TW200300798A - Mutatant CYP2D6 genes - Google Patents

Abstract

It is intended to identify the polymorphisms of mutatant CYP2D6 genes and clarify the relationship between the thus identified gene polymorphisms and drug-reactivity (drug-metabolic ability). Namely, nucleic acids composed of from 10 to 100 bases which contain at least 10 consecutive nucleotides in the base sequence represented by SEQ ID NO:1 that is characterized at having a characteristic mutation site selected from the substitution of G to A at the base of position 125, the substitution of C to A at the base of position 1858, the substitution of T to C at the base of position2874, and the substitution of C at the base of position 2875.

Description

200300798 A7 B7 V. Description of the invention (1) 1. Technical field to which the invention belongs (please read the notes on the back before filling out this page) The present invention is a novel polymorphism of the CYP2D6 gene encoding a drug metabolic enzyme. In addition, the present invention relates to a method for determining the metabolic activity of a drug by using a polymorphic variation of the CYP2D6 gene. 2. Prior art CYP2D6 is a gene of about 4.5 kbp formed by nine exons. Most of the reported molecular types of genetic polymorphisms (Nelson, DR, Pharmacogenetics, 6, 1-42, 1996; and Daly , Α · Κ.,

Pharmacogenetics, 6, 193-201, 1996). About 7% of white races, or about 0.5% of their own lack of enzymes (poor metabolizer: PM), have been reported (Dahl, ML ·, J. Pharmacol. Exp. Ther., 274, 5 1 6-5 20,1 995 pointed out that for the white race, the haploid gene (fragment chromosome; n. General somatic chromosome 2η) has 13 repeats of the CYP2D6 multi-enzyme owner (or supermetabolizer: SM). The diversity of information and the individual difference in metabolism. However, the detailed relationship between drug metabolism and the polymorphism of the CYP2D6 gene is unknown. Printed by the Consumer Cooperatives of the Intellectual Property Bureau of the Ministry of Economic Affairs III. Summary of the Invention The CYP2D6 variant is determined to solve the problem Polymorphism of genes, to elucidate the relationship between the determined genotypes and drug reactivity (metabolic energy) or the metabolic activity of pharmaceutical agents. To solve the above problems, the inventor first examined the polymorphism of the CYP2D6 gene. For this inspection, the accuracy of the results is the most important. Therefore, the size of this paper applies the Chinese National Standard (CNS) A4 specification (210X297 mm 5. ~~~ 200300798) Cooperatives printed A7 B7_ _ V. Description of the invention (2) Direct sequencing using genomic-PCR. Also, for CYP2D6 * 5, which is completely deficient in genes, the Long-PCR method is used for testing in principle, and for positive samples, The detection was performed using the southern hybridization method. As a result, the present inventors successfully sequenced a novel multiple variant gene of the CYP2D6 gene. The present invention was completed based on this knowledge. That is, the present invention provides the following inventors. (1 ) A nucleic acid having at least 10 consecutive nucleotides in the base sequence described in SEQ ID NO: 1 and characterized in that it contains a variation in which G is replaced by A including an arbitrary 125th base selected from the base sequence The nucleoside at the polymorphic site of the 1 85 base 8 mutation C substitution of T, the 2874 base T substitution of C substitution, or the 2875 base C substitution of T substitution T A nucleic acid of 100 to 100 bases of acid. (2) A nucleic acid such as (1), which is DNA. (3) A nucleic acid such as (1), which is RNA. (4) A dual gene-specific oligo A nucleotide characterized by containing a base sequence arbitrarily selected from SEQ ID NO: 1 A variation of G from A at base 125, a variation of C from T at 1858, a variation from T at C of 2874, or C from 2875 at T The sequence of the substituted polymorphic site, or a hybrid with its complement. (5) A dual gene-specific oligonucleotide such as (4), which can be used as a probe or primer. (6) An individual Genetic polymorphism analysis method, which is characterized by containing (1) a step of preparing a nucleic acid from an individual; and (2) for the nucleic acid prepared by step (1), arbitrarily selected from the sequence number 1 Standard (milk milk) 8 4 specifications (210 father 297 mm) _6_ " (Please read the precautions on the back before filling out this page) 200300798 A7 B7 Printed by the Intellectual Property Bureau of the Ministry of Economic Affairs Consumer Cooperatives V. Invention Description (3 ) The variation of the 125th base from G to A, the 1858th base from C to T, the 2874th base from T to C, or 2875 The step of identifying the test base of the mutated polytype site in which the C of base No. is substituted by T. (7) A method for judging the metabolic activity of a drug, which is characterized by containing (1) a step of preparing a nucleic acid from an individual; (2) The nucleic acid prepared by step (1) is arbitrarily selected from the sequence number 1 Variation of the 125th base with G replaced by A, the mutation of 1 85th base with C replaced by T, the mutation of the 2874th base with T replaced by C, or 2875 Steps of sequencing the bases of the mutated polymorphic site where the C of bases is replaced by T; and (3) a step of predicting the metabolic activity of the agent based on the base information determined in step (2). (8) A gene characterized by having a base sequence arbitrarily described in SEQ ID NOs: 2 to 5. (9) A protein characterized by having an amino acid sequence described in any one of SEQ ID NOs: 2 to 5. (10) A method for detecting a gene having a base sequence described in SEQ ID NO: 2 or SEQ ID NO: 5, which is characterized in that it contains a nucleic acid prepared by an individual, and is 100th in the base sequence described in SEQ ID NO: 1. A step of detecting the presence or absence of a variation in which C is replaced by T at base 2850, C is replaced by T at base 2850, and G is replaced by C at base 4180. Fourth, implementation mode ---- *-* ----- (Please read the precautions on the back before filling in this page)

、 1T This paper size is in accordance with Chinese National Standard (CNS) A4 specification (210 X 297 mm L 7 200300798 A7 ___—___ B7__ V. Description of the invention (4) The best form for implementing the invention (I) The nucleic acid of the invention (please (Please read the notes on the back before filling in this page) The present invention provides a nucleic acid whose at least 10 consecutive nucleotides in the base sequence described in SEQ ID NO: 1 are characterized by containing The variation of G at base 125 is replaced by A, the variation of C at base 1858 is replaced by T, the variation at base 2874 is replaced by C, or the variation at base 2875 is replaced by T A nucleic acid of 10 to 100 bases of the nucleotide of the substituted polymorphic site. For this nucleic acid, the base at the polymorphic site is selected from the base sequence described in SEQ ID NO: 1 Variation of G from A to 25 in base sequence, mutation of C from T to 1 85 base, mutation of T from C to base 2874, or 2875 Variation of base C with T replaced. The polymorphic site contains bases related to the metabolic activity of the drug. Printed by the Consumer Cooperative of the Intellectual Property Bureau of the Ministry of Economic Affairs The present invention further provides a dual gene-specific oligonucleotide, which is characterized by containing a G selected from bases 1 to 25 of the base sequence described in SEQ ID NO: 1 Polymorphisms with A substitutions, 1 858 bases with C replaced by T, 2 874 bases with T replaced by C, or 2875 bases with C replaced by T The sequence of the site, or a person who hybridizes with its complement. Such oligonucleotides can be used as probes or primers. The so-called nucleic acid in the description of the present invention may be DNA or RNA, single-stranded or double-stranded. Nucleotides are those that exist in nature or synthesizers, but they are generally synthesizers. The preferred nucleic acid of the present invention is a ^ 's Zhang scale containing any of the sequences sequenced by the present invention above 1 applicable to the Chinese National Standard (CNS) A4 specifications (210X297 mm;) _ 8 _ ~ 200300798 A7 ____B7 V. Description of the invention (5) (Please read the precautions on the back before filling this page) DN A fragment of the polytype site or the complement, the length is generally It is 10 to 100 consecutive bases. Many The site can exist anywhere in the fragment. For sequence number 1, the symbol T is used for both thymine in DNA and uracil in RNA, so the symbol T in RNA oligonucleotides is interpreted as uracil. Residues. Hybridization probes can be specifically bonded to the complementary strand of a nucleic acid. Examples of such probes include nucleic acids and peptide nucleic acids. The so-called primers have four phases in an appropriate buffer and temperature. In the case of heteronucleoside triphosphates and nucleic acid polymerases (DNA polymerase, RNA polymerase, reverse transcriptase, etc.), they can be single-stranded oligonucleotides that act as specific starting positions for DNA synthesis on the template. The proper length of the primer varies depending on the purpose of use, but is generally in the range of 10 to 50 nucleotides. The so-called polytype is one in which two or more gene-determined replacement sequences or dual genes exist in one group. Polymorphic markers or sites indicate the location of genes that are diverse. The preferred markers have at least two dual genes, which are more frequent than the selected group 1%, respectively, and are preferably present at a frequency of 10% or more. Polymorphic gene positions may also be 1 base pair. Printed by the Consumer Cooperative of the Intellectual Property Bureau of the Ministry of Economic Affairs The nucleic acid of the present invention as described above can be easily synthesized based on the base sequence information described in SEQ ID NO: 1 disclosed in this specification by a well-known nucleic acid synthesis method. The present invention further provides a kit comprising at least one dual gene-specific oligonucleotide as described above. The kits were hybridized under polymorphic heterogeneity and containing one or more pairs of pair-specific oligonucleotides. For several sets, dual-gene specific oligonucleotides can be immobilized on the support. The paper size is applicable to China National Standards (CNS) A4 (210X297 mm) g 200300798 A7 Employees ’Cooperatives, Intellectual Property Bureau, Ministry of Economic Affairs Printing _B7_ V. Description of the invention (6) Those with status. As any ingredient contained in the kit, such as restriction enzymes, reverse transcriptase or polymerase or matrix nucleoside triphosphates, methods used for labeling (such as avidin-enzyme binding bodies and enzyme substrates and labeling as biotin Examples include chromosomes), and suitable buffers for reverse transcription, PCR, or hybridization reactions. It also includes a kit and instructions for performing the method. (II) The method for analyzing genotypes of the present invention The present invention provides a method for analyzing the genotype of individuals, which is characterized by containing (1) a step of preparing a nucleic acid from an individual; and (2) For step (1), The prepared nucleic acid 'is arbitrarily selected from the variation in which the G at 125th base is replaced by A, the mutation at 1 85th base C is replaced by T, and the base 2874 A sequence in which the T of the base is replaced by C, or the sequence of the base of the polytype site of the mutation in which the C of T at 2875 is replaced by T. A specific form of the analysis method of the present invention will be described below. (A) Nucleic acid modulation The polymorphism in the present invention is detected by the target nucleic acid prepared by the analyzed individual. For genomic DNA analysis, any biological sample (except pure red blood cells) is suitable. For example, as the tissue sample, whole blood, semen, saliva, tears, urine, feces, sweat, buccal, skin, and hair can be used. Tissue samples for cDNA or mRNA analysis can be obtained from organs expressing the target nucleic acid. This paper size applies Chinese national standard (milk) 8 4 specifications (210 '/ 297 mm) _1〇_ one (please read the precautions on the back before filling this page) 200300798 Α7 Β7 V. Description of the invention (7) ( (Please read the notes on the back before filling this page.) When implementing the method of the present invention, DNA amplification must be performed on the target sample, which can be performed by, for example, PCR. Literature on PCR can be found in Principles and Applications for DNA Amplification (HA Edited by Erlich, Freeman Press, NY, NY, 1 992); PCR Protocols: A Guide to Methods and Applications (Edited by Innis, Academic Press, San Diego, CA, 1990); Mattila, Nucleic Acids Res. 19, 4967 (1 99 1); Eckert, PCR Methods and Applications 1, 17 (1991); PCR (edited by McPhersons, IRL Press, Oxford); and U.S. Patent No. 4,683,202. As other amplification methods can be cited, connected Enzyme chain reaction (LCR) (see Wu and Wallace, Genomics 4,560 (1989), Landegren's,

Science 241, 1 077 (1 988)), transcription amplification (Kwoh's, Proc. Natl. Acad. Sci. USA 86, 1 1 73 (1989)), and self-sustained sequence replication (Guatelli's, Proc. Natl. Acad. Sci. USA, 87, 1 874 (1990)), and nucleic acid-based sequence amplification (NASBA). (B) Detection of the polymorphism of the target DNA. The base sequence described in Sequence No. 1 printed by the Employees' Cooperative of the Intellectual Property Bureau of the Ministry of Economic Affairs was selected from the 125th base in which G is replaced by A. The polymorphic site of the variation in which C is replaced by T at base No. 2874, the mutation at which T is replaced by C at base 2874, or the mutation at which C is replaced by T at base 2875 is in an individual (for example, the analyzed Patients) can be sequenced by any of the following methods (i) to (vi), for example. (i) Dual gene-specific probes The dimensions of this paper are in accordance with Chinese National Standard (CNS) A4 (210X297 mm) _ ^ " ~ " ~ 200300798 A7 B7 V. Description of Invention (8) (Please read the back Please note this page, please fill in this page) Analysis of the design and use of dual gene-specific probes for the polymorphism of the present invention, such as Saiki et al., Nature 324, 163-166 (1986); Dattagupta, EP235,726; Saiki, WO89 / 1 1 548. Designed to hybridize on DNA fragments that are targeted by a certain body, but not on corresponding fragments from other individuals (the reason is that there are different polymorphic forms on the fragments from the two individuals) Dual gene-specific probes. The hybridization condition is that the intensity of hybridization between the dual genes is significantly poor, so the probe must be very rigorous to hybridize to only one of the dual genes. Several probes were hybridized with the target DNA fragment. As a result, the polymorphic site was designed to be arranged at the center of the probe (for example, 15 at 7th position; 16 at either 8 or 9) position). Due to the proper design of probes, hybridization between disparate dual genotypes can be easily discerned. Dual gene-specific probes are intended for users. One side of the pair shows a complete agreement with the control group of the target sequence, and the other side shows a complete agreement with the altered type. For simultaneous analysis of plural polymorphisms in the same target sequence, the plural pairs of probes can be immobilized on the same support. Printed by the Consumer Cooperatives of the Intellectual Property Bureau of the Ministry of Economic Affairs (ii) Tile-like arrays The polymorphism of the present invention can be sequenced by hybridization of nucleic acid arrays. Specific examples include those described in International Publication No. WO95 / 1 1 995. The subarray optimized for the detection of polymorphism in International Publication WO 95/1 1995 can also be used. (Hi) Dual gene-specific primers The paper size applies the Chinese National Standard (CNS) A4 specification (210X297 mm) _ 12-200300798 A7 B7 Printed by the Consumer Cooperative of the Intellectual Property Bureau of the Ministry of Economic Affairs 5. Description of the invention (9) Dual gene Specific primers that hybridize to the DN A site of the polymorphic repeat target, causing an increase in the dual genotype (which shows complete complementarity to this primer) (see Gibbs, Nucleic Acid Res. 17, 2427-2448 (1 989) ). This primer can be used in combination with a second primer that is distantly hybridized. The increase is the production of a detectable product in the presence of a specific dual genotype, starting with 2 primers. Generally use the second primer pair for control. One side of this pair shows a 1-base miss-match at the polytype site, and the other side shows complete complementation with the distant site. A mismatch of 1 base prevents the amplification from proceeding and cannot form a detectable product. In this method, the best effect is obtained when the mismatch is contained at the 3'-position of the polymorphic and aligned oligonucleotides. See, for example, International Publication W093 / 22456. (iv) Direct sequence sequencing The polymorphism of the present invention can be detected by direct sequence sequencing using a well-known sequence sequencing method such as the dideoxy chain termination method or the Maxam-Gilbert method. (v) Denaturing gradient colloid electrophoresis The amplified products produced by polymerase chain reaction can be analyzed by denaturing gradient colloid electrophoresis. At this time, the different dual genes are identified based on the melting characteristics and the electrophoretic mobility of DNA determined by the different sequences in the solution. Erlich, PCR Technology, Principles and Applications for DNA Amplification (W. H. Freeman and Co., New York, 1992), Chapter 7. (Please read the notes on the back before filling this page)

