TWI250209B - A novel G protein-coupled receptor, GAVE8 - Google Patents

Abstract

Novel GAVE8 polypeptides, proteins, and nucleic acid molecules are disclosed. In addition to isolated, full-length GAVE8 proteins, isolated GAVE8 fusion proteins, antigenic peptides and anti-GAVE8 antibodies are provided. The invention also provides GAVE8 nucleic acid molecules, recombinant expression vectors containing the nucleic acid molecule, host cells into which the expression vectors have been introduced and nonhuman transgenic animals in which a GAVE8 gene has been introduced or disrupted are described. Diagnostic, screening, and therapeutic methods utilizing GAVE8 compositions or molecules binding GAVE8 also are provided. Methods for identifying GAVE8 agonists, antagonists, inverse agonists and the like are described.

Description

1250209 A7 B7 Ministry of Economic Affairs Intellectual Property Office Staff Consumer Cooperative Printed V. Inventive Note (彳) Background of the Invention The G protein-coupled receptor (GPCR) is a group of intact membrane proteins responsible for cellular signaling. G P C R reacts to a variety of extracellular signals, including: neurotransmitters, hormones, odorants, and light, and transduces signals across cell membranes, triggering a secondary messenger response within the cell. Many of the currently used therapeutic drugs calibrate GPCRs because the receptor can indirectly mediate a variety of physiological responses including: vasodilation, heart rate, bronchodilation, endocrine secretion, and bowel movements. GPCRs are characterized by an extracellular ligand binding domain, a structural region that traverses the cell membrane, and an intracellular domain that interacts with signal-related cellular elements. The function of the receptor (e.g., binding ligand to G protein) is associated with amino acids at certain critical positions. A G P C R subgroup can be identified by molecular selection, which is a known endodermal differentiation gene (Edg). The amino acid sequence of the Edg GPCR subgroup has 31% to 34% similarity to this group, but contains two structural regions with similar structures and functions. One of the homologous structural regions, including Edg-2, Edg-4, Edg-7, and Edg-8 proteins, binds to lysophosphatidic acid (LP A) but does not bind to sphingolipids. The second homologous structural region, including Edg-1, Edg-3, and Edg-5, binds to sphingosine-1-phosphate (SIP or SPP) but does not bind to LPA (Goetzl et a I., Ad v Exp Med Biol (1 999) 469: 259-264) 〇s IP is a potent, extracellular lysate lipid phosphate mediator that is released during, for example, platelet activation (Moser et al., J Cell Bi) 01(1 992)1 1 6 : 1 51 7-1 526). SIP can cause various reactions of cells. (Please read the notes on the back and fill out this page.) Packing and P. This paper scale applies to Chinese National Standard (CNS) A4 size (210X 297 mm) -4- 1250209 A7 B7 V. INSTRUCTIONS (2) The most notable of which is cell proliferation (Zhang et a I., J Cell B i ο I (1 9 9 1 ) 1 1 4 : 1 5 5-1 67; Bornfeldt et a I., J Cell Bi 〇 1 (1995) 130 : 1 93-206; and Berger et a I., Mol

Pharmaco 丨 (1996) 50: 451-457) and anti-apoptosis (Cuvillier et a I., Nature (1 996) 381 800-803; Edsall et al., J Neurosci (1 997) 167: 6952- 6960) 〇 Various studies have shown that differences in GPCR amino acid sequences can cause differences in affinity for natural ligands or small molecule agonists or antagonists. In other words, small sequence differences can cause binding affinity and activity.

Different. (See, for example: Meng et al., J Bio C hem (1996) 271(50): 32016-20; B urd et a I., J Bio C hem (1 998) 273(51): 34488-95; And Hurley eta I ·, J 'r-I (please read the notes on the back and fill out this page). Printed by the Intellectual Property Office of the Ministry of Economic Affairs.

Neurochem (1999) 72(1): 413-21). In particular, it has been shown that differences in amino acid sequences in the structural regions within the third cell can result in different activities. The release of gonadotropin-releasing hormone receptors by Myburgh et al. showed that the intracellular loop M1 of the receptor, alanine 261, is important for G protein coupling and receptor uptake (Biochem J (1 998) 331 (Part 3) : 8 9 3 - 6). W ο neroweta I. Studies on thyroid stimulating hormone receptors indicate that the deletion of the circuit within the third cell results in constitutive activation of the receptor (J Bio Chem (1 99 8) 27 3(1 4): 790 0-5). This report is the first to mention a receptor that is coupled to a G protein, and the deletion of an amino acid in its third intracellular loop results in constitutive activation of the receptor. Since GPCR plays an important role in disease and regulates GPCR activity and has the ability to treat diseases, it is not known by the identification and characterization of the paper. The Chinese National Standard (CNS) A4 specification (210X 297 mm) -5- 1250209 Ministry of Economic Affairs Intellectual Property Office Staff Consumer Cooperatives Print A7 ___B7 V. Inventive Note (3) GPCRs can provide new compositions and methods to treat disease states involving GPCR activity. The present invention recognizes and characterizes the performance of novel GPCRs (GA\/E8) and applies the above findings to provide compositions and methods for identifying and treating diseases associated with them. SUMMARY OF THE INVENTION The present invention relates to newly recognized receptors coupled to G proteins. Specifically, the system of the present invention coupled to the G protein is particularly expressed in the spleen and the brain. Further, it is expressed in the white matter of the brain. This and other sources suggest that this novel receptor (GAVE8) is involved in a variety of inflammatory diseases including: multiple sclerosis and various immune system disorders and disorders of the nervous system, especially the central nervous system. In one of the features, the present invention relates to an isolated nucleic acid selected from the group consisting of a nucleic acid encoding a vertebrate sequence confirmation number: 2 protein amino acid, variants, mutations and fragments thereof, and a sequence confirmation number comprising: An isolated nucleic acid of the nucleotide sequence of 1, a variant, a mutation, and a fragment. Furthermore, the present invention relates to a hybridization probe and a complementary fragment of a nucleic acid hybridization probe and a complementary fragment or a nucleic acid which binds to a sequence identification number: 2 amino acid sequence. Furthermore, the present invention relates to nucleic acids having about 90%-99% similarity to sequence confirmation number: 1, including isolated nucleic acids having about 90% 99% similarity to the amino acid sequence encoding the sequence confirmation number: 2. . In a related feature, an oligonucleotide comprising at least 8 nucleotides and a method of hybridization comprises contacting a complementary oligonucleotide with a nucleoside comprising a sequence confirmation number of 1: under conditions allowing hybridization of the complement to the nucleic acid. The paper size of the acid is applicable to China National Standard (CNS) Α4 specifications (2) 0Χ 297 mm) ' -6 - (Please read the back note and then fill out this page) Pack * Ρ. 1250209 A7 B7 Ministry of Economics The Property Bureau employee consumption cooperative printed five, the invention instructions (4) nucleic acid steps. Furthermore, the complementary fragment can be an antisense oligonucleotide used in a method of inhibiting GAVE8 expression in vivo and in vitro. The method comprises the steps of: providing an oligonucleotide sequence complementary to the sequence confirmation number: 1, providing a human cell comprising a signaling sequence comprising a nucleotide sequence of sequence number: 1 and introducing the oligonucleotide into the cell The internal cells inhibit the expression of GAVE8 via mechanisms including inhibition of translation, formation of triple helix and/or activation of nucleases resulting in degradation of the signaling RNA. The invention also relates to an isolated multi-peptide selected from the group consisting of: a purified polypeptide of the sequence confirmation number 2 amino acid sequence, a variant thereof, a mutation and a fragment, and a functional property of the polypeptide and an additional amino acid residue Purified Polypeptides of the Invention The invention further relates to nucleic acids that are operably linked to a performance control element, including vectors comprising isolated nucleic acids. Further, the present invention relates to a method of transforming into a cultured cell comprising the nucleic acid of the present invention and comprising the step of growing a transformed cell comprising the nucleic acid of the present invention, and a method of expressing and purifying the polypeptide from the culture medium of the cell or cultured cell to produce a polypeptide. . Further features of the invention include isolated antibodies 'incorporated with single and multiple strains of antibodies to the polypeptides of the invention. In a related feature, the invention further discloses methods for producing antibodies and treatment with antibodies that bind to GAVE8 A method of disease associated with GAVE8. Another feature of the invention includes a method for determining the presence or absence (providing diagnostic use) of GAVE8 in a biological and/or tissue sample. In another feature of the invention, a method of treating GAVE8 signaling is disclosed, including for a patient in need thereof Use peptides, agonists, antagonists (please read the notes on the back and fill out this page). It is installed on P. This paper scale applies to Chinese National Standard (CNS) A4 specification (2丨0X 297 mm) 1250209 A7 B7 Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative printed five, invention instructions (5), reverse agonists and / or antibodies. In another feature of the invention, a method of confirming a GAVE8 modulator is disclosed which comprises providing a chemical moiety, providing a step of expressing GAVE 8 cells and determining whether the chemical moiety can modulate GAVE8 signaling activity, including endogenous ligands The presence or absence of this determines whether this regulation occurs. Among the relevant features, chemical moieties can include, but are not limited to, peptides, antibodies, agonists, retro-activators, and antagonists. Another feature of the invention includes a therapeutic composition comprising: a nucleic acid, an antibody, a multi-peptide, an agonist, a retro-activator, and an antagonist. Further methods of the invention also include methods of treating disease states and modulating GAVE8 signaling activity, which are compositions for administering the treatment to a patient in need thereof. Since GAVE8 is present in blood cells and nerve cells, the present invention relates to methods for testing peripheral blood cells to infer and determine the G A V E 8 structure and function of nerve cells. The various features of the invention are set forth in the detailed description and the drawings. Further, certain procedures or compositions will be described in more detail in the following various references. The entire contents of each of the references are hereby incorporated by reference. BRIEF DESCRIPTION OF THE FIGURES Figure 1 shows the nucleic acid sequence of GAVE8 (sequence confirmation number: 1). Figure 2 is the amino acid sequence of GAVE8 (sequence confirmation number: 2). Figure 3 is a computerized northern ink dot map from various human tissue RNAs (please read the back note first and then fill out this page) b Binding paper scale applies to China National Standard (CNS) A4 specification (2(1)χ297 mm) - 8- 1250209 A7 B7 Ministry of Economic Affairs Intellectual Property Bureau Staff Consumer Cooperative Printed 5, Invention Description (6) Figure 4 shows the computerized northern ink dot map of each part of the brain showing GAVE8. Figure 5a shows the GAVE8TaqMan in various regions of the brain. ® Performance Profile. Figure 5b is a GAVE8TaqMan® performance profile for the lower view. Figure 6a is a GAVE8TaqMan® performance profile for surrounding tissue. Figure 6b is a GAVE8TaqMan® performance profile for surrounding tissue. Figure 7 depicts the use of cells expressing GAVE 8 (Η EK transformant, a modified 293 cell line expressing Gqi5 protein and luciferase reporter gene) via S1P (SPP) (first set of bars, S1P) Changes in luciferase reading were determined after stimulation at concentrations ranging from 0.01 to 10) and LPA (second group of bars, LPA concentrations between 0.26 and 26-26). LPA is the negative control group. DETAILED DESCRIPTION OF THE INVENTION The present invention is based on the discovery of a complementary DNA molecule encoding human GAVE8, a member of the G protein-coupled receptor supergroup, which binds to the endogenous ligand sphingosine-1 -phosphate (SIP, SPP -1 or SSP). The nucleotide sequence encoding the human GAVE8 protein is shown in Figure 1 (sequence confirmation number: 1). The amino acid sequence of the GAVE8 protein is shown in Figure 2 (SEQ ID NO: 2) GAVE8 complementary DNA of Figure 1 (sequence confirmation number: 1), including untranslated regions of approximately 2589 nucleotides in length, encoding the protein Its molecular weight is approximately 41.8 kDa (not included in the post-translational modification). Using TaqMan® assay, specific subscript RNA fragments can be expressed in the month and spleen. In addition, GAVE8 is known to be expressed in peripheral blood cells. National Standard (CNS) A4 Specification (210X297 mm) (Please read the note on the back and fill out this page) b Installation · Order -9- 1250209 A7 B7 Ministry of Economic Affairs Intellectual Property Bureau Staff Consumer Cooperative Printed 5, Invention Description ( 7) GAVE8 is present in this and other tissues. GAVE8 is involved in various disease states including immune function and diseases in which various nerves occur with age or organ degeneration, such as multiple sclerosis. GAVE8 is identified in such tissues. The selection of the gene encoding GAVE8 provides various treatments to regulate the performance and activity of GAVE8 and to provide a treatment for diseases involving GAVE8. Human GAVE8 A related molecule having a conserved structure and functional characteristics of certain Edg groups. The term "group" as used herein refers to two or more proteins or nucleic acid molecules which are collectively common when used in the proteins and nucleic acid molecules of the present invention. Constructing structural regions and having sufficient similarity to the amino acid or nucleotide sequences defined herein. Members of this group may be native proteins and nucleic acid molecules and may be derived from the same or different species. For example, such a group may contain Humans and the first protein derived from murine protein homologs, and another human-derived protein and a second protein derived from murine homologs. Group members also share common functional properties. Hla &amp Maciag was the first to stimulate human endothelial cell lines with sterol esters and confirmed the genes involved in cell differentiation, and isolated the Edg-1 receptor (J Biol Chem (1 990) 265: 9308-931 3). Less than the ligand of the activatable receptor, Edg-1 was classified as an orphan GPCR. Subsequently Lee et al confirmed that the phospholipid (called sphingosine-1-phosphate) in blood sputum is Edg- 1 Endogenous ligand of the receptor (Science (1 998) 279: 1 552-1 555). The remarkable tissue distribution of GAVE8 directly shows its function in the immune system and nervous system. The receptor is also conserved. And found in, for example, humans, mice and large (please read the notes on the back and fill out this page) -

, 1T Ρ This paper scale applies to China National Standard (CNS) Α 4 specifications (210X297 mm) -10- 1250209 A7 B7 Ministry of Economic Affairs Intellectual Property Bureau staff levy cooperatives printed five, invention description (8) white rat. Certain structural areas are also conserved across germ lines. GAVE8 also regulates the expression of related apoptosis. Therefore, GAVE8 can induce or remove programmed cell death. In one embodiment, the GAVE 8 protein comprises a circular structural region within a third cell, from about amino acid 214 to about 252, and at least about 65 of the circular structural region within the third cell of sequence confirmation number: %, preferably less than about 75%, and more preferably about 85%, 95% or 98% amino acid sequence similarity. The term "equivalent amino acid residue 11" as used herein refers to an amino acid that substantially occupies the same position within a protein sequence when two or more sequences have been subjected to the alignment assay. Preferred amine groups of the GAVE8 polypeptide of the present invention The acid sequence is sufficiently similar to the consensus amino acid sequence of the circular structural region in the third cell in the sequence confirmation number: 2. The "sufficient similarity" of the term means the first amino acid or nucleotide. The sequence and the second amino acid or nucleotide sequence contain a sufficient or minimum number of amino acid residues or nucleotides of the same or equivalent (eg, similar side chains), thereby allowing the first and second amine groups The acid or nucleotide sequence has a commonly constructed structural region and/or a common functional activity. For example, a sufficiently similar amino acid or nucleotide sequence as defined herein has a common structural region containing about 65% similarity. Sexuality, preferably 75% similarity, better 85%, 95% or 98% similarity. Other important structural regions include (but are not limited to): the structural region across the cell membrane (TM) ( In sequence confirmation number: 2, TM 1 is approximately from amino acid From about 3 to about 62; TM2 is from about 70 to about 95 amino acid residues; TM3 is from about 112 amino acid residues to about 131; TIVI4 is from about 151 to about 176 amino acid residues (please read the back) Note: Please fill out this page again) • Install the paper size for the Chinese version (CNS) A4 size (210X297 mm) -11 - 1250209 A7 B7 V. Invention description (9) ; T Μ 5 About from amino acid residue 1 9 3 to about 2 1 3 ; T Μ 6 from about 2 53 to about 273; and ΤΜ7 from about amino acid residue 290 to about 310); cytoplasmic (intracellular ring) structure The region (in sequence confirmation number: 2, from about amino acid residue 63 to about 69; from about amino acid residue 1 32 to about 1 50; from about amino acid residue 214 to about 252; From amino acid residue 31 1 to about 398); and extracellular structural regions (in sequence number: 2, from about 1 to about 37 amino acid residues; from about 96 amino acid residues to about amino acid residues) 1 1 1 ; about from amino acid 177 to about 192; and about from amino acid residue 274 to about 290). Among related features, important structural regions also include, but are not limited to, consistent glycosylation. Site of action, lipid binding site and Acidification site. In this article, "GAVE8 activity", "GAVE8 biological activity" or "GAVE8 functional activity" is interoperable, meaning GAVE8 protein, polypeptide measured in vivo or in vitro according to standard techniques Or the activity exhibited by the nucleic acid molecule in GAVE8-reactive cells. The GAVE8 activity can be direct activity, e.g., can bind or activate a second protein, or indirect activity, e.g., cellular signaling activity mediated by interaction of a G A V Ε 8 protein with a second protein. In a preferred embodiment, the activity of GAVE8 comprises at least one or more of the following activities: (i) interaction with a protein of the GAVE8 signaling pathway; (ii) interaction with a GAVE8 ligand; and (Mi) It can interact with target proteins in cells. For example, the activity of GAVE8 includes, but is not limited to, binding to sphingosine-1 - p-acid salt (S-1 - P), which can be determined, for example, by a change in fluorescence (FUPR® analysis), or It was determined by binding to the labeled S-1 -P in cells transformed with the expression vector containing the sequence confirmation number: 1. Therefore, another specific embodiment of the present invention is a GAVE 8 paper size applicable to the National Standard (CNS) A4 specification (210X297 mm) (please read the back note first and then fill out this page) _ Ministry of Economic Affairs Intellectual Property Bureau Employees Consumption Cooperative Printed -12-1250209 A7 B7 V. INSTRUCTION DESCRIPTION (Gorg8 protein and its multi-peptide) isolated. The various features of the present invention will be further detailed in the following sections. I. Isolated Nucleic Acid Molecules One of the features of the invention relates to an isolated nucleic acid molecule encoding a GAVE8 protein or a biologically active portion thereof; and a nucleic acid molecule sufficient to recognize a GAVE8 nucleic acid (eg, GAVE8 signaling RNA) as a hybridization probe, and Amplification or mutation of a fragment of a GAVE8 nucleic acid molecule as a PCR primer. The term "nucleic acid molecule" includes DNA molecules (such as complementary DNA or genomic DNA) and RNA molecules (such as signaling RNA) and the use of nucleotide analogs. DNA or RNA analogs. Nucleic acid molecules can be single or double stranded molecules. "Isolated" nucleic acid molecules are isolated from other natural nucleic acid molecules. Preferably, the π-isolated nucleic acid does not contain a nucleic acid adjacent to the organism's native gene DNA sequence (preferably a protein-encoding sequence) (ie, a sequence located at the 5' and 3' ends of the nucleic acid). In various embodiments, an isolated GAVE8 nucleic acid molecule can contain less than about 5 仟 test, 4 仟 test, 3 仟 test, 2 仟 base, 1 仟 base, 0.5 仟 base, or 0.1 仟 base. a nucleotide sequence adjacent to a natural genomic DNA nucleic acid molecule. Further, an "isolated" nucleic acid molecule, such as a complementary DNA molecule made by recombinant techniques, may be substantially free of other intracellular substances, or culture fluids, or The chemically synthesized molecule may be substantially free of chemical precursors or other chemicals. The nucleic acid molecule of the invention 'for example, a nucleic acid molecule having a nucleotide sequence of sequence number: 1, or any complement of those nucleotide sequences, This paper scale can be applied to China National Standard (CNS) Α4 specifications (2) 0X 297 mm) (please read the notes on the back and fill out this page) - Installed and subscribed to the Ministry of Economic Affairs, Intellectual Property Bureau, employee consumption cooperatives -13- 1250209 A7 B7 Ministry of Economic Affairs Intellectual Property Bureau Employees Consumption Cooperatives Printed V. Description of the invention (U is separated by standard molecular biology techniques and sequence data provided herein. Sequence identification number: 1 or all of the nucleic acids can be used. Sequences are used as hybridization probes to isolate GAVE8 nucleic acid molecules using standard hybridization and selection techniques (eg, f-miao described in Sambrook et al., eds., " Molecular Cloning: A Laboratory Manual, " 2nd ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, 1989). The nucleic acid molecules of the present invention can be amplified using standard DNA amplification techniques using complementary DNA, signaling RNA or genetic DNA as a template with appropriate oligonucleotide primers. For example, the primer may include, but is not limited to, 5'-CCATGGAGTCGGGGCTGC-3' (sequence 歹!J confirmation number: 3) and 5·-TCAGTCTGCAGCCGGTTC-3 (sequence 歹U confirmation number A). The amplified nucleic acid can be optionally ligated into an appropriate vector and characterized by DNA sequence analysis. Furthermore, oligonucleotides corresponding to the GAVE8 nucleotide sequence can be prepared using standard synthetic techniques, for example using an automated DNA synthesizer. In another preferred embodiment, the isolated nucleic acid molecule of the present invention comprises a nucleic acid molecule complementary to the nucleotide sequence of Sequence Number: 1 or a portion thereof. A nucleic acid molecule complementary to a given nucleotide sequence is sufficiently complementary to a given nucleotide sequence to hybridize to a given nucleotide sequence to form a stable double-stranded complement. Furthermore, the nucleic acid molecule of the present invention may comprise only a portion of the nucleic acid sequence encoding GAVE8, such as a fragment which may serve as a probe or primer, or a fragment encoding a biologically active portion of GAVE8. For example, the fragment may comprise, but is not limited to, a region of the amino acid residue 1 - 398 in the coding sequence confirmation number: 2. The nucleotide sequence can be generated and designed from the selected human GAVE8 gene. (Please read the back note and fill out this page.) This paper size applies to the Chinese National Standard (CNS) A4 specification (210X29*7). • 14- 1250209 A7 B7 V. INSTRUCTIONS (d probes and primers for the identification and/or selection of other cell types (eg other tissues) and other mammalian GAVE8 homologs. Probes/introductions usually A relatively pure oligonucleotide is included. The oligonucleotide comprises at least hybridization to about 12 in an urgent condition, preferably about 25, more preferably about 50, 75, 10, 125, 150, 175, 200, 250, 300, 350 or 400 consecutive sequence confirmation numbers: 1 nucleotide sequence of the same strand or counter-strand or sequence confirmation number: 1 natural mutation. Based on human GAVE 8 nucleotide sequence The probe can be used to detect a transcription product or a gene sequence encoding a similar or identical protein. The probe can comprise a labeling group attached thereto, such as a radioisotope, a fluorescent compound, an enzyme or an enzyme cofactor. Needle As part of a diagnostic test set that identifies inappropriate GAVE8 proteins in cells or tissues, for example, measuring the amount of GAVE8 nucleic acid encoded in a cell sample from a patient, such as detecting the amount of GAVE8 signaling RNA or determining whether the GAVE8 gene has been mutated Or deletion. A nucleic acid fragment encoding the "biologically active portion" of GAVE8, which is prepared by isolating a sequence confirmation number encoding a portion of the polypeptide having the biological activity of GAVE8: 1, representing a portion encoded by the GAVE8 protein (e.g., in vitro) Recombinant performance) and assessment of the activity of the GAVE8 coding portion. For example, a nucleic acid fragment encoding a biologically active portion of GAVE8 can comprise a circular structural region within a third cell, such as an amino acid residue at sequence confirmation number: 2 from about 2 1 4 Up to about 2 52. The present invention further comprises a nucleic acid sequence which differs from the sequence confirmation number of the nucleotide sequence due to degradation of the genetic code, but which encodes the same GAVE8 protein as the nucleotide sequence of the sequence confirmation number: Molecular. This paper scale applies to China National Standard (CNS) A4 specification (210X297 mm) (Please read the note on the back and fill out this page) *Installation · Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative printing -15- 1250209 A7 B7 Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative printing 5, invention description (j In addition to the human G Av E 8 nucleotide sequence of sequence confirmation number: 1, those skilled in the art will recognize that polymorphisms in the DNA sequence in a population (e.g., a human population) can result in changes in the GAVE8 amino acid sequence. The genetic polymorphism of the GAVE8 gene can occur in individuals of the ethnic group due to variations in the natural dual gene. A dual gene is another gene at the locus of a given gene. The term "n gene" and "recombinant gene" as used herein mean a nucleic acid molecule comprising an open-reading architecture encoding a GAVE8 protein, preferably a mammalian GAVE8 protein. "The variant of a dual gene" Refers to the nucleotide sequence of a polypeptide that occurs at the GAVE8 locus or nucleotide sequence. Another dual gene can be confirmed by sequencing the genes important to many different individuals. It can easily confirm the locus of the same gene in individual individuals using hybridization probes. Any and all of this nucleotide variation in GAVE8 and the resulting amino acid polymorphism or variation, resulting in a natural dual gene mutation that does not alter the functional activity of GAVE8, is within the scope of the invention. Furthermore, other species encoding a GAVE8 protein nucleic acid molecule (GAVE8 homolog) having a nucleotide sequence different from human G A V E 8, are also within the scope of the invention. Using the characteristics of the human GAVE 8 nucleic acid disclosed herein, using human complementary DNA (or a portion thereof) as a hybridization probe, the variant corresponding to the natural dual gene of the present invention and the GAVE8 complementary DNA can be isolated under stringent hybridization conditions according to standard hybridization techniques. A nucleic acid molecule of a homologue. Accordingly, in another embodiment, the isolated nucleic acid molecule of the invention has a length of at least 300, 325, 350, 375' 400, 425, 450, 500, 550, 600, 650, 700, 800, 900, 1000 or 1100. The nucleotide size of this paper is applicable to the Chinese National Standard (CNS) A4 specification (210X 297 mm) " ' -16- (Please read the note on the back and fill out this page) - Decoration. 1250209 ΑΊ B7 Ministry of Economic Affairs The Intellectual Property Office employee consumption cooperative printed five, the invention description (^ ' and under urgent conditions can be hybridized with a nucleic acid molecule comprising a nucleotide sequence, preferably a sequence of the coding sequence confirmation number: 1, or its complementary sequence The term "under urgent conditions" as used herein means that at least 60% (65%, 70%, preferably 75% or more) of the same nucleotide sequences remain hybridized to each other under rinsing. Conditions. This urgent condition is well known to those skilled in the art and can be found in "Current Protocols in Molecular Biology"" John Wiley & Sons, NY (1 989), 6.3.1 -6.3.6. Urgent hybridization A preferred non-limiting example of the conditions is in 6X sodium chloride / Sodium citrate (SSC), hybridization at about 45 ° C, followed by one or more washes with 0.2X SSC, 0.1% SDS at 50-65 ° C. Preferably, the isolated nucleic acid molecule of the invention can be Under stringent conditions, it hybridizes to a sequence of sequence confirmation number: 1 or a naturally occurring nucleic acid molecule corresponding to its complement. The "naturally occurring π nucleic acid molecule" herein refers to an RNA or DNA molecule having a native nucleotide sequence (eg Encoding natural proteins). In addition to the naturally occurring variant of the GAVE8 sequence dual gene in the population, those skilled in the art will further recognize that mutations can be made in the nucleotide sequence of sequence confirmation number: 1, which can alter the amino acid sequence encoding the GAVE8 protein. Without altering the biological activity of the GAVE8 protein. For example, a nucleotide substitution on an "non-essential" amino acid residue results in the substitution of an amino acid. "Non-essential " amino acid residues refer to the wild type GAVE 8 sequence 歹U ( For example, the sequence confirmation number: the sequence of 2) changes without changing the residue of the living property, and the "must" amino acid residue is the residue required for biological activity. For example, each of the non-conserved or only semi-conserved amino acid residues in G A V E8 is a non-essential residue which may be the target residue of the above alteration. In addition, the paper size applies to the Chinese National Standard (CNS) A4 specification (2丨0X 297 mm) (please read the notes on the back and fill out this page) • Install P. -17- 1250209 Α7 Β7 Ministry of Economics The property bureau employee consumption cooperative issued 5, the invention description (d in the GAVE8 protein, each of the conserved amino acid residues may be the residue required for the activity, so it will not be the target of the change. Accordingly, the present invention Another feature relates to a nucleic acid molecule encoding a GAVE8 protein that contains residues that alter the activity that are not essential. The GAVE8 protein differs from the amino acid sequence of sequence number: 2 but retains the activity of the organism. In one embodiment, the isolated nucleic acid molecule comprises at least 87% identical, 90%, 93%, 95%, 98% or 99% identical nucleotide sequence to the amino acid sequence encoding the protein of sequence number:2. Introducing one or more nucleotides in sequence confirmation number: 1 for substitution, addition or deletion to replace, add or delete one or more amino acids in the encoded protein to create a sequence confirmation number 2 different isolated nucleic acid molecules encoding GAVE8 proteins. Mutations can be made by standard techniques, such as site-directed mutagenesis and PCR-mediated mutagenesis. Preferably, one or more predicted non-essential amino acid residues can be used. A retentive amino acid substitution is made. "Reserved amino acid substitution" is the substitution of an amino acid residue with an amino acid residue having a similar side chain. A group of amino acid residues having similar side chains has been defined. In this art, the group includes amino acids of a basic side chain (eg, aminic acid, arginine, histidine), amino acids of an acidic side chain (eg, aspartic acid, glutamic acid) ), uncharged polar side chain amino acids (eg, glycine, aspartic acid, glutamine proline, serine, threonine, tyrosine, cysteine), non Polar side chain amino acids (eg, alanine, valine, leucine, isoleucine, valine, phenylalanine, methionine, tryptophan), amine groups of the β-branched side chain Acids (eg, sulphate, valine, isoleucine) and aromatic side chain amino acids ( For example, this paper scale applies Chinese National Standard (CNS) Α4 Specifications (2) 0X297 mm) ~ ' •18- (Please read the back note first and then fill out this page) 1250209 A7 B7 V. Invention Description (β Read the precautions on the back and fill out this page. •• tyrosine, phenylalanine, tryptophan, histidine. Therefore, the better one is the same side of the non-essential amino acid residue predicted by GAVE8. Another amino acid residue of the chain group is substituted. In addition, mutations can be introduced randomly along all or part of the GAVE8 coding sequence, for example, by saturation mutagenesis, the resulting mutant can be confirmed by GAVE8 biological activity screening. Active mutants. After mutagenesis, the encoded protein can be expressed recombinantly and the activity of the protein can be determined. In one preferred embodiment, the mutant GAVE8 protein can be assayed: (1) the ability to form a protein:protein interaction with a protein of the GAVE8 signaling pathway; (2) the ability to bind to a GAVE8 ligand (eg, SIP); (3) The ability to bind to a target protein in a cell. In another preferred embodiment, the ability of the mutant GAVE8 to modulate cell proliferation or cell differentiation can be determined. Printed by the Ministry of Economic Affairs Intellectual Property Office Employees' Consumption Cooperative The present invention comprises an anti-nuclear nucleic acid molecule, i.e., a nucleic acid molecule complementary to the positive strand of the encoded protein, e.g., a crypto strand complementary to a double-stranded complementary NA molecule or complementary to a subsense RNA sequence. Accordingly, the anti-strand nucleic acid can be bonded to the positive-nuclear acid via a hydrogen bond. The anti-strand nucleic acid can be complementary to all of the GAVE8 cryptophores, or only to portions thereof, such as all or part of the protein coding region (or open-architecture architecture). The anti-strand nucleic acid molecule can be an anti-nuclear nucleotide sequence encoding a non-coding region of a crypto-bank of GAVE8. Non-coding regions ("5· and 3, untranslated or flanking regions Ί are contiguous coding regions and non-translated to amino acids 5 and 3, sequences. Given a crypto-cap sequence encoding GAVE8 (eg sequence confirmation number:

