6 and/or 12-14 hours, as indicated. The following cytokines were used; IL-1 beta at approximately l-5ng/ml, TNF alpha at approximately 5-lOng/ml, LFN gamma at approximately 20-50ng/ml, IL-4 at approximately 5-10ng/ml, IL-9 at approximately 5-10ng/ml, IL-13 at approximately 5- lOng/ml. Endothelial cells were sometimes starved for various times by culture in the basal media from Clonetics with 0.1% serum.

Mononuclear cells were prepared from blood of employees at CuraGen Corporation, using Ficoll. LAK cells were prepared from these cells by culture in DMEM 5% FCS (Hyclone), lOOμM non essential amino acids (Gibco/Life Technologies, Rockville, MD), ImM sodium pyruvate (Gibco), mercaptoethanol 5.5xlO^"5M (Gibco), and lOmM Hepes (Gibco) and Interleukin 2 for 4-6 days. Cells were then either activated with 10-20ng/ml

PMA and l-2μg/ml ionomychϊ, IL-12 at 5-10ng/ml, LFN gamma at 20-50ng/ml and IL-18 at 5-10ng/ml for 6 hours. In some cases, mononuclear cells were cultured for 4-5 days in DMEM 5% FCS (Hyclone), lOOμM non essential amino acids (Gibco), ImM sodium pyruvate (Gibco), mercaptoethanol 5.5xlO^"5M (Gibco), and lOmM Hepes (Gibco) with PHA (phytohemagglutinin) or PWM (pokeweed mitogen) at approximately 5 μg/ml. Samples were taken at 24, 48 and 72 hours for RNA preparation. MLR (mixed lymphocyte reaction) samples were obtained by taking blood from two donors, isolating the mononuclear cells using Ficoll and mixing the isolated mononuclear cells 1:1 at a final concentration of approximately 2xl0⁶cells/ml in DMEM 5% FCS (Hyclone), lOOμM non essential amino acids (Gibco), ImM sodium pyruvate (Gibco), mercaptoethanol (5.5xlO^"5M) (Gibco), and lOmM Hepes (Gibco). The MLR was cultured and samples taken at various time points ranging from 1- 7 days for RNA preparation.

Monocytes were isolated from mononuclear cells using CD 14 Miltenyi Beads, +ve VS selection columns and a Vario Magnet according to the manufacturer's instructions. Monocytes were differentiated into dendritic cells by culture in DMEM 5% fetal calf serum (FCS) (Hyclone, Logan, UT), lOOμM non essential amino acids (Gibco), ImM sodium pyruvate (Gibco), mercaptoethanol 5.5xl0^"5M (Gibco), and lOmM Hepes (Gibco), 50ng/ml GMCSF and 5ng/ml IL-4 for 5-7 days. Macrophages were prepared by culture of monocytes for 5-7 days in DMEM 5% FCS (Hyclone), lOOμM non essential amino acids (Gibco), ImM sodium pyruvate (Gibco), mercaptoethanol 5.5xlO^"5M (Gibco), lOmM Hepes (Gibco) and 10%) AB Human Serum or MCSF at approximately 50ng/ml. Monocytes, macrophages and dendritic cells were stimulated for 6 and 12-14 hours with lipopolysaccharide (LPS) at lOOng/ml. Dendritic cells were also, stimulated with anti-CD40 monoclonal antibody (Pharmingen) at lOμg/ml for 6 and 12-14 hours. CD4 lymphocytes, CD8 lymphocytes and NK cells were also isolated from mononuclear cells using CD4, CD8 and CD56 Miltenyi beads, positive VS selection columns and a Vario Magnet according to the manufacturer's instructions. CD45RA and CD45RO CD4 lymphocytes were isolated by depleting mononuclear cells of CD8, CD56, CD14 and CD19 cells using CD8, CD56, CD14 and CD19 Miltenyi beads and positive selection.

CD45RO beads were then used to isolate the CD45RO CD4 lymphocytes with the remaining cells being CD45RA CD4 lymphocytes. CD45RA CD4, CD45RO CD4 and CD8 lymphocytes were placed in DMEM 5% FCS (Hyclone), lOOμM non essential amino acids (Gibco), ImM sodium pyruvate (Gibco), mercaptoethanol 5.5xlO^"5M (Gibco), and lOmM Hepes (Gibco) and plated at 10⁶cells/ml onto Falcon 6 well tissue culture plates that had been coated overnight with 0.5 μg/ml anti-CD28 (Pharmingen) and 3ug/ml anti-CD3 (OKT3, ATCC) in PBS. After 6 and 24 hours, the cells were harvested for RNA preparation. To prepare chronically activated CD8 lymphocytes, we activated the isolated CD8 lymphocytes for 4 days on anti-CD28 and anti-CD3 coated plates and then harvested the cells and expanded them in DMEM 5% FCS (Hyclone), lOOμM non essential amino acids (Gibco), ImM sodium pyruvate (Gibco), mercaptoethanol 5.5xlO^"5M (Gibco), and lOmM Hepes (Gibco) and IL-2. The expanded CD8 cells were then activated again with plate bound anti- CD3 and anti-CD28 for 4 days and expanded as before. RNA was isolated 6 and 24 hours after the second activation and after 4 days ofthe second expansion culture. The isolated NK cells were cultured in DMEM 5% FCS (Hyclone), lOOμM non essential amino acids (Gibco), ImM sodium pyruvate (Gibco), mercaptoethanol 5.5xl0^"5M (Gibco), and lOmM Hepes (Gibco) and IL-2 for 4-6 days before RNA was prepared.

To obtain B cells, tonsils were procured from NDRI. The tonsil was cut up with sterile dissecting scissors and then passed through a sieve. Tonsil cells were then spun down and resupended at 10⁶cells/ml in DMEM 5% FCS (Hyclone), lOOμM non essential amino acids (Gibco), ImM sodium pyruvate (Gibco), mercaptoethanol 5.5xl0^"5M (Gibco), and lOmM Hepes (Gibco). To activate the cells, we used PWM at 5 μg/ml or anti-CD40 (Pharmingen) at approximately lOμg/ml and IL-4 at 5-10ng/ml. Cells were harvested for RNA preparation at 24,48 and 72 hours. To prepare the primary and secondary Thl/Th2 and Trl cells, six-well Falcon plates were coated overnight with lOμg/ml anti-CD28 (Pharmingen) and 2μg/ml OKT3 (ATCC), and then washed twice with PBS. Umbilical cord blood CD4 lymphocytes (Poietic Systems, German Town, MD) were cultured at 10⁵-10⁶cells/ml in DMEM 5% FCS (Hyclone), lOOμM non essential amino acids (Gibco), ImM sodium pyruvate (Gibco), mercaptoethanol 5.5x10^{" 5}M (Gibco), lOmM Hepes (Gibco) and IL-2 (4ng/ml). IL-12 (5ng/ml) and anti-IL4 (lμg/ml) were used to direct to Thl, while IL-4 (5ng/ml) and anti-IFN gamma (lμg/ml) were used to direct to Th2 and IL-10 at 5ng/ml was used to direct to Trl. After 4-5 days, the activated Thl, Th2 and Trl lymphocytes were washed once in DMEM and expanded for 4-7 days in DMEM 5% FCS (Hyclone), lOOμM non essential amino acids (Gibco), ImM sodium pyruvate (Gibco), mercaptoethanol 5.5xl0^"5M (Gibco), lOmM Hepes (Gibco) and LL-2 (lng/ml). Following this, the activated Thl, Th2 and Trl lymphocytes were re-stimulated for 5 days with anti-CD28/OKT3 and cytokines as described above, but with the addition of anti- CD95L (lμg/ml) to prevent apoptosis. After 4-5 days, the Thl, Th2 and Trl lymphocytes were washed and then expanded again with IL-2 for 4-7 days. Activated Thl and Th2 lymphocytes were maintained in this way for a maximum of three cycles. RNA was prepared from primary and secondary Thl, Th2 and Trl after 6 and 24 hours following the second and third activations with plate bound anti-CD3 and anti-CD28 mAbs and 4 days into the second and third expansion cultures in Interleukin 2. The following leukocyte cells lines were obtained from the ATCC: Ramos, EOL-1,

KU-812. EOL cells were further differentiated by culture in O.lmM dbcAMP at 5xl0⁵cells/ml for 8 days, changing the media every 3 days and adjusting the cell concentration to 5xl0⁵cells/ml. For the culture of these cells, we used DMEM or RPMI (as recommended by the ATCC), with the addition of 5% FCS (Hyclone), lOOμM non essential amino acids (Gibco), ImM sodium pyruvate (Gibco), mercaptoethanol 5.5xlO^"5M (Gibco), lOmM Hepes (Gibco). RNA was either prepared from resting cells or cells activated with PMA at lOng/ml and ionomycin at lμg/ml for 6 and 14 hours. Keratinocyte line CCD106 and an airway epithelial tumor line NCI-H292 were also obtained from the ATCC. Both were cultured in DMEM 5% FCS (Hyclone), lOOμM non essential amino acids (Gibco), ImM sodium pyruvate (Gibco), mercaptoethanol 5.5xl0^"5M (Gibco), and lOmM Hepes (Gibco).

CCD1106 cells were activated for 6 and 14 hours with approximately 5 ng/ml TNF alpha and lng/ml IL-1 beta, while NCI-H292 cells were activated for 6 and 14 hours with the following cytokines: 5ng/ml IL-4, 5ng/ml IL-9, 5ng/ml IL-13 and 25ng/ml IFN gamma.

For these cell lines and blood cells, RNA was prepared by lysing approximately IO⁷ cells/ml using Trizol (Gibco BRL). Briefly, 1/10 volume of bromochloropropane (Molecular Research Corporation) was added to the RNA sample, vortexed and after 10 minutes at room temperature, the tubes were spun at 14,000 rpm in a Sorvall SS34 rotor. The aqueous phase was removed and placed in a 15ml Falcon Tube. An equal volume of isopropanol was added and left at -20°C overnight. The precipitated RNA was spun down at 9,000 rpm for 15 min in a Sorvall SS34 rotor and washed in 70% ethanol. The pellet was redissolved in 300μl of RNAse-free water and 35μl buffer (Promega) 5μl DTT, 7μl RNAsin and 8μl DNAse were added. The tube was incubated at 37°C for 30 minutes to remove contaminating genomic DNA, extracted once with phenol chloroform and re-precipitated with 1/10 volume of 3M sodium acetate and 2 volumes of 100% ethanol. The RNA was spun down and placed in RNAse free water. RNA was stored at -80°C.

AI_comprehensive panel_vl.0

The plates for AI_comprehensive panel_vl.O include two control wells and 89 test samples comprised of cDNA isolated from surgical and postmortem human tissues obtained from the Backus Hospital and Clinomics (Frederick, MD). Total RNA was extracted from tissue samples from the Backus Hospital in the Facility at CuraGen. Total RNA from other tissues was obtained from Clinomics.