、 1T This paper size applies Chinese National Standard (CNS) A4 specifications (210X297 mm j 13 200300798 A7 B7 Printed by the Consumer Cooperatives of the Intellectual Property Bureau of the Ministry of Economic Affairs 5. Description of the invention (10) (vi) Multiple single-conformation (conformation) Genotyping analysis classifies the dual genes of the target sequence using single-strand conformation polymorphism analysis. This method is described in Orita et al., Proc. Natl. Acad. Sci. 86, 2766-2770 (1 989). The change in electrophoretic mobility of strand PCR products can identify base differences and form single-stranded amplification products. Single-stranded nucleic acids will again fold or form partly depending on the secondary structure of the base sequence. Different from single-stranded amplification products The electrophoretic mobility reflects the difference in the base sequence between the dual genes of the target sequence. (III) Utilization of the method for analyzing polymorphism in the present invention The present invention relates to a novel polymorphism of the CYP2D6 gene encoding a drug metabolic enzyme The detection of the polymorphism of the CYP2D6 gene can determine the metabolic activity of the agent. That is, in the present invention, the method for analyzing the polymorphism of the present invention as described above further includes the method according to step (2) Sequenced base information can predict the metabolic activity of the individual and provide a method for determining the metabolic activity of the pharmaceutical. The polymorphism of the present invention is used in general applications (for example, forensic science, blood identification, linkage analysis, and (Position selection). &Amp; Next 'will make a detailed description of the analysis method using the gene polymorphism of the present invention. (I) The relationship with the metabolic activity of the drug $ The invention provides a CYP2D6 that can predict the metabolic activity of the drug ---- .—— * --- I — (Please read the notes on the back before filling this page)

、 1T This paper size applies Chinese National Standard (CNS) M specification (21GX297 mm) 200300798 A7 B7 V. Description of invention (11) (Please read the precautions on the back before filling this page) Gene novel polymorphic site. Specific examples of the medicament prepared from the CYP2D6 matrix of the polymorphism predictive of metabolic activity of the present invention include the following. By analyzing the subject of the polymorph of the present invention, the metabolic activity of the following agents can be judged. Arrhythmia therapeutics · Na channel blockers (eg, propafenone HC1) A rapid heart rate arrhythmia (eg, flee a nide acetate) Arrhythmia (eg, amarin, mexlitine HC1, cibenzoline citric acid ) / 5-Receives beneficial blocking drugs (eg, timolol maleate, propranolol HCl, penbuolol sulfate, bupranolol HC1, alprenolol HC1) cardiac selective A 1-blocking agents (eg, metopr ο 1 ο 1 tartrate) 々-Blocker (for example, malonic acid p 〇pind ο 1 ο 1) a, / 5-blocker (for example, carvedi 1 ο 1) improvement in urination hindering • hypotensive agent (for example, urapidil) 嗓 _ throat Psychiatric neuroleptics (eg, perphenazine) Discretionary _ Psychopsychiatric agents (eg, fluphenazine, thioridazine HC1) Printed bromide antipsychotics (eg, ha 1 operid ο 1) ) Bicyclic agonist modulators (eg, nortriptyline HC1) Bicyclic antidepressants (eg, amitriptyline HC1) Anxiety • Enuresis Therapeutics (eg, clomiprami ne HCl, imipramine HC1) Depressive medicine (for example, mainserin HC1) Anesthetic cough suppressant (for example, phosphate codeine) This paper size applies Chinese National Standard (CNS) A4 specifications (210X297 mm: [15 ^ 200300798 A7 ____ B7 _ 5. Description of the invention (12) Central acute cough (eg, dextromethorphan hydrobromide) Cerebral circulation and metabolism improving agent (eg, nicergorine) (Please read the precautions on the back before filling this page) Selective heparin Recycling inhibitors (eg, fluboxamine maleate) glaucoma • IOP treatments (eg, tim ο 1 ο 1 ma 1 eate) Antihistamines (eg, promethazine HC1) (ii) Association with various diseases Concerning the polymorphism of the present invention, various diseases whose genes are not identified (e.g., agammaglobulinemia, diabetes insipidus, Lech-Niegen syndrome, myodystrophy, Wiskott-Aldrich syndrome, Fabry's disease, familial high cholesterol Disease, polycystic kidney disease, genetic spherocytosis, Willebrand's disease, ductal sclerosis, genetic hemorrhagic peripheral vasodilation, Ethnicity meat hysteria colon disease, Ehlers-Danlos syndrome, skeletal development is not the whole disease, and acute intermittent film slope disease) were tested. Printed as a phenotype by the Consumer Cooperative of the Intellectual Property Bureau of the Ministry of Economic Affairs, including symptoms of or related to multifunctional diseases (eg, autoimmune diseases, inflammation, cancer, and infection by pathogenic microorganisms) that may be genetically related Susceptibility, etc. Examples of autoimmune diseases include chronic rheumatoid arthritis, general sclerosis, diabetes (insulin-dependent and non-dependent) generalized lupus, and hyperthyroidism. Examples of cancer include cancers of the bladder, brain, breast, large intestine, esophagus, kidney, leukemia, liver, lung, oral cavity, ovary, pancreas, prostate, skin, stomach, and uterus. Phenotypes can be characterized by longevity, appearance (eg, baldness, obesity), strength, agility, endurance, reproductive ability, and so on. This paper size applies to China National Standard (CNS) A4 specifications (210X297 mm; L 16 _ 200300798 A7 _ B7 V. Description of invention (13) (iii) Forensic science (please read the precautions on the back before filling this page)) at Identification of the polymorphism of the polymorphic site setting of an individual can identify the polymorphism setting that distinguishes the individual. Generally refer to the National Research Council, The Evaluation of Forensic DNA Evidence (edited by Pollars, National Academy Press, DC , 1996). The more analysis sites, the lower the probability of the same type of irrelevant individuals for the polymorphism setting of a certain body. Therefore, the polymorphism of the present invention can be used in combination with the polymorphism of distant genes. The better polymorphism used in forensic science is the dual dual gene. The reason is that the group frequency of the two polymorphs is generally more accurately detected than those with multiple polymorphs at the position of the dual dual genes. The identification capability set by forensic labeling printed by individuals in the Consumer Cooperatives of the Intellectual Property Bureau of the Ministry of Economic Affairs can be used for forensic analysis. For example, testing blood samples obtained from suspicious criminals. Whether it is consistent with the blood or other tissue samples at the crime scene is determined by the polytype setting occupying the selected polytype site to determine whether the suspect is the same as the sample. The polytype marker is set between the suspect and the sample. When they are the same, it can be judged that the suspect is not the source of the sample. If the setting is the same as the marker, it can be judged that the DNA obtained by the suspect is the same as that seen at the crime scene. Judgment (for example, group analysis suitable for individuals) ) The polymorphic frequency of the tested gene positions can be statistically analyzed to determine whether the suspect and the crime scene samples are accidental probabilities. (Iv) Paternity tests The purpose of paternity tests is generally to determine whether the male is the father of the child. Paper size applies Chinese National Standard (CNS) A4 specification (210X297 mm) _ '200300798 A 7 B7 Printed by the Consumer Cooperatives of the Intellectual Property Bureau of the Ministry of Economic Affairs 5. Description of the invention (under normal circumstances, the mother of the child is known and therefore traceable The genotype of the child from the mother. The paternal test is the genotype of the child that is not from the mother. The investigation is made up by the father. The paternity test can be carried out by analyzing the polymorphism setting of the father and the child. If the polymorphism setting of the child from the father is not consistent with the presumption that it is a father, there can be no test error. The conclusion is that the presumption is that the father is not a true father. If the polymorphism of the child from the father is set to be 'consistent with the presumption as a father, the chance coincidence probability can be calculated statistically. Several polytype gene positions include At the time of this analysis, the 'excluded cumulative probability of a randomly selected male was very high. This probability' could be used to assess the responsibility of a father who has a polytype identification setting that is consistent with the child polytype identification setting from the child's father. (v) Phenotypic shape gene map Using the polymorphism of the present invention, it is possible to establish a polymorphic identity of gene positions that are related to the target shape (although this is not related to shape, but is connected to physical gene positions that serve the shape and are isolated at the same time) ) Physical relationship. In this analysis, it is useful to map the position of a chromosome and the position of a gene related to a phenotype, and cloning of the morphogene can be performed through this. With reference to Lander et al., Proc. Natl. Acad. Sci. (USA) 83,7353-7357 (1986); Lander et al., Proc. Natl. Acad. Sci. (USA) 84, 2363-2367 (1 987 ); Donis-Kell et al., Cell 5 1,3 1 9-337 (19 87); Lander et al., Genetics 1 21. 1 8 5- 1 99 (1989). Related configuration genes can be selected by directional cloning. Refer to (Please read the precautions on the back before filling this page) This paper size applies Chinese National Standard (CNS) A4 specification (210X297 mm) _ 18 200300798 A7 B7 Printed by the Consumers ’Cooperative of Intellectual Property Bureau of the Ministry of Economic Affairs (15) Wainwright, Med. J. Australia 159, 170-174 (1993); Collins, Nature Genetics 1, 3-6 (1992) 0 Linkage studies can be performed, for example, in families. As to whether the family members have phenotypic characteristics, the setting of multi-type logos can be investigated. Second, analyze the distribution of meiotic polymorphic markers to determine what polymorphic markers are separated from the phenotype at the same time. See, for example, Kerem et al., Science 245, 1 073-1 080 (1 989); Monaco et al., Nature 316, 842 (1 985); Yamoka et al., Neurology 40, 222-226 (1 990) Rossiter et al., FASEB Journal 5, 21 -27 (1991). IV. Genes and Proteins of the Present Invention The present invention provides genes comprising a test sequence of any of SEQ ID NOs: 2 to 5. Moreover, the gene of the present invention can be expressed naturally or in a recombinant expression vector that can be linked to other promoters. Generally, the promoter is a eukaryotic promoter that makes it behave in mammalian cells. As the transcription regulating sequence, a heterologous promoter violet, if necessary, an enhancer recognized by the host, and the like can be used. Depending on the host selected, an appropriate promoter can be selected (e.g., trp, lac, phage promoter, glycolytic promoter, and tRN A promoter). Commercially available expression vectors can be used. The expression vector may contain, for example, identification of the host's replication line, genes that can be amplified, selective identification, useful host sequences inserted into the host gene group, and the like. The method of introducing the recombinant surface vector into a host cell can be appropriately selected according to the expression vector and the host cell. Specific examples of the introduction method include fusion method, conjugation, transfaction, transformation, and electricity (please read the precautions on the back before filling this page) This paper size applies the Chinese National Standard (CNS) A4 specification (210X 297 mm) _ 19-200300798 A7 _B7_ 5. Explanation of the invention (ie) Penetration or injection. (Please read the notes on the back before filling out this page) Various host cells can be used for the expression of the gene of the present invention. Examples of suitable host cells include bacteria such as Escherichia coli, yeasts, filamentous fungi, insect cells, and mammalian cells (representatively, immortalized, for example, mouse, CHO, human, and monkey cells). Strains), and their derivatives. The transformed host cell under the recombinant expression vector containing the gene of the present invention is cultured under appropriate conditions, and the produced protein is isolated according to a method known to a person skilled in the art to obtain a product that is actually purified. In the case of protein secreted extracellularly, the protein is extracted from the culture solution of the host cell. The protein is not secreted outside the cell, and the protein is isolated from the lysate of the host cell. Printed by the Consumer Cooperative of the Intellectual Property Bureau of the Ministry of Economic Affairs as shown in Example 2 below, the enzyme activity of CYP2D6unkn0wS3 with the present invention is 5% or less when compared with the wild-type enzyme activity. Individuals with enzyme genes have shown to be less active than CYP2D6. It is presumed that CYP2D6 is a compound responsible for the enzyme responsible for metabolism. Compared with individuals with wild-type genes, it has a considerable relationship that it can remain in the body for a long time. In addition, when three amino acid residues are substituted in the comparison between the polytype gene and the wild type, it can be presumed that the matrix variability will behave differently from the wild type. Therefore, microstereometry can be used as a protein of the present invention, and its medicinal potency and side effects can be predicted by measuring the drug's metabolic energy. V. The size of the genetic test with the base sequence described in SEQ ID No. 2 or SEQ ID No. 5 is based on the Chinese National Standard (CNS) A4 (210X297 mm) _ ~ '200300798 A 7 B7 Consumption by the Intellectual Property Bureau of the Ministry of Economic Affairs Printed by the cooperative V. Description of the invention (17) The detection method detects whether the base No. 100 in the base sequence described in SEQ ID NO: 1 is replaced by C to τ in the nucleic acid prepared by the solid in the present invention. For the mutation, whether the base of No. 2850 is replaced by C and the mutation of T, and whether the base of No. 41 80 is replaced by G for C. Design appropriate primers. Use primers to perform PCR. The design of the primer was performed as follows. The translation initiation site of the gene containing the intron of the CYP2D6 gene is set as the first position, and the 100th base toward the end of the translation site is created as the end, and the 20-30 base site is designed toward the end of the translation site. Cheng Zhi primers are two types of upstream primers. One is a 3'-terminus as a wild-type sequence (100fw), and the other is a variant sequence (100fw). Two types of primers made from 20 to 30 bases designed as the 3, -end towards the start of translation were created at the 2850th position as downstream primers. One is a 3'-end as a wild type (2850rw), and the other is a variant sequence (2850m). The 2850th position is used as the 3, -end towards the 20 to 30 bases designed for translation. Two types of upstream primers. One is the 3'-end as the wild type (2850fw), and the other is the variant sequence (2850fm). The 41st and 80th positions are made as 20, 30 bases designed for the 3, -end towards the start of translation. Primers are two types of downstream primers. One is the 3'-end as a wild type (4180 rw) and the other is a variant sequence (4180 rm). The primers described above are as follows. 100fw 5 'acgctgggctgcacgctacc 3' (serial number 45) 100fm 5 'acgctgggctgcacgctact 3' (serial number 46) 2850fw 5 'agcttcaatgatgagaacctgc 3' (serial number 47) (Please read the notes on the back before filling in this page) China National Standard (CNS) A4 specification (210X297 mm) _ 21 200300798 A7 B7 V. Description of the invention (is) 2850 fm 5 'agcttcaatgatgagaacctgt 3' (serial number 48) 2850rw 5 'aggtcagccaccactatgcg 3' (serial number 49) (Please ask Read the notes on the back before filling this page} 2850rm 5 'aggtcagccaccactatgca 3' (serial number 50) 4180rw 5 'aagctcatagggggatgggc 3, sequence number 51) 4180rm 5' aagctcatagggggatatgggg 3 '(sequence number 52) As a model, combine the above Primers are used for PCR reactions. The combination is as follows. ① 100fw and 2850 rw ② 100fw and 2850rm ③ 1OOfm and 2850rw ④ 100fm and 2850rm ⑤ 2850fw and 4180rw ⑥ 2850fw and 41 80rm ⑦ 2850fm and 4180rw ⑧ 2850fm and 4180rm The Ministry of Economic Affairs Bureau of Intellectual Property Bureau has at least one side of the consumer cooperative to print the genome DNA. In the case of a gene having the base sequence described in SEQ ID NO: 2 (unk η ω 0 η 3 in the following examples), the combination ④ and ⑧ increase simultaneously. When a gene having at least one side of the genome DN A having the test sequence described in SEQ ID NO: 2 (unknown6 in the following example), the combination ③ and ⑤ are increased simultaneously. Therefore, the novel gene of the present invention can be detected by analyzing the amplified product by electrophoresis or the like. The paper size of the Japanese patent application based on the priority claimed in the present invention applies to the Chinese national standard (CNS) A4 specification (210X297 mm) _ 22 200300798 Μ B7_ one _ five, the invention's special wish (19) All the contents described in the specification No. 200 1 -372548 are incorporated in the specification of the present invention as a part of the specification of the present invention. (Please read the notes on the back before filling out this page) The following describes the present invention in more detail through examples, but it is not limited to the examples of the present invention. Examples Example 1: (I) Materials and methods (1) Test materials As test samples, frozen blood was used. (2) Main materials and reagents used