1) The anti-stock nucleic acid of the present invention can be designed according to the base pairing rules of Watson and Crick. Anti-nuclear nucleic acid molecules can be complementary to GAV E 8 communication RNA. This paper scale is applicable to China National Interpolation (CMS) A4 specification (2) 297 297 麓) -19- 1250209 A7 B7 V. Invention description (d (please read first) Note on the back side of the full coding area of this page, but better anti-strand oligonucleotides are only complementary to the GAVE 8 signaling RN A coding or non-coding region. For example, anti-strand oligonucleotides can be complementary to GAVE8 The region surrounding the start site of the translation of the RNA, such as the sequence 歹IJ 5 ' - GCAGCAGCCCCGAC TC C AT G - 3 ' (preface j [j confirm, number Shima·· 5) and 5'-(:0 octa 000(:0〇06(:(:(:0八八60-3'(序歹[" confirmation number: 6)) oligonucleotide. The length of the anti-strand oligonucleotide can be, for example, about 5, 〇, 15, 20, 25, 30, 35, 40, 45 or 50 nucleotides. The anti-nucleic acid of the present invention can be constructed using chemical synthesis and enzyme binding reactions known in the art. For example, anti-nucleic acid Natural nucleotides or various modified nucleotides (eg, phosphonothioate derivatives) can be used (eg, anti-strand oligonucleotides) Chemical synthesis of thiocyanate derivatives and acridine-substituted nucleotides to increase the biostability of the molecule or increase the physical stability of the formation of double-stranded nucleic acids between the anti-strand and the nucleus. The Property Bureau Staff Consumer Cooperative prints examples of modified nucleotides that can be used to generate anti-nuclear nucleic acids, including 5-fluorouracil, 5-bromouracil, 5-chlorouracil, 5-iodouracil, and hypoxanthine. , Astragalus, 4-ethionylcytosine, 5-(carboxylhydroxymethyl)uracil' 5-carboxymethylaminemethyl-2-thiouridine, 5-carboxymethylaminemethyluracil, II Hydrouracil, p-D-galactosyl quinosine, inosine, |6-isopentenyl adenine, 1-methylguanine, 1-methylinosine 2,2- Dimethylguanosper, 2-methyladenine, 2-methylguanine, 3-methylcytosine, 5-methylcytosine, N6-adenine, 7-methylguanine, 5 -Methylaminomethyluracil, 5-methoxyaminemethyl-2-thiouracil, β-D-mannosyl quinosine, 5-methoxycarboxymethyluracil, 5- Methoxyuracil, 2-methylthio -N6-isopentenyl adenine, uracil-5-oxyacetic acid, Dingben paper scale applicable to China National Standard (CNS) A4 specification (2) 0X297 mm) -20- 1250209 Μ Β7 Ministry of Economics intellectual property Bureau employee consumption cooperative printing 5, invention instructions (d oxin (butoxosine), pseudo-uracil, quesine (queosine), 2-thiocytosine, 5-methyl-2-thiouracil, 2-thiouracil , 4-thiouracil, 5-methyluracil, methyl uracil-5-oxyacetate, 5-methyl-2-thiouracil, 3-(3-amino-3-N-2- Carboxypropyl) uracil and 2,6-diaminoguanidine. In addition, anti-nuclear nucleic acids can be made biologically using a vector that exhibits anti-stranding (i.e., transcribed RNA after insertion of a counter-stranded target nucleic acid, as further described in the next subsection). The anti-stranded nucleic acid molecule of the present invention is generally administered to a patient or to in situ hybridization or binding of a cell-sending RNA and/or a genotype DNA encoding a GAVE8 protein, thereby inhibiting protein expression, such as inhibition of transcription and/or translation. effect. Hybridization can be a complementary nucleotide complement to form a stable duplex, or for example, a counter-nucleic acid molecule can bind to a DNA duplex via a particular interaction in the major groove of the duplex. Examples of the administration route of the anti-nucleic acid molecule of the present invention include direct injection at a tissue site. In addition, anti-nuclear acid molecules can be modified to calibrate selected cells and then administered systemically. For example, when administered systemically, the anti-strand molecule can be modified such that it can specifically bind to a receptor or antigen present on the surface of the selected cell, for example, the anti-strand nucleic acid molecule can be linked to a cell surface receptor or antigen. An antibody to a peptide. Anti-nuclear nucleic acid molecules can also be delivered to cells using the vectors described herein. In order to achieve sufficient concentration of the anti-strand molecule within the cell, the anti-nuclear nucleic acid molecule in the vector construct is preferably placed under the control of a strong RNA polymerase (po丨) or pol III promoter. The anti-nucleic acid molecule of the present invention may be a mutameric amino acid molecule. The α-raceomeric nucleic acid molecule can be parallel to the complementary RNA (please read the back of the note first and then fill out this page) - Loading · f This paper size applies to the Chinese National Standard (CMS) A4 specification (210 X 297 mm) -21 - 1250209 A7 B7 Ministry of Economic Affairs Intellectual Property Bureau Employees Consumption Cooperative Printed 5, Invention Description (d Specific twin crosses (Gaultier et a丨., Nucleic Acids Res (1 987) 1 5: 6625- 6641). The anti-nuclear nucleic acid molecule may also comprise methylribose nuclear acid (Inoue et a I., (1987) Nucleic Acids Res 15: 6131-6148) or chimeric RNA-DNA analog (Inoue et a I. (1 9 8 7 ) FEBS Left 215: 327-330) The present invention also encompasses a ribozyme which is a catalytic activity of having a ribonuclease activity and capable of cleaving a single-stranded nucleic acid having a complementary region (for example, a signaling RNA). RNA molecules. Thus, ribozymes (such as hammerhead ribozymes, described in Haselhoff et al., Nature (1988) 334: 585-591) can be used to catalyze the breakdown of GAVE8 signaling RNA transcripts, thereby inhibiting the translation of GAVE8 signaling RNA. Based on the GAVE8 complementary DNA nucleotide sequence disclosed herein (eg sequence confirmation number: 1) designed with a ribozyme encoding a GAVE 8 nucleic acid specificity. For example, a Tetrahymena L-1 9 IVS_ RNA derivative can be constructed in which the nucleotide sequence of the active site is complementary to the coding GAVE8 message. For the decomposed nucleotide sequence in the RNA, see, for example, U.S. Patent No. 4,987,07, to Cech et al., and U.S. Patent No. 5,116,742 to Cech et al. In addition, GAVE8 signaling RNA can be used from RNA molecules. A catalytic RNA having specific ribonuclease activity is selected from the population. See, for example, Bartel et al., Science (1993) 261: 1411-1418 ° The present invention also encompasses nucleic acid molecules that form a triple helical structure. For example, The targeting nucleotide sequence complementary to the GAVE8 regulatory region (eg, the GAVE8 promoter and/or enhancer) forms a triple-coiled construct to prevent transcription of the GAVE8 gene in the target cell and inhibits GAVE8 gene expression. See Helene, Anticancer Drug D is (1 991) 6(6): 569; This & Zhang scale applies Chinese National Standard (CNS) A4 specification (210X297 mm) — -22- (Read the first note on the back and refill Write this page) 1250209 A7 B7 Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative printing

V. INSTRUCTIONS (j Helene, Ann NY Acad Sci (1 992) 660:27; and Maher, Bioassays (1 992) 1 4(1 2): 807 较佳 In a preferred embodiment, the nucleic acid of the invention The molecule may modify the base moiety, the sugar side bond or the phosphate backbone to improve, for example, the stability, hybridization or solubility of the molecule. For example, the deoxyribose phosphate backbone of the nucleic acid may be modified to produce a peptide nucleic acid (see Hyrup et Al·, Bioorganic & Medicinal Chemistry (1 996) 4:5). The term "peptide nucleic acid" or "PNA" in this context means a nucleic acid mimicking agent, such as a DNA mimetic, wherein the deoxyribose phosphate backbone is Pseudopeptide backbone substitution, retaining only four natural nucleic acid bases. Neutral PNA backbone has been shown to allow DNA-RNA specific hybridization at low ionic strength. Synthetic PNA oligomers can be synthesized using standard solid phase peptides The guidelines are described in H y "upeta丨. (1 9 9 6), supra; Perry-OKeefe et al., Proc Natl Acad Sci USA (1 996) 93: 14670° PNAized GAVE 8 for treatment and diagnosis Use, for example, p NA can be used as a sequence specific An anti-femoral or antigenic agent that modulates gene expression, for example, inducing transcription or suppressing translation or inhibiting replication. The pNA-derived GAV Ε8 can also be used, for example, to analyze single base pair mutations of a gene, such as PNA-directed PCR clamps; Used to bind other enzymes (such as s 1 nuclease) as an artificial restriction enzyme (H yrup (1 9 9 6 ), supra), or as a probe or primer for d NA sequences and hybridization (Hyrup (1 996), sUpra Per "y-O'Keefe et al. (1 996), supra). In another embodiment, PNA-formed GAVE8 may be modified, for example by attaching a lipophilic or other ancillary group on the PNA to form a pma; -DNA This paper size is applicable to China National Standard (CNS) A4 specification (210X297 mm) (please read the note on the back and fill out this page). Attack P. -23- Printed by the Intellectual Property Office of the Ministry of Economic Affairs 1250209 kl B7 V. INSTRUCTIONS (2) Chimeras, or techniques known in the art of using lipid globules or other drugs to enhance their stability, specificity or cellular uptake. Techniques for synthesizing PNA-DNA chimeras Described in Hyrup (1 996), supr a, Finn et al., Nucleic Acids Res (1 996) 2 4(1 7): 3357-63, Mag et al., Nucleic Acids Res (1 989) 1 7:5973; and Peterser et a I., Bioorganic Med C. hem Lett (1975) 5:1119. II. Isolation of GAVE8 Protein and Anti-GAVE8 Antibody One of the features of the present invention relates to the isolation of the GAVE8 protein, and its biologically active portion, with appropriate polypeptide fragments, e.g., as an immunogen to elicit an anti-GAVE8 antibody. In one embodiment, the native GAVE8 protein can be isolated from cells or tissues by appropriate purification using standard protein purification techniques. In another embodiment, the GAVE8 protein can be produced by recombinant DNA techniques. In addition to recombinant expression, GAVE8 proteins or polypeptides can be chemically synthesized using standard peptide synthesis techniques. The 'isolated" or "purified" GAVE8 protein or a biologically active portion thereof is substantially free of cellular material or other cellular or tissue proteins, or substantially free of contamination by chemical precursors or other chemicals upon synthesis. "Substantially cell-free material" includes GAVE8 protein preparations isolated from cellular components or cells that recombinantly make proteins. Thus, GAVE 8 proteins of substantially cell-free materials, including less than about 30%, 20% The GAVE 8 protein of 10% or 5% (dry weight) of non-GAVE8 protein (also referred to herein as "contaminating protein") is prepared. When the GAVE 8 protein or its biologically active 咅5 line is recombinantly produced, it is preferable that the medium is substantially free of the medium, that is, the medium (please read the note on the back and then fill in the page). The bound paper size applies to the Chinese country. Standard (CNS) A4 specification (210X297 public Chu) -24 - !25〇2〇9 A7 B7 Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative printing 5, invention description (2) less than about 20%, 1 〇 ° /. Or a protein preparation volume of 5 ° / ◦. When the GAVE8 protein is produced by chemical synthesis, it is preferably substantially free of chemical precursors or other chemicals, indicating the separation of chemical precursors or other chemicals that synthesize proteins. Accordingly, the GAVE8 protein formulation is less than about 30%, 20%, 10% or 5% (dry weight) of chemical precursors or non-GAVE8 chemicals. The portion of the biological activity of the GAVE8 protein comprises a peptide comprising substantially the same or derived from the GAVE8 protein amino acid sequence (eg, the amino acid sequence shown in SEQ ID NO: 2), which may comprise less than the full length GAVE8 protein. Amino acid and exhibiting at least one GAVE8 protein activity. The biologically active portion typically comprises at least one structural region or motif of GAVE8 protein activity. The portion of the biological activity of the GAVE8 protein may be a polypeptide, i.e., an amino acid of, for example, 10, 25, 50, 100 or more. Preferred bioactive peptides include one or more structural regions of a confirmed GAVE8 construct, such as a circular structural region within a third cell (e.g., sequence confirmation number: 2). In addition, other biologically active portions of other regions of the protein can be deleted and the activity of one or more functions of the native GAVE8 protein can be prepared and evaluated using recombinant techniques. A preferred GAVE8 protein has its amino acid sequence shown in the sequence confirmation number. Other suitable GAVE8 proteins are proteins whose amino acid sequence differs from the sequence confirmation number: 2 due to mutation or mutagenesis of the native dual gene but retains protein function substantially identical to the sequence confirmation number ':2 . For example, the GAVE8 protein as well as the multi-peptide have at least one biological activity described herein. (Please read the notes on the back and fill out this page.) This paper scale applies to China National Standard (CNS) A4 specification (210X 297 mm) -25- 1250209 Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative printed kl B7 V. Invention Description (According to this, a suitable GAVE8 protein comprising an amino acid having an amino acid sequence of at least about 8 8 %, preferably 90%, 93%, 95 % or 99% identical to the sequence confirmation number: Sequence and retention sequence confirmation number: 2 GAVE8 protein functionally active protein. In other examples, 55%, 65% of the amino acid sequence with the circular structural region (sequence confirmation number: 2) in the third cell of GAVE8 75%, 85%, 95% or 99% identical GAVE8 protein. In a preferred embodiment, the GAVE8 protein retains the functional activity of the sequence confirmation number: 2 of the GAVE8 protein. To determine the two amino acid sequences or two A similar percentage of nucleic acid, the sequences can be aligned for optimal alignment (eg, 'the optimal alignment can be performed with the second amino group or nucleic acid sequence in the first amino acid sequence or the nucleic acid sequence introduction gap.) Comparison An amino acid residue or nucleotide at the amino acid position or nucleotide position. When the first sequence position has the same amino acid residue or nucleotide as the corresponding second sequence position, The molecules are identical at this position. The percent similarity between the two sequences is a function of the number of identical positions shared in the sequence (ie, similar percentage = number of identical positions / total number of positions (eg, overlapping positions) x 1 00). In one of the specific embodiments The lengths of the two sequences are the same. The mathematical algorithm can be used to determine the similarity percentage between the two sequences. The preferred non-limiting example of the mathematical calculation is to use Ka "lin et al., Proc Natl Acad Sci USA (1 990) 87:2264, Modified Karlin et al., Proc Natl Acad Sci USA (1993) 90:5873-5877 Calculate the two sequences. The algorithm has been incorporated into the NBLAST and XBBLAST programs, see Altschul et al. J Mol Bio (1990) 215: 403 〇BLAST nucleotides This paper scale applies to Chinese national standards (CNsYa4 specification (210X 297 mm) ~ ' -26- (please read the back note before filling this page). 125〇2〇9 kl B7 V. Invention (2) The search can be performed with the NBLAST program (score=100, word length=12) to obtain the nucleotide sequence homologous to the GAVE8 nucleic acid molecule of the present invention. The BLAST protein search can be performed by the XBLAST program (score=50, word length=3) To obtain the amino acid sequence homologous to the GAVE8 protein molecule of the present invention. In order to obtain the correction of the gap, B LAST can be used for gap detection, described in Altschul et al., Nucleic Acids Res (1 997) 25:3389 〇In addition, 'PSI-BLAST can be used for repeated searches to detect intermolecular The relationship between the two. Altschul et al., (1 997) supra. When using BLAST, gap BLAST, and PS BuLAST programs, various programs (such as XBLAST and NBLAST) can use preset parameters. See http : "w w w. n c b i · η 丨 m . n i h · g 〇 v. Another preferred non-limiting embodiment of the mathematical algorithm is to compare sequences using Myers et al., CABI(R) S (1 988) 4: 1 1-17. The algorithm has been incorporated into the calibration program (version 2.0) as part of the GCG sequence calibration package. When using the calibration procedure to compare amino acid sequences, the PAM 120 weighted residue table can be used with a gap length penalty of 12 and a gap penalty of 4. The percent similarity between the two sequences can be determined using techniques similar to those described above (with or without gaps). Calculate the percent similarity and count only the exact pairings. 〇 The invention also provides GAVE8 chimeric or fusion proteins. The GAVE8 "chimeric protein" or "fusion protein" herein comprises a GAVE8 polypeptide operably linked to a non-GAVE8 polymorph. "GAVE8 polypeptide•• means a polypeptide having a corresponding amino acid sequence to GAVE8, and “non-GAVE8 polypeptide" means having multiple paper sizes corresponding to amino acid sequences substantially different from GAVE8 protein for China National Standard (CNS) A4 Regulation (210X 297 mm) (Please read the note on the back and fill out this page) ip Packing · Ministry of Economic Affairs Intellectual Property Bureau Staff Consumer Cooperative Print -27-1250209 A7 B7 V. Invention Description (Peptide, for example a protein different from the GAVE8 protein, which may be derived from the same or different organisms. In the GAVE8 fusion protein, the GAVE8 polypeptide may correspond to all or part of the GAVE8 protein, preferably at least one GAVE8 protein organism Active moiety. In a fusion protein, the term "operably linked" means that the GAVE8 polypeptide and the non-GAVE8 polypeptide are fused to each other on the same framework. The non-GAVE8 polypeptide can be fused to the N-terminus or C-terminus of the GAVE8 polypeptide. A suitable fusion protein is a GST-GAVE8 fusion protein in which the GAVE8 sequence is fused to the C-terminus of the glutathione-s-transferase (GST) sequence. Purification of recombinant GAVE8 can be enhanced. In a preferred embodiment, the third intracellular loop of the invention (丨L3) (i.e., from about 214 to about 252 of sequence confirmation number 2) is amplified by PCR. And subsequencing the product into a vector (eg, pGEX-2T) to fuse with GST. The resulting construct can be introduced into a host cell (eg, E. coli), which can be passed through a suitable small molecule (eg, isoform) Propyl-1 -thio-cold·D-galactopyranoside is induced and expressed and purified (see, for example, Lee et al., J Biol C he m (1 996 ) 2 7 1 (1 9) : 1 1272-1 1279). In certain host cells (eg, mammalian host cells), the heterologous signal sequence can increase GAVE8 expression and/or secretion. For example, glutamate envelope protein gpe® can be used. Secretory sequences serve as signal sequences for heterosexuality IJ (Current Protocols in Molecular Biology, Ausubel et al., eds., John Wiley & Sons, 1992). Examples of other eukaryotic heterosexual signal sequences include: melittin and human placenta Secretion sequence of alkaline phosphatase (Stratagene; La Jolla, California). Another embodiment of this paper scale applies to the Chinese National Standard (CNS) A4 specification (210X297 mm) (please read the note on the back and fill out this page) im, install the Ministry of Economic Affairs Intellectual Property Office staff fee Co-operative Printing -28-1250209 A7 V. INSTRUCTIONS (in the case, suitable prokaryotic signal sequences include PhoA secretory signals (Sambrook et al., supra) and protein A secretory signals (Pharmacia Biotech; Piscataway, New Jersey) . In another specific embodiment, the fusion protein is a GAVE8-immunoglobulin fusion protein in which all or a portion of the GAVE8 line is fused to a protein group sequence derived from an immunoglobulin. The GAVE8-immunoglobulin fusion protein of the present invention can be incorporated into a pharmaceutical composition and administered to a patient to inhibit the interaction between the GAVE8 ligand on the cell surface and the GAVE8 protein, thereby inhibiting GAVE8-regulated signaling in vivo. The GAVE8-immunoglobulin fusion protein can be used to affect the bioavailability of GAVE8 cognate ligands. Inhibition of the interaction of GAVE8 ligand-GAVE8 is therapeutically suitable for treating proliferative and differentiated disorders and modulating (e.g., promoting or inhibiting) cell survival. In addition, the GAVE8-immunoglobulin fusion protein of the present invention can be used as an immunogen to enable an anti-GAVE8 antibody to be produced in a patient, and is used for purifying a GAVE8 ligand and a molecular assay which confirms the interaction between the GAVE8 ligand molecule and GAVE8. Preferred GAVE8 chimeric or fusion protein lines of the invention can be made using standard recombinant DNA techniques. For example, according to conventional techniques, DNA fragments encoding different polypeptide sequences are linked to a common framework, for example, using a blunt end or a viscous end to bind to limit enzymatic hydrolysis to provide a suitable binding end, filling the viscous end, Alkaline phosphatase treatment avoids unsatisfactory linkages and the binding of enzymes. In another embodiment, the fusion gene can be synthesized using conventional techniques, including automated DNA synthesizers. In addition, when using PD R for amplification of gene fragments, it can be used in two consecutive gene fragments. The Chinese National Standard ( ^NS ) A4 specification (210X297^7 - -29- (Read the back) Precautions to fill out this page) - Loading · Booking