Joint tissues including synovial fluid, synovium, bone and cartilage were obtained from patients undergoing total knee or hip replacement surgery at the Backus Hospital. Tissue samples were immediately snap frozen in liquid nitrogen to ensure that isolated RNA was of optimal quality and not degraded. Additional samples of osteoarthritis and rheumatoid arthritis joint tissues were obtained from Clinomics. Normal control tissues were supplied by

Clinomics and were obtained during autopsy of trauma victims.

Surgical specimens of psoriatic tissues and adjacent matched tissues were provided as total RNA by Clinomics. Two male and two female patients were selected between the ages of 25 and 47. None ofthe patients were taking prescription drugs at the time samples were isolated.

Surgical specimens of diseased colon from patients with ulcerative colitis and Crohns disease and adjacent matched tissues were obtained from Clinomics. Bowel tissue from three female and three male Crohn's patients between the ages of 41-69 were used. Two patients were not on prescription medication while the others were taking dexamethasone, phenobarbital, or tylenol. Ulcerative colitis tissue was from three male and four female patients. Four ofthe patients were taking lebvid and two were on phenobarbital.

Total RNA from post mortem lung tissue from trauma victims with no disease or with emphysema, asthma or COPD was purchased from Clinomics. Emphysema patients ranged in age from 40-70 and all were smokers, this age range was chosen to focus on patients with cigarette-linked emphysema and to avoid those patients with alpha- lanti-trypsin deficiencies.

Asthma patients ranged in age from 36-75, and excluded smokers to prevent those patients that could also have COPD. COPD patients ranged in age from 35-80 and included both smokers and non-smokers. Most patients were taking corticosteroids, and bronchodilators.

In the labels employed to identify tissues in the AI_comprehensive panel_vl.0 panel, the following abbreviations are used: Al = Autoimmunity

Syn = Synovial

Normal = No apparent disease

Rep22 /Rep20 = individual patients

RA = Rheumatoid arthritis Backus = From Backus Hospital

OA = Osteoarthritis

(SS) (BA) (MF) = Individual patients

Adj = Adjacent tissue

Match control = adjacent tissues -M = Male

-F = Female

COPD = Chronic obstructive pulmonary disease

Panels 5D and 51 The plates for Panel 5D and 51 include two control wells and a variety of cDNAs isolated from human tissues and cell lines with an emphasis on metabolic diseases. Metabolic tissues were obtained from patients enrolled in the Gestational Diabetes study. Cells were obtained during different stages in the differentiation of adipocytes from human mesenchymal stem cells. Human pancreatic islets were also obtained. In the Gestational Diabetes study subjects are young (18 - 40 years), otherwise healthy women with and without gestational diabetes undergoing routine (elective) Caesarean section. After delivery ofthe infant, when the surgical incisions were being repaired/closed, the obstetrician removed a small sample (<1 cc) ofthe exposed metabolic tissues during the closure of each surgical level. The biopsy material was rinsed in sterile saline, blotted and fast frozen within 5 minutes from the time of removal. The tissue was then flash frozen in liquid nitrogen and stored, individually, in sterile screw-top tubes and kept on dry ice for shipment to or to be picked up by CuraGen. The metabolic tissues of interest include uterine wall (smooth muscle), visceral adipose, skeletal muscle (rectus) and subcutaneous adipose.

Patient descriptions are as follows:

Patient 2 Diabetic Hispanic, overweight, not on insulin

Patient 7-9 Nondiabetic Caucasian and obese (BMI>30) Patient 10 Diabetic Hispanic, overweight, on insulin Patient 11 Nondiabetic African American and overweight Patient 12 Diabetic Hispanic on insulin Adipocyte differentiation was induced in donor progenitor cells obtained from Osirus (a division of Clonetics/BioWhittaker) in triplicate, except for Donor 3U which had only two replicates. Scientists at Clonetics isolated, grew and differentiated human mesenchymal stem cells (HuMSCs) for CuraGen based on the published protocol found in Mark F. Pittenger, et al., MuRilineage Potential of Adult Human Mesenchymal Stem Cells Science Apr 2 1999: 143-147. Clonetics provided Trizol lysates or frozen pellets suitable for mRNA isolation and ds cDNA production. A general description of each donor is as follows:

Donor 2 and 3 U: Mesenchymal Stem cells, Undifferentiated Adipose

Donor 2 and 3 AM: Adipose, AdiposeMidway Differentiated Donor 2 and 3 AD: Adipose, Adipose Differentiated

Human cell lines were generally obtained from ATCC (American Type Culture Collection), NCI or the German tumor cell bank and fall into the following tissue groups: kidney proximal convoluted tubule, uterine smooth muscle cells, small intestine, liver HepG2 cancer cells, heart primary stromal cells, and adrenal cortical adenoma cells. These cells are all cultured under standard recommended conditions and RNA extracted using the standard procedures. All samples were processed at CuraGen to produce single stranded cDNA. Panel 51 contains all samples previously described with the addition of pancreatic islets from a 58 year old female patient obtained from the Diabetes Research Institute at the

University of Miami School of Medicine. Islet tissue was processed to total RNA at an outside source and delivered to CuraGen for addition to panel 51.

In the labels employed to identify tissues in the 5D and 51 panels, the following abbreviations are used:

GO Adipose = Greater Omentum Adipose SK= Skeletal Muscle UT = Uterus PL = Placenta AD = Adipose Differentiated

AM = Adipose Midway Differentiated U = Undifferentiated Stem Cells

Panel CNSD.01 The plates for Panel CNSD.01 include two control wells and 94 test samples comprised of cDNA isolated from postmortem human brain tissue obtained from the Harvard

Brain Tissue Resource Center. Brains are removed from calvaria of donors between 4 and 24 hours after death, sectioned by neuroanatomists, and frozen at -80°C in liquid nitrogen vapor. All brains are sectioned and examined by neuropathologists to confirm diagnoses with clear associated neuropathology.

Disease diagnoses are taken from patient records. The panel contains two brains from each ofthe following diagnoses: Alzheimer's disease, Parkinson's disease, Huntington's disease, Progressive Supemuclear Palsy, Depression, and "Normal controls" . Within each of these brains, the following regions are represented: cingulate gyrus, temporal pole, globus palladus, substantia nigra, Brodman Area 4 (primary motor strip), Brodman Area 7 (parietal cortex), Brodman Area 9 (prefrontal cortex), and Brodman area 17 (occipital cortex). Not all brain regions are represented in all cases; e.g., Huntington's disease is characterized in part by neurodegeneration in the globus palladus, thus this region is impossible to obtain from confirmed Huntington's cases. Likewise Parkinson's disease is characterized by degeneration ofthe substantia nigra making this region more difficult to obtain. Normal control brains were examined for neuropathology and found to be free of any pathology consistent with neurodegeneration. In the labels employed to identify tissues in the CNS panel, the following abbreviations are used:

PSP = Progressive supranuclear palsy Sub Nigra = Substantia nigra Glob Palladus= Globus palladus Temp Pole = Temporal pole

Cing Gyr = Cingulate gyrus BA 4 = Brodman Area 4

Panel CNS_Neurodegeneration_V1.0 The plates for Panel CNS_Neurodegeneration_Vl .0 include two control wells and 47 test samples comprised of cDNA isolated from postmortem human brain tissue obtained from the Harvard Brain Tissue Resource Center (McLean Hospital) and the Human Brain and Spinal Fluid Resource Center (VA Greater Los Angeles Healthcare System). Brains are removed from calvaria of donors between 4 and 24 hours after death, sectioned by neuroanatomists, and frozen at -80°C in liquid nitrogen vapor. All brains are sectioned and examined by neuropathologists to confirm diagnoses with clear associated neuropathology. Disease diagnoses are taken from patient records. The panel contains six brains from Alzheimer's disease (AD) patients, and eight brains from "Normal controls" who showed no evidence of dementia prior to death. The eight normal control brains are divided into two categories: Controls with no dementia and no Alzheimer's like pathology (Controls) and controls with no dementia but evidence of severe Alzheimer's like pathology, (specifically senile plaque load rated as level 3 on a scale of 0-3; 0 = no evidence of plaques, 3 = severe

AD senile plaque load). Within each of these brains, the following regions are represented: hippocampus, temporal cortex (Brodman Area 21), parietal cortex (Brodman area 7), and occipital cortex (Brodman area 17). These regions were chosen to encompass all levels of neurodegeneration in AD. The hippocampus is a region of early and severe neuronal loss in

AD; the temporal cortex is known to show neurodegeneration in AD after the hippocampus; the parietal cortex shows moderate neuronal death in the late stages ofthe disease; the occipital cortex is spared in AD and therefore acts as a "control" region within AD patients.

Not all brain regions are represented in all cases. In the labels employed to identify tissues in the CNS_Neurodegeneration_Vl .0 panel, the following abbreviations are used:

AD = Alzheimer's disease brain; patient was demented and showed AD-like pathology upon autopsy

Control = Control brains; patient not demented, showing no neuropathology Control (Path) = Control brains; pateint not demented but showing sever AD-like pathology

SupTemporal Ctx = Superior Temporal Cortex Inf Temporal Ctx = Inferior Temporal Cortex

A. CG57883-01: Ring finger protein Expression of gene CG57883-01 was assessed using the primer-probe set Ag3352, described in Table AA. Results ofthe RTQ-PCR runs are shown in Table AB.

Table AA. Probe Name Ag3352

Table AB. General_screening_panel_vl.4

Liver ca. HepG2 0.2 (spinal Cord Pool 0.0

Kidney Pool 0.5 Adrenal Gland jo.o

Fetal Kidney 0.0 (Pituitary gland Pool jo.o

Renal ca. 786-0 0.2 (Salivary Gland jo.o

Renal ca. A498 0.3 jThyroid (female) jo.o

Renal ca. ACHN 0.0 (Pancreatic ca. CAPAN2 Jo.o

Renal ca. UO-31 1.4 [Pancreas Pool jo.i

CNS_neurodegeneration_vl.0 Summary: Ag3352 - Expression of CG57883-01 gene is low/undetectable (CTs > 35) across all ofthe samples on this panel (data not shown).

General_screening_panel_vl.4 Summary: Ag3352 - Highest expression ofthe CG57883-01 gene is seen exclusively in the thalamus (CT=27.4). Therefore, expression of this gene could be used to distinguish this sample from other samples used in this panel.

In addition, low to moderate expression of this gene is seen in lung, colon, CNS, renal, ovarian and melanoma. Therefore, therapeutic modulation ofthe activity of this gene or its protein product, through the use of small molecule drugs, protein therapeutics or antibodies, might be beneficial in the treatment of these cancers.

Among tissues with metabolic or endocrine function, this gene is expressed at low levels in adipose, and liver (CTs=34-34.8). Therefore, therapeutic modulation ofthe activity of this gene may prove useful in the treatment of endocrine/metabolically related diseases, such as obesity and diabetes.