TaqDNA polymerase (Brom Mecca) Deoxy ATP, GTP, TTP, CTP (Amacherfushi Company) Agarose (Takara Shuzo)

EtBr Staining Solution (Wako Pure Chemical Industries) Circulation Sequencing Kit (Amarchevlucia) Printed by the Consumer Cooperative of the Intellectual Property Bureau of the Ministry of Economic Affairs (3) The main electrophoresis device used by the company (Xabio) Heating cycle device (PE Ableite Biochemical Co., Ltd.) ABI automatic sequencer 373 (PE Ableite Biochemical Co., Ltd.) Cooling centrifuge (Sakuma Seisakusho) (4) Primer synthesis Primers: Based on CYP2D6 gene information, design primer sequences and perform After searching for the sameness using well-known data, the paper standard used by Semedia is applicable to the Chinese National Standard (CNS) A4 specification (210X297 mm) _ 200300798 A7 B7 Printed by the Employees ’Cooperative of the Intellectual Property Bureau of the Ministry of Economic Affairs V. Description of the invention (20) Completion. Primer sequence (5% 3 ') CYP2D6elf gcaaaggccatcatcagctcc (order 歹 U number 6) CYP2D6elr tctggtaggggagcctcagc (order 歹 J number 7) CYP2D6e2f taggacctggg (sequence number 8) CYP2cc2D6e6 ) CYP2D6e3r ggtgggagatgcgggtaagg (sequence bad Shu J number 11) CYP2D6e4f aaagcgggaactgggaaggc (sequence bad [J number 12) CYP2D6e4r tgctcttccgaggccccagt (sequence bad [1 number 13) CYP2D6e5f gagatggctggggcctgaga (sequence number 14) CYP2D6e5r ccaaccttttgcccagcctc (sequence bad [J number 15) CYP2D6e6f cccgttctgtcccgagtatg (sequence number 16) CYP2D6e6r ggtgtcccagcaaagttcat (sequence bad ij number 17) CYP2D6e7f taagcctgacctcctccaacat (sequence number 18) CYP2D6e7r agtgggcccacctggcagta (sequence bad [J number 19) CYP2D6e8f ccccgtgtgtttggtggcag (sequence number 20) CYP2D6e8r cgtcagtgagcctggctcct (sequence bad [J number 21) CYP2D6e9f tcttgcaggggtatcaccca (Sequence (J number 22) CYP2D6e9r-l ttgccctgaggaggatgatc (serial number 23) CYP2D6e9r-2 ctcagcctcaacgtacccct (serial number 24) CYP2D6 elf-2 catttggtagtgaggcaggt (serial number 25) CYP2D6elr_2 ctccaggacctcctccctc (order ij number 26) CYP2D6e2f_2 ggccctgaccctccctctctgc (serial number 27) (Please read the notes on the back before filling out this page)-Binding and ordering

This paper size is applicable to Chinese National Standard (cns) A4 (210X297mm L

200300798 A7 B7 V. Description of the invention (21) CYP2D6e2r-2 ctctgtccccaccgctgctt (serial number 28) CYP2D6e3f-2 tgcccgtcccaccccc (serial number 29) (Please read the precautions on the back before filling out this page) CYP2D6e3r-2 gccctcctgccccatccc CYP2D6e4f-2 cccgcatctcccaccccc (sequence number 31) CYP2D6e4r-2 cctcggtctctcgctccgc (sequence number 32) CYP2D6e5f-2 gaccccgttctgtctggtgt (sequence number 33) CYP2D6e5r-2 ccgtggcagccactctc (sequence number 34) CYP2D6e6f_2 gtatgctctcggccctgctc (sequence number 35) CYP2D6e6r-2 actgtttcccagatgggctc (sequence number 36) CYP2D6e7f-2 gtggggacgcatgtctgtcc (Serial number 37) CYP2D6e7r-2 ggagggcgccaggcct (Serial number 38) CYP2D6e8f-2 tcaccctgcatctcctgccc (Serial number 39) CYP2D6e8r-2 ccDccccccccgg3ccccccgggcc tggggactaggtaccccatt (serial number 42) CYP2D6 * 5f gcgacaattcaagtgtggtga (serial number 43) CYP2D6 * 5r gcatgagctaaggcacccaga (serial number 44) Printed by the Intellectual Property Office Staff Consumer Cooperative (II) Method (1) Gene Preparation of group DNA 5ml of rapid dissolution sample was divided into 200 // L and 4ml each into test tubes of appropriate volume. 200 // L of sample was added with 3 times the volume of PBND buffer (PCR buffer with Nonionic Detergents: 50mmol / L KC1, 10mmol / L Tris-HCl (pH8.3), 2.5mmol / L MgCh, O.lml / mL gelation , This paper size applies the Chinese National Standard (CNS) A4 specification (210X297 mm) _ 25 200300798 A7 B7 Printed by the Consumer Cooperatives of the Intellectual Property Bureau of the Ministry of Economic Affairs 5. Description of the invention (22) 0.45% NR40, 0.45% Tween20) After that, 8 // L of 10 mg / mL proteinase K was added, and it was completely dissolved by warming to 50-60. The same amount of TE saturated phenol was added, and after mixing and centrifugation, the aqueous layer was recovered. The recovered sample was added with 1/10 times the capacity of 3 mol / L sodium acetate (pH 5.3) and an equivalent amount of isoacetone, and the DNA was recovered by centrifugation. Wash with 70% ethanol, dry and dissolve in 50 // L TE (0.1 mol / L Tris-HCl (pH7.5), 1mmol / L EDTA (ρΗ8 · 0), use 2 · 5 // L in PCR Use a template. The unused part of the reaction is stored at 4 ° C. For a 4 mL sample, add 3 times the volume of SDS buffer for DNA preparation (150 mmol / L NaCl, 10 mmol / L Tris-HCl (pH8.0), After 10 mmol / L EDTA (ρΗ8 · 0), 1% SDS), add about 160 / L of 10mg / mL proteinase K, and completely dissolve it at 50 ~ 60t after heating. Add an equal amount of TE saturated phenol to remove After the protein, centrifugal separation was performed to recover the aqueous layer. 1 mol of the recovered sample was added with 3 mol / L sodium acetate (ρΗ5.3) and an equivalent amount of isopropanol to precipitate and recover the DNA. It was washed with 70% ethanol, dried and dissolved In 800 // L TE, as a DNA sample. The obtained sample is divided into 400 // LX 2 branches, 40 // L 3mol / L sodium acetate (ρΗ5.3) and 1mL of ethanol are mixed, and Stored at ° C. DNA samples prepared with SDS buffer solution for DNA preparation are used as sample preparation for PCR-level Southern Hybridization (if necessary, storage). (2) CYP2D6 gene amplification per 2.5 / zL DNA samples Add to 200 // L column, add the primers as shown in Table 1, and use PCR to amplify the CYP2D6 full exon (Long PCR). The bow of CYP2D6 * 5f and CYP2D6 * 5r [子 糸 和合, from CYP2D6 * 5 The detection of about 1.5kbp fragments is used for the purpose. CYP2D6elf-2 and (please read the precautions on the back before filling this page) This paper size applies the Chinese National Standard (CNS) A4 specification (210X297 mm 26-200300798)

A B V. Description of the invention (23) The combination of CYP2D6e6f is for the purpose of increasing the exon 1 ~ 6 area of CYP2D6, and part of it is used as the grinding plate to independently increase the exon 1 ~ 5. The combination of CYP2D6e6f and CYP2D9e9f-2 is for the purpose of increasing the CYP2D6 exon 6 ~ 9 region. A * part is used as a template to independently increase the exon 6 ~ 9 (2 nd PCR) ° The increase of each outer silk page, such as The combination of the first choice primer shown in Table 2 was performed, and the exon with unconfirmed increase was subjected to the increase of the second choice primer. (Please read the notes on the back before filling out this page) Printed by the Consumer Cooperatives of the Intellectual Property Bureau of the Ministry of Economic Affairs Table 2: Amplification of primers for each exon of CYP2D6 First selection primer pair Second selection primer pair an outer reverse exon 1 CYP2D6elf-2 CYP2D6elr-2 CYP2D6elf CYP2D6elr exon 2 CYP2D6e2f-2 CYP2D6e2r-2 CYP2D6e2f CYP2D6e2r exon 3 CYP2D6e3f CYP2D6e3r CYP2D6e3f-2 CYP2D6e3r_2 exon 4 CYP2D6e4f-2 CYP2D6e4r-2 CYP2D6e4f CYP2D6e4r exon 5 CYP2D6e5f CYP2D6e5r CYP2D6e5f-2 CYP2D6e5r-2 exon 6 CYP2D6e6f CYP2D6e6r CYP2D6e6f-2 CYP2D6e6r-2 exon 7 CYP2D6e7f CYP2D6e7r CYP2D6e7f-2 CYP2D6e7r-2 exon 8 CYP2D6e8f CYP2D6e8r CYP2D6e8f-2 CYP2D6e8r-2 exon 9 CYP2D6e9f CYP2D6e9r-l CYP2D6e9f-2 CYP2D6e9r-3 Table 1: Primer combinations that increase the total exon region of CYP2D6_ Target forward and reverse exons 1 ~ 6 CYP2D6elf-2 CYP2D6e6f Exon 6 ~ 9 CYP2D6e6f6f 6r CYP2D6 * 5f CYP2D6 * 5r This paper size is applicable to China National Standard (CNS) A4 (210X 297 mm) _ 27 200300798 Printed by the Consumer Cooperative of the Intellectual Property Bureau of the Ministry of Economic Affairs, A7 ___ B7__ V. Description of the invention (24) (3) Purification of each exon fragment amplified by PCR using the genotype analysis method, using a circular sequencing kit for sequencing reactions . After the reaction was completed, the ABI373 sequencing device (PE Abrite Biochemical) was used to analyze the base sequence. The obtained base sequence data is confirmed and corrected based on the peak waveform data. Data were analyzed using GeneWorks (intelligenetics Inc.) and well-known genetic data. The heterozygote was identified by the peaks obtained from ABI373 and the sense and antisense ends were identified. Mitsubishi Chemical BCL Co., Ltd. was entrusted to carry out tests on samples that were judged to be necessary for southern hybrid branches. Judgment criteria for samples: Southern hybridization is performed if the following conditions are met. When Long PCR using primers of CYP2D6 * 5f and CYP2D6 * 5r detected about 1.8 kbp fragment from CYP2D6 * 5; When genes classified as active class 3 contained CYP2D6 * 2 in the dual gene; and judged as Table 3 shows the results of the novel genotype (II) and the gene surveillance results of each sample. (Please read the precautions on the back before filling this page) This paper size applies the Chinese national standard ((: Can) 8 4 specifications (210 '/ 297 mm) -28_ 200300798 Printed by the Consumers ’Cooperative of Intellectual Property Bureau of the Ministry of Economic Affairs A7 B7 、 Explanation of the invention (Table 3 25) Dual gene frequency (%) Phenotype (this study) 4 2 (%) Phenotype (say the person) n = 324 (%) Phenotype (white race) n = 1344 (%) 1A 61 42.96 45.77 40.1 33.7 1E 1 0.70 ID 2 1.41 1 (unk η ow η 5) 1 0.70 2Α 6 4.23 5.63 12.9 26.9 2Α or Β 2 1.41 3 0 0.00 0.00 0.00 1.7 4 0 0.00 0.00 0.00 17.1 5 4 2.82 2.82 6.2 6.9 10A 23 16.20 38.03 38.6 1.4 10B 31 21.83 14 0 0.00 0.00 2.2 0.0 22 1 0.70 0.70 0.00 0.2 35 0 0.00 0.00 0.00 6.7 unk η ow η 1 1 0.70 0.70 Zhaounk η ow η 2 1 0.70 0.70 _ unknown3 6 4.24 4.23 • unknown4 1 0.70 0.70 • unknown6 1 0.70 0.70 Others 5.4 This paper size applies to China National Standard (CNS) A4 specification (210X297 mm) _ 29 (Please read the precautions on the back before filling this page) 200300798 A 7 B7 V. Description of the invention (26) (Please read the notes on the back before filling out this page) For the 142 dual genes that have been detected, the detected mutation sites are 1 3 (of which the novel mutation sites are 4), and the allelic valiant is 14 (in which the novel dual gene is 6), a total of 17 genotypes. The predicted amino acids translated by the dual gene can be reported from the literature (Kubota, T. British J. Clin. Pharmacol., 50, 3 1 -34, 2000 ; Marez, D., Pharmacogenetics, 7, 193-202, 1997; Johansson, I., Mol.

Pharmacol., 46, 452-459, 1994; Sachse, C. Am. J. Hum. Genet. 60, 284-295, 1997; Printed by Masuda Fukuda, Xenobio. Metabol.