P Ministry of Economic Affairs Intellectual Property Bureau Staff Consumer Cooperatives Printed 1250209 A7 B7 Ministry of Economic Affairs Intellectual Property Bureau Staff Consumer Cooperatives Printed V. Invention Description (2) A fixed primer that produces complementary protruding parts, which are subsequently annealed and re-amplified. A chimeric gene sequence can be generated (see, for example, Ausubel et al., supra). In addition, a number of expression vectors encoding a fusion moiety (e.g., a GST polypeptide) are commercially available. A nucleic acid encoding GAVE8 can be optionally incorporated into the expression vector, and the fusion moiety can be linked to the framework to the GAVE8 protein. The invention also relates to a GAVE8 protein variant (i.e., a protein whose sequence differs from the GAVE8 amino acid sequence). This variant functions as a GAVE8 agonist or a GAVE8 antagonist. GAVE8 protein variants can be produced by mutagenesis, such as point mutations that are not ligated or truncated GAVE8 proteins. The GAVE8 protein agonist retains substantially the same biological activity (or its secondary activity) as the native form of the GAVE8 protein. Antagonists of the GAVE8 protein inhibit the activity of one or more native form of the GAVE8 protein. For example, a competitive binding to a downstream or upstream element of a tandem reaction of a cell signal comprising a GAVE8 protein. Therefore, a specific biological effect can be caused by the modification of the restriction function. Treatment of a patient with a modification of the sub-biological activity of the native form of the protein has a lower side effect than treating the patient with the native form of GAVE8 protein. A GAVE 8 protein variant having a function as a GAVE8 agonist or a GAVE8 antagonist is identified as a GAVE8 protein having a GAVE8 protein agonist or antagonist activity in a combinatorial gene pool of a selectable mutant (e.g., a truncation mutant). In one embodiment, a library of various GAVE8 variants can be generated by combinatorial mutagenesis at the nucleic acid level and encode various gene repertoires. Methods for making various gene repertoires of the GAVE8 variant are, for example, the use of enzyme-linked oligonucleotide mixtures and gene sequences, thereby producing a set of degenerate GAVE8 sequences that are used to represent individual multi-peptides, or to express the inclusions herein ( Please read the notes on the back and fill out this page. Loading - This paper scale applies to China National Standard (CNS) A4 specification (2) OX 297 mm) -30- Ministry of Economic Affairs Intellectual Property Office Staff Consumer Cooperative Printed 1250209 A7 _____B7 V. INSTRUCTIONS (a large fusion protein of a group of j GAVE8 sequences (for example, shown by phage). There are various methods for generating a gene bank of GAVE 8 variants from degenerate oligonucleotide sequences. Chemical synthesis degradation The gene sequence can be performed using an automated DNA synthesizer, and the synthetic gene is then ligated into an appropriate expression vector. The advantage of using a degenerate genome is that all sequences encoding the desired GAVE8 sequence set can be found using only one mixture. Degenerate oligonucleotides The synthesis method is a method known in the art (see for example: Narang, Tetrahedron (1 983) 39:3; Itakura eta I., Ann Rev Biochem (1984) 53:323; Itakura et al., Science (1 984) 1 98:1 056; Ike et al., Nucleic Acid Res (1983) 11:477) In addition to the 'GAVE8 protein coding sequence fragment The gene pool can be used to generate a variety of GAVE8 fragment populations for screening and subsequent selection of GAVE8 protein variants. In one embodiment, the method for encoding a gene bank that produces sequence fragments is to double-strand the PCR fragments of the GAVE8 coding sequence with a nuclease. The reaction is carried out under the condition that only one gap occurs per molecule, and the double-stranded DNA is denatured to separate the DNA to restore the nature to form double-stranded DNA (which may include a positive/anti-strand pair from different gap products), which will be reformed. The double-strand is treated with S1 nuclease to remove the single-stranded portion, and the resulting fragment gene pool is linked to the expression vector. In this method, the phenotype gene library can be derived from the GAVE8 protein encoding N-terminal and various internal Sequences of Size Fragments There are a number of techniques known in the art for screening gene products of combinatorial gene banks made by point mutation or truncation, as well as for screening for complementary DNA libraries. Gene product. This technology can quickly screen GAVE8 protein (please read the back note first and then fill out this page) This paper scale applies to Chinese National Standard (CMS) A4 specification (210X 297 mm) -31 · Ministry of Economics intellectual property Bureau employee consumption cooperative printed 1250209 A7 ___B7 V. Invention description (gene gene library generated by combined mutagenesis. The most commonly used techniques are capable of performing high-performance analysis, screening large gene banks for gene library selection into replicable expression vectors, translating gene banks generated by vectors into appropriate cells, and performing combinations The desired activity is detected under the conditions of the gene to enhance the isolation of the vector encoding the detected product. Total Recursive Mutagenesis (REM) is a technique for increasing the frequency of functional mutants in a gene bank and can be used in conjunction with screening assays to confirm GAVE8 variants (Arkin et al., Proc Natl Acad Sci USA (1 992) 8 9:781 1 -781 5; Delgrave et a I., Protein Engineering (1993) 6(3): 327-331) 〇Isolated GAVE8 protein (or a portion or fragment thereof) can be used as an antigen, using multiple strains and monoclonal antibodies Standard skill to produce antibodies that bind to GAVE8. The full-length GAVE8 protein can be used, or the immunopeptide fragment of GAVE8 can be used as an immunogen in the present invention. The GAVE8 antigenic peptide comprises at least 8 (preferably 10, 15, 20, or 30) amino acid residues such as the amino acid sequence of the sequence confirmation number: 2, and comprises an epitope of GAVE8, thus resulting in The antibody forms a specific immune complex with GAVE8. In one embodiment, the epitope can be an 8-mer comprising a sequence confirmation number: 2 residues. In a related feature, the antigenic peptide comprises an epitope which is located in a region of the surface of the GAVE8 protein, such as a hydrophilic region. The results of water repellency analysis by human GAVE8 protein sequence showed sequence number: 2, especially between about 1 to about 37 of about amino acid, between about 96 to about 11 of about amino acid, between The region between 1 77 and about 1 92 of about amino acid and between about amino acid 274 and about 29 is a hydrophilic region 'so (please read the back note first and then fill out this page)

, 1T mr. This paper scale applies to China National Standard (CNS) A4 specifications (2) 〇 X 297 mm) -32- 1250209 A7 _____B7

V. Description of the invention (3 (J can encode surface residues for antibody production. (Please read the back note first and then fill out this page) GAVE8 is usually used as an antigen to immunize appropriate animals with antigen (eg rabbit, Antibodies may be prepared by goats, mice or other mammals. Suitable formulations for producing immunity may contain, for example, recombinantly expressed GAVE8 protein or chemically synthesized GAVE8 polypeptide. The formulation may further comprise an adjuvant, such as complete or incomplete F "eund adjuvant, or a similar immunostimulant. Immunization of an appropriate animal with an immunogenic GAVE8 preparation can induce multiple anti-GAVE8 antibody responses. Printed by the Ministry of Economic Affairs, Intellectual Property Office, Staff Consumer Cooperative, accordingly, the present invention Another feature relates to anti-GAVE 8 antibodies. The term "antibody" as used herein refers to immunoglobulin molecules and immunologically active portions of immunoglobulin molecules, ie, molecules that contain a specific binding site for the antigen binding of GAVE8. The molecule of GAVE8 is GAVE8 which can be bound to a sample (for example, a biological sample containing natural GAVE8), but does not substantially bind Molecules of other molecules in the sample. Examples of immunologically active portions of immunoglobulin molecules include F(ab) and F(ab^2 fragments, which are produced by reacting an antibody with an enzyme (eg, pepsin). Providing a multi-strain and monoclonal antibody that binds GAVE8. The term "monoclonal antibody" or "monoclonal antibody composition i" as used herein means an antibody that contains only one antigen-binding site and can immunoreact with a specific epitope of GAVE8. Therefore, the monoclonal antibody composition usually exhibits a single binding affinity to a specific GAVE8 protein at its immunoreactive site. The anti-GAVE8 antibody of the above-mentioned multiple strains can be prepared by immunizing an appropriate animal with the GAVE8 antigen. The potency of the anti-GAVE8 antibody in animals can be monitored by standard techniques for a period of time, for example, using the immobilized GAVE8 with the paper size applicable to the Chinese National Standard (CNS) A4 specification (2) 0X297 mm) -33- 1250209 A7 B7 Ministry of Economic Affairs Intellectual Property Bureau Staff Consumption Cooperatives Printing and Burning Five, Invention Description (y enzyme-linked immunosorbent assay (enzyme-linked immunosorbent assay) As needed, antibody molecules directed against GAVE8 can be isolated from mammals (eg, blood) and further purified using known techniques (eg, protein A chromatography) to obtain immunoglobulin G lysates. After the time, for example, when the titer of the anti-GAVE8 antibody is highest, the antibody-producing cells can be obtained automatically, and monoclonal antibodies are prepared using standard techniques, such as fusion tumor technology, originally described in K 〇h I ereta I. , Nature (1 975) 256:495-497, Human B Cell Fusion Tumor Technology (Kohler et a I., Immunol Today (1 983) 4:72), EBV-fusion tumor technology (Cole eta I., Monoclonal Antibodies And Cancer Therapy, (1 985), Alan R. Liss, Inc., pp. 77-96) or Trioma skills. The art of producing fusion tumors is a well-known technique (see Current Protocols in Immunology (1 994) Coligan et al., (eds.) John Wiley & Sons, I nc., New York, NY). Briefly, an immortalized cell line (generally a bone marrow cancer cell line) is fused to lymphocytes (generally spleen cells) of a mammal immunized with the above GAVE8 antigen, and the resulting culture suspension of the fusion tumor cells is screened. A fusion tumor producing a monoclonal antibody that binds to GAVE8 was confirmed. Anti-GAVE8 monoclonal antibodies can be produced using well-known experimental guidelines for any of a number of lymphocytes and immortalized cell lines (see, for example, Current Protocols in Immunology, supra; G a If re et a I., Nature (1977) 266:55052 Kenneth, in Monoclonal Antibodies: A New Dimension In Biological Analyses, Plenum Publishing Corp., New York, NY (1 980); and Lerner, (please read the notes on the back and fill out this page)

P This paper scale applies to China National Standard (CMS) A4 specification (210X297 mm) -34- Ministry of Economic Affairs Intellectual Property Office Staff Consumer Cooperative Printed 1250209 A7 __ B7 V. Invention Description (3) Yale J Biol M ed (1 9 8 1 ) 54 : 3 8 7-4 02 ). Moreover, those skilled in the art will recognize that many variations of the method can be applied. Usually, immortalized cell lines (e.g., myeloid cancer cell lines) are derived from lymphocytes of the same mammal. For example, a murine fusion tumor can use a lymphocyte of a mouse immunized with an immunogenic preparation of the present invention and an immortalized mouse cell strain (for example, a culture solution containing hypoxanthine, aminopterin, and thymine) (" Hat culture medium") a sensitive bone marrow cancer cell line). Many bone marrow cancer cell lines can be used as fusion partners according to standard techniques, for example: P3-NS1/1-Ag4-1, P3-x63-Ag8.653 Or Sp2/0-Ag14 bone marrow cancer cell line. The above bone marrow cancer cell line can be purchased from ATCC. Generally, mouse bone marrow cancer cells sensitive to HAT can be fused to mouse spleen cells using polyethylene glycol ("PEG") The fusion-derived tumor cells can be selected using Η AT broth, which kills unfused and non-productive fused bone marrow cancer cells (unfused spleen cells die for several days because they have not been transformed) Thereafter, for example, a fusion cell of a single cell strain producing the antibody of the present invention can be detected by screening a fusion culture suspension of GAVE8 using a standard enzyme-linked immunosorbent assay. In the method of secreting a fusion cell antibody, the recombinant immunoglobulin gene library (for example, an antibody phage display gene library) is screened by GAVE8, thereby isolating the immunoglobulin gene library member that binds to GAVE 8, and the individual plant can be confirmed and isolated. Anti-GAVE8 antibody. A set of phage display gene banks for production and screening is commercially available (eg Pharmacia recombinant phage antibody system, catalog number 27-9400-01; and Stratagene SurfZAP® Phage Display Kit, catalog number 24061 2 Bu (please read the note on the back and fill in this page) This paper scale applies to China National Standard (CNS) A4 specification (210X297 mm) -35- 1250209 A7 B7 Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative printed five Description of the Invention (3$ In addition, examples of methods and reagents that are particularly applicable to the production and screening of antibody display gene banks can be found, for example, in U.S. Patent No. 5,223,409; PCT Publication No. WO 92/1 861 9; PCT Publication No. W0 91/1 7271 ; PCT publication number WO92/20791 ; PCT publication number W0 92/1 5679 ; PCT publication number W0 93/01 288 ; PCT Cloth number W092/01 047; PCT publication number W092/09690; PCT publication number W0 90/02809; Fuchs et a I., B io/Technology (1991) 9:1 370-1 372; Hay eta I., Hum Anti Bod Hybridomas (1 992) 3:81 -85; Huse et a I., Science (1 989) 246:1 275-1281; and Griffiths et al., EMBO J (1 993) 25 1 2:725-734 c In addition, recombinant anti-GAVE8 antibodies (e.g., chimeric and humanized monoclonal antibodies) comprising human and non-human portions can be made using standard recombinant DNA techniques, and are within the scope of the invention. The chimeric and humanized monoclonal antibodies can be made using recombinant DNA techniques known in the art, for example, as described in PCT Publication No. WO 87/0267 1; European Patent Application No. 1 84,1 87; European Patent Application No. 1 71,496; European Patent Application No. 1 73,494; PCT Publication No. WO 86/01 533; US Patent No. 4,81 6,567; European Patent Application No. 1 25,023; Better et a I., Science ( 1988) 240: 1041-1043; Liu et a I., Proc Natl Acad Sci USA (1987) 84 · 3439-3443; Lin et a I., J Immunol (1 987) 1 39:3521 -3526; Sun et a I., Proc Natl Acad Sci USA (1 987) 84:214-21 8; Nishimura eta i., Cane Res (1 987) 47:999-1 005; Wood et al., Nature (please read the back) Note: Please fill in this page again) ip Packing · Book size is applicable to China National Standard (CNS) Α4 size (210X297 mm) -36- 1250209 A7 B7 Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative printing 5, invention description ( 3) (1985) 314:446-449; Shaw et a I., J Natl Cancer Inst (1988) 80:1 553-1559; Morrison, Science (1 985) 229:1202-1207; Ieta I. , Bio/Techniques (1 9 86) 4:21 4; US Patent No. 5,225,539; Jones eta I., Nature (1 986) 321:552-525; Verhoeyan et a I., Science (1 988) 239:1 534; and Beidler et al., J Immunol (1 988) 1 41:4053-4 0 60 〇 Complete human antibodies are especially suitable for the treatment of human patients. This anti-system was produced using a mouse that does not express endogenous immunoglobulin heavy and light chain genes, but which can express foreign heavy and light chain genes and introduce foreign genes. Mice that are introduced with a foreign gene are immunized in a normal manner with a selected antigen (for example, all or part of GAVE8). Monoclonal antibodies directed against the antigen can be obtained using the fusion fusion technique of the prior art. During B cell differentiation, the human immunoglobulin-introduced gene can be recombined into a mouse into which a foreign gene is introduced, followed by class switching and mutation of a somatic chromosome. Therefore, it can be confirmed using the antigen-determining site (e.g., an antibody that inhibits GAVE8 activity). The heavy and light chains of non-human antibodies have been cloned and used to create phage-displayed Fab fragments. For example, the heavy chain gene can be optionally ligated into a plastid vector such that the heavy chain can be secreted from the bacterium. The light chain gene can optionally be introduced into the phage coat protein gene, and the light chain can be expressed on the surface of the phage. Bacterial flora (random collection) fused to the human light chain is used to infect bacteria that exhibit non-human heavy bonds. The resulting phage can display hybrid antibodies (human light chain/non-human heavy chain). Phages that bind to selective antigens are screened using a selective antigen panning. The phage can be confirmed after several selections. Next, the humans who select the antigen will be combined (please read the notes on the back and fill out this page). 0, Packing and binding f The paper size applies to the Chinese National Standard (CNS) A4 specification (210X 297 mm) -37 - χ25 〇2〇9 Α7 Β7 V. Description of the invention (35) (Please read the notes on the back and fill out this page) The light chain gene is isolated from the selected phage. Such selected human light chain genes can then be used to select human heavy chain genes. It inserts the selected human light chain gene into a bacterial expression vector. Bacteria expressing the selected human light chain are infected with a phage population that is fused to the human heavy chain. The resulting progeny phage can display human antibodies (human light chain/human heavy chain). Next, the phage that binds to the selected antigen is screened using the selected antigen panning. The phage selected at this step (showing intact human antibodies) confirmed the same epitope as the original selective non-human monoclonal antibody. The genes encoding the heavy and light chains are isolated and can be further manipulated to produce human antibodies. This technique is described in Jespers et al. (Bio/Technology (1 994) 12: 899-903). GAVE8 can be isolated by anti-GAVE8 antibodies (e.g., monoclonal antibodies) using standard techniques (e.g., affinity chromatography or immunoprecipitation). The anti-GAVE8 antibody enhances the purification of the recombinant GAVE8 produced by the natural GAVE8 and host cell expression of the cells. In addition, anti-GAVE8 antibodies can be used to detect proteins (such as cell lysates or cell sputum) printed by the GAVE8 Ministry of Economic Intelligence, Inc., and to assess the biological abundance and pattern of G A V E 8 protein expression. Anti-GAVE8 antibodies can be used to diagnostically monitor the amount of protein in a tissue that is part of a clinical testing procedure', e.g., to determine the efficacy of a therapeutic regimen. Coupling antibodies to detectable substances facilitates detection. Examples of detectable substances include various enzymes, prosthetic groups, fluorescent materials, luminescent materials, biological luminescent materials, and radioactive materials. Examples of suitable enzymes include: horseradish peroxidase, alkaline phosphatase, half? L-glycosidase or acetamidine test; appropriate examples of prosthetic complexes, including streptavidin/biotin and avidin/biological paper scales applicable to China National Standard (CNS) A4 Specifications (2j〇X29^i7-----38- 1250209 Α7 Β7 V. Inventive Note (30>Prime; Examples of suitable fluorescent materials include umbelliferone, luciferin, luciferin Isothiocyanate, rhodamine, dichlorotrimethylamine luciferin, dansyl chloride, a given fluorescent protein or phycoerythrin; examples of luminescent materials include Rummin; Examples of biological luminescent materials include: luciferase, luciferin, and aequorin. Examples of suitable radioactive materials include 1251, 1311, 35S, or 3H. 111. Recombinant Expression Vectors and Host Cells of the Invention Another feature relates to a vector, preferably an expression vector containing a GAVE8 nucleic acid (or a portion thereof). The term "vector" as used herein means a nucleic acid molecule capable of carrying another linked nucleic acid. One of the types of vectors is n plastid ", which can link additional DNA fragments Round double-stranded circular DNA. Another type of vector is a viral vector in which additional DNA fragments can be linked to viral genomes. Certain vectors (eg bacterial vectors and gene-containing organisms with bacterial origins of replication) The mammalian vector can replicate autonomously in the host cell into which it is introduced. Other vectors (e.g., non-gene-available mammalian vectors) can be inserted into the genome of the host cell upon introduction into the host cell, thereby replicating along with the host genome. In addition, certain vectors (expression vectors) are operatively linked to express genes directly. In general, expression vectors for recombinant DNA techniques are often in the form of plastids (vectors). However, the invention includes other forms. A performance vector, such as a functionally applicable viral vector (eg, a replication-defective retrovirus, an adenovirus, and an adenovirus-associated virus). The recombinant expression vector of the present invention is suitable for expressing a native paper scale in a host cell. Applicable to China National Standard (CNS) A4 specification (210Χ 29Τ^"ϊ ' ---- -39- (Please read the back In addition, please fill out this page. Packing · Ministry of Economic Affairs Intellectual Property Bureau Staff Consumer Cooperative Printed 1250209 A7 B7 Ministry of Economic Affairs Intellectual Property Bureau Staff Consumer Cooperative Printed 5, Invention Description (37) Invented nucleic acid form. Including one or more regulatory sequences (depending on the host cell used for expression) operably linked to the nucleic acid of expression. In a recombinant expression vector, "operably linked" means an important nucleoside The acid sequence is linked to a regulated sequence in a manner that allows expression of the nucleotide sequence (eg, an in vivo transcription/translation system or a host cell into which the vector is introduced). The term "regulated sequence" includes promoters, enhancers And other performance control elements (eg, polyadenylation signals). The sequence of this regulation 描述 ij describes the order such as: Goeddel, Gene Expression Technology: Methods in Enzymology Vo 1. 185, Academic Press, San Diego, CA (1 990). Regulatory sequences include nucleotide sequences that are directly constitutive in many types of host cells (e.g., tissue-specific regulatory sequences). Professionals familiar with this skill are aware of the factors that characterize the design performance carrier: the choice of the transformed host cell, the level of performance of the desired protein, and the like. The expression vector of the present invention can be introduced into a host cell to produce a protein or peptide encoded by the nucleic acid described herein (e.g., GAVE8 protein, mutant form of GAVE8, fusion protein, etc.). The recombinant expression vector of the present invention can be designed to express GAVE8 in prokaryotic or eukaryotic cells, for example, bacterial cells such as Escherichia coli, insect cells (using a baculovirus expression vector), yeast cells or mammalian cells. . Suitable host cells are further discussed in G 〇 e d d e丨, s u p r a. In addition, recombinant expression vectors can be transcribed and translated in vitro, for example, using the T7 promoter-regulated sequence and T7 polymerase. Proteins expressed in prokaryotes, mostly using a constitutive or inducible promoter containing a control for fusion or non-fusion proteins, are applicable to the Chinese National Standard (CNS) A4 specification (210X297 mm) on this paper scale. ) ~ -40- (Read the first note on the back and fill out this page) 1250209 A7 B7 Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative printing 5, invention instructions (38) E. coli. The fusion vector is typically encoded by a number of amino acids at the amine end of the recombinant protein. The general purpose of the fusion vector is to: 1) increase the expression of the recombinant protein; 2) increase the solubility of the recombinant protein; and 3) act as a ligand in the affinity purification to assist in the purification of the recombinant protein. In the fusion expression vector, a proteolytic excision site is often introduced at the junction between the fusion portion and the recombinant protein, and the recombinant protein can be isolated from the fusion portion after purification of the fusion protein. The enzyme, and its homologous sequence of recognition, includes Factor Xa, thrombin, and enterokinase. Representative fusion expression vectors include: pGEX (Pharmacia Biotech Inc.; Smith et al., Gene (1988) 67: 31-40), pMAL (New England Biolabs, Beverly, MA) and pRITS (Pharmacia, Piscataway, NJ). It is a fusion of glutathione 5-transferase (GST), maltose E-binding protein or protein A to recombinant protein, respectively. Examples of suitable inducible non-fusion E. coli manifestations include: pTrc (Amann et al., Gene (1 988) 69:301 -31 5) 3⁄4 pET 11d (Studier et a I., Gene Expression Technology: Methods in E nzymology, Academic Press, San Diego, California (1 990) 1 85:60-89). When the target gene is expressed from the pTrc vector, it is the initiation of transcription of the sin-primary R N A polymerase transcription and the t r p -丨a c fusion promoter. When the target gene is expressed from the P E T 1 1 d vector, the T7 gnMac fusion promoter is transcribed by a co-presented viral RNA polymerase (T7 gnl). The viral polymerase can be supplied by the host strain BL21 (DE3) or HMS 174 (DE3) inserted under the lacUV 5 promoter-controlled transcription of the λ spp phage Τ7 gnl gene. (Please read the notes on the back and fill out this page.) This paper scale applies to China National Standard (CMS) A4 specification (210X 297 mm) -41 - 1250209 kl B7 Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative printing 5, invention Description (g) One of the strategies for optimizing the expression of recombinant proteins in E. coli is to express proteins in host bacteria that impair the ability to hydrolyze recombinant proteins (Gottesman, Gene Expression Technology: Methods in Enzy mology, Academic Press, San Diego, California (1 990) 1 85: 1 1 9-1 28). Another strategy is to change the nucleic acid sequence of the nucleic acid inserted into the expression vector, so that the codons of each amino acid are codons that are preferentially utilized by E. coli (Wads et a I., Nucleic Acids Res (1 992) 212111-2118). The alteration of the nucleic acid sequence of the present invention can be carried out by standard DNA synthesis techniques. In another embodiment, the expression vector of GAVE8 is a yeast expression vector. Examples of vectors for expression in Saccharomyces cerevisiae include: · pYepsecl (Baldari et al., EMBO J (1 987) 6: 229-234), pMFa (Kurjan et al., Cell (1 982) 30 933-943), pJRY88 (Schultz et al., Gene (1 987) 54: 113-123), pYES2 (lnvitrogen Corporation, San Diego, CA), and pPicZ (lnvitrogen Corp, San Diego, CA) GAVE8 is expressed in insect cells using a baculovirus expression vector. Baculovirus vectors expressing proteins in cultured insect cells (eg, Sf9 cells) include: pAc series (Smith et al., Mol Cell Biol (1 983) 3: 2 1 56-21 65) and the pVL series (Lucklow et al., Virology (1 989) 1 70: 31-39. In another specific embodiment, mammalian expression vectors can be used to express the expression in mammalian cells. Nucleic acid of the invention. Examples of mammalian expression vectors, including: pC DM8 (Seed, Nature (1 987) 329: 840) and I-------- ί ^-- (read first Read the notes on the back and fill out this page.) "The standard of the paper size is applicable. National Standard (CNS) A4 Specification (2IOX 297 mm) -42 - 1250209 A7 B7 Ministry of Economic Affairs Intellectual Property Bureau Staff Consumer Cooperative Printed Five , Invention Description (4C) pMT2PC (Kaufman et al., EMBO J (1 987) 6: 1 87-1 95. When mammalian cells are used, regulatory elements that control the pathogenesis of the function of the vector are often used. For example, commonly used promoters are derived from: polyoma, adenovirus 2, cytomegalovirus, and monkey virus 40. Suitable profiling systems for other prokaryotic and eukaryotic cells can be found in Chapters 16 and 17 of Sam Brook et al., supra. In another embodiment, a recombinant mammalian expression vector can preferentially perform nucleic acid expression directly on a particular cell type (e.g., a tissue-specific regulatory element for expressing a nucleic acid). Organizing a specific regulatory element is a skill known in the art. Non-limiting examples of appropriate tissue-specific promoters include: albumin promoter (with liver specificity); Pinkert et al., Genes Dev (1 987) 1:268-277), lymphoid-specific promoter ( Calame et al., Adv Immunol (1 988) 43:235-275), in particular the T cell receptor promoter (Winoto et al., EMBO J (1 989) 8: 729-733) and immunoglobulin promoters (Banerji et al., Cell (1 983) 33: 729-740; Queen et al., Cell (1 983) 33: 741-748), a neuron-specific promoter (eg neurofilament promoter; Byme) Et al., Proc Natl Acad Sci USA (1989) 86: 5473-5477), a pancreatic-specific promoter (Edlund et al., Science (1 985) 230·912-916) and a breast-specific promoter ( For example: milk chyle promoter; US Patent No. 4, 8 7 3, 3 16 and European Application Publication No. 264, 1 66). It may also contain a promoter that regulates development, such as the mux promoter of the murine (Kessel et al., Science (1 990) 249: 374-379) and the alpha-fetoprotein promoter (Campes et al., Genes). Dev (1 989) 3:537-54). (Please read the precautions on the back and fill out this page) • Loading and setting the paper size for the Chinese National Standard (CNS) A4 specification (2) 0X 297 mm) -43- 1250209 A7 ___B7 V. Invention description ( (please The invention of the present invention further provides a recombinant expression vector comprising a DN A molecule of the present invention comprising a counter-stranding direction, that is, to allow expression (transcription of D ΝΑ molecules) GAVE8 signaling RNA anti-strand The RNA molecule is operatively linked to the DNA molecule of the regulatory sequence. In various cell types, the sequence that can be operably linked to the nucleic acid in the anti-strand direction to express the regulation of the anti-strand RNA molecule is, for example: a viral promoter and/or enhancer Alternatively, a direct or constitutive, tissue-specific or cell type-specific expression of the anti-strand RNA may be selected. The anti-strand expression vector may be in the form of recombinant plastid, phage or attenuating virus. The production of nucleic acids is controlled by highly efficient regulatory regions, and the activity can be measured after the cells are introduced into the vector. For discussion of the use of antisense genes to regulate gene expression, see Weintraub et a l.( Re views-T rends in Genetics, Vol. 1 (1 ) 1 986. Printed by the Ministry of Economic Affairs, Intellectual Property Office, Staff Consumer Cooperatives Another feature of the present invention relates to host cells into which the recombinant expression vector of the present invention has been introduced. The terms "host cell" and "recombinant host cell" are mutually versatile. It is understood that the term refers not only to a particular cell, but also to the progeny or progeny of the cell. Due to mutation or environmental influence, Some modifications may occur in the offspring, in fact the progeny may be different from the parent cell, but still fall within the scope of the term. The host cell may be any prokaryotic or eukaryotic cell. For example, the GAVE8 protein may be expressed in the cells of the bacteria, For example: E. coli, insect cells, yeast or mammalian cells (e.g., Chinese hamster ovary cells (C Η 0) or C 0 S cells). Other suitable host cells are known to those skilled in the art. The vector D Ν 引入 can be introduced into prokaryotic or eukaryotic cells by the conventional transformation or transfection technique. The term "transformation" "&&q Uot; This paper scale applies to China National Standard (CNS) A4 specification (210X 297 mm) -44 - 1250209 A7 ____ B7 V. Invention Description (Transfection effect) means various nucleic acids that have been confirmed to be introduced into foreign countries ( Techniques such as DNA) to host cells include: calcium phosphate or calcium chloride co-precipitation, DEAE-dextran-mediated transfection, lipofection or electroporation, for stable transfection of mammalian cells It is currently known that depending on the performance vector and the transfection technique used, only a small fraction of the cells can invade the foreign DNA into the genome. To identify and select invading cells, genes encoding alternative markers (e.g., antibiotic resistance) can be introduced into host cells with important genes. Preferred selectable markers include markers that confer resistance to drugs (e.g., G4 18, hygromycin, and methotrexate). The nucleic acid encoding the selectable marker can be introduced into the host cell either as the vector encoding GAVE8 or as an isolated vector. Cells that are stably transfected with the introduced nucleic acid can be identified by drug selection (for example, cells that enter the selectable marker gene will survive and other cells will die). A host cell of the invention, e.g., a cultured prokaryotic or eukaryotic host cell, can be used to produce (i.e., express) a GAVE8 protein. Accordingly, the present invention further provides a method of producing a GAVE8 protein using the host cell of the present invention. In one embodiment, the method comprises culturing a host cell of the invention in a suitable culture medium in which the G A V E 8 protein is produced (a recombinant expression vector encoding G A V E 8 has been introduced). In another embodiment, the method further comprises isolating GAVE8 from the culture broth or host cell. Host cells of the invention can also be used to produce non-human gene transgenic animals. For example, in one of the specific embodiments, the host cell of the present invention is a stem cell of a fertilized oocyte or embryo that has been introduced into the GAVE8-encoding sequence. However, the paper size is applicable to China National Standard (CNS) A4 specification (2) 0X 297 mm) " ' One-45 - (Please read the note on the back and fill in this page) - Install·