Panel 4D Summary: Ag3352 - Expression of CG57883-01 gene is low/undetectable (CTs > 35) across all ofthe samples on this panel (data not shown).

B. CG57881-01: Ring finger protein

Expression of gene CG57881-01 was assessed using the primer-probe set Ag3353, described in Table BA. Results ofthe RTQ-PCR runs are shown in Tables BB.

Table BA. Probe Name Ag3353

Table BB. Panel 4D

CNS__neurodegeneration_vl.0 Summary: Ag3353 - Expression ofthe CG57881- 01 gene is low/undetectable (CTs > 35) across all ofthe samples on this panel (data not shown).

General_screening_panel_vl.4 Summary: Ag3353 - Results from one experiment are not included. The amp plot indicates that there were experimental difficulties with this run.

Panel 4D Summary: Ag3353 - Highest expression ofthe CG57881-01 gene is seen in ionomycin treated Ramos B cells (CT=31.7). Interestingly, there is low/undectable expression of this gene in untreated Ramos B cells (CT=35.7). Therefore, expression of this gene can be used to distinguish ionomycin treated Ramos B cells from the untreated cells and other samples used in this panel. In addition, expression of this gene in ionomycin stimulated B cells suggests that this gene may be involved in rheumatic disease including rheumatoid arthritis, lupus, osteoarthritis, and hyperproliferative B cell disorders.

Low expression of this gene is also detected in a liver cmhosis sample (CT = 34.4). The presence of this gene in liver cirrhosis (a component of which involves liver inflammation and fibrosis) suggests that antibodies to the protein encoded by this gene could also be used for the diagnosis of liver cirrhosis. Furthermore, therapeutic agents involving this gene may be useful in reducing or inhibiting the inflammation associated with fibrotic and inflammatory diseases.

C. CG57407-01: BRAIN CDNA, CLONE MNCB-2146

Expression of gene CG57407-01 was assessed using the primer-probe set Ag3227, described in Table CA. Results ofthe RTQ-PCR runs are shown in Tables CB

Table CA. Probe Name Ag3227

Table CB. General screening panel vl.4

CNS_neurodegeneration_vl.0 Summary: Ag3227 - Expression of this gene is low/undetectable (CTs > 35) across all ofthe samples on this panel (data not shown).

General_screening_panel_vl.4 Summary: Ag3227 Highest expression of this gene is detected in gastric cancer cell line NCI-N87 (CT=33.7). Significant expression of this gene is also seen small intestine (CT=34.8), and ovarian cancer cell line SK-OV-3 (CT=34.4).

Therefore, expression of this gene can be used to distinguish these samples from other samples on this panel. In addition, therapeutic modulation ofthe activity of this gene or its protein product, through the use of small molecule drugs, protein therapeutics or antibodies, might be beneficial in the treatment of gastric cancer or ovarian cancer.

Panel 2.2 Summary: Ag3227 - Results from one experiment are not included. The amp plot indicates that there were experimental difficulties with this run.

Panel 4D Summary: Ag3227 - Expression of this gene is low/undetectable (CTs > 35) across all ofthe samples on this panel (data not shown).

D. CG58651-02: SM-20

Expression of gene CG58651-02 was assessed using the primer-probe set Ag3388, described in Table DA. Results of the RTQ-PCR runs are shown in Tables DB, DC and DD.

Table DA. Probe Name Ag3388

Table DB. CNS_neurodegeneration_vl.0

Table DC. General_screening_panel_vl.4

Table DD. Panel 4D

CNS_neurodegeneration_vl.0 Summary: Ag3388 This panel confirms the expression ofthe CG58651-02 gene at low levels in the brains of an independent group of individuals. However, no differential expression of this gene was detected between Alzheimer's diseased postmortem brains and those of non-demented controls in this experiment. Please see Panel 1.4 for a discussion ofthe potential utility of this gene in treatment of central nervous system disorders.

General_screening_panel_vl.4 Summary: Ag3388 Highest expression ofthe CG58651-02 gene is seen in a breast cancer cell line T47D (CT=27.3). Therefore, expression of this gene can be used to distinguish this sample from other samples on this panel. In addition, high expression of these gene is also detected in ovarian, breast, CNS, lung, gastric and colon cancer. Therefore, therapeutic modulation ofthe activity of this gene or its protein product, through the use of small molecule drugs, protein therapeutics or antibodies, might be beneficial in the treatment of these cancers.

Among tissues with metabolic or endocrine function, this gene is expressed at high to moderate levels in pancreas, adipose, adrenal gland, thyroid, pituitary gland, skeletal muscle, heart, liver and the gastrointestinal tract. Therefore, therapeutic modulation ofthe activity of this gene may prove useful in the treatment of endocrine/metabolically related diseases, such as obesity and diabetes.

Interestingly, this gene is expressed at much higher levels in fetal (CT = 31) when compared to adult liver (CT = 34). This observation suggests that expression of this gene can be used to distinguish fetal from adult liver.

In addition, this gene is expressed at high levels in all regions ofthe central nervous system examined, including amygdala, hippocampus, substantia nigra, thalamus, cerebellum, cerebral cortex, and spinal cord. Therefore, this gene may play a role in central nervous system disorders such as Alzheimer's disease, Parkinson's disease, epilepsy, multiple sclerosis, schizophrenia and depression.

CG58651-02 codes for a variant of EGLN2 protein. EGLN2 is a homologue of Caenorhabditis elegans gene egl-9. See, Online Mendelian Inheritance in Man (OMIM) accno. 606424. Recently, it has been shown that C. elegans EGL-9 and mammalian homologs define a family of dioxygenases that regulate HLF by prolyl hydroxylation.. See, e.g., Epstein et al. (2001) Cell 107(l):43-54. HLF is a transcriptional complex that plays a central role in mammalian oxygen homeostasis. It is activated in a broad array of ischemic/hypoxic and neoplastic diseases. Therefore, therapeutic modulation ofthe activity of the protein encoded by CG58651 -02 gene may be beneficial in the treatment ischemic/hypoxic and neoplastic diseases.

Panel 4D Summary: Ag3388 Highest expresion ofthe CG58651-02 gene is seen in PWM treated B lymphocytes (CT=29.4). However, this gene is expressed at high to moderate levels in a wide range of cell types of significance in the immune response in health and disease. These cells include members ofthe T-cell, B-cell, endothelial cell, , macrophage/monocyte, and peripheral blood mononuclear cell family, as well as epithelial and fibroblast cell types from lung and skin, and normal tissues represented by colon, lung, thymus and kidney. This ubiquitous pattern of expression suggests that this gene product may be involved in homeostatic processes for these and other cell types and tissues. This pattern is in agreement with the expression profile in General_screenmg_panel_vl .4 and also suggests a role for the gene product in cell survival and proliferation. Therefore, modulation ofthe gene product with a functional therapeutic may lead to the alteration of functions associated with these cell types and lead to improvement ofthe symptoms of patients suffering from autoimmune and inflammatory diseases such as asthma, allergies, inflammatory bowel disease, lupus erythematosus, psoriasis, rheumatoid arthritis, and osteoarthritis.

E. CG59574-01: Synaptonemal complex protein

Expression of gene CG59574-01 was assessed using the primer-probe set Ag3477, described in Table EA. Results ofthe RTQ-PCR runs are shown in Tables EB, and EC.

Table EA. Probe Name Ag3477

Table EB. General_screening_panel_vl.4

Table ED. Panel 4D

CNS_neurodegeneration_vl.0 Summary: Ag3477 Expression ofthe CG59574-01 gene is low/undetectable in all samples on this panel (CTs>34.3). (Data not shown.) General_screening_panel_vl.4 Summary: Ag3477 Expression ofthe CG59574-01 gene is restricted to the testis (CT=30). Thus, expression of this gene could be used to differentiate between this sample and other samples on this panel and as a marker for testicular tissue. Furthermore, therapeutic modulation ofthe expression or function of this gene may be effective in the treatment of male infertility and hypogonadism.

Panel 4D Summary: Ag3477 Highest expression ofthe CG59574-01 gene is seen in dermal fibroblasts treated with TNF alpha (CT=31.7). The significant levels of expression in this sample suggests that this gene product may be involved in skin disorders, including psoriasis. In addition, this transcript is expressed in T cells, LAK cells, macrophages, and dendritic cells. In addition to these hematopoietic cell types, expression is also seen in the PMA/ionomycin treated basophil cell line KU-812, small airway epithelium treated with TNF-alpha and IL-1 beta, and treated and untreated samples from the mucoepidermoid cell line H292. Thymus and kidney also express low levels ofthe transcript. Thus, this transcript or the protein it encodes could be used to design therapeutics that could be important in the regulation of the function of antigen presenting cells (macrophages and dendritic cells)or T cells and be important in the treatment of asthma, emphysema, psoriasis, arthritis, and IBD.

F. CG59536-01: Paraneoplastic Antigen

Expression of gene CG59536-01 was assessed using the primer-probe set Ag3458, described in Table FA. Results ofthe RTQ-PCR runs are shown in Tables FB, FC, FD, and FE.

Table FA. Probe Name Ag3458

Table FB. CNS_neurodegeneration_vl.0

Table FC. General_screening_panel_vl.4

(Renal ca. A498 jo.o jThyroid (female) |θ.4

JRenal ca. ACHN M jPancreatic ca. CAPAN2 jθ.4

Renal ca. UO-31 |8-9 Pancreas Pool J14.9

Table FD. Panel 2.2

Table FE. Panel 4D

CNS_neurodegeneration_vl.0 Summary: Ag3458 This panel confirms the expression of CG59536-01 gene at significant levels in the brains of an independent group of individuals. However, no differential expression of this gene was detected between Alzheimer's diseased postmortem brains and those of non-demented controls in this experiment. Please see Panel 1.4 for a discussion ofthe potential utility of this gene in treatment of central nervous system disorders.

General_screening_panel_vl.4 Summary: Ag3458 Expression ofthe CG59536-01 gene is widespread throughout this panel, with highest expression in the fetal heart (CT=29.6). In addition, expression of this gene appears to be higher in fetal heart when compared to expression in the adult heart (CT=34.5). Thus, expression of this gene could be used to differentiate between adult and fetal heart.

This gene is also expressed at moderate levels in samples derived from colon cancer and lung cancer cell lines. Thus, expression of this gene could be used to differentiate between these samples and other samples on this panel and as a marker to detect the presence of colon and lung cancer. Furthermore, therapeutic modulation of the expression or function of this gene may be effective in the treatment of colon and lung cancer.

Among tissues with metabolic or endocrine function, this gene is expressed at low to moderate levels in pancreas, adrenal gland, pituitary gland, fetal skeletal muscle, heart, and the gastrointestinal tract. Therefore, therapeutic modulation ofthe activity of this gene may prove useful in the treatment of endocrine/metabolically related diseases, such as obesity and diabetes.