Dispos., 15, 1 65- 1 70, 2000; and Wang, S.L., Drug Metabol. Disp., 27, 3 85-3 8 8, 1 998) estimated the enzyme activity of each amino acid type. The expression of each dual gene is considered to be the same, and the enzyme activity of each genotype is estimated. CYP2D6 氺 1A (wild-type homo) activity was used as an indicator of the active class classification. Specimens with strong enzyme activity (Class 1; equivalent to 氺 1A / * 1 A activity) were 24 examples (33.8%). Specimens that are relatively weak but not problematic (Class 2; * IA / * 1A activity is more than 50%) are 25 examples (35.2%), and are considered to be less active samples (Class 3; 氺 1A / * 1A activity was less than 50%) was 22 cases (30.9%). No sample presumed to be completely inactive was detected (Class 4). The frequency of dual genes is shown in Table 3 and Figure 1. CYP2D6 is not a gene necessary for sustaining life in Yujin Chemical. It is presumed to be redundant and fail to function through environmental selection. As a result, 71 gene polytypes have been reported (Cytochrome 450 (P450) Nomenclature Committee, http://www.imm.ki.se/CYP Dual Genes / cyp2d6.htm), and the paper size of each pair is applicable to China Standard (cns) A4 specifications (210X297 mm) _ 3〇. 200300798 A7 B7 V. Description of invention (27) (Please read the precautions on the back before filling this page) The frequency of gene occurrence varies from person to person. The frequency of dual genes in this sample was similar to that reported by healthy Japanese (Kubota, T. et al., British J. Clin. Pharmacol., 50, 31-34, 2000), and was detected in most yellow races. CYP2D 6 * 10 is 38.03% (38.6% in healthy Japanese, 1.4% in white people), and there is no enzyme activity-deficient CYP2D6 * 4 (17.1% in white people) detected in most white people. Same results as healthy Japanese. The frequency of fully defective CYP2D6 * 5 gene was 2.82%, showing a lower tendency compared with 6.2% of healthy Japanese and 6.9% of white race, but this is considered to be less than the number of samples tested Therefore, the frequency of occurrence of printed variation detection sites in the Consumer Cooperatives of the Intellectual Property Bureau of the Ministry of Economic Affairs is shown in Figure 1. The most confirmed site is 1661 (> (49.30%) (1661〇) (: indicates that the G of the base No. 1661 is replaced by C, the same applies hereinafter). This site is not substituted by amino acids The mutation (silent mutatin) has the same result as the frequency of occurrence of white races (52.6%). This can be presumed to be 1661 G > C mutation occurred before races were divided into yellow and white races. 100C > T (The amino acid is replaced by proline (P) to serine (S)) and 4 1 80G > C (the substitution of serine (S) to threonine (P)) is 44.37, respectively. % And 48.59%, this result reflects the results of the majority of CYP2D6 * 10 containing the elements that constitute the area (Table 3). For whites, the frequency of 100C > T and 4180G > C mutations is 18 · 4% and 52.9%, especially for 100C > T. A large frequency difference is confirmed. The generation period of 100C > T mutation must be discussed in detail, at least 100C > T and 4180G > C as constituent elements. The retained CYP2D6 * 10 is a dual gene that has occurred in yellow races since human differences. I paper size applies Chinese national standards CNS) A4 specification (210X 297 mm) _ 31 _ '200300798 A7 Printed by the Consumers ’Cooperative of the Intellectual Property Bureau of the Ministry of Economic Affairs B7 V. Description of the invention (2δ) There are reports that CYP2D6 * 10 is the metabolic ratio (MR) There are significant differences under * 1 / * 1, 1 / * 10 and * 10 / * 10 (Johansson, I. et al., Mol. Pharmacol., 46, 452-459, 1994), and matrix specificity has also been reported Sexual changes (Fukuta Takeshi, Xenobio. Metabol. And Dispos, 15, 165-170, 2000). In the past, the metabolizable energy of CYP2D6 as the main metabolic enzyme failed to show an extensive metabolizer (EM ) And clear 2-peak nature from PM, confirmed as intermediate metabolize (r (IM)) with metabolizable energy intermediate to EM and PM or hypermetabolism with several to dozens of times of metabolizable energy of EM (Super metabolizer (SM)). According to the results of this test, 30.9% of those who can be presumed to be IM and classified as Class 3 have more than 90% of their dual genes as * 10. From the above, when administering a drug metabolized by CYP2D6, considering that CYP2D6 * 10 is important, the data analysis and appropriate administration amount can be set according to the activity level. In the detection of 142 dual genes in 71 samples, three dual genes (unknownl, 2 and 4) with novel mutations and novel three dual genes composed of existing mutations were detected. The unknown genes of unknownl and 2 are assigned to undefined genotypes from the same sample. That is, for a sample, 100C > T, 125G > A, 1039C > T, 1661G > C, 1858C > T, 418 0G > C (where 125G > eight and 1 85 8C > T is a novel mutation site At the same time, the novel mutation site caused the amino acid substitution of G42E and R173C respectively) are all isoforms. Unknown4's variants are 100C > T, 1039C > T, 1661G> C, 2874TC > CT, 4180G > C (where 2874TC > CT is new). The paper size of this paper applies Chinese National Standard (CNS) A4 specifications ( 210 X 297mm L 32. &Quot; " (Please read the precautions on the back before filling this page) 200300798A7B7 Printed by the Consumer Cooperatives of the Intellectual Property Bureau of the Ministry of Economic Affairs 5. Description of the invention (29) Amino acid variation to P34S, S304L and S486T (of which S304L is new). As other novel dual genes, unknown3 (100C > T, 1661G > C, 2850C > T, 3790C > T, 4180C > T amino acid mutations are P34S, R29 6C and S48 6T) were 6 samples and 6 dual genes, and the frequency of occurrence was 4.23%. Also, unknown5 (1661G> C silent) 1 sample, 1 dual gene was detected, and the frequency of occurrence was 0.7%. Unknown3, unknown4, unknown5 The base sequence and amino acid sequence of cDNA of unknown6 and amino acid are respectively listed in the sequence listing sequence numbers 2, 3, 4 and 5. In vitro experiments using complex CYP2D6, the introduction of a 100C > T variant mutant will cause an enzyme of CYP2D6 * 1A live It was reduced to about 1/20 (Wang, SL et al., Drug Metabol. Disp., 27, 3 85-3 88, 1 998), and unknown 3 and 6 were predicted to have the same or lower activity as CYP2D6 * 10. Particularly, unknown3 has a high frequency of 4.23%, which becomes a necessary test item. 〇 Example 2: Expression and activity of a novel CYP2D6 protein using a baculovirus expression system In the enzyme activity of the gene, a novel polymorphic gene was produced using the following method based on CYP2D6 * 2 cloned from the human liver cDNA gene library. (I) Materials and methods The synthesis center part has a mutation introduction part. The following primers are made by a sequence of 10 to 30 bases before and after. This paper size applies the Chinese National Standard (CNS) A4 specification (210X297 mm) _ 33 '(Please read the precautions on the back before filling this page) 200300798 A7 B7 V. Description of the invention (30) Bow [Child 1 atggggctagaagcactggt (Order 歹! 1 number 53) Primer 2 ggcagggggcctggtgagtagcgtg (Serial number 54) Bow [Child 3 ctgggctgcacgctactcaccacacac (Child 3 j! J Number 55) Bow [子4 ctagcggggcacagcacaaag (Order No. [J No. 56) Using primers 1 and 2 to make a slice 2 using the following method using fragment 1 and paragraph 4 as described below. Template Fragment 3 and Slice

Primer 1 (or primer 3) Primer 2 (or primer 4) 10x buffer (buffer) 20mmol / L dNTPs AdvantageHF2 DNA polymerase (Clontech) Adjusted to dH2〇 to a total volume of 50 // L

l〇ng 20pmole 20pmole 5 β L 5 β L 1 β L (Please read the notes on the back before filling out this page) Printed at 94 ° C (thermal degeneration) 57 ° C ( Calcination) 68 ° C (primer extension reaction) 30 seconds 30 seconds 3 minutes The above was performed as one cycle, and 32 cycles were performed. The calcination temperature varies depending on the primer sequence. The target fragment was cut out and purified after agarose electrophoresis. The purified fragments 1 and 2 were mixed at a 1: 1 molar ratio, and the following reactions were performed. Fragment 1 50ng Fragment 2 5 0 n g This paper size is applicable to Chinese National Standard (CNS) A4 (210X297 mm L 34 200300798 A7 B7 Printed by the Consumer Cooperatives of the Intellectual Property Bureau of the Ministry of Economic Affairs V. Invention Description (31)

10x buffer buffer (Clontech) 20mmol / L dNTPs (Clontech) AdvaiitageHF2 DNA polymerase (Clontech) Adjust the total volume to 50 // L with dH2〇

5 β L 5 β L 1 (I L reaction conditions 94 ° C (thermal denaturation) 68t (primer extension reaction) 1 minute 3 minutes (Please read the precautions on the back before filling this page)

The above was performed as one cycle, and the following reaction was performed in a part of 25 cycles. Template primer 1 Primer 4 1 0 X buffer buffer (C1 ο ntech) 20mmol / L dNTPs (Clontech) AdvantageHF2 DNA polymerase (Clontech) Adjust the total amount to 50 with dH2〇 // L reaction conditions 94 ° C (thermal denaturation) 57 ° C (calcined) 68 ° C (primer extension response) The paper size applies to the Chinese National Standard (CNS) A4 specification (210X297 mm) _ 35

10ng 20pmole 20pmole 5 β L 5 β L 1 β L using its reaction solution 30 seconds 30 seconds 3 minutes 200 300 798 A7 Printed by the Consumer Cooperatives of the Intellectual Property Bureau of the Ministry of Economic Affairs _ B7 V. Description of the invention (32) The above is a cycle, Implement 32 cycles. The target fragment was cut out and purified after agarose electrophoresis (fragment 3). Fragment 3 was inserted into pT7Blue vector (Novagen) cut with EcoRV and transformed into coliform. The plastids were prepared from clones with the correct base sequence inserted, and the inserts were cut out using the restriction enzymes κρηΙ and Bglll, and then cloned into the baculovirus transfer vector pPSC8. The above-mentioned transfer vector is used to obtain the target protein according to a conventional method. The concentration of the target protein was obtained by Western blot method. Enzyme activity was measured using 3- [2- (N, N-diethylamine) ethyl] -7-methoxy-4-methylcoumarin (hereinafter referred to as AMMC: GENTES) as the substrate fluorescence Method to do the determination. 9 6-channel microtiter dishes add 1 0 0 // L of NADPH-Cofactor mix (1.75 // L Cofactors (GENTEST), 1 // L GAPDH, 0.67V // L control protein (GENTEST ) And 96.5 8 // LH20). After pre-heating at 37 ° C for 10 minutes, add 100 // L of warmed enzyme / matrix mixture (69.25 // L H20, 75 // L 2 // 1 15nmol / L AMMC) per groove ). After reacting at 37 ° C for 30 minutes, a 75 / L reaction stop solution (0.5 mmol / L Tris Base) was added, and the fluorescence intensity (Ex 3 90 nm, Em 460 nm) was measured. (Π) Results When CYP2D6 氺 1 (wild type) is 100, the enzyme activity of CYP2D6unknown3 is less than 5%. It is presumed that the solid with this gene is compared with individuals with wild-type genes, and the metabolism of CYP2D6 as a responsible metabolic enzyme will be delayed. In addition, the amino acid of this polytype gene is substituted at three positions compared with the wild type, and its effect is different from that of the wild type in terms of substrate specificity. Therefore, this paper size applies the Chinese National Standard (CNS) A4 specification (210X 297 mm) _ 36 (Please read the notes on the back before filling this page) 200300798 A7 B7 V. Description of the invention (33) When it comes to efficacy and side effects, it is necessary to investigate the genotype on the genome and observe the effect of drugs using microsomes and combined proteins. Industrial Applicability A novel polytype of the CYP2D6 gene is identified in the present invention. Using the information of the polymorphic genes identified by the present invention, it is possible to judge the metabolic activity of the subject to which the drug is administered, or to search for novel drugs. V. Brief description of the diagram Figure 1 shows the detected CYP2D6 mutation site and the frequency of occurrence ° -------- β-pack-(Please read the precautions on the back before filling this page), 1T Ministry of Economic Affairs wisdom The paper size printed by the Property Cooperative's Consumer Cooperative is applicable to the Chinese national standard (..) 8 4 specifications (210 parent 297 mm) _37- v _ —— 200300798 A7 B7 V. Description of the invention (

SEQUENCE LISTING < 110 > Tsumura Inc. < 120〉 CYP2D6 mutant gene < 130 > FA2566A / TW < 160 > 56 < 210〉 1 < 211〉 4500 < 212 > DNA < 213〉 Human < 400 > 1 atggggctag aagcactggt gcccctggcc gtgatagtgg ccatcttcct gctcctggtg 60 gacctgatgc accggcgcca acgctgggct gcacgctact caccaggccc cctgccactg 120 cccgggctgg gcaacctgct gcatgtggac ttccagaaca caccatactg cttogaccag 180 gtgagggagg aggtcctgga gggcggcaga ggtgctgagg ctcccctaco agaagcaaac 240 atggatggtg ggtgaaacca caggctggac cagaagccag gctgagaagg ggaagcaggt 300 ttgggggacg gccaaggaag tcctggagaa gggcatttat acatggcatg aaggactgga ttttccaaag 360 agtagggcaa gggcctggag gtggagctgg acttggcagt gggcatgcaa 420 gcccattggg caacatatgt tatggagtac aaagtccctt ctgctgacac cagaaggaaa 480 ggccttggga atggaagatg agttagtcct gagtgccgtt taaatcacga aatcgaggat 540 gaagggggtg cagtgacccg gttcaaacct tttgcactgt gggtcctcgg gcctcactgc 600 ctcaccggca tggaccatca tctgggaatg ggatgctaac tggggcctct cggcaatttt 660 ggtgactctt gcaaggtcat acctgggtga cgcatccaaa ctgagttcct ccatcacaga 720 aggtgtgacc cccacccccg ccccacgatc aggaggctgg gtctcctcct tccacctgct 780 cactcctggt agccccgggg gtcgtccaag gttcaaatag gactaggacc tgtagtctgg 840 ggtgatcctg gcttgacaag aggccctgac cctccctctg cagttgcggc gccgcttcgg 900 ggacgtgttc agcctgcagc tggcctggac gccggtggtc gtgctcaatg ggctggcggc 960 Ben paper scale applicable to Chinese National Standard (CNS) A4 size (210 x 297 mm) . -38-(Please read the notes on the back before filling this page)

, 1T Ministry of Economic Affairs Intellectual Property Office employees consumer cooperatives printed 200300798 A7 B7 V. description of the invention (cgtgcgcgag gcgctggtga cccacggcga ggacaccgcc gaccgcccgc ctgtgcccat 1020 cacccagatc ctgggttttg ggccgcgttc ccaaggcaag cagcggtggg gacagagaca 1080 gatttccgtg ggacccgggt gggtgatgac cgtagtccga gctgggcaga gagggcgcgg 1140 ggtcgtggac atgaaacagg ccagcgagtg gggacagcgg gccaagaaac cacctgcact 1200 agggaggtgt gagcatgggg acgagggcgg ggcttgtgac gagtgggcgg ggccactgcc 1260 gagacctggc aggagcccaa tgggtgagcg tggcgcattt cccagctgga atccggtgtc 1320 gaagtggggg cggggaccgc acctgtgctg taagctcagt gtgggtggcg cggggcccgc 1380 ggggtcttcc ctgagtgcaa aggcggtcag ggtgggcaga gacgaggtgg ggcaaagcct 1440 gccccagcca agggagcaag gtggatgcac aaagagtggg ccctgtgacc agctggacag 1500 agccagggac tgcgggagac cagggggagc atagggttgg agtgggtggt ggatggtggg 1560 gctaatgcct tcatggccac gcgcacgtgc ccgtcccacc cccaggggtg ttcctggcgc 1620 gctatgggcc cgcgtggcgc gagcagaggc gcttctccgt ctccaccttg cgcaacttgg 1680 gcctgggcaa gaagtcgctg gagcagtggg tgaccgagga ggccgcct gc ctttgtgccg 1740 ccttcgccaa ccactccggt gggtgatggg cagaagggca caaagcggga actgggaagg 1800 cgggggacgg ggaaggcgac pccttacccg catctcccac ccccaggacg cccctttcgc 1860 cccaacggtc tcttggacaa agccgtgagc aacgtgatcg cctccctcac ctgcgggcgc 1920 cgcttcgagt acgacgaccc tcgcttcctc aggctgctgg acctagctca ggagggactg 1980 aaggaggagt cgggctttct gcgcgaggtg cggagcgaga gaccgaggag tctctgcagg 2040 gcgagctccc gagaggtgcc ggggctggac tggggcctcg gaagagcagg atttgcatag 2100 atgggtttgg gaaaggacat tccaggagac cccactgtaa gaagggcctg gaggaggagg 2160 ggacatctca gacatggtcg tgggagaggt gtgcccgggt cagggggcac caggagaggc 2220 caaggactct gtacctccta tccacgtcag agatttcgat tttaggtttc tcctctgggc 2280 aaggagagag ggtggaggct ggcacttggg gagggacttg gtgaggtcag tggtaaggac 2340 aggcaggccc tgggtctacc tggagatggc tggggcctga gacttgtcca ggtgaacgca 2400 gagcacagga gggattgaga ccccgttctg tctggtgtag gtgctgaatg ctgtccccgt 2460 cctcctgcat atcccagcgc tggctggcaa ggtcctacgc ttccaaaagg ctttcctgac 2520 ccagctggat gagctgctaa ctgagcacag gatgacctgg gacccagccc agc ccccccg 2580 This paper size is applicable to China National Standard (CNS) A4 specification (210X297 mm) (Please read the precautions on the back before filling this page).