P Ministry of Economic Affairs Intellectual Property Bureau Staff Consumer Cooperative Printed 1250209 A7 B7 Ministry of Economic Affairs Intellectual Property Bureau 8 Workers Consumption Cooperative Printed 5, Invention Description (The host cell can be used to create a genome to introduce an exogenous GAVE8 sequence of non-human gene-transgenic animals or A homologous recombinant animal that has altered the endogenous GAVE8 sequence. The animal can be used to study the function and/or activity of GAVE8 and to confirm and/or evaluate the modulator of GAVE8 activity. "The animal that introduces the foreign gene" Preferably, the mammal is a mammal, and more preferably a rodent, such as a rat or a mouse, wherein one or more cells of the animal include an introduced gene. Other embodiments of the genetically modified animal include: non-human primates, sheep , dog 'bovine, goat, chicken, amphibian, etc.. The introduced gene is inserted into exogenous DNA in the genome of the cell, developed into an animal that introduces a foreign gene and maintains the genome in a mature animal, thereby introducing one of the foreign gene animals or The gene product encoded directly in a variety of cell types or tissues. The homologous recombinant animal Preferably, it is a mammal, and more preferably a mouse, wherein the endogenous GAVE8 gene is homologously recombined with an endogenous gene and an introduced DNA molecule (for example, an animal embryo cell before the animal is formed). Alteration. Introduction of the nucleic acid encoding GAVE8 to the male pronuclei of the fertilized oocyte (eg, microinjection, anti-viral infection) and allowing oocytes to occur in pseudopregnant female nourishment animals, may create the present invention An animal into which a foreign gene is introduced. The complementary DNA sequence of GAVE8 (for example, sequence confirmation number: 1) can be introduced into the genome of a non-human animal as an introduced gene. Further, based on the property of hybridization with human GAVE8 complementary DNA, the non-human human GAVE8 gene can be isolated. A homologue (such as the GAVE8 gene of mouse) is used as an introduced gene. The introduced gene may also include a sequence of a neutron and a polyadenylation signal to increase the efficiency of the introduced gene. Organization-specific adjustments-- Read the notes on the back and fill out this page. The standard paper size applies to the Chinese National Standard (CNS). Α4 Specifications (2!0Χ 297 mm) -46 - 1250209 A7 B7 Ministry of Economic Affairs Zhici Property Bureau Staff Consumer Cooperative Printed 5, Invention Description (Controlled sequence operability linked to GAVE8 import gene to specific cells The GAVE8 protein is directly expressed in the medium. The method of producing a genetically transgenic animal (especially an animal such as a mouse) by embryo manipulation and microinjection is a conventional technique, and is described, for example, in U.S. Patent No. 4,7 3 6,8 6 U.S. Patent No. 4,873,191 and Hogan, Manipulating the Mouse Embryo, (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, 1986). A similar method can be used to generate other genetically transformed animals. The animal carrying the additional introduced gene can then be bred with the starting base animal into which the foreign gene is introduced. In addition, gene-transforming animals carrying the GAVE8-introduced gene can be further mated with other gene-transforming animals carrying other introduced genes. To create a homologous recombinant animal, a GAVE 8 gene (eg, a human or non-human homologous GAVE 8 gene, such as the GAVE8 gene of a murine) can be prepared for deletion, addition or substitution. Thus, the vector of the GAVE8 gene (for example, disrupting its function) is altered. In a preferred embodiment, when a homologous recombination vector is designed, it functionally disrupts the endogenous GAVE8 gene (i.e., no longer encodes a functional protein; also known as "knock out" animals). In addition, when the homologous recombination vector is designed, the endogenous GAVE8 gene is mutated or otherwise altered but the expression of the functional protein is retained (for example, the upstream regulatory region can be altered to alter the expression of the endogenous GAVE8 protein). In the vector of homologous recombination, the additional GAVE8 gene nucleic acid can be contiguous at the 5' and 3' ends of the GAX/E8 gene-changing portion to allow the vector to carry the endogenous GAVE8 gene and the endogenous GAVE 8 of the embryonic stem cell. Homologous recombination between genes (please read the notes on the back and fill out this page) This paper scale applies to the Chinese National Standard (CN'S) A4 specification (210X 297 mm) -47 - 1250209 A7 B7 Ministry of Economic Affairs Intellectual Property Bureau Employee consumption cooperatives printed five, invention instructions (y role. Additional nucleic acids adjacent to GAVE8 should be of sufficient length to successfully carry out with endogenous genes Homologous recombination. The vector contains contiguous DNA (5' and 3' ends) which are generally several lengths. For a description of homologous recombination vectors, see, for example, Thomas et al., Cell (1987) 51 : 503). Introduce the vector into an embryonic stem cell line (for example, electroporation) and select the cell into which the introduced GAY/E8 gene is homologously recombined with the endogenous GAVE8 gene (see, for example, Li et a I., Cell (1). 992) 69:915). The selected cells are then injected into the blastocyst of an animal (eg, a mouse) to form an agglutination chimera (see, for example, Bradley in Teratocarcinomas and Embryonic Stem Cells: A Practical Approach, Robertson, ed., IR L, Oxford, (1 987) ρρ·1 1 3-152). The chimeric embryos can then be implanted into appropriate pseudo-pregnant female fostering animals for pregnancy production. Available reproductive fine cells with the same source and source recombination The progeny breeding of D Ν 产生 produces the introduced animal progeny of all animal cells containing homologous recombination DNA. Methods for constructing homologous recombination vectors and homologous recombinant animals are further described in Brad I ey , Current Op Inion in Bio/Technology (1 991) 2:823-829 and PCT Bulletin W〇 90/11354, W〇 91/01140, W〇 92/0968 and W〇 93/041 69. In another embodiment, a non-human animal containing a foreign gene introduced to allow selection of a system for regulating the expression of the introduced gene can be made. One of the examples of the system is the c"c /1 ο XP recombinase system of the bacterium P 1 . The description of the cre /1 ο XP recombinase system can be found, for example: La aksoeta I., P roc N at丨Acad Sci USA (1992) 89: 6232-6 236. Another example of a recombinant enzyme system is the FLP recombinase system of Saccharomyces cerevisiae ((VGorman et al., this paper scale applies to the Chinese National Standard (CNS) A4 specification). (2]0X 297 mm) ~ -48- (Please read the note on the back and fill out this page) - Binding