In addition, this gene is expressed at high to moderate levels (CTs=30-33) in all regions ofthe central nervous system examined, including amygdala, hippocampus, substantia nigra, thalamus, cerebellum, cerebral cortex, and spinal cord. Therefore, this gene may play a role in central nervous system disorders such as Alzheimer's disease, Parkinson's disease, epilepsy, multiple sclerosis, schizophrenia and depression.

CG59536-01 codes for protein similar to paraneoplastic cancer-testis-brain antigen. Proteins belonging to paraneoplastic cancer-testis-brain antigen are known to be associated with paraneoplastic syndrome, an immune-mediated disorder. The tumor expression of these proteins normally restricted to neurons (or other immunoprivileged sites, such as testis) but ectopically expressed in some cancers results in an immunological response characterized by high titers of antibodies targeting the "onconeuronal" antigen. A T-cell response is also elicited in some paraneoplastic syndromes and may be the cause of neuronal destruction. In some individuals with cancer but no paraneoplastic syndrome, low titers of antibody can be identified in the serum. Low titers of antibody are associated with a better prognosis ofthe cancer. Experimental animals immunized against a paraneoplastic antigen are partially protected against tumors that express that antigen. See, e.g., Posner et al, (2000) Clin Chem Lab Med 38(2): 117-22. Panel 2.2 Summary: Ag3458 Expression ofthe CG59536-01 gene is restricted to a sample derived from normal lung (CT=28.4). Thus, expression of this gene could be used to differentiate between this sample and other samples on this panel and as a marker to detect the presence of lung cancer. Furthermore, therapeutic modulation ofthe expression or function of this gene may be effective in the treatment of lung cancer. Panel 4D Summary: Ag3458 Highest expression ofthe CG59536-01 gene is seen in the colon (CT=31.1). Therefore, expression of this gene could potentially be used to distinguish colon from the other tissues on this panel. Furthermore, expression of this gene is decreased in colon samples from patients with IBD colitis and Crohn's disease relative to normal colon. Therefore, therapeutic modulation ofthe activity ofthe protein encoded by this gene may be useful in the treatment of inflammatory bowel disease.

In addition, this gene is expressed at moderate levels in a wide range of cell types of significance in the immune response in health and disease. These cells include members of the T-cell, B-cell, endothelial cell, macrophage/monocyte, and peripheral blood mononuclear cell family, as well as epithelial and fibroblast cell types from lung and skin. This widespread pattern of expression suggests that this gene product may be involved in homeostatic processes for these and other cell types and tissues. This pattern is in agreement with the expression profile in General_screening_panel_vl.4 and also suggests a role for the gene product in cell survival and proliferation. Therefore, modulation ofthe gene product with a functional therapeutic may lead to the alteration of functions associated with these cell types and lead to improvement ofthe symptoms of patients suffering from autoimmune and inflammatory diseases such as asthma, allergies, inflammatory bowel disease, lupus erythematosus, psoriasis, rheumatoid arthritis, and osteoarthritis.

Panel CNS_1 Summary: Ag3458 Results from one experiment with the CG59536- 01 gene are not included. The amp plot indicates that there were experimental difficulties with this run.

G. CG59299-01: Q9U I0 C380A1.1B (NOVEL PROTEIN)

Expression of gene CG59299-01 was assessed using the primer-probe set Ag3535, described in Table GA. Results ofthe RTQ-PCR runs are shown in Tables GB, GC and GD.

Table GB. CNS_neurodegeneration_vl.0

Table GC. General_screeningjpanel_vl.4

Table GD. Panel 4.1D

EOL-1 dbcAMP PMA/ionomycin 0.0 jDermal fibroblast IFN gamma 0.8

Dendritic cells none 0.0 ! Dermal fibroblast IL-4 j 1.6 _

Dendritic cells LPS 0.0 JDermal Fibroblasts rest |4.1

Dendritic cells anti-CD40 0.0 JNeutrophils TNFa+LPS |θ.O

Monocytes rest 0.0 JNeutrophils rest JO.O

Monocytes LPS 0.0 (Colon jθ.4

Macrophages rest 0.0 (Lung |0.0

Macrophages LPS 0.0 |Thymus J2.8

HUVEC none 2.9 (Kidney JO.O

HUVEC starved 1.6 1 !

CNS_neurodegeneration_vl.O Summary: Ag3535 This panel confirms the expression ofthe CG59299-01 gene at high levels in the brain in an independent group of individuals. However, no differential expression of this gene was detected between Alzheimer's diseased postmortem brains and those of non-demented controls in this experiment. Please see Panel 1.4 for a discussion ofthe potential utility of this gene in treatment of central nervous system disorders.

General_screeningjpanel_vl.4 Summary: Ag3535 High expression ofthe CG59299-01 is detected throughout the CNS, including in amygdala, substantia nigra, thalamus, cerebellum, cerebral cortex, spinal cord, and CNS cancer samples (CTs=24.9-27). Therefore, this gene may play a role in central nervous system disorders such as Alzheimer's disease, Parkinson's disease, epilepsy, multiple sclerosis, schizophrenia and depression.

Significant expression of this gene is also detected in testis, prostate, one melenoma cell line, prostate cancer, ovarian cancer, breast cancer, lung cancer, renal cancer, gastric cancer and colon cancer (CTs=26.8-28). Therefore, therapeutic modulation ofthe activity of this gene or its protein product, through the use of small molecule drugs, protein therapeutics or antibodies, might be beneficial in the treatment of these cancers.

Panel 4.1D Summary: Ag3535 Highest expression ofthe CG59299-01 gene is detected in TNFalpha + LLlbeta treated astrocytes (CT=30). Significant expression of this gene is also detected in resting secondary Th2, TNFalpha + LLlbeta treated bronchial epithelium, astiocytes, small airway epithelium, CCD 1106 (keratinocytes), IL-9 and IL-13 treated lung fibroblast, LL treated NCI-H292, and dermal fibroblast. Therefore, therapeutic modulation of this gene or its protein product may be beneficial in the treatment of general autoimmunity, asthma and parasitic disease, psoriasis and emphysema. H. CG59632-01 and CG59632-01: Novel expressed mitochondrial protein

Expression of gene CG59632-01 was assessed using the primer-probe set Ag3426, described in Table HA. Please note that CG59632-02 represents a full-length physical clone ofthe CG59632-01 gene, validating the prediction ofthe gene sequence.

Table HA. Probe Name Ag3426

SEQ ID

Primers Sequences Length Start Position NO.

Forward 5 ' -tgagtaagatggcgtccaag-3 ' 120 78

Probe TET-5 ' -gtgctggtggatgacaccagcagt-3 89 TAMRA 24 110

Reversej5 ' -tacagctcatccagcacctc-3 ' 90 20 134

CNS_neurodegeneration_vl.O Summary: Ag3426 Expression ofthe CG59632-01 gene is low/undetectable (CTs > 34.5) across all ofthe samples on this panel (data not shown).

General_screening_panel_vl.4 Summary: Ag3426 Results from one experiment with the CG59632-01 gene are not included. The amp plot indicates that there were experimental difficulties with this run.

Panel 4.1D Summary: Ag3426 Expression ofthe CG59632-01 gene is low/undetectable (CTs > 34.5) across all ofthe samples on this panel (data not shown).

I. CG59303-01: CAC15523 DJ697K14.9.1 (NOVEL PROTEIN) Expression of gene CG59303-01 was assessed using the primer-probe set Ag3537, described in Table LA. Results ofthe RTQ-PCR runs are shown in Tables IB, IC and ID.

Table LA. Probe Name Ag3537

Table IB. CNS_neurodegeneration_vl.0

Table IC. General_screening_panel_vl.4

Table ID. Panel 4D

CNS_neurodegeneration_vl.O Summary: Ag3537 This panel confirms the expression ofthe CG59303-01 gene at low levels in the brain in an independent group of individuals. However, no differential expression of this gene was detected between Alzheimer's diseased postmortem brains and those of non-demented controls in this experiment. Please see Panel 1.4 for a discussion ofthe potential utility of this gene in treatment of central nervous system disorders.

General_screening_panel_vl.4 Summary: Ag3537 Highest expession ofthe CG59303-01 gene is detected in sample derived from the breast cancer cell line T47D (CT=29.3). However, similar high expression of this gene is also seen in a cluster of lung, ovarian, breast, colon, gastric and CNS cancer cell lines. Therefore, therapeutic modulation ofthe activity of this gene or its protein product, through the use of small molecule drugs, protein therapeutics or antibodies, might be beneficial in the treatment of these cancers. Among tissues with metabolic or endocrine function, this gene is expressed at low to moderate levels in pancreas, adipose, thyroid, skeletal muscle, heart, and the gastrointestinal tract. Therefore, therapeutic modulation ofthe activity of this gene may prove useful in the treatment of endocrine/metabolically related diseases, such as obesity and diabetes. Low expression is also detected in fetal liver. Interestingly, this gene is expressed at much higher levels in fetal (CT = 34) when compared to adult liver (CT = 37). This observation suggests that expression of this gene can be used to distinguish fetal from adult liver.

In addition, this gene is expressed at moderate levels in all regions ofthe central nervous system examined, including amygdala, hippocampus, substantia nigra, thalamus, cerebellum, cerebral cortex, and spinal cord. Therefore, this gene may play a role in central nervous system disorders such as Alzheimer's disease, Parkinson's disease, epilepsy, multiple sclerosis, schizophrenia and depression.

Panel 4D Summary: Ag3537 Highest expression ofthe CG59303-01 gene is detected in samples derived from colon (CT=29.5). Thus expression of this gene can be used to distinguish this sample from other samples in the panel. Furthermore, expression of this gene is decreased in colon samples from patients with IBD colitis and Crohn's disease relative to normal colon. Therefore, therapeutic modulation ofthe activity ofthe protein encoded by this gene may be useful in the treatment of inflammatory bowel disease. In addition, this gene is expressed at low to moderate levels in a wide range of cell types of significance in the immune response in health and disease. These cells include members ofthe T-cell, B-cell, endothelial cell, macrophage/monocyte, and peripheral blood mononuclear cell family, as well as epithelial and fibroblast cell types from lung and skin, and normal tissues represented by colon, lung, thymus and kidney. This ubiquitous pattern of expression suggests that this gene product may be involved in homeostatic processes for these and other cell types and tissues. This pattern is in agreement with the expression profile in General_screening_panel_vl .5 and also suggests a role for the gene product in cell survival and proliferation. Therefore, modulation ofthe gene product with a functional therapeutic may lead to the alteration of functions associated with these cell types and lead to improvement ofthe symptoms of patients suffering from autoimmune and inflammatory diseases such as asthma, allergies, inflammatory bowel disease, lupus erythematosus, psoriasis, rheumatoid arthritis, and osteoarthritis. J. CG59719-01: Novel GAP protein

Expression of gene CG59719-01 was assessed using the primer-probe set Ag3654, described in Table JA. Results ofthe RTQ-PCR runs are shown in Tables JB, JC and JD.