, 1T Ministry of Economic Affairs Intellectual Property Office employees consumer cooperatives printed -39- 200300798 A7.. B7 V. description of the invention (agacctgact gaggccttcc tggcagagat ggagaaggtg agagtggctg ccacggtggg 2640 gggcaagggt ggtgggttga gcgtcccagg aggaatgagg ggaggctggg caaaaggttg 2700 gaccagtgca tcacccggcg agccgcatct gggctgacag gtgcagaatt ggaggtcatt 2760 tgggggctac cccgttctgt cccgagtatg ctctcggccc tgctcaggcc aaggggaacc 2820 ctgagagcag cttcaatgat gagaacctgt gcatagtggt ggctgacctg ttctctgccg 2880 ggatggtgac cacctcgacc acgctggcct ggggcctcct gctcatgatc ctacatccgg 2940 atgtgcagcg tgagcccatc tgggaaacag tgcaggggcc gagggaggaa gggtacaggc 3000 gggggcccat gaactttgct gggacacccg gggctccaag cacaggcttg accaggatcc 3060 tgtaagcctg acctcctcca acataggagg caagaaggag tgtcagggcc ggaccccctg 3120 ggtgctgacc cattgtgggg acgcatgtct gtccaggccg tgtccaacag gagatcgacg 3180 acgtgatagg gcaggtgcgg cgaccagaga tgggtgacca ggctcacatg ccctacacca 3240 ctgccgtgat tcatgaggtg cagcgctttg gggacatcgt ccccctgggt gtgacccata 3300 tgacatcccg tgacatcgaa gtacagggct tccgcatccc taaggtaggc ctggcgccct 3360 cctcacccca gctcagcacc agcacctggt gatagcccca gcatggctac tgccaggtgg 3420 gcccactcta ggaaccctgg ccacctagtc ctcaatgcca ccacactgac tgtccccact 3480 tgggtggggg gtccagagta taggcagggc tggcctgtcc atccagagcc cccgtctagt 3540 ggggagacaa accaggacct gccagaatgt tggaggaccc aacgcctgca gggagagggg 3600 gcagtgtggg tgcctctgag aggtgtgact gcgccctgct gtggggtcgg agagggtact 3660 gtggagcttc tcgggcgcag gactagttga cagagtccag ctgtgtgcca ggcagtgtgt 3720 gtcccccgtg tgtttggtgg caggggtccc agcatcctag agtccagtcc ccactctcac 3780 cctgcatctc ctgcccaggg aacgacactc atcaccaacc tgtcatcggt gctgaaggat 3840 gaggccgtct gggagaagcc cttccgcttc caccccgaac acttcctgga tgcccagggc 3900 cactttgtga agccggaggc cttcctgcct ttctcagcag gtgcctgtgg ggagcccggc 3960 tccctgtccc cttccgtgga gtcttgcagg ggtatcaccc aggagccagg ctcactgacg 4020 cccctcccct ccccacaggc cgccgtgcat gcctcgggga gcccctggcc cgcatggagc 4080 tcttcctctt cttcacctcc ctgctgcagc acttcagctt ctcggtgccc actggacagc 4140 cccggcccag ccaccatggt gtctttgctt tcctggtgac cccat ccccc tatgagcttt 4200 — -----——-— --- This paper size is applicable to China National Standard (CNS) A4 (21〇X 297 mm) (Please read the precautions on the back before filling this page) ·

, 1T Ministry of Economic Affairs Intellectual Property Office employees consumer cooperatives printed -40- 200300798 A7 B7 V. invention is described in (gtgctgtgcc ccgctagaat ggggtaccta gtccccagcc tgctccctag ccagaggctc 4260 taatgtacaa taaagcaatg tggtagttcc aactcgggtc ccctgctcac gccctcgttg 4320 ggatcatcct cctcagggca accccacccc tgcctcattc ctgcttaccc caccgcctgg 4380 (please read the back of the Please fill out this page again) ccgcatttga gacaggggta cgttgaggct gagcagatgt cagttaccct tgcccataat 4440 cccatgtccc ccactgaccc aactctgact gcccagattg gtgacaaggaga ctacattgtc 4500 < 210〉 2 < 211〉 1567 < 212> > 212> DNA gca ctg gtg ccc ctg gee gtg ata gtg gee ate ttc 48

Met Gly Leu Glu Ala Leu Val Pro Leu Ala Val He Val Ala He Phe 1 5 10 15 ctg etc ctg gtg gac ctg atg cac egg ege caa ege tgg get gca ege 96

Leu Leu Leu Val Asp Leu Met His Arg Arg Gin Arg Trp Ala Ala Arg 20 25 30 tac tea cca ggc ccc ctg cca ctg ccc ggg ctg ggc aac ctg ctg cat 144

Tyr Ser Pro Gly Pro Leu Pro Leu Pro Gly Leu Gly Asn Leu Leu His 35 40 45 Printed by the Consumer Cooperatives of the Intellectual Property Bureau of the Ministry of Economic Affairs gtg gac ttc cag aac aca cca tac tgc ttc gac cag ttg egg ege ege 192

Val Asp Phe Gin Asn Thr Pro Tyr Cys Phe Asp Gin Leu Arg Arg Arg 50 55 60 ttc ggg gac gtg ttc age ctg cag ctg gee tgg aeg ccg gtg gtc gtg 240

Phe Gly Asp Val Phe Ser Leu Gin Leu Ala Trp Thr Pro Val Val Val 65 70 75 80 etc aat ggg ctg geg gee gtg ege gag geg ctg gtg acc cac ggc gag 288 This paper size applies to China National Standard (CNS) A4 specifications ( 210x 297 mm) -41-200300798 A7 B7 V. Description of the invention (

Leu Asn Gly Leu Ala Ala Val Arg Glu Ala Leu Val Thr His Gly Glu 85 90 95 (Please read the notes on the back before filling in this page) gac acc gcc gac cgc ccg cct gtg ccc ate acc cag ate ctg ggt ttc 336 Asp Thr Ala Asp Arg Pro Pro Val Pro lie Thr Gin lie Leu Gly Phe 100 105 Π0 ggg ccg cgt tcc caa ggg gtg ttc ctg geg cgc tat ggg ccc geg tgg 384 Gly Pro Arg Ser Gin Gly Val Phe Leu Ala Arg Tyr Gly Pro Ala Trp 115 120 125 cgc gag cag agg cgc ttc tcc gtc tcc acc ttg cgc aac ttg ggc ctg 432 Arg Glu Gin Arg Arg Phe Ser Val Ser Thr Leu Arg Asn Leu Gly Leu 130 135 140 ggc aag aag teg ctg gag accg gag gag gcc gcc tgc ett 480 Gly Lys Lys Ser Leu Glu Gin Trp Val Thr Glu Glu Ala Ala Cys Leu 145 150 155 160 tgt gcc gcc ttc gcc aac cac tcc gga cgc ccc ttt cgc ccc aac ggt 528 Cys Ala Ala Ala Phe His Ser Gly Arg Pro Phe Arg Pro Asn Gly 165 170 175 etc ttg gac aaa gcc gtg age aac gtg ate gcc tcc etc acc tgc ggg 576

Leu Leu Asp Lys Ala Val Ser Asn Val lie Ala Ser Leu Thr Cys Gly 180 185 190 Printed by the Consumer Cooperative of the Intellectual Property Bureau of the Ministry of Economic Affairs cgc cgc ttc gag tac gac gac cct cgc ttc etc agg ctg ctg gac eta 624

Arg Arg Phe Glu Tyr Asp Asp Pro Arg Phe Leu Arg Leu Leu Asp Leu 195 200 205 get cag gag gga ctg aag gag gag teg ggc ttt ctg cgc gag gtg ctg 672

Ala Gin Glu Gly Leu Lys Glu Glu Ser Gly Phe Leu Arg Glu Val Leu 210 215 220 aat get gtc ccc gtc etc ctg cat ate cca geg ctg get ggc aag gtc 720 This paper is in accordance with the Chinese National Standard (CNS) A4 specifications (21 〇 > < 297 mm) -42- 200300798 A7 B7 V. Description of the invention (

Asn Ala Val Pro Val Leu Leu His lie Pro Ala Leu Ala Gly Lys Val 225 230 235 240 eta ege ttc caa aag get ttc ctg acc cag ctg gat gag ctg eta act 768 (Please read the notes on the back before filling this page)

Leu Arg Phe Gin Lys Ala Phe Leu Thr Gin Leu Asp Glu Leu Leu Thr 245 250 255 gag cac agg atg acc tgg gac cca gcc cag ccc ccc ega gac ctg act 816

Glu His Arg Met Thr Trp Asp Pro Ala Gin Pro Pro Arg Asp Leu Thr 260 265 270 gag gcc ttc ctg gca gag atg gag aag gcc aag ggg aac cct gag age 864

Glu Ala Phe Leu Ala Glu Met Glu Lys Ala Lys Gly Asn Pro Glu Ser 275 280 285 age ttc aat gat gag aac ctg tgc ata gtg gtg get gac ctg ttc tet 912

Ser Phe Asn Asp Glu Asn Leu Cys lie Val Val Ala Asp Leu Phe Ser 290 295 300 gcc ggg atg gtg acc acc teg acc aeg ctg gcc tgg ggc etc ctg etc 960

Ala Gly Met Val Thr Thr Ser Thr Thr Leu Ala Trp Gly Leu Leu Leu 305 310 315 320 atg ate eta cat ccg gat gtg cag ege cgt gtc caa cag gag ate gac 1008

Met He Leu His Pro Asp Val Gin Arg Arg Val Gin Gin Glu lie Asp 325 330 335 Printed by the Consumer Cooperative of the Intellectual Property Bureau of the Ministry of Economic Affairs gac gtg ata ggg cag gtg egg ega cca gag atg ggt gac cag get cac 1056

Asp Val lie Gly Gin Val Arg Arg Pro Glu Met Gly Asp Gin Ala His 340 345 350 atg ccc tac acc act gcc gtg att cat gag gtg cag ege ttt ggg gac Π04

Met Pro Tyr Thr Thr Ala Val lie His Glu Val Gin Arg Phe Gly Asp 355 360 365 ate gtc ccc ctg ggt gtg acc cat atg aca tee cgt gac ate gaa gta 1152 This paper is in accordance with the Chinese National Standard (CNS) A4 specification (2l 〇x 297 mm) -43- 200300798 A7, _B7____ Master and invention description (40 lie Val Pro Leu Gly Val Thr His Met Thr Ser Arg Asp lie Glu Val 370 375 380 cag ggc ttc cgc ate cct aag gga aeg aca etc ate acc aac ctg tea 1200 (Please read the notes on the back before filling this page)

Gin Gly Phe Arg He Pro Lys Gly Thr Thr Leu lie Thr Asn Leu Ser 385 390 395 400 teg gtg ctg aag gat gag gee gtc tgg gag aag ccc ttc cgc ttc cac 1248

Ser Val Leu Lys Asp Glu Ala Val Trp Glu Lys Pro Phe Arg Phe His 405 410 415 ccc gaa cac ttc ctg gat gee cag ggc cac ttt gtg aag ccg gag gee 1296

Pro Glu His Phe Leu Asp Ala Gin Gly His Phe Val Lys Pro Glu Ala 420 425 430 ttc ctg cct ttc tea gca ggc cgc cgt gca tgc etc ggg gag ccc ctg 1344

Phe Leu Pro Phe Ser Ala Gly Arg Arg Ala Cys Leu Gly Glu Pro Leu 435 440 445 gee cgc atg gag etc ttc etc ttc ttc acc tee ctg ctg cag cac ttc 1392

Ala Arg Met Glu Leu Phe Leu Phe Phe Thr Ser Leu Leu Gin His Phe 450 455 460 age ttc teg gtg ccc act gga cag ccc egg ccc age cac cat ggt gtc 1440

Ser Phe Ser Val Pro Thr Gly Gin Pro Arg Pro Ser His His Gly Val 465 470 475 480 Printed by the Consumer Cooperative of the Intellectual Property Bureau of the Ministry of Economy ttt get ttc ctg gtg acc cca tee ccc tat gag ett tgt get gtg ccc 1488

Phe Ala Phe Leu Val Thr Pro Ser Pro Tyr Glu Leu Cys Ala Val Pro 485 490 495 cgc tag aat ggg gta cct agt ccc cag cct get cct age cca gag get 1536 Arg eta atg tac aat aaa gca atg tgg tag ttc c 1567 Paper size applies Chinese National Standard (CNS) A4 specification (210x297 mm) -44- 200300798 A7 B7 V. Description of invention (4) < 210〉 3 < 211 > 1567 < 212〉 DNA < 213〉 I human l < 400 > 3 atg ggg eta gaa gca ctg gtg ccc ctg gee gtg ata gtg gee ate ttc 48

Met Gly Leu Glu Ala Leu Val Pro Leu Ala Val He Val Ala lie Phe 15 10 15 ctg etc ctg gtg gac ctg atg cac egg ege caa ege tgg get gca ege 96