P 1250209 Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative printed A7 _ B7___ V. Description of invention ( Science (1991) 251: 1351-1355). If the c"e/loxP recombinase system is used to regulate the expression of the introduced gene, the animal into which the gene is introduced may contain a c"e recombinase and a selected protein. For example, the introduced gene encoding the selected protein and the intron coding recombination After the two genes of the enzyme-introduced gene are transferred to the animal, a "double" gene-transforming animal can be constructed. The pure lines of the non-human gene-transforming animals described herein can also be made according to the methods described in Wilmut et al., Nature (1 997) 385:81 0-81 3 and PCT Publication WO 97/07668 and WO 97/0 7669. . Briefly, cells (e.g., somatic cells) of an animal into which a foreign gene has been introduced are isolated, induced to escape from the growth cycle and enter the G phase. This inactive cell is then fused to the nucleus-depleted oocytes isolated from the inactive and inactive cell-like animals (e.g., via the use of electrical pulses). The reconstructed oocytes are then cultured to form morula or blasts, which are then transferred to pseudopregnant female nourishment animals. The cloned line is isolated from cells (e.g., somatic cells) of progeny produced by female nourishing animals. IV. Pharmaceutical Compositions The GAVE 8 nucleic acid molecules, GAVE8 proteins, and anti-GAVE8 antibodies (also referred to herein as "active compounds" of the invention can be incorporated into pharmaceutical compositions suitable for administration. The compositions typically comprise nucleic acid molecules, proteins or An antibody and a pharmaceutically acceptable carrier. The "pharmaceutically acceptable carrier" herein includes any and all solvents, dispersion cultures, coatings, antibacterial and antifungal compatible with pharmaceutical administration. Medicament, isotonic agent, absorption delaying agent, etc. The medicinal active substance uses the culture solution and the drug is already (CNS) (23 0 X 297^^ ) ~~一-49 - (please read the precautions on the back first) Fill in this page) - Binding P. 1250209 A7 B7 V. Description of the invention ((Please read the notes on the back and fill out this page) Known techniques. Any known culture solution other than the range incompatible with the active compound Or a pharmaceutical agent may be considered for use in the composition. The composition may also be incorporated with a supplementary active compound. The pharmaceutical composition of the present invention may be adjusted to a composition compatible with the intended route of administration. Examples of routes include: parenteral, for example, intravenous, intradermal, subcutaneous, oral (e.g., inhalation), transdermal (coated), transmucosal, and rectal administration. The solution or suspension for enteral, intradermal or subcutaneous administration may comprise the following ingredients: a sterile diluent such as water for injection, physiological saline solution, non-volatile oil, polyethylene glycol, glycerol, propylene glycol. Or other synthetic solvents; antibacterial agents such as: benzyl alcohol or methyl paraben; antioxidants, such as: ascorbic acid or sodium bisulfite; chelating agents, such as E DTA; buffer solutions, such as: acetate, lemon An acid or phosphate; and an agent that adjusts the osmotic pressure, such as sodium chloride or dextrose. An acid or base that adjusts the pH, such as HCI or NaOH. The parenteral preparation can be sealed in ampoules and discarded when used. Syringe or multi-dose vial made of glass or plastic. Ministry of Economic Affairs Intellectual Property Bureau g (Medical Consumer Cooperatives printed pharmaceutical composition suitable for injection may include sterile aqueous solution When it is water-soluble) or the dispersion and the sterilized powder which can be prepared immediately by using the sterilized injection solution or dispersion. Suitable carriers include intravenous saline, bacteriostatic water, Cremophor EL ® (BASF; Parsippany, when administered intravenously). NJ) or phosphate buffered saline (PBS). In all cases, the composition must be sterilized and should be a fluid that is easy to inject. It must be stable under the conditions of manufacture and storage, and under storage. It must be resistant to contamination by microorganisms such as bacteria and fungi. The carrier can be a solvent or a dispersion culture solution, which can be used inside - ___ This paper scale applies to the Chinese National Standard (CNS) A4 specification (210X 297 mm) -50-1250209 A7 B7 V. Description of the invention (d contains, for example, water, ethanol, polyol (for example: glycerin, propylene glycol, liquid polyethylene: 3⁄4 (B) glycol, etc.) and suitable mixtures thereof. The use of a coating agent such as lecithin can maintain the required particle size when dispersed and the use of a surfactant can maintain proper fluidity. The action of preventing microorganisms can be achieved by various antibacterial and antifungal agents, for example, p-oxybenzoic acid, chlorobutanol, phenol, ascorbic acid, topical antibacterial agents and the like. Preferred in many cases will include: isotonic agents 'e.g., sugars, polyalcohols such as mannitol, sorbitol, sodium chloride. The composition includes delayed absorption, for example, an agent such as aluminum monostearate or gelatin can be used as a long-term absorption injection composition. The sterilized injection solution can be prepared by injecting the desired amount of the active compound (e.g., GAVE8 protein or anti-GAVE8 antibody) into one or a group of suitable solvents containing the above listed ingredients, followed by filter sterilization. In general, the preparation of the dispersion is carried out by introducing the active compound into a sterile carrier containing the alkaline dispersion culture solution and the other ingredients listed above. When preparing a sterilized powder of a sterilized injection solution, the preferred method is to prepare a powder of the active ingredient produced by vacuum drying and lyophilization with a powder from a previously sterilized solution of any additional desired ingredients. In general, oral compositions can include inert diluents or edible carriers. It can be sealed in gelatin capsules or compressed into tablets. In the case of oral administration, the active compound can be incorporated into the excipient and used in the form of tablets, troches or capsules. Oral compositions can also be prepared using a fluid carrier as a preparation, wherein the compound in the fluid carrier can be administered orally, inhaled, and coughed or swallowed. The composition may include a pharmaceutically compatible binder, and/or an adjuvant material. The paper size is Chinese National Standard (CNS) A4 specification (23 0x 297 mm) ~ " 十' -51· (Read first Note on the back of the page. • Installed -, 11 Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative printed 1250209 A7 B7 Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative printing 5, invention description (d material. Pills, medicine IX, Capsules, guilloche and the like may contain any of the following ingredients: a similar compound: a binding agent such as microcrystalline cellulose, Tragacons gum or gelatin; an excipient such as starch Or lactose; disintegrating agents, for example: alginic acid, skin glue (Prim〇ge|) or corn starch; lubricants, such as: magnesium stearate or sterotes; sliding 'for example, monoxide a colloid; a sweetener such as sucrose or saccharin; or a flavoring agent such as peppermint, methyl salicylate or an orange flavoring agent. In the case of inhalation administration, the compound is in the form of an aerosol from a pressurized container or contains Proper propellant, such as a gas For example, a drug dispenser or a sprayer can be sprayed and transported. It can also be administered systemically via mucosal or transdermal methods. When administered by mucosal or transdermal administration, a suitable penetrating agent that can pass through the barrier can be used in the formulation. In general, the penetrant is a known art 'including, for example, an amphoteric surfactant, a bile salt, and a fusidic acid derivative when administered via a mucosa. Transmucosal administration can be accomplished by using a nasal spray or a suppository. When the transdermal administration is carried out, the active compound can be adjusted into a generally known ointment ointment, gel or cream. The ointment can also be prepared in the form of a suppository (for example, a conventional suppository such as cocoa butter and other glycerides). Or retention of the enema to carry out the rectal delivery. In one embodiment, the active compound can be prepared with a carrier that protects against rapid elimination of the body, such as a controlled release formulation, including implants and microencapsulated Transport system. Biodegradable, biocompatible polymers can be used, for example: ethylene vinyl acetate, polyanhydride, polyglycolic acid, collagen Protein, poly-orthoester and polylactic acid. Preparation (please read the back note and fill out this page) This paper scale applies to Chinese National Standard (CNS) A4 specification (210X 297 mm) -52- 1250209 A7 B7 Ministry of Economic Affairs Intellectual Property Bureau Staff Consumer Cooperative Printed 5, Invention Description (5 (^ The method of the formulation is well known to professionals familiar with this technique. The material is also commercially available from Alza Corporation and Nova Pharmaceuticals, I nc. A suspension of granules (including lipid globules that administer infected cells with a single antibody against a viral antigen) can also be used as a pharmaceutically acceptable carrier. It can be prepared according to methods well known to those skilled in the art, for example as described in U.S. Patent No. 4,522,81. Oral or parenteral compositions are especially advantageous for forming dosage unit forms which are easy to administer and have a uniform dosage. Dosage unit form as used herein refers to a single dose formed by physically unconnected units, each unit containing a predetermined amount of active compound being capable of producing the desired therapeutic effect with a carrier which is computationally pharmaceutically required. Whether administered in one or more separate doses or continuously, depending on the type and severity of the disease, antibodies from about 1 μg/kg to 15 mg/kg (eg 〇 1 to 20 mg/kg) are administered to The patient's starting candidate dose. Depending on the factors mentioned above, representative daily doses range from about 1 microgram/kg to 100 mg/kg or more. When administered repeatedly for several days or longer, depending on the symptoms, the treatment may be prolonged until the symptoms of the disease are suppressed. However, other dosing regimens are also applicable. The progress of treatment can be easily monitored and measured using the techniques of the experience. A typical administration regimen is disclosed in WO 94/CM188. The specification of the dosage unit form of the present invention is directly dependent on the unique characteristics of the active compound and the particular therapeutic effect desired to be achieved, as well as the limitations of the treating individual in the art of the active compound formulation and in the art. The nucleic acid molecule of the invention can be inserted into a vector and used as a vector for gene therapy. The vector of gene therapy can be delivered to the patient, for example, intravenously, topically (US Patent No. 5,328,470) or stereotactic injection (see (Please read the back note first and then fill out this page). China National Standard (CNS) A4 Specification (210X29? mm) -53> 1250209 A7 B7 Ministry of Economic Affairs Zhici Property Bureau Staff Consumer Cooperative Printed 5, Invention Description (51) For example: Chen et al., Proc Natl Acad Sci USA (1 994) 91 : 3 0 54-30 57). The pharmaceutical preparation of the gene therapy vector may comprise a gene therapy carrier and an acceptable diluent, or may comprise a sustained release matrix encapsulating the gene delivery vehicle. The recombinant gene can be used to make a complete gene delivery vector, such as a retrovirus vector, and the pharmaceutical preparation can include one or more cells that produce a gene delivery system. The pharmaceutical composition can be included in the container together with the administration instructions. In the form of a mask or dispenser. V. Uses and Methods of the Invention Nucleic acid molecules, proteins, protein homologs, and antibodies described herein may be used in one or The following methods are performed: a) screening assays; b) detection assays (eg, chromosome maps, tissue classification, forensic biology); c) predictive drugs (eg, diagnostic assays, predictive assays, clinical trials) And pharmacogenetic monitoring); and d) treatment (eg, therapeutic and prophylactic). The G A V E 8 protein that interacts with other cellular proteins can be used to (i) regulate cell proliferation; (Μ) regulate cell differentiation; and (j j j) regulate cell remnant. The isolated nucleic acid molecule of the present invention can be used to express a GAVE 8 protein (for example, expressed in a host cell in a gene therapy by a recombinant expression vector), and can be used to detect (eg, in a biological sample) GAVE 8 signaling RNA or GAVE8 gene. Genetic damage, and regulation of GAVE8 activity. In addition, the GAVE8 protein can be used to screen for drugs or compounds that modulate GAVE8 activity or expression and to treat conditions characterized by insufficient or excessive production of GAVE 8 protein or GAVE8 wild-type protein, GAVE8 eggs (read first read on the back) Note: Please fill out this page again. This paper scale applies to China National Standard (CMS) A4 specification (210><297 public) -54- 1250209 A7 B7 Ministry of Economic Affairs Zhici Property Bureau employee consumption cooperative printing 5, invention description (A drug or compound having a reduced or abnormal white matter activity. Furthermore, the anti-GAVE8 antibody of the invention can be used to detect and isolate the GAVE8 protein and modulate G AV E 8 activity. The invention further relates to the above described herein. Screening assays for novel agents identified and using their treatment or testing. A. Screening assays The present invention provides methods (also known as "screening assays") to confirm modulators that bind to GAVE8 protein or have the effect of stimulating or inhibiting GAVE8 protein Candidates or test compounds or agents (eg, ···peptides, eg, GAVE 8 or GAVE 8 activity) Classes, peptidomimetics, small molecules or other drugs. In one of the embodiments, the invention provides a candidate or test compound for screening for activity of a GAVE 8 protein or polypeptide or a biologically active portion thereof that binds to or modulates a membrane-bound form. Assay method. The test compound of the present invention can be obtained from a combinatorial gene pool known in the art, including: a biological gene pool; a spatially located parallel solid phase or lytic gene library; a deconvolution synthetic gene library; a gene library of "a bead-compound"; and a synthetic gene library selected using affinity chromatography. The gene library of the organism is limited to the peptide gene pool, while the other four gene banks are peptides, non-peptide oligomers or Small molecule compound gene library (Lam, Anticancer Drug Des (1 997) 1 2:1 45). Examples of methods for synthesizing molecular gene pools can be found, for example, in DeWitt et al., Proc Natl Acad Sci USA (1 993) 90 : 6909 ; Erb et al · , Proc Natl Acad Sci US A ( 1 994 ) 91 : 1 1 422 ; Zuckermann et al . , J Med Chem (1 994 ) 37 : 2678 ; Cho et (please read the notes on the back first) Fill in again Page) - Loading · This paper scale applies to China National Standard (CNS) Α4 Specifications (2) 297 297; PCT) -55 - 1250209 A7 B7 Ministry of Economic Affairs Intellectual Property Bureau Employees Consumption Cooperative Printed 5, Invention Description (53) 3 I., Science (1 993) 261: 1 3 0 3 ; Carrel I eta I., Angew Chem Int Ed Engl (1994) 33: 2059; Care II et al., Angew Chem Int Ed Engl (1994) 33: 2061; And Gallop et al., J Med Chem (1 994) 37: 1 233 〇 compound gene library can be present in solution (eg: ought oughten, B io/Techniques (1 992 ) 1 3 : 4 1 2-42 1 ) , or ball (La m, N at ure (1991) 354: 82-84), wafer (Fodor, Nature (1 993) 3 6 4: 555-556), bacteria (US Patent No. 5, 223, 409), spores ( U.S. Patent Nos. 5,571,698; 5,403,4 84; and 5,223,409), plastids (Cull et a I., Proc Natl Acad Sci USA (1 992) 89: 1 865-1 869) or phage (Scott et al., Science) (1 990) 249: 386-390; Devlin, Science (1990) 249: 4 0 4-406; C wir I a et a I., Proc Natl Acad Sci USA (1 990) 87: 6378-6382; and Felici J Mol B iol (1991) 222 : 301-310 Bu Since the GAVE8 ligand is SIP, it is possible to investigate the specific portion of the action of S I P and G A V E 8 by a known method. This particular region can be synthesized by conventional biosynthetic methods, combined with carbohydrate synthesis methods and enzymatic reaction synthesis. This SIP portion corresponds to an "antigenic epitope". The GAVE8 antigenic epitope can be modified with other monomeric or non-carbohydrate moieties to produce a modified epitope that enhances properties (e.g., blood half-life, GAVE8 binding constant, etc.). Any suitable epitope variant can be assayed using the binding and screening assays taught herein. In one embodiment, the assay is a cellular assay wherein the cell surface-expressing membrane-bound form of the GAVE 8 protein, or a biologically active portion thereof, is applied to the Chinese National Standard (CNS) A4 specification (210 x 297 mm). (Please read the precautions on the back and fill out this page) - Packing, v, f-56- 1250209 A7 Ministry of Economic Affairs, Intellectual Property Bureau, Staff Consumer Cooperative, Printed B7 V. Invention Description (The cells are in contact with test compounds, The ability of the test compound to bind to the GAVE8 protein. The cell can be a yeast cell or a mammalian-derived cell. The ability of the test compound to bind to the GAVE8 protein is determined, and the test compound can be coupled to the label of the radioisotope or enzyme to detect the complex. a compound labeled to determine the binding of a test compound to a GAVE 8 protein or a biologically active portion thereof. For example, a test compound can be directly or indirectly labeled 125, 35S, 14C or 3H, and the radioisotope can directly count radioactivity or count scintillation. In addition, test compounds can be labeled with enzymes, such as: Root peroxidase, alkaline phosphatase, or luciferase, and can be transformed with a suitable substrate to detect the labeled enzyme. In one of the preferred embodiments, the assay comprises a cell surface representation membrane. - binding of the G AV E8 protein, or a biologically active portion thereof, to a conventional GAVE8 binding compound to form a assay mixture, contacting the assay mixture with the test compound, and determining the interaction of the test compound with the GAVE8 protein. The ability to determine the ability of a test compound to interact with a GAVE8 protein comprises determining the ability of the test compound to preferentially bind to GAVE8 or a biologically active portion thereof as compared to conventional compounds. In another embodiment, the assay is a cellular assay, the system Contacting a cell that expresses a membrane-bound form of the GAVE8 protein, or a biologically active portion thereof, with a test compound determines the ability of the test compound to modulate (eg, stimulate or inhibit) the activity of the GAVE8 protein or a biologically active portion thereof. The ability of a compound to modulate the activity of GAVE8 or its biologically active portion , can be determined by the ability to determine the binding or interaction of GAVE8 protein and GAVE8 target molecule. The "target molecule" in this article is compatible with (please read the back note before filling this page) _ this paper scale Applicable to China National Standard (CNS) A4 Specification (210X 297 mm) -57- 1250209 A7 B7 Ministry of Economic Affairs Intellectual Property Bureau Staff Consumer Cooperative Printed 5, Invention Description (y GAVE8 protein binding or interaction molecule, such as cell surface expression The GAVE 8 protein molecule, the molecule on the second cell surface, and the molecule in the extracellular environment are 'molecular or cytoplasmic molecules associated with the inner surface of the cell membrane. The GAVE8 target molecule can be a non-GAVE8 molecule or a GAVE8 protein or polypeptide of the invention. In a specific embodiment, the GAVE8 target molecule is a component of a message delivery pathway that enhances extracellular signals (e.g., signals produced by binding of the compound to the membrane-bound GAVE8 molecule) via the cell membrane into the cell. For example, the target molecule may be a protein between catalytically active secondary cells or a protein that enhances downstream signaling molecules associated with GAVE8. In another embodiment, GAVE 1 can be made using known techniques to perform signal transduction in a target cell as taught herein, see for example: W0 00/22131 and W0 00/22129, and then expose the cells. To various candidate modulators to determine whether signal activity is enhanced (disclosed as a candidate agonist), or to reduce (disclose it as a candidate antagonist), or to reduce activity below baseline (which is a candidate retro-acting agent) ). The ability to bind to or interact with a GAVE8 target molecule can be determined by direct binding of the methods described above. In a preferred embodiment, the ability to determine whether a GAVE8 protein binds to or interacts with a GAVE8 target molecule can be determined by the activity of the target molecule. For example, it can detect secondary messengers of cells induced by target molecules (including, but not limited to, intracellular Ca2+, dimercaptoglycerol, and IP3), detect the target molecule's catalysis/activity against appropriate receptors, and detect Detecting reporter genes (eg, operably linking GAVE8-reactive regulatory elements to this paper size encoding a detectable marker (eg, luciferin) using Chinese National Standard (CNS) A4 specification (210X297 mm) (please Read the precautions on the back and fill out this page) - Install ' 订 f -58- 1250209 Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative printing 5, invention instructions (enzyme nucleic acid) induced, or detect cell response (such as cells Differentiation or cell proliferation) to determine the activity of a target molecule. In another embodiment, the assay of the invention is a cell-free assay comprising contacting a GAVE8 protein or a biologically active portion thereof with a test compound, determining binding of the test compound to GAVE8 The ability of the protein or its biologically active portion. As described above, the test compound can be directly or indirectly assayed for binding to the GAVE8 protein. In a specific embodiment, the assay comprises contacting a GAVE8 protein or a biologically active portion thereof with a conventional GAVE8-binding compound to form an assay mixture, contacting the assay mixture with the test compound, and determining the ability of the test compound to interact with the GAVE8 protein, wherein The ability of a test compound to interact with a GAVE 8 protein comprises determining the ability of the test compound to preferentially bind to GAVE8 or a biologically active portion thereof as compared to conventional compounds. In another embodiment, the assay is a cell-free assay comprising Contacting a GAVE8 protein or a biologically active portion thereof with a test compound and determining the ability of the test compound to modulate (eg, stimulate or inhibit) the activity of the GAVE8 protein or a biologically active portion thereof. The ability of the test compound to modulate GAVE8 activity is determined by the above description The method of binding determines the ability of the GAX/E8 protein to bind to the GAVE8 target molecule. In another embodiment, the ability of the test compound to modulate GAVE8 activity is determined, and the GAVE8 protein can be further regulated to control the GAVE8 target molecule. The ability to complete, for example, the catalysis/activity of the target molecule to the appropriate substrate can be determined by the methods previously described. In another embodiment, the cell-free assay comprises exposure to the GAVE8 protein (please read the back note before completing this page) * Packing and ordering ip-· This paper scale is applicable to China National Standard (CNS) A4 specification (210X 297 mm) -59- 1250209 A7 B7 V. Description of invention (57) Combination of quality or biological activity with conventional knowledge The GAVE 8 compound forms the assay mixture, contacts the assay mixture and the test compound, and determines the ability of the test compound to interact with the GAVE8 protein, wherein determining the ability of the test compound to interact with the GAVE8 protein comprises determining that the GAVE8 protein preferentially binds to or modulates the GAVE8 target molecule. The ability to activate. The cell-free assay of the present invention may use a soluble or membrane-bound GAVE8. The membrane-bound GAVE8 can be maintained in a solution state by using a lysing agent when performing cell-free assay using membrane-bound GAVE8. Examples of the solubilizing agent include a nonionic amphoteric surfactant, such as η-octyl glucoside, η-dodecyl glucoside, η-dodecyl glucoside, octyl decyl-hydrazine- Methyl glucosamine, decyl-hydrazine-methyl glucosamine, Triton Χ-100, Triton X-1 14, Thesit®, isotridecyl poly(ethylene (ethylene) glycol-ether) n, 3-[ (3-Chloroaminopropyl)dimethylamino]-1 -propane sulfonate (CHAPS), 3-[(3-chloroguanidinopropyl)dimethylamino]-2-hydroxyl 1-propane sulfonate (CHAPSO), or N-dodecyl-N,N-dimethyl-3-ammonio-1·propane sulfonate. In one or more specific embodiments of the above assays of the present invention, GAVE8 or a target molecule can be satisfactorily immobilized to enhance the separation of the complex from an uncomplexed form of one or both proteins, and to provide automated assays. The binding of the test compound to GAVE8, or the interaction of GAVE8 with the target molecule in the presence and absence of the candidate compound, can be accomplished in any suitable container containing the reactants. Examples of the container include: micro test trays, test tubes, and microcentrifuge tubes. In one embodiment, the fusion protein provides an additional structural region that allows one or two proteins to bind to the matrix. The paper size applies to the Chinese National Standard (CNS) A4 specification (2) 0X 297 mm) ( Read the first note on the back and then fill out this page. Installed. Ministry of Economic Affairs Intellectual Property Bureau Staff Consumer Cooperative Printed -60 - 1250209 A7 _____ B7 V. Invention Description (d (Please read the note on the back and fill out this page) For example, glutathione-S-transferase/GAVE8 fusion protein or glutathione-S-transferase/target fusion protein, which can be adsorbed to glutathione agarose beads (Sigma Chemical) , St. Louis, MO) or a glutathione-derived micro-test disc, which can then be combined with a test compound or test compound and a non-adsorbed target protein or GAVE8 protein, and the mixture is reacted under conditions for complex formation (eg Salts and pH under physiological conditions. After the reaction, wash the beads or micro-test disc holes, remove any unbound components, directly or indirectly measure the formed complex, for example, as explained above In addition, the 'complex can be dissociated from the matrix, and the level or activity of GAVE8 binding can be determined using standard techniques. The Ministry of Economic Affairs, Intellectual Property Office, the employee consumption cooperative, the printing technique of other immobilized proteins on the substrate can also be used in the present invention. Screening assays. For example, GAVE8 or its target molecule can be immobilized using the conjugation of biotin and streptavidin. Biotinylated GAVE8 or target molecules can be used in techniques known in the art (eg biotinylation group) The kit, Pierce Chemicals, Rockford, IL) was prepared from biotin-NHS (N-hydroxy-succinimide) and fixed in a 96-well test dish coated with streptavidin (Pierce Chemicals). An antibody that reacts with GAVE8 or a target molecule but does not interfere with binding of the GAVE8 protein to the target molecule can be conjugated to the well of the plate to capture the unbound target or GAVE8 in the well with the antibody. - In addition to the method of immobilizing the complex, it may also include the use of an antibody immunodetection complex that reacts with GAVE8 or a target molecule, and with GAVE8 or a target Molecular activity-related enzyme binding assay. In another specific example, the method can be used to confirm the GAVE8 performance modifier. The paper size is applicable to the Chinese National Standard (CNS) A4 specification (210X 297 mm) -61 - 1250209 A7 B7 Ministry of Economics Intellectual Property Bureau employee consumption cooperatives printed five, invention instructions (d, which is to contact the cells with candidate compounds, and determine the GAVE8 signaling RNA or protein expressed in the cells. Compare the performance level of GAVE 8 signaling RNA or protein in the presence of candidate compounds and lack GAVE8 signals the level of RNA or protein expression when a candidate compound is present. Based on the alignment results, candidate compounds of the GAVE8 performance modulator can then be confirmed. For example, when the expression of GAVE8 signaling RNA or protein in the presence of a candidate compound is greater than (generally significantly greater than) the absence of a candidate compound, then the candidate compound is confirmed to be a stimulator of GAVE8 signaling RNA or protein expression. Furthermore, when the GAVE8 signaling RNA or protein exhibits less than (statistically significantly less than) the candidate compound in the absence of the candidate compound, it can be confirmed that the candidate compound is an inhibitor of GAVE8 signaling RNA or protein expression. The level of GAVE8 signaling RNA or protein expression in the cells can be determined by the method described herein for detecting GAVE8 signaling RNA or protein. In another feature of the invention, the GAVE8 protein can be used as a "bait protein" in a two-hybrid assay or a three-hybrid assay (see, e.g., U.S. Patent No. 5,283,31 7; Zervos et al., Cell (1) 993) 72 : 2 2 3-232 ; Madura et al., J Biol C hem (1 993) 268: 1 2046-1 2054; Bartel et a I., Bio/Techniques (1 993) 14: 920-924; Iwabuchi et a I., Oncogene (1 993) 8:1693-1696; and PCT publication number W094/1 0300) to confirm that it can bind to or interact with GAVE8 ("GAVE8-binding protein" or "GAVE8-bp&quot ;) and other proteins that regulate GAVE8 activity. Such GAVE8 binding proteins may also be involved in the propagation of GAVE8 protein signals, such as elements upstream or downstream of the GAVE8 pathway. (Please read the precautions on the back and fill out this page.) Loading · The size of the paper is applicable to the Chinese National Standard (CNS) A4 specification (210X 297 mm) -62- 1250209 A7 B7 V. Invention Description ( Further invention For the novel agents identified in the above screening assays and the treatments or tests described therewith (please read the notes on the back and fill out this page) B. Detect the complementary DNA sequence portions or fragments identified herein. (and the corresponding complete gene sequence) can be used as a polynucleotide reagent for many purposes. For example, the sequence can be used to: (i) map a gene on a chromosome and thus locate a gene region associated with a congenital disease; (M) Confirmation of individuals (tissue classification) from small biological samples; and (Mi) Samples of assisted forensic identification of organisms. This use is described in the following subsections. Ministry of Economic Affairs Intellectual Property Office Staff Consumer Cooperatives Printed 63 - 1250209 A7 B7 Economy Ministry of Intellectual Property Bureau employee consumption cooperative printing 5, invention description (61) hybrid cells containing individual human somatic chromosomes The somatic cells of different mammals (for example, human and mouse cells) can be used. When the hybrid cells of human and mouse cells grow and divide, the cells gradually and randomly lose the human chromosome, but still maintain the mouse chromosome. A culture medium that cannot grow but human cells can grow (due to the lack of specific selection enzymes) will retain a human chromosome containing the enzyme gene encoding it. Various culture media can be used to create panels of various hybrid cell lines. A cell line can contain a single human chromosome or a small number of human chromosomes, as well as the entire set of mouse chromosomes, and can map individual gene-specific maps on human chromosomes (UEustachio et al., Science (1 983) 220: 9 1 9-924). It is also possible to use translocations and hybrid somatic cells containing human chromosome fragments after deletion of human chromosomes. PC R map localization of hybrid somatic cells is a rapid procedure for localizing specific sequences to specific chromosomes. GAVE8 sequence design oligonucleotide primer, which can be dyed with specific on the panel The chromosomal fragment achieves a sublocalization. Other strategies for locating the GAVE8 sequence map on the chromosome, including in situ hybridization (described in Fan et al., Proc Natl Acad Sci USA (1 990) 87: 622 3 -27), the pre-screening marker for the flow classification chromosome method and the hybridization to the chromosome-specific complementary DNA gene pool pre-selection method. The fluorescence in situ hybridization reaction (FISH) of the DNA sequence is carried out on the chromosomes dispersed in the middle of the cell division. Further for precise chromosome localization, chemicals such as colchicine can be used to destroy the mitotic spine and stop cell division in the middle of cell division, so that scattered chromosomes can be obtained. The chromosome can be treated with trypsin for a short time, then ------------- (Please read the back note first and then fill out this page), \sx» This paper scale applies to the national standard ( CNS ) A4 size (2) 0X297 mm) -64 - 1250209 A7 B7 V. Description of invention (d (please read the notes on the back and fill out this page)

Giemsa staining. Each chromosome produces a bright and slightly black band pattern and confirms individual chromosomes. FISH technology can be used for DNA sequences of 500 or 600 base pairs. However, when it is larger than 1,000 base pairs, it can be easily detected in combination with a unique chromosomal locus and has a high sufficient signal intensity. The better one is the 〇〇〇 base pair, and the better is 2,000 base pairs, which will give good results in a reasonable time. This technique can be found in Verma et al., (Human Chromosomes: A Manual of Basic Techniques (Pergamon Press, New York, 1 988)). The reagents for performing the chromosome mapping test can be individually labeled on a single chromosome or on a single chromosome, or multiple sites and/or multiple chromosomes can be labeled with panel reagents. Reagents corresponding to the non-coding regions of the gene can also be used for mapping purposes. Some coding sequences within the gene family are conserved sequences, thus increasing the chance of sporadic hybridization when the chromosome map is located. Printing by the Intellectual Property Office of the Ministry of Economic Affairs, the Consumer Cooperatives Once the sequence is mapped to the exact location on the chromosome, the physical location of the sequence on the chromosome can be compared to the spectral data of the gene. (See, for example, V. McKusick, Mendel ian Inheritance in Man, available on-line through Johns Hopkins University, Welch Medical Library). The relationship between the gene (located in the same chromosomal region) and the disease can then be confirmed by linkage analysis (common inheritance of physically adjacent genes) as described, for example, in Egeland et al., Nature (1 987) 325:783- 787 ° In addition, differences in the GAVE8 gene DNA sequence between individuals affected by the disease and not affected by fe can be determined. If a mutation is observed in some or all of the affected individuals, but no mutation is observed in the unaffected one, the paper size applies to the National Standard (CNS) A4 specification (2) 〇 X 297 PCT) -65- 1250209 A7 B7 V. INSTRUCTIONS (It can be inferred that the mutation may be the cause of a particular disease. The more affected and unaffected individuals generally include first-inspected changes in chromosome structure, such as from scattered chromosomes. Observe the visible deletion or translocation or use pc R to detect the DNA sequence. Finally, complete genetic sequencing of several individuals can confirm the presence of mutations and distinguish between mutations and polymorphisms. The GAVE8 sequence of the present invention can also confirm an individual from a biological sample. For example, the US military uses a polymorphic technique (RFLP) of restriction enzyme fragment length to confirm a person. In this technique, the individual's genetic DNA is restricted by one or more types. Enzymatic hydrolysis, the detection of southern ink spots, the identification of the unique strips produced by the personnel. This method has no current n Dog Tags" restrictions, will not be lost, lost Packing or theft creates difficulties in positive identification. Furthermore, the sequences of the invention are applicable to DNA markers as RFLPs (described in U.S. Patent No. 5,272,057). Furthermore, the sequences of the invention can be used in another technique for determining genomes. The DNA sequence of the selected portion is actually a base-to-base arrangement. Therefore, the GAVE8 sequence described herein can be used to prepare PCR primers for the two 5' and 3' sequences, and then the primer can be used to amplify the genome. DNA and its sequence 〇 Because of the difference in the dual gene, each person's DNA sequence has a unique genome, so the method can prepare a panel corresponding to the DNA sequence, and can provide a unique personal identification. The sequence of the present invention is available. The identification sequence is obtained from the individual and from the organization. The unique paper size of the GAVE8 sequence of the present invention is applicable to the Chinese National Standard (CNS) A4 specification (210X 297 mm) (please read the back note first and then fill out this page)