Table JA. Probe Name Ag3654

Table JB. CNS_neurodegeneration_vl.0

Table JC. General_screeningjpanel_vl.4

Table JD. Panel 4.1D

CNS_neurodegeneration_vl.O Summary: Ag3654 This panel confirms the expression ofthe CG59719-01 gene at low to moderate levels in the brain in an independent group of individuals. However, no differential expression of this gene was detected between Alzheimer's diseased postmortem brains and those of non-demented controls in this experiment. Please see Panel 1.4 for a discussion ofthe potential utility of this gene in treatment of central nervous system disorders.

General_screening_panel_vl.4 Summary: Ag3654 Highest expression ofthe CG59719-01 gene is detected in a sample derived from one ofthe colon cancer cell line (CT=26.51). Thus, expression of this gene can be used to distinguish this sample from other samples in the panel. In addition, low levels of expression of this gene is also associated with colon cancer, ovarian cancer, breast cancer, and CNS cancer cell lines. Therefore, therapeutic modulation ofthe activity of this gene or its protein product, through the use of small molecule drugs, protein therapeutics or antibodies, might be beneficial in the treatment of these cancers. Interestingly, this gene is expressed at much higher levels in fetal (CT=27.7) when compared to adult lung (CT=32). This observation suggests that expression of this gene can be used to distinguish fetal from adult lung.

Among tissues with metabolic or endocrine function, this gene is expressed at moderate levels in pancreas, adipose, adrenal gland, thyroid, pituitary gland, skeletal muscle, heart, liver and the gastrointestinal tract. Therefore, therapeutic modulation ofthe activity of this gene may prove useful in the treatment of endocrine/metabolically related diseases, such as obesity and diabetes. In addition, this gene is expressed at significant levels in all regions ofthe central nervous system examined, including amygdala, hippocampus, substantia nigra, thalamus, cerebellum, cerebral cortex, and spinal cord. Therefore, this gene may play a role in central nervous system disorders such as Alzheimer's disease, Parkinson's disease, epilepsy, multiple sclerosis, schizophrenia and depression.

CG59719-01 codes for a protein with Ran-GAP domain. The Rap/ran-GAP domain is found in the GTPase activating protein (GAP) responsible for the activation of nuclear Ras- related regulatory proteins Rapl, Rsrl and Ran in vitro converting it to the putatively inactive GDP-bound state. See, e.g., Rubinfeld et al. (1991) Cell 65(6): 1033-1042; Hattori et al. (1995) Mol. Cell. Biol. 15(1): 552-560.

Panel 4.1D Summary: Ag3654 The CG59719-01 gene is expressed at high to moderate levels in a wide range of cell types of significance in the immune response in health and disease. These cells include members ofthe T-cell, B-cell, endothelial cell, macrophage/monocyte, and peripheral blood mononuclear cell family, as well as epithelial and fibroblast cell types from lung and skin, and normal tissues represented by colon, lung, thymus and kidney. This ubiquitous pattern of expression suggests that this gene product may be involved in homeostatic processes for these and other cell types and tissues.

Therefore, modulation ofthe gene product with a functional therapeutic may lead to the alteration of functions associated with these cell types and lead to improvement ofthe symptoms of patients suffering from autoimmune and inflammatory diseases such as asthma, allergies, inflammatory bowel disease, lupus erythematosus, psoriasis, rheumatoid arthritis, and osteoarthritis.

K. CG59777-01: GTPase activator protein

Expression of gene CG59777-01 was assessed using the primer-probe set Ag3582, described in Table KA. Results of he RTQ-PCR runs are shown in Tables KB, and KC.

Table KA. Probe Name Ag3582

Table KB. CNS_neurodegeneration_yl.O

Tissue Name Rel. Exp.(%) Ag3582, Run Rel. Exp.(%) Ag3582, Run

Tissue Name 211006253 211006253

Table KC. Panel 4.1D

CNS_neurodegeneration_vl.0 Summary: Ag3582 This panel demonstrates the expression ofthe CG59777-01 gene at moderate levels in the brains of several individuals. However, no differential expression of this gene was detected between Alzheimer's diseased postmortem brains and those of non-demented controls in this experiment. The CG59777-01 gene encodes a protein with homology to vascular Rab-GAP/TBC domain-containing protein (VRP), a protein identified in a screen for angiogenesis-related proteins. See, e.g., Yonekura (2001) Ann N Y Acad Sci 947:382-6.

General_screening_panel_vl.4 Summary: Ag3582 Results from one experiment with the CG59777-01 gene are not included. The amp plot indicates that there were experimental difficulties with this run.

Panel 4.1D Summary: Ag3582 Expression ofthe CG59777-01 gene is up regulated in LPS-stimulated monocytes (CT = 26.2), compared to resting monocytes (CT = 28.4). Thus, expression of this gene may be used as a marker for activated monocytes. The putative GTPase activating protein encoded by this gene may therefore be involved in the activation of monocytes in their function as antigen-presenting cells. This suggests that inhibition ofthe activity of this gene or its protein product may be useful as anti-inflammatory therapeutics for the treatment of autoimmune and inflammatory diseases.

This gene is also expressed at moderate levels in a number of other samples on this panel, including dermal fibroblasts, lung fibroblasts, keratinocytes, astrocytes, bronchial and small airway epithelium, dendritic cells, and macrophages as well as normal tissues represented by colon, lung, thymus and kidney. This ubiquitous pattern of expression suggests that this gene product may be involved in homeostatic processes for these and other cell types and tissues. Furthermore, therapeutic modulation ofthe activity of this gene or its protein product may lead to the alteration of functions associated with these cell types and lead to improvement ofthe symptoms of patients suffering from autoimmune and inflammatory diseases such as asthma, allergies, inflammatory bowel disease, lupus erythematosus, psoriasis, rheumatoid arthritis, and osteoarthritis.

L. CG59658-01: F-Box

Expression of gene CG59658-01 was assessed using the primer-probe sets Ag3359 and Ag3765, described in Tables LA and LB. Results ofthe RTQ-PCR runs are shown in Tables LC, LD, and LE.

Table LA. Probe Name Ag3359

Table LB. Probe Name Ag3765

Table LD. General _screening_panel_vl.4

Rel. Exp.(%) Rel. Exp.(%)

Tissue Name Ag3359, Run Tissue Name Ag3359, Run

216523479 216523479

Adipose 5.1 Renal ca. TK-10 15.7

Melanoma* Hs688(A).T 7.9 Bladder 10.4

Melanoma* Hs688(B).T 6,7 _ Gastric ca. (liver met.) NCI-N87 48.6

Melanoma* M14 12.9 Gastric ca. KATO III 94.0

Melanoma* LOXIMVI 29.3 Colon ca. SW-948 15.8

Melanoma* SK-MEL-5 21.2 Colon ca. SW480 57.4

Squamous cell carcinoma SCC-4 15.0 Colon ca.* (SW480 met) SW620 47.3

Testis Pool 12.0 Colon ca. HT29 13.6

Prostate ca.* (bone met) PC-3 18.6 Colon ca. HCT-116 43.2

Prostate Pool 5.4 Colon ca. CaCo-2 41.2

Placenta 8.0 Colon cancer tissue 21.5

Uterus Pool 3.1 Colon ca. SW1116 9.9

Ovarian ca. OVCAR-3 40.6 Colon ca. Colo-205 14.7

Ovarian ca. SK-OV-3 23.3 Colon ca. SW-48 11.0

Ovarian ca. OVCAR-4 110.2 Colon Pool 9.2

Table LE. Panel 4D

CNS_neurodegeneration_vl.O Summary: Ag3359/Ag3765 This panel confirms the expression ofthe CG59658-01 gene at low levels in the brain in an independent group of individuals. However, no differential expression of this gene was detected between Alzheimer's diseased postmortem brains and those of non-demented controls in this experiment. Please see Panel 1.4 for a discussion ofthe potential utility of this gene in treatment of central nervous system disorders.

General_screening_panel_yl.4 Summary: Ag3359 Highest expression ofthe CG59658-01 gene is seen in one ofthe CNS cancer (glio/astro) U-118-MG cell line (CT=28). In addition, expression of this gene is up-regulated in lung cancer, CNS cancer, colon cancer, gastric cancer, pancreatic cancer, ovarian cancer, breast cancer and melanoma cell-lines.

Thus, expression of this gene can be used to distinguish cancer cells from normal tissue used in this panel. Furthermore, therapeutic modulation ofthe activity ofthe protein encoded by this gene may be beneficial in the treatment of these cancers.

Among tissues with metabolic or endocrine function, this gene is expressed at low to moderate levels in pancreas, adipose, adrenal gland, thyroid, pancrease, pituitary gland, skeletal muscle, heart, liver and the gastrointestinal tract. Therefore, therapeutic modulation ofthe activity of this gene may prove useful in the treatment of endocrine/metabolically related diseases, such as obesity and diabetes.

In addition, this gene is expressed at high levels in all regions ofthe central nervous system examined, including amygdala, hippocampus, substantia nigra, thalamus, cerebellum, cerebral cortex, and spinal cord. Therefore, this gene may play a role in central nervous system disorders such as Alzheimer's disease, Parkinson's disease, epilepsy, multiple sclerosis, schizophrenia and depression.

Panel 4.1D Summary: Ag3765 Results from one experiment with the CG59658-01 gene are not included. The amp plot indicates that there were experimental difficulties with this run.

Panel 4D Summary: Ag3359 Highest expression ofthe CG59658-01 gene is seen in ionomycin treated Ramos (B cell) sample (CT=27). Lower but still significant levels of expression of this gene are seen in untreated Ramos B cells. In addition, expression of this gene is up-regulated in PWM treated - PBMC and B lymphocytes (CTs=28). B cells represent a principle component of immunity and contribute to the immune response in a number of important functional roles, including antibody production. Production of antibodies against self-antigens is a major component in autoimmune disorders. Since B cells play an important role in autoimmunity, inflammatory processes and inflammatory cascades, therapeutic modulation of this gene product may reduce or eliminate the symptoms of patients suffering from asthma, allergies, chronic obstructive pulmonary disease, emphysema, Crohn's disease, ulcerative colitis, rheumatoid arthritis, psoriasis, osteoarthritis, systemic lupus erythematosus and other autoimmune disorders.