Leu Leu Leu Val Asp Leu Met His Arg Arg Gin Arg Trp Ala Ala Arg 20 25 30 tac tea cca ggc ccc ctg cca ctg ccc ggg ctg ggc aac ctg ctg cat 144 Tyr Ser Pro Gly Pro Leu Pro Leu Pro Gly Leu Gly Asn Leu Leu His 35 40 45 gtg gac ttc cag aac aca cca tac tgc ttc gac cag ttg egg ege ege 192 Val Asp Phe Gin Asn Thr Pro Tyr Cys Phe Asp Gin Leu Arg Arg Arg 50 55 60 ttc ggg gac gtg ttc age ctg cag gee tgg aeg ccg gtg gtc gtg 240 Phe Gly Asp Val Phe Ser Leu Gin Leu Ala Trp Thr Pro Val Val Val 65 70 75 80 etc aat ggg ctg geg gee gtg ege gag geg ctg gtg ace cac ggc gag 288 Leu Asn Gly Leula Ala Val Arg Glu Ala Leu Val Thr His Gly Glu 85 90 95 gac acc gee gac ege ccg cct gtg ccc ate acc cag ate ctg ggt ttt 336 Asp Thr Ala Asp Arg Pro Pro Val Pro lie Thr Gin lie Leu Gly Phe 100 105 110 ggg ccg cgt tee caa ggg gtg ttc ctg geg ege tat ggg ccc geg tgg 384 This paper size applies to China National Standard (CNS) A4 specification (21〇 > < 297 mm) (Please read the notes on the back before filling in (This page) Economic Affairs Intellectual Property Office employees consumer cooperatives printed -45- 200300798 A7 B7 five - Ministry of Economic Affairs Intellectual Property Office employees consumer cooperatives printed description of the invention (4)

Gly Pro Arg Ser Gin Gly Val Phe Leu Ala Arg Tyr Gly Pro Ala Trp 115 120 125 cgc gag cag agg cgc ttc tcc gtc tcc acc ttg cgc aac ttg ggc ctg 432 (Please read the notes on the back before filling this page)

Arg Glu Gin Arg Arg Phe Ser Val Ser Thr Leu Arg Asn Leu Gly Leu 130 135 140 ggc aag aag teg ctg gag cag tgg gtg acc gag gag gcc gcc tgc ett 480

Gly Lys Lys Ser Leu Glu Gin Trp Val Thr Glu Glu Ala Ala Cys Leu 145 150 155 160 tgt gcc gcc gcc ttc gcc aac cac tcc gga cgc ccc ttt cgc ccc aac ggt 528

Cys Ala Ala Phe Ala Asn His Ser Gly Arg Pro Phe Arg Pro Asn Gly 165 170 175 etc ttg gac aaa gcc gtg age aac gtg ate gcc tcc etc acc tgc ggg 576

Leu Leu Asp Lys Ala Val Ser Asn Val lie Ala Ser Leu Thr Cys Gly 180 185 190 cgc cgc ttc gag tac gac gac cct cgc ttc etc agg ctg ctg gac eta 624

Arg Arg Phe Glu Tyr Asp Asp Pro Arg Phe Leu Arg Leu Leu Asp Leu 195 200 205 get cag gag gga ctg aag gag gag teg ggc ttt ctg cgc gag gtg ctg 672

Ala Gin Glu Gly Leu Lys Glu Glu Ser Gly Phe Leu Arg Glu Val Leu 210 215 220 aat get gtc ccc gtc etc ctg cat ate cca geg ctg get ggc aag gtc 720

Asn Ala Val Pro Val Leu Leu His lie Pro Ala Leu Ala Gly Lys Val 225 230 235 240 eta cgc ttc caa aag get ttc ctg acc cag ctg gat gag ctg eta act 768

Leu Arg Phe Gin Lys Ala Phe Leu Thr Gin Leu Asp Glu Leu Leu Thr 245 250 255 gag cac agg atg acc tgg gac cca gcc cag ccc ccc ega gac ctg act 816 This paper size applies to the Chinese National Standard (CNS) A4 size (x 297 mm) -46- 200300798 A7 '* B7 V. Description of Invention (43

Glu His Arg Met Thr Trp Asp Pro Ala Gin Pro Pro Arg Asp Leu Thr 260 265 270 gag gcc ttc ctg gca gag atg gag aag gcc aag ggg aac cct gag age 864 (Please read the notes on the back before filling this page)

Glu Ala Phe Leu Ala Glu Met Glu Lys Ala Lys Gly Asn Pro Glu Ser 275 280 285 age ttc aat gat gag aac ctg ege ata gtg get gac ctg ttc ett 912

Ser Phe Asn Asp Glu Asn Leu Arg He Val Val Ala Asp Leu Phe Leu 290 295 300 gcc ggg atg gtg acc acc teg acc aeg ctg gcc tgg ggc etc ctg etc 960

Ala Gly Met Val Thr Thr Ser Thr Thr Leu Ala Trp Gly Leu Leu Leu 305 310 315 320 atg ate eta cat ccg gat gtg cag ege cgt gtc caa cag gag ate gac 1008

Met lie Leu His Pro Asp Val Gin Arg Arg Val Gin Gin Glu lie Asp 325 330 335 gac gtg ata ggg cag gtg egg ega cca gag atg ggt gac cag get cac 1056

Asp Val He Gly Gin Val Arg Arg Pro Glu Met Gly Asp Gin Ala His 340 345 350 atg ccc tac acc act gcc gtg att cat gag gtg cag ege ttt ggg gac 1104

Met Pro Tyr Thx Thr Ala Val lie His Glu Val Gin Arg Phe Gly Asp 355 360 365 Printed by the Consumer Cooperative of the Intellectual Property Bureau of the Ministry of Economic Affairs ate gtc ccc ctg ggt gtg acc cat atg aca tee cgt gac ate gaa gta 1152 lie Yal Pro Leu Gly Val Thr His Met Thr Ser Arg Asp lie Glu Val 370 375 380 cag ggc ttc ege ate cct aag gga aeg aca etc ate acc aac ctg tea 1200

Gin Gly Phe Arg lie Pro Lys Gly Thr Thr Leu lie Thr Asn Leu Ser 385 390 395 400 teg gtg ctg aag gat gag gcc gtc tgg gag aag ccc ttc ege ttc cac 1248 This paper applies the Chinese National Standard (CNS) A4 specifications ( 210X mm) -47-200300798 A7 B7 V. Description of the invention (d

Ser Val Leu Lys Asp Glu Ala Val Trp Glu Lys Pro Phe Arg Phe His 405 410 415 ccc gaa cac ttc ctg gat gcc cag ggc cac ttt gtg aag ccg gag gcc 1296

Pro Glu His Phe Leu Asp Ala Gin Gly His Phe Val Lys Pro Glu Ala 420 425 430 ttc ctg cct ttc tea gca ggc ege cgt gca tgc etc ggg gag ccc ctg 1344

Phe Leu Pro Phe Ser Ala Gly Arg Arg Ala Cys Leu Gly Glu Pro Leu 435 440 445 gcc ege atg gag etc ttc etc ttc ttc acc tee ctg ctg cag cac ttc 1392

Ala Arg Met Glu Leu Phe Leu Phe Phe Thr Ser Leu Leu Gin His Phe 450 455 460 age ttc teg gtg ccc act gga cag ccc egg ccc age cac cat ggt gtc 1440

Ser Phe Ser Val Pro Thr Gly Gin Pro Arg Pro Ser His His Gly Val 465 470 475 480 ttt get ttc ctg gtg acc cca tee ccc tat gag ett tgt get gtg ccc 1488

Phe Ala Phe Leu Val Thr Pro Ser Pro Tyr Glu Leu Cys Ala Val Pro 485 490 495 ege tag aat ggg gta cct agt ccc cag cct get cct age cca gag get 1536 Arg eta atg tac aat aaa gca atg tgg tag ttc c 1567

< 210〉 4 < 211〉 1567 < 212〉 DNA < 213〉 human < 400〉 4 atg ggg eta gaa gca ctg gtg ccc ctg gcc gtg ata gtg gcc ate ttc 48 This paper is in accordance with Chinese national standards ( CNS) A4 specification (210X 297 public envy) I --.—— ^ ---- I (Please read the precautions on the back before filling this page), 11 Printed by the Consumer Cooperatives of the Intellectual Property Bureau of the Ministry of Economic Affairs • 48- 200300798 A7 B7 V. Description of the invention (

Met Gly Leu Glu Ala Leu Val Pro Leu Ala Val lie Val Ala lie Phe 1 5 10 15 ctg etc ctg gtg gac ctg atg cac egg ege caa ege tgg get. Gca ege 96 (Please read the notes on the back before filling this page )

Leu Leu Leu Val Asp Leu Met His Arg Axg Gin Arg Trp Ala Ala Arg 20 25 30 tac cca cca ggc ccc ctg cca ctg ccc ggg ctg ggc aac ctg ctg cat 144

Tyr Pro Pro Gly Pro Leu Pro Leu Pro Gly Leu Gly Asn Leu Leu His 35 40 45 gtg gac ttc cag aac aca cca tac tgc ttc gac cag ttg egg ege ege 192

Val Asp Phe Gin Asn Thr Pro Tyr Cys Phe Asp Gin Leu Arg Arg Arg 50 55 60 ttc ggg gac gtg ttc age ctg cag ctg gee tgg aeg ccg gtg gtc gtg 240

Phe Gly Asp Val Phe Ser Leu Gin Leu Ala Trp Thr Pro Val Val Val 65 70 75 80 etc aat ggg ctg geg gee gtg ege gag geg ctg gtg acc cac ggc gag 288

Leu Asn Gly Leu Ala Ala Val Arg Glu Ala Leu Val Thr His Gly Glu 85 90 95 gac acc gee gac ege ccg cct gtg ccc ate acc cag ate ctg ggt ttc 336

Asp Thr Ala Asp Arg Pro Pro Val Pro lie Thr Gin lie Leu Gly Phe 100 105 110 Printed by the Consumer Cooperative of the Intellectual Property Bureau of the Ministry of Economic Affairs ggg ccg cgt tee caa ggg gtg ttc ctg geg ege tat ggg ccc geg tgg 384

Gly Pro Arg Ser Gin Gly Val Phe Leu Ala Arg Tyr Gly Pro Ala Trp 115 120 125 ege gag cag agg ege ttc tee gtc tee acc ttg ege aac ttg ggc ctg 432

Arg Glu Gin Arg Arg Phe Ser Val Ser Thr Leu Arg Asn Leu Gly Leu 130 135 140 ggc aag aag teg ctg gag cag tgg gtg acc gag gag gee gee tgc ett 480 This paper is in accordance with the Chinese National Standard (CNS) A4 specification (210X297 (Mm)-49- 200300798 A7, B7____ 5. Description of the invention (θ

Gly Lys Lys Ser Leu Glu Gin Trp Val Thr Glu Glu Ala Ala Cys Leu 145 150 155 160 tgt gcc gcc gcc ttc gcc aac cac tcc gga cgc ccc ttt cgc ccc aac ggt 528 (Please read the notes on the back before filling in this page)

Cys Ala Ala Phe Ala Asn His Ser Gly Arg Pro Phe Arg Pro Asn Gly 165 170 175 etc ttg gac aaa gcc gtg age ^ ac gtg ate gcc tcc etc acc tgc ggg 576

Leu Leu Asp Lys Ala Val Ser Asn Val lie Ala Ser Leu Thr Cys Gly 180 185 190 cgc cgc ttc gag tac gac gac cct cgc ttc etc agg ctg ctg gac eta 624

Arg Arg Phe Glu Tyr Asp Asp Pro Arg Phe Leu Arg Leu Leu Asp Leu 195 200 205 get cag gag gga ctg aag gag gag teg ggc ttt ctg cgc gag gtg ctg 672

Ala Gin Glu Gly Leu Lys Glu Glu Ser Gly Phe Leu Arg Glu Val Leu 210 215 220 aat get gtc ccc gtc etc ctg cat ate cca geg ctg get ggc aag gtc 720

Asn Ala Val Pro Val Leu Leu His lie Pro Ala Leu Ala Gly Lys Val 225 230 235 240 eta cgc ttc caa aag get ttc ctg acc cag ctg gat gag ctg eta act 768

Leu Arg Phe Gin Lys Ala Phe Leu Thr Gin Leu Asp Glu Leu Leu Thr 245 250 255 Printed by the Consumer Cooperatives of the Intellectual Property Bureau, Ministry of Economic Affairs gag cac agg atg acc tgg gac cca gcc cag ccc ccc ega gac ctg act 816

Glu His Arg Met Thr Trp Asp Pro Ala Gin Pro Pro Arg Asp Leu Thr 260 265 270 gag gcc ttc ctg gca gag atg gag aag gcc aag ggg aac cct gag age 864

Glu Ala Phe Leu Ala Glu Met Glu Lys Ala Lys Gly Asn Pro Glu Ser 275 280 285 age ttc aat gat gag aac ctg cgc ata gtg gtg get gac ctg ttc tet 912 This paper size applies the Chinese National Standard (CNS) A4 specification (210x297) Mm) -50- 200300798 A7 _ B7 ____ V. Description of the invention (θ

Ser Phe Asn Asp Glu Asn Leu Arg lie Val Val Ala Asp Leu Phe Ser 290 295 300 (Please read the notes on the back before filling in this page) gcc ggg atg gtg acc acc teg acc aeg ctg gec tgg ggc etc ctg etc 960

Ala Gly Met Yal Thr Thr Ser Thr Thr Leu Ala Trp Gly Leu Leu Leu 305 310 315 320 atg ate eta cat ccg gat gtg cag ege cgt gtc caa cag gag ate gac 1008

Met He Leu His Pro Asp Val. Gin Arg Arg Val Gin Gin Glu He Asp 325 330 335 gac gtg ata ggg cag gtg egg ega cca gag atg ggt gac cag get cac 1056

Asp Val lie Gly Gin Val Arg Arg Pro Glu Met Gly Asp Gin Ala His 340 345 350 atg ccc tac acc act gee gtg att cat gag gtg cag ege ttt ggg gac 1104

Met Pro Tyr Thr Thr Ala Val lie His Glu Val Gin Arg Phe Gly Asp 355 360 365 ate gtc ccc ctg ggt gtg acc cat atg aca tee cgt gac ate gaa gta 1152 lie Val Pro Leu Gly Val Thr His Met Thr Ser Arg Asp lie Glu Val 370 375 380 cag ggc ttc ege ate cct aag gga aeg aca etc ate acc aac ctg tea 1200

Gin Gly Phe Arg lie Pro Lys Gly Thr Thr Leu lie Thr Asn Leu Ser 385 390 395 400 Printed by the Consumer Cooperatives of the Intellectual Property Bureau of the Ministry of Economic Affairs teg gtg ctg aag gat gag gee gtc tgg gag aag ccc ttc ege ttc cac 1248

Ser Val Leu Lys Asp Glu Ala Val Trp Glu Lys Pro Phe Arg Phe His 405 410 415 ccc gaa cac ttc ctg gat gee cag ggc cac ttt gtg aag ccg gag gee 1296

Pro Glu His Phe Leu Asp Ala Gin Gly His Phe Val Lys Pro Glu Ala 420 425 430 ttc ctg cct ttc tea gca ggc ege cgt gca tgc etc ggg gag ccc ctg 1344 This paper size applies to the Chinese National Standard (CNS) A4 specification (210x297 (Mm) -51-200300798 A7, B7______ 5. Description of Invention

Phe Leu Pro Phe Ser Ala Gly Arg Arg Ala Cys Leu Gly Glu Pro Leu 435 440 445 gcc cgc atg gag etc ttc etc ttc ttc acc tcc ctg ctg cag cac ttc 1392 (Please read the notes on the back before filling this page)

Ala Arg Met Glu Leu Phe Leu Phe Phe Thr Ser Leu Leu Gin His Phe 450 455 460 age ttc teg gtg ccc act gga cag ccc egg ccc age cac cat ggt gtc 1440