P Pack· Ministry of Economic Affairs Intellectual Property Bureau g (Working Consumer Cooperatives Printed -66 - 1250209 A7 B7 Ministry of Economic Affairs Intellectual Property Bureau Staff Consumer Cooperative Printed 5, Invention Description (64) Represents a part of the human genome. The sequence coding area can Some mutations in the dual gene occur, and the non-coding region can undergo a large degree of variation. It is estimated that the frequency of mutation of the even gene between humans occurs approximately 500 bases. The sequences described herein are to some extent. It can be used as a standard sample for the identification of individual DNA. Because a large number of polymorphisms can occur in non-coding regions, fewer sequences can be used to distinguish individuals. Sequence confirmation number: 1 non-coding sequence can provide a group of 1〇 to 1,000 Primers that can be amplified to produce 100 kb bases of uncoded sequences for positive personal identification. If a predicted coding sequence, such as sequence confirmation number: 1, is used, it is more appropriate to perform positive individual identification. The number of primers is 500-2,000. If a set of reagents in the GAVE8 sequence described herein is used to generate a database for personal identification, then the same test It can be used later to identify individual organizations. Using a unique identification database, positive identification of individuals (whether dead or alive) can be performed from very small tissue samples. 3. Use of GAVE8 sequences in forensic biology. A-identification techniques can also be used in forensic biology. Forensic biology is the scientific field of positive confirmation (such as the perpetrator) using biological evidence of the genetics found in crimes. This identification can be performed using criminal techniques from crime.埸 Find samples of very small organisms collected, such as tissues, such as hair or skin, or body fluids, such as blood, saliva, or semen, to amplify the DNA sequence. The amplified sequence can then be compared to a standard sequence to identify Source of the biological sample. The sequence of the present invention can be used to provide a polynucleotide reagent, for example, the size of the PCR paper is applicable to China"^^ (CNS) A4 specification (2) OX 297 mm) (please read the back note first) Matters fill out this page) 0, Loading - order f -67- 1250209 A7 B7 V. Description of the invention (sub, calibrating the human genome-specific locus, which can enhance D The reliability of NA forensic identification, for example, provides another "identification mark" (ie, a unique DNA sequence of another particular individual). As described above, the actual base sequence data can be formed by restriction enzymes producing fragments. Accurate another pattern for identification. Calibration sequence confirmation number: 1 sequence of non-coding regions, especially suitable for generating a large number of polymorphisms, so it is easier to distinguish individuals using the technology of this non-coding region. Examples of reagents include the GAVE8 sequence or a portion thereof, such as a fragment having a non-coding region of at least 20 or 30 bases in length from the sequence confirmation number: 1. The GAVE8 sequences described herein can be used to further provide polynucleotide reagents, such as labeled or labelable probes, which can be used, for example, in situ hybridization techniques to identify a particular tissue, such as brain tissue. This technique is very useful when forensic pathologists face organizations of unknown origin. The GAN/E8 probe can confirm tissue via a panel of species and/or organ type. In a similar method, contaminated tissue cultures can be screened with reagents (e. g., G A V E 8 primers or probes) (i.e., screened in culture mixtures of different cell types). B. Predictive Drugs The present invention is also directed to the field of predictive drugs in which individual testing can be performed using diagnostic assays, predictive assays, pharmacogenetics, and clinical monitoring. Accordingly, one of the features of the present invention relates to the determination of a diagnosis to determine the activity of GAVE8 protein and/or nucleic acid and the activity of GAVE8 'from biological samples (eg, blood, urine, sputum, blood stasis, cells, This paper scale applies to China National Standard (CNS) A4 specification (210X297 mm) (please read the note on the back and fill out this page) - Install - , 11 Ministry of Economic Affairs Intellectual Property Bureau 8 Workers Consumption Cooperative Printed -68- 1250209 A7 B7 V. Description of the invention (^ tissue) Determination of whether an individual has a disease or condition associated with abnormal GAVE8 expression or activity, or a risk of developing a disorder. The present invention also provides predictive assays to determine whether an individual has development and The risk of a GAVE8 protein, nucleic acid expression or activity-related disorder. For example, a mutation in the G.AVE8 gene can be determined in a biological sample. This assay can be used for the purpose of prediction, and thus the individual is characterized in or associated with GAVE8 protein, nucleic acid. Preventive treatment is performed prior to the onset of a manifestation or activity of the disorder. Another feature of the invention is to provide a method for determining GAVE8 protein in an individual. The nucleic acid is expressed or the activity of GAVE8, thereby selecting an appropriate therapeutic or prophylactic agent (herein referred to as "pharmaceutical genetics"). Pharmacogenetics can select a therapeutic or prophylactic agent for treating a patient based on the genotype of the patient. (For example, examining a patient's genotype and determining the patient's ability to respond to a particular agent.) Another feature of the invention relates to monitoring the effect of a clinically tested agent (e.g., drug or other compound) on GAVE8 performance or activity. The agents and other agents of the present invention are described in further detail in the next chapter. 1. Diagnostic assays A typical method for detecting the presence or absence of GAVE8 in a biological sample comprises: obtaining a biological sample from the test patient, and applying the biological sample to the detectable A GAVE8 protein or a compound or agent that encodes a nucleic acid encoding a GAVE8 protein (eg, a signaling RNA, a genomic DNA) is contacted to detect GAVE8 present in a biological sample. A preferred agent for detecting GAVE8 signaling RNA or genetic DNA is capable Labeled nucleic acids that hybridize to GAVE8 signaling RNA or genomic DNA. This paper scale applies to the Chinese National Standard (CNS) A4 specification. (210X297 mm) (Please read the note on the back and fill out this page) Packing · 1 Economic Intelligence Bureau Intellectual Property Office Staff Cooperatives Printed -69- 1250209 A7 B7 Ministry of Economic Affairs Intellectual Property Bureau Staff Consumer Cooperative Printed V. DESCRIPTION OF THE INVENTION (Probe. The nucleic acid probe may be, for example, a full-length GAVE8 nucleic acid, such as a nucleic acid of sequence number: 1 or a portion thereof, for example at least 15, 5, 50, 100 ' 2 50 or 500 nucleotides in length Oligonucleotides, and under stringent conditions, are sufficient to specifically hybridize to GAVE8 signaling RNA or genomic DNA. Other suitable probes to which the present invention is applied for diagnostic assays will be described herein. A preferred agent for detecting the GAVE8 protein is an antibody that binds to the GAVE8 protein. Preferably, the antibody has a detectable label. The antibody may be a plurality of antibodies or a better monoclonal antibody. An intact antibody, or a fragment thereof (e.g., Fab or F(at)2) can be used. "Labeled" with respect to a probe or antibody, as used herein, includes a coupled (ie, physically linked) detectable substance to a probe or antibody directly labeled probe or antibody, and reacts with another reagent that is directly labeled. An inscribed labeled probe or antibody. Examples of indirect labeling include the use of fluorescently labeled secondary antibodies to detect primary antibodies and DNA probes that use terminally labeled biotin, and thus can be detected using fluorescently labeled streptavidin. The term "biological sample" as used herein includes: body fluids isolated from the tissues, cells, and organisms of the patient, as well as tissues, cells, and body fluids in the patient's body. The detection method of the present invention can be used to detect G A V E 8 in the living body and in vitro to detect R N A, protein or gene 〇 N A . For example, the technique of detecting G AV E 8 signaling R N A in vivo includes: northern hybridization and in situ hybridization. The technology for detecting GAVE8 protein in vitro includes: enzyme-linked immunosorbent assay (enzyme-linked immunosorbent assay), Western blotting, immunoprecipitation, and immunofluorescence. The in vitro technique for detecting the GAVE8 gene D N A includes Southern hybridization. In addition, the in vivo technique of detecting GAVE8 protein includes the introduction of labeled anti-resistance to patients (please read the note on the back and fill out this page). - _ f This paper scale applies to Chinese National Standard (CNS) A4 specification (210X) 297 public f ) -70- 1250209 A7 B7 Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative printing 5, invention description (d GAVE8 antibody. For example, the antibody can label the radioactive label and use standard imaging techniques to detect the position in the patient In one of the specific embodiments, the biological sample containing the protein molecule can be obtained from the test patient. Further, the biological sample containing the signaling RNA molecule can be obtained from the test patient or the genetic DNA molecule can be obtained from the test patient. The biological sample is a peripheral blood leukocyte sample isolated from the patient by a general method. In another specific embodiment, the method further comprises obtaining a biological sample of the control group from the control group, contacting the control sample and detecting the GAVE 8 protein. a compound or agent that signals RNA or genomic DNA to detect GAVE8 protein and signaling RNA present in biological samples. Or genomic DNA, comparing the GAVE8 protein, the signaling RNA or the genomic DNA present in the control sample and the test sample. The invention also includes detecting a set of GAVE8 in the biological sample. The set can be used to determine whether the patient has a disease. A condition associated with abnormal GAVE8 expression (eg, an immune condition) may increase the risk of developing the condition. For example, the set may include a compound or agent that detects a marker of GAVE8 protein or signaling RNA in a biological sample and determines the sample. a method for the content of GA\/E 8 (for example: an anti-GAVE8 antibody or an oligonucleotide probe that binds to DNA encoding GAVE8) such as sequence confirmation number: 1). The kit may also include an explanation of the test patient suffering from or Description of the risk of developing a condition associated with GAVE8 abnormality (if the level of GAVE8 protein or signaling RNA is above or below normal). In an antibody set, the set may contain, for example: (1) binding to GAVE8 (Please read the notes on the back and fill out this page) 0, Install f This paper scale applies to Chinese National Standard (CNS) A4 specification (210X 297 mm) -71 - 1250209 ΑΊ V. Description (the first antibody to the protein (eg, attached to the solid support); and (as needed) (2) the second antibody that binds to the detectable agent to the different antibody or first antibody of the GAVE8 protein. In a set of nucleotides, the set may comprise, for example: (1) an oligonucleotide that hybridizes to a GAVE8 nucleic acid sequence (eg, a detectably labeled oligonucleotide) or (2) to amplify a GAVE8 nucleic acid molecule. The kit may also contain, for example, a buffering agent, a preservative or a protein stabilizer. The kit can also contain the necessary ingredients to detect detectable agents such as enzymes or receptors. The kit may also contain a control sample or a control sample that can measure and compare a series of test samples. The components of the kit are typically sealed in separate containers, and a single package can contain all of the various containers and instructions for testing whether the patient has or is at risk of developing a condition associated with abnormal G A V E 8 performance. 2. Predictive assays The methods described herein are further useful as diagnostic or predictive assays to identify patients with or at risk of developing a disorder associated with abnormal GAVE8 performance or activity. For example, assays described herein, such as pre-diagnostic assays or post-assays, can be used to identify patients suffering from or at risk of developing a disorder associated with a disordered GAVE8 protein, nucleic acid expression or activity, such as a condition of the immune system or the nervous system, For example, an immune response associated with the onset of multiple sclerosis. In addition, predictive measures can be used to identify patients who are at risk of developing or having a condition associated with abnormal G A/E 8 performance or activity. Therefore, the present invention provides a method in which a test sample is obtained from a patient and the paper size is applied to the Chinese National Standard (CNS) A4 specification (2] 0×297 mm) ^~ -72- (please read the notes on the back first) Fill in this page) - Install - Ministry of Economic Affairs Intellectual Property Bureau Staff Consumer Cooperative Printed 1250209 A7 B7 Ministry of Economic Affairs Intellectual Property Bureau Staff Consumer Cooperative Printed 5, Invention Description (g Detects GAVE8 protein or nucleic acid (eg, communication RNA, genomic body) DNA), if present in the presence of a GAVE8 protein or nucleic acid, can diagnose a patient with or at risk of developing a condition associated with abnormal GAVE 8 performance or activity. "Test sample" in this article means a patient-derived organism. Samples. For example, the test sample can be a biological fluid (eg, blood sputum), cell sample, or tissue of a living being. In addition, the predictive assays described herein can be used to determine whether a patient can administer an agent (eg, an agonist, antagonist, Peptides, proteins, peptides, nucleic acids, small molecules or other drug candidates) treat conditions associated with abnormal GAVE8 expression or activity. For example, the method is available Determining whether a patient can be effectively treated with a specific agent or group of agents (eg, an agent that reduces GAVE8 activity). Accordingly, the present invention provides a method (a test sample, a GAVE8 protein or a nucleic acid is detected) to determine whether a patient is effectively available. Treating a condition associated with abnormal GAVE8 expression or activity, (eg, if a G AV E 8 protein or nucleic acid is present, it can be diagnosed that the patient can administer the agent to treat a condition associated with abnormal GAVE 8 expression or activity). The method of the invention can also be used to detect genetic damage or mutations in the GAVE8 gene to determine whether a patient having a damaged gene has a risk of a disorder characterized by abnormal cell proliferation and/or differentiation. In a preferred embodiment, the method Including detecting whether a cell sample of a patient has at least one gene damage or mutation characterized by affecting the integrity of the gene encoding the GAVE8-protein or causing the GAVE8 gene to be abnormal. For example, confirmation of damage or mutation of the gene The method is to detect whether there is at least one of the following: 1) one of the GAVE8 genes Or deletion of multiple nucleotides; 2) addition of one or more nucleotides to the GAVE8 gene; 3) substitution of one of the GAVE 8 genes or the paper size applicable to the Chinese National Standard (CNS) A4 specification (210X297 mm) ( Please read the notes on the back and fill out this page. mr Pack. Order -73- 1250209 Ministry of Economic Affairs Intellectual Property Office Staff Consumer Cooperative Print A7 ___B7__ V. Invention Description (71) Multiple Nucleotides; 4) GAVE8 Gene Chromosomal rearrangement; 5) altering the content of GAVE8 gene signaling RNA transcript; 6) modification of GAVE8 gene abnormality, such as methylation pattern of genomic DNA; 7) GAVE8 gene signaling RNA transcript has non-wild type The presence of the junction pattern; 8) non-wild type GAVE8-protein; 9) loss of the GAVE8 gene dual gene; and 10) inappropriate post-translational modification of the GAVE8 protein. As described herein, there are a number of techniques known in the art for detecting damage to the GAVE8 gene. A preferred biological sample is a peripheral blood leukocyte sample isolated from a patient. In some embodiments, the technique for detecting damage comprises polymerase chain reaction (PC R) using a probe/introduction (see, for example, U.S. Patent No. 4,683,1 95 and 4,683,202), such as anchor PCR or RACE PCR or linkage chain reaction (LCR) (see for example: Landegran et al., Science (1 988) 241:1 077-1 080; and Nakazawa et al· Proc Natl Acad Sci USA (1994) 91:360-364), the latter especially for point mutations that can be used to detect the GAVE 8 gene (see for example: abravayeta I., Nucleic Acids Res (1 995) 23: 675-682) . The steps of the method comprise collecting a cell sample from a patient, and isolating the nucleic acid (eg, gene, signaling RNA or one) from the cell sample. The contact nucleic acid sample can specifically hybridize with one or more of the hybridization and GAVE8 gene amplification conditions. To the primer of the GAVE8 gene, the presence or absence of the amplified product is detected, or the size of the amplified product is detected and compared with the length of the control sample. It is contemplated that P C R and/or L C R can be used in the initial amplification step and used in conjunction with any of the techniques described herein for detecting mutations. (Please read the precautions on the back and fill out this page.) The paper size applies to the Chinese National Standard (CNS) A4 specification (210X 297 mm) -74 - 1250209 Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative printed A7 ____ B7 V. DESCRIPTION OF THE INVENTION (Another method of amplification includes: self-sustained sequence replication (Guate||i et al., Proc Natl Acad Sci USA (1990) 87: 1874-1878), transcriptional amplification system (Kwoh et a|., PrC) C Natl Acad Sci USA (1 9 89) 86 : 1 1 73-1 1 77), Q-Wan replicase (Uzardi et al., Bio/Technology (1 988) 6:1197), or any other nucleic acid amplification The method then detects the amplified molecules using techniques well known to those skilled in the art. Detection techniques are especially useful for detecting nucleic acid molecules if the amount of the molecule is very low. In another embodiment, the mutation of the GAVE 8 gene in the sample can be confirmed by limiting the change in the enzyme excision pattern. For example, the sample and the control group D N A are separated, amplified (as needed), hydrolyzed with one or more restriction nucleases, and the fragment length is determined and compared by colloidal electrophoresis. The difference in the length of the fragment between the sample and the control group D N A represents the presence of a mutation in the sample D N A . In addition, the presence of a specific mutation can be determined by using a sequence-specific ribozyme (see, e.g., U.S. Patent No. 5,498,531) to examine whether a ribozyme excision site is produced or lost. In other embodiments, mutations in the GAVE8 gene can be confirmed by hybridization of a sample and control nucleic acid (eg, DNA or RNA) to a high density array containing hundreds or thousands of oligonucleotide probes (Cronin et al. , Human Mutation (1 996) 7: 244-255; Kozal et al., Nature Medicine (1996) 2: 753-759). For example, a genetic mutation in GAVE8 can be confirmed using a two-dimensional array of D N A probes that produce light (described in Cmniri et al., supra). Briefly, the first hybridization probe array can be used to scan the elongated DNA in the sample and apply the Chinese National Standard (CNS) A4 specification (210X 297 mm) via the generated line paper size with continuous overlapping probes. Read the notes on the back and fill out this page) - Install _

, 1T f -75- 1250209 A7 B7 Ministry of Economic Affairs Intellectual Property Bureau Staff Consumer Cooperative Printed 5, Invention Description (Sexual alignment confirms the change between bases. This step can identify point mutations. This step is followed by complementary to all Smaller, specialized probe arrays that detect variants or mutations perform a second hybridization sequence to identify specific mutations. Each mutant array consists of a combination of parallel probes, one probe complementary to the wild-type gene, and the other One complementary to the mutated gene. In another embodiment, the GAVE8 gene can be directly sequenced using any of a variety of sequencing reactions known in the art and the GAVE8 sequence of the sample can be compared to the corresponding wild type control sequence to detect the mutation. The implementation of the reaction includes the development of Maxim & Gilbert (Proc Natl Acad Sci USA (1977) 74: 560) or Sanger (Proc Natl Acad Sci U SA (1 977) 74: 5463). Any various automation can also be considered. The sequencing procedure for the determination of the diagnosis (Bio/Techniques (1 995) 1 9 · 448), including the mass spectrometry legal sequence (see, for example, PCT publication number WO94/16101; Cohen et a I., Ad v Chromatogr (1996) 36: 127-162; and Griffin et a I., App I Biochem Biotechnol (1 993) 38: 1 47-159). Other methods for detecting mutations in the GAVE8 gene include: A method of protection against excision to detect mismatched bases in RNA/RNA or RNA/DNA heteroduplexes (Myers et al., Science (1985) 230: 1 242). In general, ' The 'pairing error resection" technique can hybridize a (marked) RNA or DNA containing a wild-type GAVE8 sequence to a possibly mutated RNA or DNA derived from a tissue sample to form a heteroduplex. It is possible to react the single-stranded region between the control group and the sample strands due to the mismatch of the salt base pair in the split duplex. RN A/D NA double strand (please read the back note first and then fill in the page) )

This paper scale applies to the Chinese National Standard (CNS) A4 specification (210X 297 mm) -76- 1250209

The RNase can be used to hydrolyze and cleave the mismatched region, dna/dna hybrids. S彳 nuclease can be used to hydrolyze and cleave the mismatched region. In other embodiments, the DNA/DNA or RNA/DNA duplex can be hydrolyzed to cut the mismatched region by hydroxylamine or tetraoxide and/or hydrogen D. After hydrolyzing the paired erroneous region, the resulting material can be separated by size using a denatured polyamidoamide gel to determine the mutation site. See, for example, Cotton et al., Proc Natl Acad Sci USA (1 988) 85:4397; Saleeba et al.,

Methods Enzymol (1 992) 21 7:286-295. In a preferred embodiment, the control D ΝΑ or RNA can be labeled for detection. In another embodiment, the paired error resection reaction in the designated system may use one or more proteins that recognize pairs of mismatched base pairs in the double-stranded DNA (so-called "DNA pairing error repair" enzymes), Measure and map the point mutations from the GAVE8 complementary DNA of the cell sample. For example, the mutY enzyme of E. coli can cut A when the G/A pairing error occurs, and the thymidine DNA glycosylase of HeLa cells can be mismatched in G/T. T (H su et a I., Carcinogenesis (1 9 9 4) 1 5 : 1657-1662). According to a typical embodiment, a probe based on the G A V E 8 sequence (e.g., the wild-type GAVE8 sequence) can hybridize to complementary DNA or other DNA products of the test cells. The DNA pairing error repairs the enzyme-treated duplex and the excised product, which can be detected by electrophoresis. See, for example, U.S. Patent No. 5,459,039. In one of the other specific embodiments, the GAVE8 gene mutation can be confirmed by a change in electrophoretic mobility. For example, single-strand conformal polymorphism (SSCP) can be used to detect differences in electrophoretic mobility between mutant and wild-type nucleic acids (Orita et al., this paper scale applies Chinese National Standard (CNS) A4 specification (210X 297 mm) (Please read the precautions on the back and fill out this page) • Installed. Ministry of Economic Affairs Intellectual Property Bureau Staff Consumer Cooperative Printed -77- Ministry of Economic Affairs Intellectual Property Bureau Staff Consumer Cooperative Printed 1250209 A7 B7 V. Invention Description (^ Proc Natl Acad Sci USA (1 989) 86:2766; see also Cotton, Mutat Res (1 993) 285: 1 25-1 44; H ayashi, Genet Anal Tech Appl (1 992) 9:7 3-7 9) The single-stranded DNA fragment of the sample and the control GAVE 8 nucleic acid are denatured and returned to the original. Different single-stranded nucleic acid secondary structures are produced depending on the sequence, even if the single base change can be changed in electrophoretic mobility. Detection can be performed by labeling DNA fragments or by using labeled probes. RNA (rather than DNA) can be used to enhance assay sensitivity because its sequence is more sensitive to changes in secondary structure. In a preferred embodiment Mainly to make Heteroduplex analysis separates double-stranded duplex molecules by electrophoretic mobility changes (Keen et al., Trends Genet (1 991 ) 7: 5) ° In another embodiment, an internal denaturing agent can be used. Gradient Polypropylene • Melamine Gel for Denaturing Gradient Colloidal Electrophoresis (DGGE) for the Determination of Mutations in Mutant or Wild Type Fragments (Myers et al., Nature (1 985) 31 3:495). When performing D GG E method analysis First, modify D ΝΑ to ensure incomplete denaturation, such as PCR to add a GC clamp of approximately 40 bases of GC-rich high melting point DNA. In a further embodiment, a temperature gradient can be used instead of the denaturation gradient to confirm the control group. And differences in DNA mobility of sample samples (Rosenbaum et al., Biophys Chem (1 987) 265:1 2753). Other examples of detection point mutation techniques include, but are not limited to, selection of oligonucleotide hybridization, Selected amplification or selection of primer extensions. For example, oligonucleotide primers can be prepared in which known mutations are placed centrally and then hybridized to the target DNA under hybridization conditions that allow ideal pairing (Saiki et al_, Nature (1 986) ) 324:1 63) ; S Aiki et al., Proc Natl Acad Sci (Please read the note on the back and fill out this page) This paper size applies to the Chinese National Standard (CNS) A4 specification (210X 297 mm) -78- 1250209 A7 B7 Ministry of Economics Intellectual Property Bureau employee consumption cooperatives printed five, invention instructions (g USA (1 989) 86: 6230). When the oligonucleotide is attached to the hybridization membrane and hybridized to the labeled target DNA, the dual gene-specific oligonucleotide can hybridize to the PCR amplified target DNA or a plurality of different mutations. In addition, a dual gene-specific amplification technique for selective P C R amplification can be used in conjunction with the present invention. Oligonucleotide primers that are specifically amplified can carry important mutations at the center of the molecule or at the 3' distal end of the primer (and amplification depends on the difference in hybridization) (Gibbs et al., Nucleic Acids Res (1989) 1 7:243 7-244 8) Under appropriate conditions, pairing errors can prevent or reduce polymerase extension (Prossner, Tibtech (1 993) 1 1 : 238). In addition, novel restriction points can be satisfactorily introduced in the mutated region for base excision detection (Gasparini et al., Mol Cell Probes (1992) 6:1). In some embodiments, Amplification was performed using Taq ligase (Barany, Proc Natl Acad Sci USA (1 991 ) 88: 189). In this case, only when there is a known mutation at a specific site, the 3' end of the 5' sequence will be ideally paired, and the linkage will occur, and the presence or absence of amplification will be detected. Performing the methods described herein, for example, using a diagnostic kit packaged with at least one nucleic acid probe or antibody reagent prior to conveniently utilizing clinical setting data for a patient having a GAVE8 gene symptom or a history of the disease or disease group for rash . In addition, the predictive assays described herein can be performed using any cell type or tissue that expresses GAVE8. 3. Pharmacogenetics (please read the precautions on the back and fill out this page) -Installation, -port f This paper scale applies to Chinese National Standard (CMS) A4 specification (210X 297 mm) -79- 1250209 A7 ____B7 V. Description of the invention ((Please read the notes on the back and then fill out this page) The screening assays described here confirm that the GAVE8 activity (such as GAVE8 gene expression) has a 朿fj or inhibitory effect [J, or Modulation ij can be administered to a patient to treat (preventively or therapeutically) a condition associated with GAVE8 activity (including, for example, but not limited to, an immune response associated with multiple sclerosis). Taking into account the individual's pharmacogenetics (ie, the relationship between the genotype of the patient's patient and the response of the patient to the foreign compound or drug). The difference in the metabolism of the therapeutic agent can cause a pharmacologically active drug relationship between the dose and the blood concentration. Changes can lead to severe toxicity or therapeutic success. Therefore, the patient's pharmacogenetics selects effective agents (eg drugs) based on the patient's genotype for prophylactic Or therapeutic treatment or testing. Further pharmacogenetics can be used to determine the appropriate dose and course of treatment. Accordingly, the activity of the patient's GAVE8 protein, the expression of the GAVE8 nucleic acid or the mutation of the GAVE8 gene can be determined to select appropriate Therapeutic or prophylactic treatment of the patient. Because pharmacogenetics can change the patient's effects on drug elimination and abnormalities, it is clinically a genetic variable with significant drug response. See for example: Under, Clin Chem (1 997) 43(2): 254-2 66. For the printing of the Consumers' Cooperatives of the Intellectual Property Office of the Ministry of Economic Affairs, the patterns of the two pharmacogenetic conditions can be distinguished. The gene transfer status of a single factor can be Changing the way a drug works on the body is called “the drug effect of change.” The gene delivery status of a single factor can change the way a drug works in the body, called “change drug metabolism.” The state of pharmacogenetics Can be a rare defect or polymorphism. For example, 6-phosphate glucose dehydrogenase deficiency (GOD or stupid bean disease) is general inheritance Enzyme disease, the main clinical manifestation of which is the intake of oxidant drugs (anti-malarial drugs, sulfonic acid paper scale applicable to China National Standard (CNS) A4 specifications (210X 297 metric tons) -80-1250209 kl B7 Ministry of Economic Affairs The Intellectual Property Bureau employee consumption cooperative printed five, invention instructions (drugs, painkillers, nitrofurans) and hemolysis after consumption of broad beans. In the specific embodiment, the activity of the drug metabolizing enzyme determines the strength and duration of drug action. The main factor during the period. The genetic polymorphism of drug metabolizing enzymes (such as N-acetyltransferase 2 (ΝΑΤ2) and cytochrome P450 enzymes, C.YP2D6 and CYP2C19) may explain why some patients have no expected drug effects or It shows severe drug toxicity and severe toxicity after taking standard and safe doses of the drug. Polymorphism is the two phenotypes expressed in the ethnic group, which are a wide range of metabolizers (ΕΕ) and poor metabolizers (ΡΜ). The prevalence of sputum is not the same among different ethnic groups. For example, the gene encoding CYP2D6 is a highly polymorphic gene, and many mutations have been identified in sputum, all of which can result in a lack of functional C 丫 Ρ 2 D 6 . When taking standard doses, poor metabolizers of CYP2D6 and CYP2C1 9 often experience an extended drug response and side effects. If the metabolite is the active therapeutic component (for example, the analgesic effect of codeine base is regulated by the metabolite (morphine) formed by CYP2D6), then sputum will show an untreated response. Other extreme examples are the so-called ultra-fast metabolizers, which do not respond to standard doses. Recently, it has been confirmed that ultra-rapid metabolism is molecularly amplified by the CYP2D6 gene. Therefore, the activity of the GAVE8 protein, the expression of the GAX/E8 nucleic acid, or the mutation amount of the patient's GAVE8 gene can be determined to select a suitable agent for therapeutic or prophylactic treatment of the patient. In addition, pharmacogenetic research can be used to classify genes that encode polymorphic dual genes for drug metabolizing enzymes to identify phenotypes of patient drug responses. Apply this knowledge to a drug or drug selection (for example, using a typical screening assay described here (please read the back note first and then fill out this page) if