Also, this gene is expressed at low to moderate levels in a wide range of cell types of significance in the immune response in health and disease. These cells include members of the T-cell, B-cell, endothelial cell, macrophage/monocyte, and peripheral blood mononuclear cell family, as well as epithelial and fibroblast cell types from lung and skin, and normal tissues represented by colon, lung, thymus and kidney. This ubiquitous pattern of expression suggests that this gene product may be involved in homeostatic processes for these and other cell types and tissues. This pattern is in agreement with the expression profile in General_screening_panel_vl.5 and also suggests a role for the gene product in cell survival and proliferation. Therefore, modulation of the gene product with a functional therapeutic may lead to the alteration of functions associated with these cell types and lead to improvement ofthe symptoms of patients suffering from autoimmune and inflammatory diseases such as asthma, allergies, inflammatory bowel disease, lupus erythematosus, psoriasis, rheumatoid arthritis, and osteoarthritis. M. CG59907-01 : Novel GAP protein

Expression of gene CG59907-01 was assessed using the primer-probe set Ag3629, described in Table MA. Results ofthe RTQ-PCR runs are shown in Tables MB, MC and MD.

Table MA. Probe Name Ag3629

Table MB. CNS neurodeeeneration vl.O

Table MD. Panel 4.1D

CNS_neurodegeneration_vl.O Summary: Ag3629 This panel confirms the expression ofthe CG59907-01 gene at low levels in the brains of an independent group of individuals. However, no differential expression of this gene was detected between Alzheimer's diseased postmortem brains and those of non-demented controls in this experiment. Please see Panel 1.4 for a discussion ofthe potential utility of this gene in treatment of central nervous system disorders.

General_screening_panel_vl.4 Summary: Ag3629 Highest expression ofthe CG59907-01 gene is seen in a breast cancer cell line (CT=26.4), with expression widespread throughout this panel. Significant levels of expression are also seen in samples derived from an ovarian cancer cell line and a pancreatic cancer cell line. Thus, expression of this gene could be used to differentiate between these samples and other samples on this panel and as a marker to detect the presence of breast, ovarian, and pancreatic cancers. Furthermore, therapeutic modulation ofthe expression or function of this gene may be effective in the treatment of breast, ovarian, and pancreatic cancers. In addition, this gene is expressed at significant levels in all regions ofthe central nervous system examined, including amygdala, hippocampus, substantia nigra, thalamus, cerebellum, cerebral cortex, and spinal cord. Therefore, this gene may play a role in central nervous system disorders such as Alzheimer's disease, Parkinson's disease, epilepsy, multiple sclerosis, schizophrenia and depression. Among tissues with metabolic function, this gene is expressed at moderate to low levels in pituitary, adipose, adrenal gland, pancreas, thyroid, fetal liver and adult and fetal skeletal muscle, and heart. This widespread expression among these tissues suggests that this gene product may play a role in normal neuroendocrine and metabolic and that disregulated expression of this gene may contribute to neuroendocrine disorders or metabolic diseases, such as obesity and diabetes.

In addition, this gene is expressed at much higher levels in fetal lung and liver tissue (CTs=29-32) when compared to expression in the adult counterpart (CTs=32-38). Thus, expression of this gene may be used to differentiate between the fetal and adult source of these tissue. Panel 4.1D Summary: Ag3629 Expression ofthe CG59907-01 gene is ubiquitous in this panel, with highest expression in the thymus (CT=31.7). In addition, this gene is expressed at high to moderate levels in a wide range of cell types of significance in the immune response in health and disease. These cells include members ofthe T-cell, B-cell, endothelial cell, macrophage/monocyte, and peripheral blood mononuclear cell family, as well as epithelial and fibroblast cell types from lung and skin, and normal tissues represented by colon, lung, thymus and kidney. This ubiquitous pattern of expression suggests that this gene product may be involved in homeostatic processes for these and other cell types and tissues. This pattern is in agreement with the expression profile in

General_screening_panel_vl.4 and also suggests a role for the gene product in cell survival and proliferation. Therefore, modulation ofthe gene product with a functional therapeutic may lead to the alteration of functions associated with these cell types and lead to improvement ofthe symptoms of patients suffering from autoimmune and inflammatory diseases such as asthma, allergies, inflammatory bowel disease, lupus erythematosus, psoriasis, rheumatoid arthritis, and osteoarthritis.

N. CG59903-01: Novel nuclear protein

Expression of gene CG59903-01 was assessed using the primer-probe set Ag3628, described in Table NA. Results ofthe RTQ-PCR runs are shown in Tables NB, NC and ND.

Table NB. CNS_neurodegeneration_vl.0

AD 3 Temporal Ctx 163 Control 4 Occipital Ctx 7.5

AD 4 Temporal Ctx 33.7 Control (Path) 1 Occipital Ctx 91.4

AD 5 Inf Temporal Ctx 97.3 Control (Path) 2 Occipital Ctx 14.4

AD 5 SupTemporal Ctx 36.9 Control (Path) 3 Occipital Ctx 6.2

AD 6 Inf Temporal Ctx 59.9 Control (Path) 4 Occipital Ctx 13.2

AD 6 Sup Temporal Ctx 753 Control 1 Parietal Ctx 19.1

Control 1 Temporal Ctx 29.1 Control 2 Parietal Ctx 49.3

Control 2 Temporal Ctx 35.1 Control 3 Parietal Ctx 12.5

Control 3 Temporal Ctx 12.5 Control (Path) 1 Parietal Ctx 100.0

Control 4 Temporal Ctx 16.5 Control (Path) 2 Parietal Ctx 44.8

Control (Path) 1 Temporal Ctx 87.1 Control (Path) 3 Parietal Ctx 8.2

Control (Path) 2 Temporal Ctx 74.2 Control (Path) 4 Parietal Ctx 34.9

Table NC. General_screening_panel_vl.4

Rel. Exp.(%) Rel. Exp.(%)

Tissue Name Ag3628, Run Tissue Name Ag3628, Run

218211823 218211823

Adipose 6.1 Renal ca. TK-10 23.2

Melanoma* Hs688(A).T 0.0 Bladder 19.1

Melanoma* Hs688(B).T 0.1 Gastric ca. (liver met.) NCI-N87 15.5

Melanoma* M14 0.0 Gastric ca. KATO III 56.3

Melanoma* LOXIMVI 0.1 Colon ca. SW-948 12.2

Melanoma* SK-MEL-5 11.0 Colon ca. SW480 253

Squamous cell carcinoma SCC-4 28.5 Colon ca.* (SW480 met) SW620 27.5

Testis Pool 7.7 Colon ca. HT29 16.7

Prostate ca.* (bone met) PC-3 14.6 Colon ca. HCT-116 23.7

Prostate Pool 0.0 Colon ca. CaCo-2 21.6

Placenta 3.0 Colon cancer tissue 13.6

Uterus Pool 3.0 Colon ca. SW1116 7.7

Ovarian ca. OVCAR-3 19.1 Colon ca. Colo-205 4.2

Ovarian ca. SK-OV-3 13.2 Colon ca. SW-48 4.0

Ovarian ca. OVCAR-4 14.3 Colon Pool 8.5

Ovarian ca. OVCAR-5 64.2 Small Intestine Pool 7.4

Ovarian ca. IGROV-1 5.1 Stomach Pool 5.4

Ovarian ca. OVCAR-8 2.1 Bone Marrow Pool 3.6

Ovary 3.3 Fetal Heart 3.9

Breast ca. MCF-7 23.8 Heart Pool 2.2

Breast ca. MDA-MB-231 31.9 Lymph Node Pool 9.0

Breast ca. BT 549 34.9 Fetal Skeletal Muscle 4.7

Breast ca. T47D 100.0 Skeletal Muscle Pool 2.0

Breast ca. MDA-N o-o Spleen Pool 34.2

Breast Pool 11.0 Thymus Pool 473

Trachea 28.1 CNS cancer (glio/astro) U87-MG 0.0

Lung 0.7 CNS cancer (glio/astro) U-118-MG 0.0

Fetal Lung 60.3 CNS cancer (neuro;met) SK-N-AS 0.0 Lung ca. NCI-N417 3.4 CNS cancer (astro) SF-539 0.1

Lung ca. LX-1 33.0 iCNS cancer (astro) SNB-75 0.5

Lung ca. NCI-H146 (8.7 CNS cancer (glio) SNB- 19 7.0 Lung ca. SHP-77 161.1 CNS cancer (glio) SF-295 7.7

Lung ca. A549 |ll.6 Brain (Amygdala) Pool 3.6

Lung ca. NCI-H526 |7.7 Brain (cerebellum) 1.5

Lung ca. NCI-H23 |8.7 Brain (fetal) 23.8

Lung ca. NCI-H460 (9.3 Brain (Hippocampus) Pool 2.9

Lung ca. HOP-62 J3.5 Cerebral Cortex Pool 2.1

Lung ca. NCI-H522 32.5 JBrain (Substantia nigra) Pool 2.5 Liver jθ.6 Brain (Thalamus) Pool 5.7

Fetal Liver J17.0 Brain (whole) 6.0

Liver ca. HepG2 14.0 Spinal Cord Pool 4.2

Kidney Pool (14.3 Adrenal Gland 2.2

Fetal Kidney (18.6 Pituitary gland Pool 3.2

Renal ca. 786-0 |4.8 Salivary Gland 8.1

Renal ca. A498 (0.6 Thyroid (female) 1.5 Renal ca. ACHN |4.9 Pancreatic ca. CAPAN2 23.7

Renal ca. UO-31 |7.4 Pancreas Pool 12.9

Table ND. Panel 4.1D

CNS_neurodegeneration_vl.O Summary: Ag3628 This panel does not show differential expression ofthe CG59903-01 gene in Alzheimer's disease. However, this expression profile confirms the presence of this gene in the brain. Please see Panel 1.4 for discussion of utility of this gene in the central nervous system.

General_screeningjpanel__vl.4 Summary: Ag3628 Highest expression ofthe CG59903-01 gene is seen in a breast cancer cell line (CT=25.5). Significant levels of expression are also seen in cell lines derived from gastric, lung, ovarian, and colon cancers. Thus, expression of this gene could be used as a marker to detect the presence of these cancers. Furthermore, therapeutic modulation ofthe expression or function of this gene may be effective in the treatment of breast, gastric, lung, ovarian, and colon cancers.

Among tissues with metabolic function, this gene is expressed at moderate to low levels in pituitary, adipose, adrenal gland, pancreas, thyroid, and adult and fetal skeletal muscle, heart, and liver. This widespread expression among these tissues suggests that this gene product may play a role in normal neuroendocrine and metabolic and that disregulated expression of this gene may contribute to neuroendocrine disorders or metabolic diseases, such as obesity and diabetes.

This molecule is also expressed at moderate levels in the CNS and may be a small molecule target for the treatment of neurologic diseases.

In addition, expression of this gene is higher in fetal lung and liver (CTs=26-28) when compared to expression in the corresponding adult tissues (CTs=32). Thus, expression of this gene may be used to differentiate between the fetal and adult sources of these tissue.

Panel 4.1D Summary: Ag3628 Highest expression ofthe CG59903-01 gene is seen in a sample derived from primary resting T cells (CT=27), with expression widespread throughout this panel.