Ser Phe Ser Val Pro Thr Gly Gin Pro Arg Pro Ser His His Gly Val 465 470 475 480 ttt get ttc ctg gtg age cca tcc ccc tat gag ett tgt get gtg ccc 1488 Phe Ala Phe Leu Val Ser Pro Ser Tyr Glu Leu Cys Ala Val Pro 485 490 495 cgc tag aat ggg gta cot agt ccc cag cct get cct age cca gag get 1536 Arg eta atg tac aat aaa gca atg tgg tag ttc c 1567 < 210〉 5 < 211〉 1567 < 212 > DNA < 213〉 Human / < 400〉 5 Printed by the Consumer Cooperatives of the Intellectual Property Bureau of the Ministry of Economic Affairs atg ggg eta gaa gca ctg gtg ccc ctg gcc gtg ata gtg gcc ate ttc 48

Met Gly Leu Glu Ala Leu Val Pro Leu Ala Val lie Val Ala lie Phe 1 5 10 15 ctg etc ctg gtg gac ctg atg cac egg cgc caa cgc tgg get gca cgc 96

Leu Leu Leu Val Asp Leu Met His Arg Arg Gin Arg Trp Ala Ala Arg 20 25 30 tac tea cca ggc ccc ctg cca ctg ccc ggg ctg ggc aac ctg ctg cat 144 This paper size applies to the Chinese National Standard (CNS) A4 specification (210X 297 mm) -52- 200300798 A7 _B7_____ 5. Description of the invention (β

Tyr Ser Pro Gly Pro Leu Pro Leu Pro Gly Leu Gly Asn Leu Leu His 35 40 45 (Please read the notes on the back before filling out this page) gtg gac ttc cag aac aca cca tac tgc ttc gac cag ttg egg ege ege 192

Val Asp Phe Gin Asn Thr Pro Tyr Cys Phe Asp Gin Leu Arg Arg Arg 50 55 60 ttc ggg gac gtg ttc age ctg cag ctg gee tgg aeg ccg gtg gtc gtg 240

Phe Gly Asp Val Phe Ser Leu Gin Leu Ala Trp Thr Pro Val Val Val 65 70 75 80 etc aat ggg ctg geg gee gtg ege gag geg ctg gtg acc cac ggc gag 288

Leu Asn Gly Leu Ala Ala Val Arg Glu Ala Leu Val Thr His Gly Glu 85 90 95 gac acc gee gac ege ccg cot gtg ccc ate acc cag ate ctg ggt ttc 336

Asp Thr Ala Asp Arg Pro Pro Val Pro lie Thr Gin lie Leu Gly Phe 100 105 110 ggg ccg cgt tee caa ggg gtg ttc ctg geg ege tat ggg ccc geg tgg 384

Gly Pro Arg Ser Gin Gly Val Phe Leu Ala Arg Tyr Gly Pro Ala Trp 115 120 125 c ^ c gag cag agg ege ttc tee gtg tee acc ttg ege aac ttg ggc ctg 432

Arg Glu Gin Arg Arg Phe Ser Val Ser Thr Leu Arg Asn Leu Gly Leu 135 140 Printed by the Consumer Cooperatives of the Intellectual Property Bureau of the Ministry of Economic Affairs ggc aag aag teg ctg gag cag tgg gtg acc gag gag gee gee tgc ett 480

Gly Lys Lys Ser Leu Glu Gin Trp Val Thr Glu Glu Ala Ala Cys Leu 145 150 155 160 tgt gee gee ttc gee aac cac tee gga ege ccc ttt ege ccc aac ggt 528

Cys Ala Ala Phe Ala Asn His Ser Gly Arg Pro Phe Arg Pro Asn Gly 165 170 175 etc ttg gac aaa gee gtg age aac gtg ate gee tee etc acc tgc ggg 576 — _ '> This paper size applies to Chinese national standards ( CNS) A4 specifications (210X 297 mm) -53- 200300798 A7 B7 One-fifth Intellectual Property Bureau of the Ministry of Economic Affairs printed a description of the invention (50 Leu Leu Asp Lys Ala Val Ser Asn Val lie Ala Ser Leu Thr Cys Gly 180 185 19〇cgc cgc ttc gag tac gac gac cct cgc ttc etc agg ctg ctg gac eta 624 Arg Arg Phe Glu Tyr Asp Asp Pro Arg Phe Leu Arg Leu Leu Asp Leu 195 200 205 get cag gag gga ctg aag gag tag ctg cgc gag gtg ctg 672 Ala Gin. Glu Gly Leu Lys Glu Glu Ser Gly Phe Leu Arg Glu Val Leu 210 215 220 aat get gtc ccc gtc etc ctg cat ate cca geg ctg get ggc aag gtc 720 Asn Ala Val Pro Val Leu His lie Pro Ala Leu Ala Gly Lys Val 225 230 235 240 eta cgc ttc caa aag get ttc ctg acc cag ctg gat gag ctg eta act 768 Leu Arg Phe Gin Lys Ala Phe Leu Thr Gin Leu Asp Glu Leu Leu Thr 245 250 255 gag c ac agg atg acc tgg gac cca gcc cag ccc ccc ega gac ctg act 816 Glu His Arg Met Thr Trp Asp Pro Ala Gin Pro Pro Arg Asp Leu Thr 260 265 270 gag gcc ttc ctg gca gag atg gag aag gcc aag ggg aac cct g age 864 Glu Ala Phe Leu Ala Glu Met Glu Lys Ala Lys Gly Asn Pro Glu Ser 275 280 285 age ttc aat gat gag aac ctg cgc ata gtg gtg get gac ctg ttc tet 912 Ser Phe Asn Asp Glu Asm Leu Arg lie Val Asp Leu Phe Ser 290 295 300 gcc ggg atg gtg acc acc teg acc aeg ctg gcc tgg ggc etc ctg etc 960 Ala Gly Met Val Thr Thr Ser Thr Thr Leu Ala Trp Gly Leu Leu Leu Leu 305 310 315 320 atg ate eta cat ccg gat gtg cag cgc cgt gtc caa cag gag ate gac 1008 (Please read the precautions on the back before filling out this page) (The size of the paper is applied to the Zhongguanjia Standard (CNS) M specification (2i〇x297 mm) -54 -200300798 A7 B7 V. Description of the invention (y

Met He Leu His Pro Asp Val Gin Arg Arg Val Gin Gin Glu He Asp 325 330 335 gac gtg ata ggg cag gtg egg ega cca gag atg ggt gac cag get cac 1056 (Please read the notes on the back before filling this page)

Asp Val lie Gly Gin Val Arg Arg Pro Glu Met Gly Asp Gin Ala His 340 345 350 atg ccc tac acc act gcc gtg att cat gag gtg cag ege ttt ggg gac 1104

Met Pro Tyr Thr Thr Ala Val lie His Glu Val Gin Arg Phe Gly Asp 355 360 365 ate gtc ccc ctg ggt gtg acc cat atg aca tec cgt gac ate gaa gta 1152 lie Val Pro Leu Gly Val Thr His Met Thr Ser Arg Asp lie Glu Val 370 375 380 cag ggc ttc ege ate cct aag gga aeg aca etc ate acc aac ctg tea 1200 Gin Gly Phe Arg He Pro Lys Gly Thr Thr Leu He Thr Asn Leu Ser 385 390 395 400 teg gtg ctg aag gat. Gag gee gtc tgg gag aag ccc ttc ege ttc cac 1248

Ser Val Leu Lys Asp Glu Ala Val Trp Glu Lys Pro Phe Arg Phe His 405 410 415 ccc gaa cac ttc ctg gat gcc cag ggc cac ttt gtg aag ccg gag gee 1296 Pro Glu His Phe Leu Asp Ala Gin Gly His Phe Val Lys Pro Glu Ala 420 425 430 Printed by the Consumer Cooperatives of the Intellectual Property Bureau of the Ministry of Economic Affairs ttc ctg cct ttc tea gca ggc ege cgt gca tgc etc ggg gag ccc ctg 1344 Phe Leu Pro Phe Ser Ala Gly Arg Arg Ala Cys Leu Gly Glu Pro Leu 435 440 445 gcc ege atg gag etc ttc etc ttc ttc acc tee ctg ctg cag cac ttc 1392 Ala Arg Met Glu Leu Phe Leu Phe Phe Thr Ser Leu Leu Gin His Phe 450 455 460 age ttc teg gtg ccc act gga cag ccc egg ccc age cat ggt gtc 1440 This paper size applies to Chinese National Standard (CNS) A4 (210X 297 mm) -55- 200300798 A7 B7

5. Description of the invention (

Ser Phe Ser Val Pro Thr Gly Gin Pro Arg Pro Ser His His Gly Val 465 470 475 480 ttt get ttc ctg gtg age cca tcc ccc tat gag ett tgt get gtg ccc 1488 Phe Ala Phe Leu Val Ser Pro Ser Tyr Glu Leu Cys Ala Val Pro 485 490 495 ege tag aat ggg gta cct agt ccc cag cct get cct age cca gag get 1536 Arg eta atg tac aat aaa gca atg tgg tag ttc c 1567 <210> 6 &lt; 211> 21 <212> DNA &lt; 213〉 Artificial sequence 1-&lt; 220〉 & &lt; 223 &gt; 1 Description of artificial sequence: synthetic DNA &lt; 400〉 6 gcaaaggcca tcatcagctc c 21 (Please read the precautions on the back before filling this page) • Binding and ordering Printed by the Consumer Cooperatives of the Intellectual Property Bureau of the Ministry of Economic Affairs &lt; 210 &gt; 7 &lt; 211〉 20 &lt; 212 &gt; DNA &lt; 213〉 Artificial Sequence 1 &lt; 220〉 &lt; 223 &gt; Description of Artificial Sequence · Synthetic DNA &lt; 400 〉 7 tctggtaggg gagcctcagc 20 This paper size is applicable to China National Standard (CNS) A4 (210X297 mm) -56- 200300798 A7 B7 V. Description of the invention (&lt; 210> 8 &lt; 211 &gt; 20

&lt; 212 &gt; DNA (please read the notes on the back before filling this page) &lt; 213〉 1 artificial sequence &lt; 220〉 &lt; 223〉 description of artificial sequence: synthetic! ^^^ &lt; 400> 8 taggacctgt agtctggggt 20

&lt; 210〉 9 &lt; 2H &gt; 20 &lt; 212 &gt; DNA &lt; 213 &gt; artificial sequence, &lt; 220〉 &lt; 223〉 description of artificial sequence: synthetic DNA &lt; 400 &gt; 9 gctcggacta cggtcatcac 20 Intellectual Property Office, Ministry of Economic Affairs Printed by Employee Consumer Cooperative

&lt; 210 &gt; 1〇 &lt; 211> 20 &lt; 212 &gt; DNA <213 &gt; .Artificial sequence &lt; 220 &gt; &lt; 223 &gt; Description of artificial sequence: Synthetic DNA &lt; 400 &gt; 10 cgggagacca gggggagcat 20 This paper is applicable to China National Standard (CNS) A4 Specification (210X297 mm) -57- 200300798 A7 Printed by the Consumer Cooperative of the Intellectual Property Bureau of the Ministry of Economic Affairs B7 V. Invention Description (54 &lt; 210〉 11 &lt; 211 &gt; 20 &lt; 212〉 DNA &lt; 213〉 artificial sequence: &lt; 220〉 &lt; 223〉 description of artificial sequence: synthetic DNA &lt; 400 &gt; Π ggtgggagat gcgggtaagg 20 &lt; 210> 12 &lt; 2Π &gt; 20 &lt; 212 &gt; DNA &lt; 213 &gt;; Artificial sequence, &lt; 220 &gt; <223 &gt; _Description of artificial sequence: synthetic DNA &lt; 400> 12 aaagcgggaa ct.gggaaggc 20 &lt; 210 &gt; 13 &lt; 211 &gt; 20 &lt; 212 &gt; DNA &lt; 213> artificial sequence 丨&Lt; 220〉 &lt; 223〉 Description of Artificial Sequences · Synthetic DNA &lt; 400 &gt; 13 tgctcttccg aggccccagt 20 (Please read the precautions on the back before filling this page)

、 1T plus This paper size applies the Chinese National Standard (CNS) A4 specification (210X 297mm) -58- 200300798 A7 B7 V. Description of the invention (5 go &lt; 210〉 14 &lt; 211〉 20

&lt; 212〉 DNA (please read the notes on the back before filling out this page) &lt; 213〉 artificial sequence &lt; 220〉

&lt; 223〉 Description of artificial sequence: synthetic DNA &lt; 400 &gt; 14 gagatggctg gggcctgaga 20 &lt; 210 &gt; 15 &lt; 211 &gt; 20 &lt; 212〉 DNA &lt; 213〉. artificial sequence &lt; 220〉

<223> Description of artificial sequence: synthetic DNA &lt; 400> 15 ccaacctttt gcccagcctc 20 Printed by the Consumer Cooperative of Intellectual Property Bureau, Ministry of Economic Affairs

&lt; 210〉 16 &lt; 211〉 20 &lt; 212〉 DNA &lt; 213〉 artificial sequence * &lt; 220〉 &lt; 223 &gt;. Description of artificial sequence: synthetic DNA &lt; 400> 16 cccgttctgt cccgagtatg 20 Applicable to paper size China National Standard (CNS) A4 Specification (· 210X 297mm) -59- 200300798 A7, Β7 V. Description of Invention (5) &lt; 210 &gt; 17

&lt; 211〉 20 &lt; 212 &gt; DNA (Please read the notes on the back before filling this page) Human sequence / Λ &lt; 220> &lt; 223 &gt; Description of the artificial sequence: synthetic DNA &lt; 400 &gt; 17 ggtgtcccag caaagttcat 20

&lt; 21〇 &gt; 18 &lt; 211〉 22 &lt; 212〉 DNA &lt; 213 &gt;. artificial sequence &lt; 220>

&lt; 223〉 Description of artificial sequence: synthetic DNA &lt; 400 &gt; 18 taagcctgac ctcctccaac at 22 Printed by the Consumer Cooperative of Intellectual Property Bureau, Ministry of Economic Affairs

&lt; 210〉 19 &lt; 211 &gt; 20 &lt; 212〉 DNA &lt; 213 &gt; artificial sequence ’&lt; 220>