'P This paper scale applies to Chinese National Standard (CNS) A4 specification (210X 297 mm) -81 - 1250209 ΑΊ _____ _Β7__ V. Invention description (d regulator), can avoid adverse reactions or treatment failure, so use GAVE8 modulators increase the efficiency of treatment or prophylaxis when treating patients. 4. Monitoring effector monitoring agents (eg, drugs, compounds) that affect GAVE8 performance or activity (eg, ability to modulate abnormal cell proliferation and/or differentiation) during clinical trials can be applied not only to basic drug screening, but also to applications In clinical trials. For example, in the screening assay described herein, an agent that measures the efficacy of the GAVE8 gene to increase the protein content or protein activity can be used for clinical trials in patients exhibiting reduced GAVE8 gene expression, protein content, or protein activity. monitor. In addition, the agent whose screening assay is effective in reducing the GAVE8 gene expression, protein content or protein activity can be monitored by a patient exhibiting an increase in GAVE8 gene expression, protein content or protein activity. In clinical trials of the performance or activity of GAVE8, it is preferred that other genes involved in proliferative disorders such as cells be used as markers for a particular cellular immune response. For example, but not limited to, treatment with an agent (eg, a compound, drug, or small molecule) that modulates GAVE8 activity (eg, the activity identified by the screening assays described herein) can identify genes that are regulated in the cell (including GA\/). E 8 ). Thus, in order to investigate the effects of agents on cell proliferative disorders (e.g., clinical trials), cells can be isolated and RNA can be prepared and analyzed for GAVE 8 expression and other gene content involved in the condition. The content of gene expression (ie, gene expression pattern) can be quantified by Northern blotting or RT-PCR, or measured by the method described here, or measured by GAVE8 activity or other genes. (CNS) A4 size (210X 297 mm) (Please read the note on the back and fill out this page) - Install - Order Ministry of Economic Affairs Intellectual Property Bureau Staff Consumer Cooperative Printed -82- 1250209 A7 B7 Ministry of Economic Affairs Intellectual Property Bureau employees The consumer cooperative prints five, the invention description (quantity. With this method, the gene expression pattern can be used as the identifier 'to indicate the physiological response of the drug to the cell. According to this, the patient can be measured before and during the treatment of the patient. Reaction state. In a preferred embodiment, the invention provides methods for monitoring drug U (eg, agonists, inhibitors, peptidomimetics, proteins, peptides, nucleic acids, small molecules, or other screening assays described herein) The drug efficacy of the drug candidate for the patient comprises the following steps: (i) obtaining a pre-dosing sample from the patient prior to administration; (N) The performance level of GAVE8 protein, signaling RNA or genomic DNA in the pre-tested sample; (iii) one or more post-administration samples from the patient; (iv) the GAVE8 protein, the signaling RNA or the genomic body in the administered sample after detection The level of DNA performance or activity; (v) comparing the level of performance or activity of GAVE8 protein, signaling RNA or genomic DNA in pre- and post-administered samples; and (vi) changing the administration of the drug to the patient accordingly. The agent can increase the performance or activity of GAVE8 to a higher content, which means increase the efficacy of the agent. In addition, reducing the dosage of the drug can reduce the performance or activity of GAVE 8 to a lower content, which means lowering the efficacy of the agent. Methods of Treatment The present invention provides prophylactic as well as therapeutic methods of treating a patient having or at risk of (or susceptible to) a disorder associated with abnormal GAVE8 expression or activity, including but not limited to ): Immune response associated with multiple sclerosis. This paper size applies to the Chinese National Standard (CNS) Α4 specification (210X297 mm) (please Read the notes on the back and fill out this page. Sticky-book-83- 1250209 A7 _____B7 V. Invention instructions ((Please read the notes on the back and fill out this page) 1. One of the preventive methods features, this The invention provides a method for preventing a disease or symptom associated with abnormal GAVE8 expression or activity in a patient, which is administered to a patient to modulate GAVE8 expression or at least one drug having GAVE8 activity. The abnormal GAV Ε 8 expression or activity of the patient The risk of the resulting disease can be confirmed by any or a combination of diagnostic or predictive assays described herein. The administration of a prophylactic agent can prevent or delay the progression of a disease or condition prior to the onset of a characteristic GAVE8 symptom. The patient may be treated with a GAVE8 agonist or a GAVE8 antagonist depending on the type of GAVE8 abnormality. Appropriate agents can also be determined based on the screening assays described herein. 2. Methods of Treatment Printed by the Ministry of Economic Affairs, Intellectual Property Bureau, Staff Consumer Cooperatives Another feature of the present invention relates to a method of controlling the treatment of GAVE8 expression or activity. The method of regulation of the present invention comprises: an agent that contacts one or more activities associated with the activity of a GAVE8 protein in a cell. An agent that modulates the activity of a GAVE8 protein can be used as an agent described herein, such as a nucleic acid or protein, a naturally occurring ligand of a GAV E8 protein, a peptide, a GAV E8 peptidomimetic or other small molecule. The agent can be an agonist, a retro agonist or an antagonist. In one embodiment, the agent stimulates the biological activity of one or more G AV E 8 proteins. Examples of such stimulating agents include: an active GAVE8 protein and a nucleic acid molecule encoding GAVE8 that has been introduced into a cell. In another embodiment, the agent inhibits the activity of one or more of the biological GAVE8 proteins. Examples of the inhibitory agent include: a counter-GAVE8 nucleic acid molecule and an anti-GAVE8 antibody. The control method can be applied to the Chinese National Standard (CNS) A4 specification (210X 297 mm) at the scale of the living paper. -84-1250209 Printed by the Intellectual Property Office of the Intellectual Property Office of the Ministry of Economic Affairs ΑΊ 五 五 Β 发明 发明 发明 发明 发明 发明 发明 发明 发明 发明 发明 发明 发明 发明The cells are cultured or administered in vivo (e.g., administration of a medicament to a patient). The invention therefore provides a method of treating a patient suffering from a disease or condition characterized by abnormalities in the performance or activity of a GAVE8 protein or nucleic acid molecule. In one embodiment, the method comprises administering an agent that modulates (eg, up-regulates or down-regulates) GAVE8 expression or activity (eg, a drug identified in the screening assays described herein) or a combination agent. In another embodiment, the method Included with the administration of a GAVE8 protein or nucleic acid molecule for treatment to correct for a decrease or abnormality in GAVE8 expression or activity. GAVE8 activity can be stimulated in the case of a downregulation of GAVE8 abnormality and/or an increase in GAVE8 activity with a beneficial effect. Conversely, In the case of an upward regulation of GAVE8 abnormality and/or a beneficial effect of reducing GAVE8 activity, The GAVE8 activity can be inhibited. The invention is further illustrated by the following examples, without limitation. All of the cited references, patents, and issued patent applications are hereby incorporated by reference in its entirety. A portion of the human GAVE8 coding region (AC02651 0) has been identified from the human genome database using the FASTA algorithm, which uses the human GAX/E8 coding region-specific primer: 5'GCGGTGCGCGCTACCAG 3' (sequence confirmation number·· 7) and 51 GCGCCTGCCAGCAGATCC 3, (Sequence 歹 [J Confirmation Number: 8) PCR, screening for colonies from the complementary DNA library of the human brain. The human complementary DNA library with primary recombinant colonies can be subdivided into 90 pools. Approximately 3 X 1〇4 colonies of each pool (P〇〇l) load to 96-well paper scale applicable to China National Standard (CNS) A4 specifications (2] 0'乂297 mm) (Please read the notes on the back first) Fill in this page) -85- 1250209 A7 B7 Ministry of Economic Affairs Intellectual Property Bureau Employees Consumption Cooperative Printed 5, Invention Description (d PCR plate for PCR amplification. PCR is used: 94 ° C, 3 minutes; 40 weeks The cycle was 94 ° C for 30 seconds, 52 ° C for 30 seconds, and 68 ° C for 45 seconds. The PCR products of each well in the 96 μL PCR plate were examined by 2% agarose gel electrophoresis. A possible GAVE8 colony subpool for the PCR fragment. The positive secondary pool was then diluted to a pool of 90 for further PCR screening. The second colony of the positive secondary pool was planted into agar plates, and the positive plastids were confirmed by PCR and subjected to DNA sequencing analysis. Example 2 - Production of HEK293 cells overexpressing hGAVE8 For further experiments in which hGAVE8 was provided in large amounts, complementary DNA encoding hGAVE8 was cloned into expression vectors and transfected into HE K2 93 cells. HEK293 producing over-expression of hGAVE8 is a 2 ml F12 HAM medium (Gibco/BRL, catalog number 1 1 765-054) containing 10% fetal calf blood (Gibco/BRL catalog number 1 600-044) E. sinensis E K293 cells were seeded into six-well 35 mm tissue culture plates (3 X 1 05 ΗΕΚ293 cells per well, ATCC catalog number CRL-1 573, Costar record number 3 5 16). The cells were then cultured in a 37 t C02 incubator until the cells reached 50-80% overfill. The selected HGAVE8 complementary DNA nucleic acid sequence was inserted into the pc DN A3.1 selection vector (Invitrogen, catalog number V7 90-20) using the procedure described above. Two micrograms of DNA was diluted to 1 microliter with a blood-free F1 2 HAM medium. Separately add 25 μl of fat (please read the note on the back and fill out this page). The paper size is applicable to the Chinese National Standard (CNS) A4 specification (210X297 mm) -86- 1250209 智慧 Ministry of Economics Intellectual Property Bureau staff consumption cooperatives India purple ___________Β7, invention instructions (Lipofectamine reagent (Life Technologies, catalog number 1 8324-020) diluted to 100 μl with blood-free F12 HAM medium. Then DNA The solution was gently mixed with a lipofectamine (li ρ 〇fectamine) solution and allowed to react at room temperature for 45 minutes to form a DNA-lipid complex. The cells were washed once with 2 ml of blood-free F 1 2 HA Μ medium. To each transfection (six transfections in six test wells), 0.8 ml of blood-free F1 2 HAM medium was added to the solution containing the DNA-lipid complex ( The total volume was 22 ml) and gently mixed. The resulting mixture (hereinafter referred to as "transfection mixture") (0.8 ml + 0.2 ml) was overlaid on the washed cells. fine The reagent of the bacteria is then transfected with the lipid-DNA complex in a C〇2 incubator at 37 ° C for 16 hours. After the reaction is completed, the transfection mixture is not removed and the cells are reconstituted. Cover 1 ml of F 1 2 HA Μ medium containing 1% fetal calf blood. After 18 hours of transfection, aspirate the culture medium covered with the cells. Then wash the cells with PBS pH 2-4 ( Gibco/BRL catalog number 10010-023) and replace the pbs with F 1 2 HA Μ culture medium containing 5% blood sputum. 72 hours after transfection, the cells contain the antibacterial agent genactin (genetecin) The selected culture medium (400 μg/ml, L if e T e ch η ο I og i es, catalog number 1 1 8 1 1 ) was diluted ten times. Example 3 - Agonist assay for screening human GAVE8 efficacies Agent, artificially couples hGAVE8 to the Gaqi 5 signal component. Activates the G q mechanism and stimulates intracellular c a2 + from the paper scale to apply Chinese National Standard (CMS) A4 specification (2}0X 297 mm)~^ ~ -87 - (Please read the notes on the back and fill out this page) i·.

1T 1250209 A7 B7 Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative printing 5, invention description (release of sarcoplasmic reticulum vesicles. Ca2+ chelate dye can be used to detect Ca2+ release to the cytoplasm. Fluorescence measurement imaging plate reading The device (or FLIPR® device, Molecular Devices) uses the luciferase reporter gene system to monitor the results of any fluorescence changes. The agonist activity can be reflected in the increase in fluorescence. HEK293 cells expressing hGAVE8 are pre-genetically engineered. The operation represented a universal form of Gq protein (Gaqi5). To prepare the cells, the expression of hGAVE8 in the cells was enhanced using Ga 16-coupled HEK293 cells and the experimental guidelines of Example 2. The cells were at 37 ° C and contained 10%. Fetal bovine blood stasis, 100 IU/ml penicillin (Gibco/BRL, catalog number 1 51 40-1 48), 100 μg/ml streptomycin (catalog number 1 51 40-1 48, Gibco/BRL), 400 μg/ ML G418 (Gibco/BRL, Cat. No. 1 01 31 -035) and F1 2 HAM medium (200 mg/ml Merlin (Invitrogen, Cat. No. 8250-05)) (Gibco/BRL, Cat. No. 1 1 765-054 ), 5% C〇2 Logarithmic growth was maintained. At 24 hours prior to assay, 1 2,500 cells/well of HEK293 cells were seeded into 96-384 Multidrop ^ g (La b systems, type 832) into a 384-well bottom assay plate, each well. The volume is 50 μl (Greiner/Marsh, catalog number N581 02). The cells are cultured in a humidified 5% C〇2 incubator at 37 ° C (Forma Scientific C〇2 water-tube thermostat, mode 3110) Prepare the following stock solutions: 1 M Hepes stock solution (pH 7.5) (Gibco/BRL, catalog number 15630-080); 250 millimolar concentration of probenicid stock solution (Sigma, catalog number P8761) 1 N Na〇H; and 1 millimol of Fluo 4-AM Dye stock solution (please read the back note first and then fill out this page) This paper scale applies to China National Standard (CNS) A4 specification (210X297 mm) -88- 1250209 A7 B7 Ministry of Economic Affairs Intellectual Property Bureau B (Ministry Consumer Cooperatives Printed 5, Inventions (d (Molecular Probes, Catalog No. FI 4202) DMS〇 (Sigma D 2 6 50). A reaction buffer solution was prepared using a 1000 halo Hank's balanced salt solution (Fisher/Mediatech, catalog number MT21023), 20 ml of 1 Μ Hepes stock solution and 10 ml of a 250 mM molar probenicid stock solution. To prepare the load buffer, 1.6 ml of a 1 mM molar Fluo 4-AM Dye stock solution was mixed with 0.32 ml of pluronic acid (Molecular Probes, Cat. No. P6866) and then with 400 ml of the above reaction buffer. The solution and 4 ml of fetal calf blood were mixed. One hour prior to the assay, 50 microliters of freshly prepared load buffer solution was added to each well of the 384-test disc using a 96/384 Multidrop apparatus. The cells were cultured in a humidified incubator at 37 °C until the optimum dye intake. Prior to the assay, the cells were rinsed twice with 90 μl of the reaction buffer solution using a 384 EMBLA cell wash (Skatron; Model 1 2386) with a suction head assembly at least 10 mm above the bottom of the plate, leaving each well Lower 45 microliters of buffer solution. The CCD camera of the FLIPR® II (Molecular Devices) instrument (Princeton Instruments) has an aperture of 2.0 and a exposure of 0.4 seconds. Monitor the cell plate with a camera to accurately load the dye. A library of compound genes containing possible agonists was tested in a physiological salt buffer solution at 10 micromolar. The change in fluorescence was measured 10 seconds before the addition of the compound. After the addition of the compound, the fluorescence was measured once per second at the first minute, then every six seconds, and the analysis time of the total experiment was three minutes. After the tenth scan, add five microliters of the same sample (please read the notes on the back and fill out this page) - loading and ordering

P This paper scale applies to China National Standard (CNS) A4 specification (210X297 mm) -89- 1250209 A7 B7 Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative printing 5, invention description (8) 1 00 micromolar concentration The starting compound, the final compound of the cell's degree is 1 0 / (the concentration of the sputum ear. Record the change of the luminescence in the first 80 scans, measure the activity of the agonist and the concentration of 1 〇 micromolar AT P (S Igma A9062) Comparison of changes induced by maximal 値 fluorescence. Example 4 - Antagonist assay Screening of human GAVE8 antagonists, artificially coupling hGAVE8 to the G q system. As in Example 3, monitoring with FL丨PR® device Any change in fluorescence produced. Any decrease in fluorescence represents the activity of the antagonist. As in Example 3, HEK293 cells expressing hGAVE8 were previously genetically engineered to express a universal form of Gq protein (G aqi 5). F12 HAM medium (Gibco/BRL, catalog number 11765-054) at 37t: and 5% CO2 containing 10% fetal calf blood, 1〇〇IU/ml penicillin (Gibco/BRL, catalog number 1 5140- 1 48), 100 μg/ml chain (Cat. No. 15140-148, Gib co/BRL), 40 μg/ml G41 8 (Gibco/BRL, catalog number 1 0 1 3 1-035) and 200 μg/ml Meeking (Invitrogen, catalog number 8250) Maintain log phase growth in -05). Twenty-four hours before the assay, HEK293 cells at 12,500 cells/well were seeded into the assay plate at the bottom of the 3 84-well Clarion using a 96/384 Multidrop device in a volume of 5 μm. l (Greiner/Marsh, catalog number N581 02). Cells were cultured in humidified 5% C〇2 at 37 ° C. Prepare the following stock solutions: 1 M Hepes stock solution (pH 7.5) (Gibco/BRL, catalogue No. 15630-080); 250 millimolar concentration of probenicid stock solution (Sigma, catalog number P876 1) (please read the notes on the back and fill out this page) - loading., - loan paper The scale applies to the Chinese National Standard (CNS) A4 specification (210X 297 mm) -90- 1250209 ΑΊ _ Β7_ V. Invention description (^ (Please read the note on the back and fill out this page) 1 N Na〇H ; 1 Millocent concentration Fluo 4-AM Dye stock solution (Molecular Probes, catalog number f 1 4202) DMSO (Sigma D2650); and 480 nM SPP-1 (BioMol) stock solution. Use 1 000 ml of Hank's balanced salt solution (Fisher/Mediatech, catalog number 210T210 23), 20 ml of 1 molar concentration of Hepes stock solution, 10 ml of 250 mM molar probenicid stock solution and 1 mmol of C a concentration A reaction buffer solution was prepared by C 1 2 . To prepare a load buffer solution, 80 μl of a 1 mM concentration of Fluo 4-AM Dye stock solution was mixed with 16 μl of pluronic acid (Molecular Probes, Cat. No. P6866) and then 20 ml of the above The reaction buffer solution was mixed with 2. 2 ml fetal calf blood. Ten microliters of freshly prepared load buffer solution was added to each well of the 384-test disc using a 96/384 Multidrop apparatus ten minutes prior to the assay. The cells were incubated at 37 ° C in a humidified C02 incubator to optimize the dye intake. Before the assay, the cells were washed three times with 1 〇〇 microliter of reaction buffer solution using a 384 EMBLA cell washer with a suction head assembly at least 40 mm above the bottom of the plate, leaving 4 5 μl per well. Buffer solution. Printed by the Intellectual Property Office of the Ministry of Economic Affairs, the Consumers' Cooperative, using a Platemate-384 absorbing device (Matrix) to add five microliters of a 100 micromolar concentration of the starting compound to the cells. The concentration of the compound during the incubation step is approximately 10 micromolar. The cells were then placed in FLIPR® II, and the fluorescence was measured once per second for the first minute, followed by every six seconds, and the total analysis time was three minutes. After the tenth scan, sphingosine-1-phosphate (10 micromolar concentration, Bio Mol SL 140) was added. After each addition, '3 8 4 pipe tips are used with 20 microliters. q 1 % d MS〇 water-based paper scale applies to China National Standard (CNS) Α4 specification (210X 297 mm) -91 - 1250209 Economy Ministry of Intellectual Property Bureau employee consumption cooperative printed kl B7 five, invention instructions (washing 1 times. Example 5 - receptor binding assay in order to prepare membrane layer containing hGAVE 8 receptor, containing 1 millimole The HEK293 cell line overexpressing hGAVE8 was harvested in phosphate-buffered physiological saline (1 ml) of EDTA. The cells were further phosphate-buffered saline containing 10 mM EDT. Dilute three times and resuspend in 5 ml of buffer solution A (50 millimolar Tris-HCI (pH 7.8) (Sigma T6791), 5 millimolar concentration MgC12 (Sigma M8266), and 1 millimolar concentration EGTA (Sigma 0396) was then disrupted by tissue homogenizer (Polytron, Kinemetica, Model PT 10/35) for 1 minute. The resulting homogenate was centrifuged at 49,000 X g, 4 ° C for 20 minutes using a Sorvall Instruments ROB centrifuge. The pellet is resuspended in 25 ml The solution A was flushed and the centrifugation step was repeated three times. After the final centrifugation, the sedimentation block was again resuspended in 5 ml of buffer solution A, and the sample was dispensed in equal amounts and stored at -7 (TC. The receptor binding assay was performed using a membrane solution. Fractions and radiolabeled sphingosine-1-phosphate (or SPP-1) as a tracer. The assay was performed in a 96-well microtiter plate (Beckman Instruments). The binding reaction consisted of 18 micrograms of HEK293 cell preparation. With radioactive SPP-1 (〇.〇1 nanomolar concentration -25 nanomolar concentration), final volume 〇.2 ml containing 〇.彳% bovine blood albumin buffer solution A (Sigma, catalogue No. 34287) Composition (see lm et al., J Biol C hem (2000) 275 (1 9) : 1 4 2 8 1 -1 4 2 8 6). This paper scale applies to the Chinese National Standard (CNS) A4 specification ( 2lOX 297 mm) (Please read the notes on the back and fill out this page) -92- 1250209 A7 B7 V. Inventions (90> (Please read the notes on the back and fill out this page) Reagents at room temperature Reaction for 1 hour. With 0.3% polyethyleneimine (Sigma, catalog number P3143) and 0.1% bovine blood albumin BSA) Whatman GF treated for 1 hour prior to the time / C filter via a multi-collector corridor (a Brandell) filtering the reaction was terminated. The mixture was applied to a filter and reacted for one hour. The filter was washed 6 times with 1 ml of ice-cold 50 mM concentration of 丨5_|_1(^|(pH 7.6). The radioactivity was measured, based on the total concentration of each tracer, combined with non-specific binding (background) The difference between the calculations is specific. It requires 8 to 16 concentration data points to determine the binding of the ligand to the receptor to achieve equilibrium between the ligand and the receptor (balance binding parameters) and non-radioactive SP P-1 The amount of competitive radioactive SPP-1 bound to the receptor (competitive binding 値). The inhibition curve was prepared and the concentration required to inhibit 50% binding (IC50) was determined. Example 6 - Northern blot analysis Northern blot analysis It is performed using RNA samples derived from several human tissues to determine whether the tissue expresses the hGAVE8 receptor gene. Commercially available (eg CI n ntech) filters containing various human tissue RNA can be used. The probe can be P32- Labeled full-length hGAVE8 complementary DNA. Method of preparation of P32-labeled hGAVE8 complementary DNA by the Ministry of Economic Affairs Intellectual Property Office Staff Consumer Cooperative Printed probes. Resuspend twenty-five ng of hGAVE8 complementary DNA in 45 μl. Moer concentration Tris-HC丨 (pH 7.5); Microfluidic tube at 1 mM concentration, heated at 95 ° C for 5 minutes. The tube was then cooled in ice for 5 minutes. After quenching, the tube was chilled. The mixture is resuspended in 45 microliters of paper. The Chinese National Standard (CNS) A4 specification (2) 0X29*7 mm) -93- 1250209 kl B7 Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative printing 5, invention Description (91) GAVE8 complementary DNA and buffer solution as described above, mixed with RTS Rad Prime Mix (supplied by RTS Rad Prime DN A-labeling system) (L if e Technologies, catalog number 1 0387-0 1 7 ). Microliters of p32-marker has been ^^dCTP (Amersham, AA0005), J: 匕活一生: 3 0 00 Ci / millimolar concentration, mild but thorough agitation. The resulting mixture is at 37 ° C The reaction was stopped for 1 minute. The reaction was stopped by adding 5 μl of 0.2 molar concentration EDTA (pH 8.0). Five microliters of the same sample was taken from the mixture and the radioactivity was counted to evaluate the radioactive α-dCTP of 倂 in the hGAVE8 complementary DNA. RNA extraction Add 1 ml of Trizol reagent to the plate ( Life Technologies, Cat. No. 1 55 9 6 ) Directly dissolve cells or tissues. Then pipette the cell lysate several times to spread the solution evenly (then transfer the cell lysate to the tube). After homogenization, the solution was completely reacted at 30 ° C for 5 minutes to completely separate the nucleoprotein complex. After the reaction, 0.2 ml of chloroform (Sigma, Cat. No. C 53 12) / 1 ml of Trizol reagent was added to the solution, and the tube was vigorously shaken for 15 seconds. The solution was then reacted at 30 ° C for 3 minutes. After the reaction, the solution was centrifuged at 1 2,0 0 X g, 4 ° C for 15 minutes. The resulting aqueous phase was transferred to a new tube and 〇. 5 ml of isopropanol / 1 ml of Trizol reagent was added. The aqueous phase sample was then reacted at 30 ° C for 10 minutes and centrifuged at 1 2,000 X g for 1 minute at 4 °C. The supernatant was removed after centrifugation. The residual RNA pellet was rinsed with 70% ethanol. The rinsed sample is then centrifuged at 4 C, 7 5 0 X g for 1 〇 minutes, and the resulting upper 淸 paper size is applicable to national standards (CNS) A4 ϋϊ〇Χ 297 mm) ---- -- -94- (Please read the notes on the back and fill out this page) 1250209 A7 B7 Ministry of Economic Affairs Intellectual Property Bureau Staff Consumer Cooperative Printed V. Inventions (92> Solution. Then dry the residual RNA pellets and resuspend In ribonucleic acid-free water (Life Technologies, catalog number 10977-015). Colloidal electrophoresis preparation of agarose gel system 2 g of agarose (Sigma, catalog number A0 169) was melted in water, 5X formaldehyde gel operation buffer Solution (see description below) and 2.2 Μ Furfural (Sigma, Cat. No. P82031). Samples for colloidal electrophoresis were prepared as follows: RNA 4.5 μl (total 5 μg) 5X formaldehyde manipulation buffer solution 2.0 μl formaldehyde 3.5 μL Indoleamine (Sigma, Cat. No. F9037) 1 0.0 μl (5X Formaldehyde Gel Solution Buffer Solution is 0.1 Μ 3-(N-morpholinyl) Propane Sulfonate (MOPS) (pH 7.0) (Sigma, Cat. No. M5162 ); 40 millimolar concentration B Sodium (Sigma, Cat. No. 57670); and 5 millimolar EDTA (Acidity 8.0) (Sigma, Cat. No. E7889). The sample was reacted at 65 ° C for 15 minutes and then snap frozen in ice. After freezing, the sample was centrifuged for 5 seconds. Then two microliters of aldehyde gel load buffer solution was added to the sample; 50% glycerol (Sigma, catalog number 〇 5516); 1 millimolar concentration 〇 六 ( (pH 8.0) ;0.25% bromophenol blue (Sigma, catalog number 18046); 0.25% xylyl cyanohydrazide "(3丨9〇13 (please read the back note first and then fill out this page) This paper scale applies to Chinese national standards (CNS A4 size (210 X 297 mm) - 95- 1250209 A7 B7 V. Invention description (93), catalog number 335940). (Please read the note on the back and fill out this page.) In addition, it contains various organ RNA samples. The hybridization filter can be purchased from the market (CI〇ntech). The gel is pre-treated for 5 minutes at 5 V/cm. The sample is loaded into the gel after pre-operation. The gel is then immersed in formaldehyde at 4 V/cm. The gel operation buffer solution was operated. The buffer solution was updated at 2 hours of operation. Transfer of A from the gel to nitrocellulose. The gel was stained with ethidium bromide (Sigma, Cat. No. 385) (0.5 μg/ml 1·1 molar concentration of ammonium acetate (Sigma, Cat. No. 09689)) for 30 minutes. To confirm that the RNA has not degraded. The RNA was then used in the experiment described in Sambrook eta I., ed s. (Molecular Cloning: A Laboratory Manual, volume 1 , pp. 7.46-7.51, Cold Spring Harbor Laboratory Press (1 989)). The glycogel is transferred to a nitrocellulose filter (Schleicher & Schuell Inc, Cat. No. 74330-026)° with the P32-labeled complementary DNA hybridization Ministry of Commerce, Intellectual Property Office, Staff Consumer Cooperative, Printing Clontech ExpressHyb Hybrid Solution (Clontech, Catalog No. 8015-1) was reacted at 68 ° C for 2 hours. After the reaction, 15 ml of the heated hybridization solution was poured onto a north (MTN) membrane of a multi-tissue sample. The MTN membrane was immersed in the hybridization solution at 68 ° C with shaking. After 1 hour, the hGAVES complementary DNA probe denatured at 95 ° C for 5 minutes was added to a concentration of 1 〇 6 counts/ml. The reacted hybridization solution was covered with gel at 68 ° C and then shaken for 2 hours to overnight. This paper size applies to the Chinese National Standard (CNS) A4 specification (210X297 mm) -96- 1250209 A7 B7 V. Inventive Note (94) Then remove the MTN film from the Clontech ExpressHyb hybridization solution and immerse the membrane in 15 ml Clontech Wash solution 1 (2X SSC; ◦ • 05% SDS), shake at room temperature for 40 minutes, and refresh the solution for 3 times every 40 minutes. Clontech rinse solution 2 (0.1X SSC; 0.1% SDS) was then heated at 60 °C for 1 hour. The membrane was then immersed in 15 ml of Clontech Wash Solution 2 (0.1X SSC; 0.1% SDS) for 6 minutes at 6 CTC and continuously washed 3 times. The rinse solution was updated every 15 minutes. Development The film was exposed to a Kodak X-GMAT AR (Kodak, Cat. No. 1 65 1 579) film plus a reinforced plate at -70° overnight. Results The high amount of hGAVE8 was expressed in human spleen, brain and PBL. The unique transcript size is approximately 2.4 仟 base. Example 7 - P C R determination