Significant levels of expression are also found in other T cells including activated primary Thl, Th2 and Trl cells, resting primary Th2 and Trl cells, CD45RO CD4 lymphocytes, resting secondary CD8 lymphocytes, and IL2+IL12 and IL2+IL18 stimulated lymphokine activated killer (LAK) cells. In additional, expression is detected in peripheral blood mononuclear cells (PBMC), polkweed mitogen stimulated B lymphocytes, normal thymus, and stimulated dermal fibroblasts. Since eosinophils B cells and T cells play an important role in lung pathology, inflammatory bowel disease and autoimmune disorders, therapeutic modulation ofthe protein encoded by this gene could block or inhibit inflammation or tissue damage due to lung conditions including asthma, allergies, hypersensitivity reactions, inflammatory bowel disease, viral infections and autoimmune diseases, such as rheumatoid arthritis, systemic lupus erythematosus. In addition, the expression of this gene in T cells, B cells, and PBMCs also suggests that therapeutic modulation of this gene product may ameliorate symptoms associated with synovitis associated with osteoarthritis. Furthermore, detection of this gene in LAK cells suggests that modulation ofthe function of this gene product may also lead to improvement of symptoms associated with tumor immunology and tumor cell clearance, as well as removal of virally and bacterial infected cells. O. CG59985-01: WD repeat protein - isoforml, submitted to study DDNPAT on 03/28/01 by cpena; clone status=FIS; noveIty=Novel; ORF start=14, ORF stop=962, frame=2; 1043 bp.

Expression of gene CG59985-01 was assessed using the primer-probe set Ag3642, described in Table OA. Results ofthe RTQ-PCR runs are shown in Tables OB, and OC.

Table OD. Panel 4.1D

CNS_neurodegeneration_vl.0 Summary: Ag3642 Results from one experiment with the CG59985-01 gene are not included. The amp plot indicates that there were experimental difficulties with this run.

General_screening_panel_vl.4 Summary: Ag3642 Expression ofthe CG59985-01 gene is ubiquitous in this panel, with highest expression in a breast cancer cell line (CT=26.6). Overall, expression of this gene appears to be higher in samples derived from cancer cell lines than in normal tissues. This widespread expression suggests that this gene product is involved in cell growth and proliferation. Thus, expression of this gene could be used as a marker to detect the presence of cancer. Furthermore, therapeutic modulation ofthe expression or function of this gene may be useful in the treatment of cancer.

Interestingly, this gene is expressed at much higher levels in fetal lung and liver (CTs=29-30) when compared to expression in the adult counterpart (CTs=33). Thus, expression of this gene may be used to differentiate between the fetal and adult sources of these tissue. Among tissues with metabolic function, this gene is expressed at moderate to low levels in pituitary, adipose, adrenal gland, pancreas, thyroid, and adult and fetal skeletal muscle, heart, and liver. This widespread expression among these tissues suggests that this gene product may play a role in normal neuroendocrine and metabolic and that disregulated expression of this gene may contribute to neuroendocrine disorders or metabolic diseases, such as obesity and diabetes.

In addition, this gene is expressed at low to moderate levels in all regions ofthe central nervous system examined, including amygdala, hippocampus, substantia nigra, thalamus, cerebellum, cerebral cortex, and spinal cord. Therefore, this gene may play a role in central nervous system disorders such as Alzheimer's disease, Parkinson's disease, epilepsy, multiple sclerosis, schizophrenia and depression.

Panel 4.1D Summary: Ag3642 The CG59985-01 gene is expressed at high to moderate levels in a wide range of cell types of significance in the immune response in health and disease. Highest expression ofthe gene is seen in activated primary T cells (CT=30.7). Significant levels of expression are also seen in members ofthe T-cell, B-cell, endothelial cell, macrophage/monocyte, and peripheral blood mononuclear cell family, as well as epithelial and fibroblast cell types from lung and skin, and normal tissues represented by colon, lung, thymus and kidney. This ubiquitous pattern of expression suggests that this gene product may be involved in homeostatic processes for these and other cell types and tissues. This pattern is in agreement with the expression profile in General_screenmg_panel_vl .4 and also suggests a role for the gene product in cell survival and proliferation. Therefore, modulation ofthe gene product with a functional therapeutic may lead to the alteration of functions associated with these cell types and lead to improvement ofthe symptoms of patients suffering from autoimmune and inflammatory diseases such as asthma, allergies, inflammatory bowel disease, lupus erythematosus, psoriasis, rheumatoid arthritis, and osteoarthritis.

Example D. Identification of Single Nucleotide Polymorphisms in NOVX nucleic acid sequences Variant sequences are also included in this application. A variant sequence can include a single nucleotide polymorphism (SNP). A SNP can, in some instances, be referred to as a "cSNP" to denote that the nucleotide sequence containing the SNP originates as a cDNA. A SNP can arise in several ways. For example, a SNP may be due to a substitution of one nucleotide for another at the polymoφhic site. Such a substitution can be either a transition or a transversion. A SNP can also arise from a deletion of a nucleotide or an insertion of a nucleotide, relative to a reference allele. In this case, the polymorphic site is a site at which one allele bears a gap with respect to a particular nucleotide in another allele. SNPs occurring within genes may result in an alteration ofthe amino acid encoded by the gene at the position ofthe SNP. Intragenic SNPs may also be silent, when a codon including a SNP encodes the same amino acid as a result ofthe redundancy ofthe genetic code. SNPs occurring outside the region of a gene, or in an intron within a gene, do not result in changes in any amino acid sequence of a protein but may result in altered regulation ofthe expression pattern. Examples include alteration in temporal expression, physiological response regulation, cell type expression regulation, intensity of expression, and stability of transcribed message.

SeqCalling assemblies produced by the exon linking process were selected and extended using the following criteria. Genomic clones having regions with 98% identity to all or part ofthe initial or extended sequence were identified by BLASTN searches using the relevant sequence to query human genomic databases. The genomic clones that resulted were selected for further analysis because this identity indicates that these clones contain the genomic locus for these SeqCalling assemblies. These sequences were analyzed for putative coding regions as well as for similarity to the known DNA and protein sequences. Programs used for these analyses include Grail, Genscan, BLAST, HMMER, FASTA, Hybrid and other relevant programs. Some additional genomic regions may have also been identified because selected

SeqCalling assemblies map to those regions. Such SeqCalling sequences may have overlapped with regions defined by homology or exon prediction. They may also be included because the location ofthe fragment was in the vicinity of genomic regions identified by similarity or exon prediction that had been included in the original predicted sequence. The sequence so identified was manually assembled and then may have been extended using one or more additional sequences taken from CuraGen Corporation's human SeqCalling database. SeqCalling fragments suitable for inclusion were identified by the CuraTools program SeqExtend or by identifying SeqCalling fragments mapping to the appropriate regions ofthe genomic clones analyzed.

The regions defined by the procedures described above were then manually integrated and corrected for apparent inconsistencies that may have arisen, for example, from miscalled bases in the original fragments or from discrepancies between predicted exon junctions, EST locations and regions of sequence similarity, to derive the final sequence disclosed herein. When necessary, the process to identify and analyze SeqCalling assemblies and genomic clones was reiterated to derive the full length sequence (Alderborn et al., Determination of Single Nucleotide Polymorphisms by Real-time Pyrophosphate DNA Sequencing. Genome Research. 10 (8) 1249-1265, 2000). Variants are reported individually but any combination of all or a select subset of variants are also mcluded as contemplated NOVX embodiments ofthe invention. NOV3 SNP data:

NOV3 has three SNP variants, whose variant positions for its nucleotide and amino acid sequences is numbered according to SEQ ID NOs:7 and 8, respectively. The nucleotide sequence ofthe NOV3 variant differs as shown in Table SNP1. Table SNP1. NOV3 variants.

NOV6a SNP data:

NOV6a has two SNP variants, whose variant positions for its nucleotide and amino acid sequences is numbered according to SEQ ID NOs:13 and 14, respectively. The nucleotide sequence ofthe NOV6a variant differs as shown in Table SNP2. Table SNP2. NOV6a variants.

NOV8 SNP data:

NOV8 has two SNP variants, whose variant positions for its nucleotide and amino acid sequences is numbered accordmg to SEQ ID NOs:19 and 20, respectively. The nucleotide sequence ofthe NOV8 variant differs as shown in Table SNP3.

Table SNP3. NOV8 variants.

NOV12 SNP data:

NOVl 2 has 2 SNP variants, whose variant positions for its nucleotide and amino acid sequences is numbered according to SEQ ID NOs:29 and 30, respectively. The nucleotide sequence ofthe NOVl 2 variant differs as shown in Table SNP4. Table SNP4. NOV12 variantsi

NOV13a SNP data: NOVl 3a has two SNP variants, whose variant positions for its nucleotide and amino acid sequences is numbered according to SEQ ID NOs:31 and 32, respectively. The nucleotide sequence ofthe NOV13a variant differs as shown in Table SNP5. Table SNP5. NOV13a variants.

NOV15 SNP data:

NOVl 5 has one SNP variant, whose variant position for its nucleotide and amino acid sequences is numbered according to SEQ ID NOs:37 and 38, respectively. The nucleotide sequence ofthe NOVl 5 variant differs as shown in Table SNP6.

Table SNP6. NOV15 variants.

NOV21 SNP data:

NOV21 has two SNP variants, whose variant positions for its nucleotide and amino acid sequences is numbered according to SEQ ID NOs:51 and 52, respectively. The nucleotide sequence ofthe NOV21 variant differs as shown in Table SNP7.

Table SNP7. NOV21 variants.

NOV22 SNP data:

NOV22 has one SNP variant, whose variant position for its nucleotide and amino acid sequences is numbered according to SEQ ID NOs:53 and 54, respectively. The nucleotide sequence ofthe NOV22 variant differs as shown in Table SNP8.

Table SNP8. NOV22 variants.

NOV24 SNP data:

NOV24 has one SNP variant, whose variant position for its nucleotide and amino acid sequences is numbered according to SEQ ID NOs:57 and 58, respectively. The nucleotide sequence ofthe NOV24 variant differs as shown in Table SNP9. Table SNP9. NOV24 variant.

NOV25 SNP data:

^' NOV25 has one SNP variant, whose variant position for its nucleotide and amino acid sequences is numbered according to SEQ ID NOs:59 and 60, respectively. The nucleotide sequence ofthe NOV25 variant differs as shown in Table SNP 10. Table SNP10. NOV25 variant.

NOV27 SNP data: NOV27 has two SNP variants, whose variant positions for its nucleotide and amino acid sequences is numbered according to SEQ ID NOs:63 and 64, respectively. The nucleotide sequence ofthe NOV27 variant differs as shown in Table SNP11. Table SNP11. NOV27 variants.

OTHER EMBODIMENTS

Although particular embodiments have been disclosed herein in detail, this has been done by way of example for purposes of illustration only, and is not intended to be limiting with respect to the scope ofthe appended claims, which follow. In particular, it is contemplated by the inventors that various substitutions, alterations, and modifications may be made to the invention without departing from the spirit and scope ofthe invention as defined by the claims. The choice of nucleic acid starting material, clone of interest, or library type is believed to be a matter of routine for a person of ordinary skill in the art with knowledge ofthe embodiments described herein. Other aspects, advantages, and modifications considered to be within the scope ofthe following claims.