TaqMan® or RT-p C R is a powerful tool for detecting interfering RNA in samples. This technique uses the 5' nuclease activity of AmpMTaqGold® DNA polymerase to decompose the TaqMan® probe during pcR. The TaqMan® probe contains a reporter dye (in this experiment: 6-FAM (6-carboxyfluorescein)) at the 5' end and a quencher dye at the 3' end (in this experiment: TAMRA (6- Carboxyl-Ν, Ν, Ν', Ν1·tetramethylrhodamine)). The TaqMan® probe system is specifically designed to work with positive and reverse primer sites. The paper size applies to the Chinese National Standard (CNS) A4 specification (2) 297 297 mm) (please read the back first) Note: Please fill out this page again} • Installed, can be edited by the Ministry of Economic Affairs, Intellectual Property Bureau, Staff Consumer Cooperatives - 97 - 1250209 A7 B7 V. Invention Description (95) Complementary DNA hybridization. When the probe system is complete, the end of the probe The let-off dye inhibits the fluorescence of the dye at the 5'-end. During PCR, the 5' to 3' activity of AmpliTaq G ο丨d ® DNA polymerase can cleave the $, _ end reporter dye and 3' end of the probe. A quencher dye that causes the replacement of the reporter dye. Once replaced, the fluorescent dye reported is no longer inhibited by the quencher dye. Therefore, monitoring the fluorescent increase of the reported dye can detect the complementary DNA template produced from the target. The PCR product was stacked. The ABI Prism Sequence Detector System (Model AB1 7700) from Perkin Elmer Applied Biosystems was used to monitor the increase in reported fluorescence during PCR. The reporter signal was normalized with a passive reference emission enthalpy. The template total RNA and several tissues of polyadenylation + RNA were purchased from Clontech (see Tables 1 and 2 for catalog numbers). (Please read the notes on the back and fill out this page)

Printed by the Intellectual Property Office of the Ministry of Economic Affairs, the Consumer Cooperatives. This paper scale applies to the Chinese National Standard (CMS) A4 specification (2j〇X 297 mm) -98- 1250209 A7

7 B V. Description of Invention (96) Ministry of Economic Affairs Intellectual Property Bureau Staff Consumption Cooperative Printed Table 1 Human Total RNA CI ο ntech Catalog Number Human Brain, All 64420-1 Human Heart 64025-1 Human Kidney 64030-1 Human Liver 64022-1 Human lung. 64023-1 Human pancreas 64031-1 Human skeletal muscle 64033-1 Human small intestine 64039-1 Human spleen 64034-1 Human stomach 64090-1 Human thymus 64029-1 (Please read the back note first) Fill in this page again) This paper size applies to Chinese National Standard (CNS) A4 specification (210X 297 mm) -99- 1250209 A7 ____B7 V. Invention description (97) Table 2 RNA sample Clontech catalogue number Human brain all 6516- 1 Human brain, tonsil 6574-1 human brain, caudate nucleus 6575-1 human brain, | cerebellum 6543-1 human brain, • corpus callosum 6577-1 human brain: ... hippocampus back 6576-1 human brain Department:, black matter 6580-1 human brain, thalamus 6582-1 human fetal brain 6525-1 (please read the back note first and then fill out this page) Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative Five micrograms of total RNA was mixed with 2 microliters (50 ng/L) of random six-body primers (L ife T ech η ο丨〇gies, catalog number 1 8 0 90) with a total reaction volume of 7 μm. Rise. The resulting mixture was heated at 70 ° C for 10 minutes and rapidly cooled in ice. Then, 4 μl of 5X first buffer solution, 2 μl of 0·1 mmol concentration DTT, 1 μl of 1 mM milliliter concentration dNTP, and 1 μl of water were added to the mixture. The mixture was gently mixed and reacted at 37 ° C for 2 minutes. Five microliters of Superscript RT-PCR reverse transcriptase (Life Technologies, catalog number 1 8090) was added after the reaction. The mixture was then reacted at 37 ° C for 60 minutes. The reaction was stopped by adding 1 μl of 2.5 mM concentration of EDTA. Then the mixture is applied to the Chinese National Standard (CNS) A4 specification (2) 0X297 mm on the paper scale. " -100- 1250209 A7 B7 5. Invention description (y 6 5 °C reaction 1 〇 min. (Please first Read the notes on the back and fill out this page. The PCR and TaqMan® assays were performed in a 96-well microtiter plate Mic “〇Amp optical tube (Perkin Elmer, Cat. No. N801 -0933). The reaction mixture for each well contained 25 μm. Ascending TaqMan® PC R mixture (Perkin Elmer, catalog number N 8 08-02 30), 1 microliter of positive bow 1 (5,-Ding 0 6 8 6 6 (: Ding Ding 0 (: D (: 0) Eight (: Ding 0 Ding-3,) (Preface 1 [1 Confirmation Number: 9), 1 μl of reversal bow scorpion (5,-TTGCCGCTCTACGCCAAGGCC-1) (sequence confirmation number: 10), 1 μl of TaqMan ® probe (5·- AGCACGCAGAAGAGCACG third-3,) (preface 确认U confirmation number··11), 1 μl of complementary DNA, and 2 μl of water. Concentration of complementary tissue template for each tissue sample: 5. Double replicate TaqMan® samples were prepared at 2, 1, 0.5, 0.25, 0.125, 0.0625 ng/μl (the template complementary DNA concentration was the final concentration). The plate was sealed with a MicroAmp optical 8-stripping cap (Perkin Elmer, catalog number N801-0935). The Ministry of Economic Affairs Intellectual Property Office employee consumption cooperative printed the human β-actin gene (P erki η EI mer, catalog number 401 846) Double-repeat production standard curve. The number of amplified molecules is obtained from the complementary DNA template concentration of the standard curve. The standard curve obtained by amplifying the known gene is used to quantify the complementary DNA molecule amplified by the unknown target gene. Internal control is regulated. The results of the above Ta q M a η® reaction are expressed in terms of an optional adjustment factor relative to the tissue (eg, cardiac GAV Ε 8 performance 値 / brain GAV Ε 8 performance 値). In the central nervous system The cerebellum is selected as the reference G of the GAVE8 performance, and the relative multiples are compared for comparison. Although the present invention has been described in detail using the above embodiments, it is understood that the paper size can be applied to the national standard (CNS) A4 specification. (210X29*7 mm) • 101 - 1250209 A7 B7 V. Description of the Invention (There are various modifications made therefrom without departing from the essence of the invention. Accordingly, the invention is to be limited only by the scope of the following claims. (Please read the note on the back and then fill out this page.) Ministry of Economic Affairs ^ Property Bureau Staff Consumer Cooperative Print This paper scale applies to China National Standard (CNS) A4 specification (210X 297 mm) -102 -

Claims (1)

1250209 A8 B8 C8 D8 If Year/Month ^曰|Amendment VII, Patent Application Scope Attachment 2A: Patent Application No. 91 1 16166 Chinese Application Patent Scope Replaced by the Republic of China on January 27, 1994 1. A code A single nucleic acid of human GAVE8 consisting of the nucleotide sequence of SEQ NO: 1, or a variant thereof, which encodes a mutant which is capable of adding, deleting or replacing human GAVE8 equivalent to human GAVES, The nucleic acid is expressed in the white blood cells of the brain, spleen and peripheral blood, but not in the heart, kidney or skeletal muscle, SEQ ID NO ·· 1 (please read the notes on the back and fill out this page) 丨41· Ministry of economic Affairs intellectual property Office employees consumer cooperatives printed in this paper scale applicable Chinese national standard (CNS) A4 size (210X297 mm) 1250209 Μ C8 D8 々, patented range GCCTAGACAGTGGTTCAAAGTTTmTCTTCCATTTCAGGTGTCGTGAAAAGCTrGAATTCCiG CGCGCCAGATATCACACGTGCCAAGGGGCTGGCTCAGGCCTGAGCCACTGCCTCAGGCCTG AGCCrGGGGGGATTACAGGCGGGGATTACAGGCCTGAGCCACCGCGCCCAGCCATGTGTGA ACCACCGCGCCCAGCCATTCATGTGTTATTAATGTCCCTTrTTTTGGGAC TATGTCACCTTGT TCCC CGTGATATTATTTGTGTGTATGTCTGTCTCCACGGACTCTATCCCTCCATCCKMACT CCCCCTTGGACACCATGTATAGGGGGTGATGGTCAGGGGGTAGGGCCCCTTTCCCACGACTT AGCCGGCTGCTGCGAGCGTGCTTACGGGGACCGCGGCCTGACACCGTATCTTGCCTCTCCAA CAGCCTTGGGGCGCGCGGCCCA Ding Ding Ding CGGGGC GGAG GC Ding Ding Ding CATCGTCCTGCATTACAAC GCGGCCGGCGCCGGTGAGCGAGG people CCGGCAAGCTCCGCGGTGCGCGC AC Ding Ding ACCAGCCGGGTGCC: GGCC GCGCGCCGACGCCGTGGTGTGCCTGGCGGTGTGCGCCTTCATCGTGCTAGAGAATCT AGCCGTG Ding Ding Ding TGTTGG GCTCGGACGCCACCCGCGCTTCCACGCTCCCATGTTCCTGCTCCTGG GCAGCCTCACGTTG CGGA Ding Ding Ding CTGC GGCAGGCGCCGCCTACGCCGCCAACATCCTACTGTCG gggccgctcacgctgAaactgtcccccgcgctctggttcgcacgggagggaggcgtcttcgt GGCACTCACTGCGTCCGTGCTGAGCCTCCTGGCCATCGCGCTGGAGCGCAGCCTCACCATGG CGCGCAGGGGGCCCGCGCCCGTCTCCAGTCGGGQGCGCACGCTGGCGA Ding GGCAGCCGCGGC CTGGGGCGTGTCGCTGCTCCTCGGGCTCCTGCCAGCGCTGGGCTGGAATTGCCTGGGTCGCC TGGACGCTTGCTCCACTGTCTTGCCGCTCTACGCCAAGGCCTACGTGCTCTTCTGCGTGCTCG CCTTCGTGGGCATCCTGGCCGCGATCTGTGCACTCTACGCGCGCATCTACTGCCAGGTACGC GCCAACGCGCGGCGCCTGCCGGCACGGCCCGGGACTGCG GGGACCACCTCGACCCGGGCGC GTCGCAAGCCGCGCTCGCTGGCCTTGCTGCGCACGCTCAGCGTGGTGCTCCTGGCCTTTGTG GCATGTTGGGGCCCCCTCTTCCTGCTGCTGTTGCTCGACGTGGCGTGCCCGGCGCGCACCTG TCCTGTACTCCTGCACKjGCGATCCCinrCCTGGGACTGGCCATGGCCAACTCACTTCTGAACC CCATCATCTACACGCTCACCAACCGCGACCTGCGCCACGCGCTCCTGCGCCTGGTCTGCTGC GGACGCCACTCCTGCGGCAGAGACCCGAGTGGCTCCCAGCAGTCGGCGAGCGCGGCTGAGG CTTCCGGGGGCCTGCGCCGCTGCCTGCCCCCGGGCCTTGATGGGAGCTTCAGCGGCTCGGAG CGCTCATCGCCCCAGCGCGACGGGCTGGACACCAGCGGCTCCACAGGCAGCCCCGGTGCAC CCACAGCCGCCCGGACTCTGGTATCAGAACCGGCTGCAGACTGACACCCTCGGCCTACGAC TGTCTTCCCAAGTnTACAGACTTGTTCmTTACA AAAGGAATTTG Ding Ding Ding AGGAAATGCAGCCA AAGG GCAG CGGAAAAGA Ding Ding Ding G0AGGGGAAA GTATTTATGCAGCGACACCCCACAATGTGA ACAAACAGACAAAAAATCTGTGCCCTCGTGGAATTGACGTTCTGCTTGGGAACACAGAAAA GAACTCGGTGATGAAATAATGGAGATGATTCCAGTGACAAACGACAGAGATGGTGATGGTG GTCAGGGAAGACCTCTCTGCAGAGGTAGTGACTTGTGATGTGAGCTGAGACCTCTGTCCTGG GAAGACCAAAAGAAAAGCATTTCAGGATGAGGGAATGGCATGCGCAAAGGCCCTGAGGCT GAAATGTGCCCATGTGTTCTAAGAAATGCAGCGATGCTGGTGTGCCTGGAGCAGGGACGGA GGGGG AGAA Ding Ding Ding GGGAGGAGACAAGGAGC butoxy GAAG butyl AGTTCCCGAAGGACC GTGGGTGATA butoxy AGAGGACTTCGCTTTTGCTCTGAATGAGGTGGGAGCCCATAGAAGCTTCTAAGCAGAAGA AGGACTTGCCCTAATTCAAGTGATCACAGGTGTCTTGTGGCCTCCATGGGAGGTTGAAAACC CAGAAAAGGTGAANGGGGGCTTGCACTTGAGCCCCAGGAACCATGATGGAGATTCCACTTA agcccaaaaccccgnggattcttagatagattttaaaggcancagaccgnaattactggag GAATrGAGNGTAAAAGTGGGAATAAGGTTTTCAAGGGCCATTGGCCAANGGTGGGGCACCC CCCAAATTTGACTTTTGGGAAAATTAGCCNAATCCNATNTGGNAANAAAATTTNTTTTTTAT ΤΤΤΠΓΝΑΑΑΑΑΑΛΑΑΑΑΑΑΑΑΑΑΛΛΛΑΑΑΑΑΑΑΑΑΑΑΑΑΑΑΑΑΑΑΑΑΑΑΑΑΑΑΑΑΑΑΑΑΑ 2. The patent application meganuclease, Paragraph 1, wherein the SEQ ID NO: The nucleotide sequence encoding SEQ 1 ID NO: 2 The amine Acid sequence, SEQ ID NO: 2 MESGLLRPAPVSEVIVL-HYNYTGKLRGARVQPGAGLRADAVVCLAVCAFIVLENLAVLLVLCR 1AJLERSL Ding MARRGPAPVSSRGRTLAMAAAAWGVSLLLGLLPALGWNCLGRLDACSTVLPLYAK AYVLFCVLAFVGILAAICALYARIYCOVRANARRLPARPGTAGTTSTRARRKPRSLALLRTLSVV 1_.1^ΑΡνΑ(:\ν6ΡυΡΙΙΧ^ί〇νΑΟΡΑΚΤΊ:Ρνυ^〇ΑΟΡ}=Ί>Χ31_ΑΜΑΝ$ϋ^ΝΡ>]1ΥΤΙΤΝΙΙΟίϊυ·1Αΐ^Κ1^ν CCGRHSCGRDPSGSQQSASAAEASGGLRRCLPPGLDGSFSGSERSSPQRDGLD Ding SGSTGSPGAH. AARTLVSEPAAD This paper scale applies to China National Standard (CNS) A4 specification (21 OX297 mm) -2 - (Please read the back note first and then fill in This page), 1T •^^1. Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative printing 1250209 Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative printing A8 B8 C8 D8 VI. Patent application scope 3. 3 kinds of isolated nucleic acids, It consists of a nucleotide sequence which hybridizes to the nucleic acid of claim 1 or its complementary sequence under the following conditions: a solution of 6X sodium chloride/sodium citrate (SSC) at about 45 ° C. The hybridization reaction was carried out, followed by rinsing in a 0.2X SSC, 0.1% SDS solution at 50 to 65QC. 4. A purified polypeptide consisting of the amino acid sequence of SEQ ID NO: 2. 5. A purified polypeptide consisting of the amino acid sequence of amino acids 214 to 252 of SEQ ID NO: 2, which forms the first intracellular domain of human GAVE8. 6. A recombinant vector comprising the nucleic acid of claim 1 of the invention, wherein the recombinant vector contains the recombinant vector of claim 6 to express and produce functional human GAVE8. 8. A method for producing a functional human GAVE8 comprising culturing a transformant strain of claim 7 in a medium temperature at a temperature of C02 and in a medium to express the human GAVE8, and recovering Collect the resulting human GAVE8. 9. An antibody that specifically binds to human GAVE8. 10. The antibody of claim 9 is a plurality of antibodies or monoclonal antibodies. 1 1. An antibody according to claim 9 or 10 which prevents binding of a ligand to human GAVE8. (Please read the notes on the back and fill out this page) L0, 1Τ f This paper scale applies to Chinese National Standard (CNS) A4 specification (210X297 mm) -3 - 1250209 Annex 3A: Patent Application No. 9H16166 Chinese Sequence Listing January 27, 1994 <11〇> Avantis Pharmaceutical Co., Ltd. <120> Novel receptor coupled with 〇 protein GAVE8 <140>TW 091116166 <141> 2002-07-19 <150>US 60/306,434 GB 0125704.7 <151> 2001-07-20 2001-10-26 <160> 11 <210> SEQ ID NO: 1 <211> 2589 nucleotide acids <212>cDNA <213>human <400>; SEQ TD NO: 1 GCCTAGACAGTGGTTCAAAGTTTTTTTCTTCCATTTCAGGTGTCGTGAAAAGCTTGAATTCGG CGCGCCAGATATCACACGTGCCAAGGGGCTGGCTCAGGCCTGAGCCACTGCCTCAGGCCTG AGCCTGGGGGGAfTACAGGCGGGGATTACAGGCCTGAGCCACCGCGCCCAGCCATGTGTGA ACCACCGCGCCCAGCCATTCATGTGTTATTAATGTCCCTTTTTTTGGGACTATGTCACCTTGT TCCCTCGTGATATTATTTGTGTGTATGTCTGTCTCCACGGACTCTATCCCTCCATCCTGAACT CCCCCTTGGAGACCATGTATAGGGGGTGATGGTCAGGGGGTAGGGCCCCTTTCCCACGACTT AGCCGGCTGCTGCGAGCGTGCTTACGGGGACCGCGGCCTGACACCGTATCTTGCCTCTCCAA CAGCCTTGGGGCGCGCGGCCCATGGAGTCGGGGCTGCTGCGGCCGGCGCCGGTGAGCGAGG TCATCGTCCTGCATTACAACTACACCGGCAAGCTCCGCGGTGCGCGCTACCAGCCGGGTGCC: GGCCTGCGCGCCGACGCCGTGGTGTGCCTGGCGGTGTGCGCCTTCATCGTGCTAGAGAATCT AGCCGTGTTGTTGGTGCTCGGACGCCACCCGCGCTTCCACGCTCCCATGTTCCrGCTCCTGG GCAGCCTCACGTTGTCGGATCtGCTGGCAGGCGCCGCCTACGCCGCCAACATCCTACTGTCG gggccgctcacgctgAaactgtcccccgcgctctggttcgcacgggagggaggcgtcttcgt GGCACTCACTGCGTCCGTGCTGAGCCTCCTGGCCATCGCGCTGGAGCGCAGCCTCACCATGG CGCGCAGGGGGCCCGCGCCCGTCTCCAGTCGGGGGCGCACGCTGGCGATGGCAGCCGCGGC CTGGGGCGTGTCGCTGCTCCTCGGGCTCCTGCCAGCGCTGGGCTGGAATTGCCrGGGTCGCC TGGACGCTTGCTCCACTGTCTTGCCGCTCTACGCCAAGGCCTACGTGCTCTTCTGCGTGCTCG CCTTCGTGGGCATCCTGGCCGCGATCTGTGCACTCTACGCGCGCATCTACTGCCAGGTACGC GCCAACGCGCGGCGCCTGCCGGCACGGCCCGGGACTGCGGGGACCACCTCGACCCGGGCGC GTCGCAAGCCGCGCTCGCTGGCCTTGCTGCGCACGCTCAGCGTGGTGCTCCTGGCCTTTGTG GCATGTTGGGGCCCCCTCTTCCTGCTGCTGTTGCTCGACGTGGCGTGCCCGGCGCGCACCTG TCCTGTACTCCTGCAGGCCGATCCCTTCCTGGGACTGGCCATGGCCAACTCACTTCTGAACC CCATCATCTACACGCTGACCAACCGCGACCTGCGCCACGCGCTCCTGCGCCTGGTCTGCTGC GGACGCCACTCCTGCGGCAGAGACCCGAGTGGCTCCCA GCAGTCGGCGAGCGCGGCTGAGG CTTCCGGGGGCCTGCGCCGCTGCCTGCCCCCGGGCCTTGATGGGAGCTTCAGCGGCTCGGAG CGCTCATOGCCCCAGCGCGACGGGCTGGACACCAGCGGCTCCACAGGCAGCCCCGGTGCAC CCACAGCCGCCCGGACTCTGGTATCAGAACCGGCTGCAGACTGACACCCTCGGCCTACGAC 1250209 TGTCTTCCCAAGTTTrACAGACTTGTTCTTTTTACATAAAGGAATTTGTAGGAAATGCAGCCA AAGGTGCAGTCGGAAAAGATGCAGGGGAAATGTATTTATGCAGCGACACCCCACAATGTGA ACAAACAGACAAAAAATCTGTGCCCTCGTGGAATTGACGTTCTCCTTGGGAACACAGAAAA GAACTCGGTGATGAAATAATGGAGATGATTCCAGTGACAAACGACAGAGATGGTGATGGTG GTCAGGGAAGACCTCTCTGCAGAGGTAGTGACTTGTGATGTGAGCTGAGACCTCTGTCCTGG GAAGACCAAAAGAAAAGCATTTCAGGATGAGGGAATGGCATGCGCAAAGGCCCTGAGGCT GAAATGTGCCCATGTGTTCTAAGAAATGCAGCGATGCTGGTGTGCCTGGAGCAGGGACGGA GGGGGAGAATGGGAGGAGACAAGGAGCTGAAGTAGTTCCCGAAGGACCTTGTGGGTGATA TAGAGGACTTCGCTTTTGCTCTGAATGAGGTGGGAGCCCATAGAAGCTTCTAAGCAGAAGA AGGACTTGCCCTAATTCAAGTGATCACAGGTGTCTTGTGGCCTCCATGGGAGGTTGAAAACC CAGAAAAGGTGAANGGGGGCTTGCACTTGAGCCCCAGGAACCATGATGGAGATTCCACTTA AGCCCAAAACCCCGNGGATTCTTAGATAGATTTTAAAGGCANCAGACCGNAATTACTGGAG GAATTGAGNGTAAAAGTG GGAATAAGGTTTTCAAGGGCCATTGGCCAANGGTGGGGCACCC CCCAAATTTGACTTTTGGGAAAATTAGCCNAATCCNATNTGGNAANAAAATTTNTTTTTTAT -TTTTNAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA < 210 > SEQIDNO: 2 < 211 > 399 th amino acid < 212> amino acid sequence < 213> Human < 400> SEQ ID NO: 2 MESGLLRPAPVSEVIVLUYNYTGKLRGARYQPGACLRADAVVCLAVCAFIVLF.NLAVLLVLGR hprfhapmflllgsltlsdllagaayaanillsgpltlklspalwfareggvfvaltasvlslla IAL £ RSLTMARRGPAPVSSRGRTLAMAAAAWGVSLLLGLLPALGWNCLGRL. DACSTVLPLYAK AYVLFCVLAFVGILAAICALVARIYCQVRANARRLPARPGTAGTTSTRARRKPRSLALLRTLSVV LLAFVACWGPLFLLLLLDVACPARTCPVLLQADPFLGLAMANSLLNPIIYTLTNRDLRJMLLRLV CGGRHSCGRDPSGSQQSASAAEASGGLRRCLPPGLDGSFSGSERSSPQRDGLDTSGSTGSPGAPT AARTl-VSEPAAD < 210 > SEQ ID NO: 3 < 211 > 18 nucleotide < 212> DNA < 213> artificial sequence < 400 > SEQ ID NO: 3 CCATGGAGTCGGGGCTGC 1250209 < 210 & gt SEQ ID NO: 4 < 211 > 18 nucleotides < 212 > DNA < 213 > artificial sequence < 400 > SEQ ID NO: 4 TCAGTCTGCAGCCGGTTC < 210 > SEQ ID NO : 5 < 211 > 20 nucleotides <212 > DNA < 213 > artificial sequence < 400 > SEQ ID NO: 5 GCAGCAGCCCCGACTCCATG < 210 > SEQ ID NO: 6 < 211 > 20 nucleotides <212> DNA <213>Artificial sequence <400> SEQ ID NO: 6 CCATGGGCCGCGCCCCAAGG <210> SEQ ID NO: 7 <211> 18 nucleotides <212>DNA <213><400> SEQ JD NO: 7 CGCGGTGCGCGCTACCAG <210>SEQ ID NO: 8 <211> 18 nucleotides <212> DNA <213>Artificial sequence <400> SEQ ID NO: 8 GCGCCTGCCAGCAGATCC <;210> SEQ ID NO: 9 <211> 19 nucleotides 1250209 <212> DNA <213>Artificial sequence <400>SEQ Π) NO: 9 TGGACGCTTGCTCCACTGC <210> SEQ ID NO: 10 <;211>21 nucleotides <212> DNA <213>Artificial sequence<400> SEQ ID NO: 10 TTGCCGCTCTACGCCAAGGCC <210>SEQ ID NO: 11 <211> 19 nucleotides <212 〉DNA <213>Artificial sequence<400>SEQ Π) NO: 11 AGCACGCAGAAGAGCACGT