The claims presented are representative ofthe inventions disclosed herein. Other, unclaimed inventions are also contemplated. Applicants reserve the right to pursue such inventions in later claims.

Claims

WHAT IS CLAIMED IS:

1. An isolated polypeptide comprising an amino acid sequence selected from the group consisting of: a) a mature form ofthe amino acid sequence selected from the group consisting of SEQ ID NO: 2n, wherein n is an integer between 1 and 33; b) a variant of a mature form ofthe amino acid sequence selected from the group consisting of SEQ ID NO: 2n, wherein n is an integer between 1 and 33, wherein any amino acid in the mature form is changed to a different amino acid, provided that no more than 15% ofthe amino acid residues in the sequence ofthe mature form are so changed; c) the amino acid sequence selected from the group consisting of SEQ ID NO: 2n, wherein n is an integer between 1 and 33; d) a variant ofthe amino acid sequence selected from the group consisting of SEQ ID NO: 2n, wherein n is an integer between 1 and 33, wherein any amino acid specified in the chosen sequence is changed to a different amino acid, provided that no more than 15% ofthe amino acid residues in the sequence are so changed; and e) a fragment of any of a) through d).

2. The polypeptide of claim 1 that is a naturally occurring allelic variant ofthe sequence selected from the group consisting of SEQ ID NO: 2n, wherein n is an integer between 1 and 33.

3. The polypeptide of claim 2, wherein the allelic variant comprises an amino acid sequence that is the translation of a nucleic acid sequence differing by a single nucleotide from a nucleic acid sequence selected from the group consisting of SEQ ID NOS: 2n, wherein n is an integer between 1 and 33.

4. The polypeptide of claim 1 that is a variant polypeptide described therein, wherein any amino acid specified in the chosen sequence is changed to provide a conservative substitution.

5. A pharmaceutical composition comprising the polypeptide of claim 1 and a pharmaceutically acceptable carrier.

6. A kit comprising in one or more containers, the pharmaceutical composition of claim

5.

7. The use of a therapeutic in the manufacture of a medicament for treating a syndrome associated with a human disease, the disease selected from a pathology associated with the polypeptide of claim 1, wherein the therapeutic is the polypeptide of claim 1.

8. A method for determining the presence or amount ofthe polypeptide of claim 1 in a sample, the method comprising:

(a) providing the sample;

(b) introducing the sample to an antibody that binds immunospecifically to the polypeptide; and

(c) determining the presence or amount of antibody bound to the polypeptide, thereby determining the presence or amount of polypeptide in the sample.

9. A method for determining the presence of or predisposition to a disease associated with altered levels ofthe polypeptide of claim 1 in a first mammalian subject, the method comprising: a) measuring the level of expression ofthe polypeptide in a sample from the first mammalian subject; and b) comparing the amount ofthe polypeptide in the sample of step (a) to the amount ofthe polypeptide present in a control sample from a second mammalian subject known not to have, or not to be predisposed to, the disease, wherein an alteration in the expression level ofthe polypeptide in the first subject as compared to the control sample indicates the presence of or predisposition to the disease.

10. A method of identifying an agent that binds to the polypeptide of claim 1 , the method comprising:

(a) introducing the polypeptide to the agent; and

(b) determining whether the agent binds to the polypeptide.

11. The method of claim 10 wherein the agent is a cellular receptor or a downstream effector.

12. A method for identifying ^ potential therapeutic agent for use in treatment of a pathology, wherein the pathology is related to aberrant expression or aberrant physiological interactions ofthe polypeptide of claim 1, the method comprising:

(a) providing a cell expressing the polypeptide of claim 1 and having a property or function ascribable to the polypeptide;

(b) contacting the cell with a composition comprising a candidate substance; and (c) determining whether the substance alters the property or function ascribable to the polypeptide; whereby, if an alteration observed in the presence ofthe substance is not observed when the cell is contacted with a composition devoid ofthe substance, the substance is identified as a potential therapeutic agent.

13. A method for screening for a modulator of activity or of latency or predisposition to a pathology associated with the polypeptide of claim 1, the method comprising: a) administering a test compound to a test animal at increased risk for a pathology associated with the polypeptide of claim 1, wherein the test animal recombinantly expresses the polypeptide of claim 1; b) measuring the activity ofthe polypeptide in the test animal after administering the compound of step (a); and c) comparing the activity ofthe protein in the test animal with the activity ofthe polypeptide in a control animal not administered the polypeptide, wherein a change in the activity ofthe polypeptide in the test animal relative to the control animal indicates the test compound is a modulator of latency of, or predisposition to, a pathology associated with the polypeptide of claim 1.

14. The method of claim 13, wherein the test animal is a recombinant test animal that expresses a test protein transgene or expresses the transgene under the control of a promoter at an increased level relative to a wild-type test animal, and wherein the promoter is not the native gene promoter ofthe transgene.

15. A method for modulating the activity of the polypeptide of claim 1 , the method comprising introducing a cell sample expressing the polypeptide ofthe claim with a compound that binds to the polypeptide in an amount sufficient to modulate the activity ofthe polypeptide.

16. A method of treating or preventing a pathology associated with the polypeptide of claim 1, the method comprising administering the polypeptide of claim 1 to a subject in which such treatment or prevention is desired in an amount sufficient to treat or prevent the pathology in the subject.

17. The method of claim 16, wherein the subject is a human.

18. A method of treating a pathological state in a mammal, the method comprising administering to the mammal a polypeptide in an amount that is sufficient to alleviate the pathological state, wherein the polypeptide is a polypeptide having an amino acid sequence at least 95% identical to a polypeptide comprising the amino acid sequence selected from the group consisting of SEQ ID NO: 2n, wherein n is an integer between 1 and 33, or a biologically active fragment thereof.

19. An isolated nucleic acid molecule comprising a nucleic acid sequence encoding a polypeptide comprising an amino acid sequence selected from the group consisting of: a) a mature form of the amino acid sequence given SEQ ID NO: 2n, wherein n is an integer between 1 and 33; b) a variant of a mature form ofthe amino acid sequence selected from the group consisting of SEQ ID NO: 2n, wherein n is an integer between 1 and 33, wherein any amino acid in the mature form ofthe chosen sequence is changed to a different amino acid, provided that no more than 15% ofthe amino acid residues in the sequence of the mature form are so changed; c) the amino acid sequence selected from the group consisting of SEQ ID NO: 2n, wherein n is an integer between 1 and 33; d) a variant ofthe amino acid sequence selected from the group consisting of SEQ ID NO: 2n, wherein n is an integer between 1 and 33, in which any amino acid specified in the chosen sequence is changed to a different amino acid, provided that no more than 15% ofthe amino acid residues in the sequence are so changed; e) a nucleic acid fragment encoding at least a portion of a polypeptide comprising the amino acid sequence selected from the group consisting of SEQ ID NO: 2n, wherein n is an integer between 1 and 33, or any variant ofthe polypeptide wherein any amino acid ofthe chosen sequence is changed to a different amino acid, provided that no more than 10% ofthe amino acid residues in the sequence are so changed; and f) the complement of any ofthe nucleic acid molecules.

20. The nucleic acid molecule of claim 19, wherein the nucleic acid molecule comprises the nucleotide sequence of a naturally occurring allelic nucleic acid variant.

21. The nucleic acid molecule of claim 19 that encodes a variant polypeptide, wherein the variant polypeptide has the polypeptide sequence of a naturally occurring polypeptide variant.

22. The nucleic acid molecule of claim 19, wherein the nucleic acid molecule differs by a single nucleotide from a nucleic acid sequence selected from the group consisting of SEQ ID NOS: 2n-l, wherein n is an integer between 1 and 33.

23. The nucleic acid molecule of claim 19, wherein the nucleic acid molecule comprises a nucleotide sequence selected from the group consisting of a) the nucleotide sequence selected from the group consisting of SEQ ID NO: 2n-l, wherein n is an integer between 1 and 33; b) a nucleotide sequence wherein one or more nucleotides in the nucleotide sequence selected from the group consisting of SEQ ID NO: 2n-l, wherein n is an integer between 1 and 33, is changed from that selected from the group consisting of the chosen sequence to a different nucleotide provided that no more than 15% ofthe nucleotides are so changed; c) a nucleic acid fragment ofthe sequence selected from the group consisting of SEQ ID NO: 2n-l, wherein n is an integer between 1 and 33; and d) a nucleic acid fragment wherein one or more nucleotides in the nucleotide sequence selected from the group consisting of SEQ ID NO: 2n-l, wherein n is an integer between 1 and 33, is changed from that selected from the group consisting of the chosen sequence to a different nucleotide provided that no more than 15% ofthe nucleotides are so changed.

24. The nucleic acid molecule of claim 19, wherein the nucleic acid molecule hybridizes under stringent conditions to the nucleotide sequence selected from the group consisting of SEQ ID NO: 2n-l, wherein n is an integer between 1 and 33, or a complement ofthe nucleotide sequence.

25. The nucleic acid molecule of claim 19, wherein the nucleic acid molecule comprises a nucleotide sequence in which any nucleotide specified in the coding sequence ofthe chosen nucleotide sequence is changed from that selected from the group consisting ofthe chosen sequence to a different nucleotide provided that no more than 15% of the nucleotides in the chosen coding sequence are so changed, an isolated second polynucleotide that is a complement ofthe first polynucleotide, or a fragment of any of them.

26. A vector comprising the nucleic acid molecule of claim 19.

27. The vector of claim 26, further comprising a promoter operably linked to the nucleic acid molecule.

28. A cell comprising the vector of claim 27.

29. A method for determining the presence or amount ofthe nucleic acid molecule of claim 19 in a sample, the method comprising:

(a) providing the sample;

(b) introducing the sample to a probe that binds to the nucleic acid molecule; and

(c) determining the presence or amount ofthe probe bound to the nucleic acid molecule, thereby determining the presence or amount ofthe nucleic acid molecule in the sample.

30. The method of claim 29 wherein presence or amount of the nucleic acid molecule is used as a marker for cell or tissue type.

31. The method of claim 30 wherein the cell or tissue type is cancerous.

32. A method for determining the presence of or predisposition to a disease associated with altered levels ofthe nucleic acid molecule of claim 19 in a first mammalian subject, the method comprising: a) measuring the amount ofthe nucleic acid in a sample from the first mammalian subject; and b) comparing the amount ofthe nucleic acid in the sample of step (a) to the amount ofthe nucleic acid present in a control sample from a second mammalian subject known not to have or not be predisposed to, the disease; wherein an alteration in the level ofthe nucleic acid in the first subject as compared to the control sample indicates the presence of or predisposition to the disease.