&lt; 223 &gt; Description of artificial sequence ·· Synthetic DNA &lt; 400 &gt; 19 agtgggccca cctggcagta 20 This paper is applicable to China National Standards (CNS) A4 specifications (210 × 297 mm) -60- 200300798 A7 B7 Intellectual Property Bureau, Ministry of Economic Affairs Printed by Employee Consumer Cooperatives, Description of Invention (&lt; 210〉 20 &lt; 211〉 20 &lt; 212 &gt; DNA &lt; 213〉 Artificial Sequence &lt; 220〉 &lt; 223〉 Description of Artificial Sequence: Synthetic DNA &lt; 400〉 20 ccccgtgtgt ttggtggcag 20 &lt; 210〉 21 &lt; 211〉 20 &lt; 212 &gt; DNA &lt; 213 &gt; artificial sequence &lt; 220> &lt; 223 &gt; Description of artificial sequence: synthetic DNA &lt; 400 &gt; 21 cgtcagtgag cctggctcct 20 &lt; 210〉 22 &lt; 211 &gt; 20 &lt; 212 &gt; DNA &lt; 213 &gt; .Artificial sequence &lt; 220〉 &lt; 223 &gt; Description of artificial sequence: synthetic DNA &lt; 400〉 22 tcttgcaggg gtatcaccca 20 (Please read the note on the back first Please fill in this page for matters) • The size of the bound and bound paper is in accordance with the Chinese National Standard (CNS) A4 specification (210X297 mm) -61-200300798 A7. · B7 Printed by the Intellectual Property Bureau of the Ministry of Economic Affairs Consumer Cooperatives 5έ &lt; 21 0〉 23 &lt; 2Π &gt; 20 &lt; 212〉 DNA &lt; 2U &gt; artificial sequence &lt; 220〉 &lt; 223 &gt; Description of artificial sequence: synthetic DNA. ________ „&lt; 400〉 23 ttgccctgag gaggatgatc 20 &lt; 21〇 &gt; 24 &lt; 211 &gt; 20 &lt; 212 &gt; DNA &lt; 213 &gt; artificial sequence &lt; 220〉 &lt; 223〉 description of artificial sequence: synthetic DNA VII &lt; 400> 24 ctcagcctca acgtacccct 20 &lt; 210 &gt; 25 &lt; 211 〉 20 &lt; 212〉 DNA &lt; 213 &gt; artificial sequence &lt; 220〉 &lt; 223〉 Description of artificial sequence: synthetic DNA v &lt; 400〉 25 catttggtag tgaggcaggt 20 (Please read the precautions on the back before filling this page) The size of the bound paper is in accordance with the Chinese National Standard (CNS) A4 specification (210X297 mm) -62- 200300798 A7 B7 Printed by the Intellectual Property Bureau Employee Consumer Cooperative of the Ministry of Economic Affairs and the invention description (5 go &lt; 210〉 26 &lt; 211〉 19 &lt; 212 &gt; DNA &lt; 213 &gt; Artificial sequence &lt; 220 &gt; &lt; 223 &gt; Description of artificial sequence: Synthetic DNA &lt; 400〉 26 ctccaggacc tcctccctc 19 &lt; 210〉 27 &lt; 211 &gt; 20 &lt; 212 &gt; DNA &lt; 213 &gt; Artificial Sequence &lt; 220 &gt; &lt; 223 &gt; Human Description of synthetic sequence: synthetic DNA &lt; 400 &gt; 27 ggccctgacc ctccctctgc 20 &lt; 210> 28 &lt; 211> 20 &lt; 212 &gt; DNA 〈213〉 artificial sequence &lt; 220 &gt; &lt; 223 &gt; description of synthetic sequence: synthetic DNA &lt; 400〉 28 ctctgtcccc accgctgctt 20 (Please read the precautions on the back before filling in this page) C. Order i # This paper size is applicable to China National Standard (CNS) A4 specification (210'〆297 mm) -63- 200300798 A7 Printed by the Consumer Cooperative of the Intellectual Property Bureau of the Ministry of Economic Affairs B7 V. Description of the invention (60 &lt; 210 &gt; 29 &lt; 211 &gt; 16 &lt; 212 &gt; DNA &lt; 213 &gt; Artificial sequence &lt; 220〉 &lt; 223〉 Artificial sequence Description: Synthetic DNA &lt; 400〉 29 tgcccgtccc accccc 16 &lt; 210〉 30 &lt; 211〉 19 &lt; 212〉 DNA &lt; 213 &gt; Artificial Sequence &lt; 220〉 &lt; 223〉 Description of Artificial Sequence: Synthetic DNA &lt; 400 &gt; 30 gcccttctgc ccatcaccc 19 &lt; 210〉 31 &lt; 211〉 18 &lt; 212〉 DNA &lt; 213 &gt; Artificial Sequence &lt; 220〉 &lt; 223 &gt; Description of Artificial Sequence: Synthetic DNA &lt; 400〉 31 cccgcatctc ccaccccc 18 (Please read the notes on the back before filling (Page) Binding. 1 # This paper size is applicable to China National Standard (CNS) A4 specification (210X297 mm) -64- 200300798 A7 B7 Printed by the Ministry of Economic Affairs, Intellectual Property Bureau X Consumer Cooperatives 5. Description of the invention (6) &lt; 210〉 32 &lt; 211〉 19 &lt; 212 &gt; DNA &lt; 213> artificial sequence &lt; 220 &gt; &lt; 223〉 description of artificial sequence: synthetic DNA &lt; 400 &gt; 32 cctcggtctc tcgctccgc 19 &lt; 210> 33 &lt; 211〉 20 &lt; 212〉 DNA &lt; 213 &gt; artificial sequence &lt; 220〉 &lt; 223〉 description of artificial sequence: synthetic DNA &lt; 400 &gt; 33 gaccccgttc tgtctggtgt 20 &lt; 210〉 34 &lt; 211 &gt; 17 &lt; 212 &gt; DNA &lt; 213〉 Artificial sequence &lt; 220〉 &lt; 223〉 Description of artificial sequence: Synthetic DNA &lt; 400 &gt; 34 ccgtggcagc cactctc 17 (Please read the precautions on the back before filling this page) -pack-

、 1T This paper size applies Chinese National Standard (CNS) A4 specification (210X 297 mm) -65- 200300798 A7 B7 Printed by the Consumer Cooperatives of the Intellectual Property Bureau of the Ministry of Economic Affairs 5. Description of the invention (&lt; 210 &gt; 35 &lt; 211〉 20 &lt; 212〉 DNA &lt; 213 &gt; artificial sequence &lt; 220〉 &lt; 223〉 description of artificial sequence: synthetic DNA &lt; 400> 35 gtatgctctc ggccctgctc 20 &lt; 210> 36 &lt; 211 &gt; 20 &lt; 212 &gt; DNA <213 &gt; Artificial Sequence &lt; 220> &lt; 223 &gt; Description of Artificial Sequence: Synthetic DNA &lt; 400 &gt; 36 actgtttccc agatgggctc 20 &lt; 210> 37 &lt; 211 &gt; 20 &lt; 212 &gt; DNA &lt; 213>, artificial sequence &lt; 220〉 &lt; 223〉 Description of artificial sequence: Synthetic DNA &lt; 400〉 37 g ^ ggggacgc atgtctgtcc 20 (Please read the notes on the back before filling out this page) The size of the binding and binding papers is in accordance with Chinese National Standards (CNS ) A4 specification (210X 297 mm) -66-200300798 A7 Printed by B7 of the Intellectual Property Bureau of the Ministry of Economic Affairs Consumer Cooperatives V. Invention description (6 knows &lt; 210 &gt; 38 &lt; 211〉 16 &lt; 212> DNA &lt; 213 &gt; Artificial sequence &lt; 220 &gt; &lt; 223〉 丨Description of synthetic sequence: synthetic DNA &lt; 400 &gt; 38 gg ^ gggcgcc ^ ggcct 16 &lt; 210 &gt; 39 &lt; 211〉 20 &lt; 212 &gt; DNA &lt; 213 &gt; artificial sequence &lt; 220 &gt; &lt; 223> artificial sequence description : Synthetic DNA &lt; 400 &gt; 39 tcaccctgca tctcctgccc 20 &lt; 210 &gt; 40 &lt; 211> 17 &lt; 212 &gt; DNA &lt; 213> Artificial sequence &lt; 220 &gt; &lt; 223 &gt; Description of artificial sequence · Synthetic DNA &lt; 400〉 40 ccgggctccc cacaggc 17 (Please read the precautions on the reverse side before filling out this page) Binding and ordering This paper size applies to China National Standard (CNS) A4 (210X297 mm) -67- 200300798 A7 B7 Intellectual Property of the Ministry of Economic Affairs Printed by the Bureau ’s Consumer Cooperatives V. Description of the invention (64 &lt; 210 &gt; 41 &lt; 211 &gt; 15 &lt; 212 &gt; DNA &lt; 213〉. Artificial sequence &lt; 220〉 &lt; 223〉 Description of artificial sequence: Synthetic DNA &lt; 400 &gt; 41 ccctcccctc cccac 15 &lt; 210〉 42 &lt; 211〉 20 &lt; 212〉 DNA &lt; 213〉 artificial sequence &lt; 220〉 &lt; 223 &gt; description of artificial sequence: synthetic DNA &lt; 400 &gt; 42 tggggactag gtaccccatt 20 &lt; 210 &gt; 43 &lt; 2Π &gt; 21 <212 &gt; DNA & l t; 213〉 Artificial sequence &lt; 220 &gt; &lt; 223〉 Description of artificial sequence: synthetic DNA &lt; 400 &gt; 43 gcgacaattc aagtgtggtg a 21 (Please read the precautions on the back before filling this page)

、 1T This paper size applies Chinese National Standard (CNS) A4 specification (210X 297 mm) -68- 200300798 A7 、 B7 Printed by the Intellectual Property Bureau of the Ministry of Economic Affairs Consumer Cooperatives V. Invention Description (6 Meetings &lt; 210〉 44 <211 &gt; 21 &lt; 212 &gt; DNA &lt; 213> Artificial sequence &lt; 220> &lt; 223> Description of artificial sequence: synthetic DNA &lt; 400 &gt; 44 gcatgagcta aggcacccag a 21 &lt; 210> 45 &lt; 211> 20 &lt; 212 &gt; DNA &lt; 213 &gt; Artificial sequence &lt; 220> <223 &gt; Description of artificial sequence: synthetic DNA &lt; 400> 45 acgctgggct gcacgctacc 20. &Lt; 210 &gt; 46 &lt; 211> 20 &lt; 212 &gt; DNA &lt; 213〉 Artificial Sequence &lt; 220〉 &lt; 223〉 Description of Artificial Sequence: Synthetic DNA &lt; 400〉 46 acgctgggct gcacgctact 20 (Please read the notes on the back before filling out this page)-Packing · 11 National Standard (CNS) A4 Specification (210X297 mm) -69- 200300798 A7 Printed by the Consumer Cooperative of the Intellectual Property Bureau of the Ministry of Economic Affairs B7 V. Invention Description (&lt; 210〉 47 &lt; 211〉 22 &lt; 212 &gt; DNA &lt; 213 &gt; Artificial sequence &lt; 220〉 • &lt; 223 &gt; Description of artificial sequence: synthetic DNA &lt; 400 &gt; 47 agcttcaatg atgagaacct go 22 &lt; 210 &gt; 48 &lt; 211> 22 &lt; 212 &gt; DNA &lt; 213 &gt; artificial sequence &lt; 220 &gt; &lt; 223 &gt; artificial Description of the sequence: synthetic DNA &lt; 400 &gt; 48 agcttcaatg atgagaacct gt, 22 &lt; 210 &gt; 49 &lt; 211 &gt; 20 &lt; 212 &gt; DNA &lt; 213 &gt; artificial sequence &lt; 220〉 &lt; 223> artificial sequence description: Synthetic DNA &lt; 400 &gt; 49 aggtcagcca ccactatgcg 20 (Please read the notes on the back before filling out this page) C · The size of the paper is applicable to the Chinese National Standard (CNS) A4 specification (210'〆297 mm) -70- 200300798 A7, 1 B7 Printed by the Consumer Cooperative of the Intellectual Property Bureau of the Ministry of Economic Affairs V. Invention Description (6 &gt; &lt; 210〉 50 &lt; 211〉 20 &lt; 212 &gt; DNA &lt; 213 &gt; Artificial Sequence &lt; 220〉 &lt; 223〉 Description of artificial sequence: synthetic DNA &lt; 400 &gt; 50 aggtcagcca ccactatgca 20 &lt; 210 &gt; 51 &lt; 211〉 20 &lt; 212〉 DNA &lt; 213 &gt; artificial sequence &lt; 220〉 &lt; 223> description of artificial sequence: synthesis DNA &lt; 400〉 51 aagctcatag ggggatgggc 20 &lt; 2 10〉 52 &lt; 211〉 20 &lt; 212 &gt; DNA &lt; 213〉 Artificial sequence &lt; 220 &gt; &lt; 223 &gt; Description of artificial sequence: synthetic DNA &lt; 400〉 52 * aagctcatag ggggatgggg 20 (Please read the note on the back first (Fill in this page again)

This paper size applies to China National Standard (CNS) A4 (210X297 mm) -71-200300798 A7 '' B7 Printed by the Consumer Cooperatives of the Intellectual Property Bureau of the Ministry of Economic Affairs 5. Description of Invention (6 & &lt; 210 &gt; 53 &lt; 211 〉 20 <212 &gt; DNA &lt; 213> Artificial sequence &lt; 220> &lt; 223> Description of artificial sequence: synthetic DNA &lt; 400 &gt; 53 atggggctag aagcactggt 20 &lt; 210> 54 &lt; 2Π &gt; 25 &lt; 212> DNA &lt; 213> Artificial sequence &lt; 220> <223 &gt; Description of artificial sequence · Synthetic DNA &lt; 400> 54 ggcagggggc ctggtgagta gcgtg 25 &lt; 210> 55 &lt; 211 &gt; 25 &lt; 212 &gt; DNA <213> artificial sequence &lt;; 220 &gt; &lt; 223〉 Description of artificial sequence: synthetic DNA &lt; '400 &gt; 55 ctgggctgca cgctactcac caggc 25 (Please read the precautions on the back before filling this page).

、 1T This paper size is applicable to China National Standard (CNS) A4 specification (210X mm) -72- 200300798 A7 * B7 _ I l. V. Description of the invention (乂 &lt; 210> 56 &lt; 211 &gt; 21

&lt; 212 &gt; DNA &lt; 213〉 artificial sequence, &lt; 220>

&lt; 223〉 Description of artificial sequence: synthetic DNA ^ etic I &lt; 400 &gt; 56 ctagcggggc acagcacaaa g 21 I -------- install-(Please read the precautions on the back before filling this page)

、 1T Printed by the Consumer Cooperatives of the Intellectual Property Bureau of the Ministry of Economic Affairs This paper size applies to China National Standard (CNS) Α4 specification (210 × 297 mm) -73-

Claims (1)

200300798 A8 B8 C8 D8 Printed by the Consumer Cooperatives of the Intellectual Property Bureau of the Ministry of Economic Affairs 6. Application for patent scope 1 1. A nucleic acid whose at least 10 consecutive nucleotides in the base sequence described in SEQ ID No. 1 is characterized by Contains a variant comprising any substitution of G from A at base 125, a variant from C at T of base 1858, a variant from T at C of base 2874, or Nucleic acid at 10 to 100 bases of the mutated polymorphic site in which the C at base 2875 is substituted by T. 2. If the nucleic acid in the scope of patent application No. 1 is DNA. 3. If the nucleic acid of item 1 of the patent application scope, it is RNA. 4. A dual gene-specific oligonucleotide, characterized in that it contains a mutation in which G is arbitrarily selected from bases 1 to 25 of base sequence described in SEQ ID NO: 1 and bases 1 85 to 8 A mutation in which C is replaced by T, a mutation in which T is replaced by C at base 2874, or a sequence of a polymorphic site in a mutation in which C is replaced by T at base 2875, or a hybrid thereof. 5. For example, dual gene-specific oligonucleotides in the scope of patent application, which can be used as probes or primers. 6. — An individual's genetic polymorphism analysis method, which is characterized by containing (1) a step of preparing a nucleic acid by the individual; and (2) for the nucleic acid prepared by step (1), arbitrarily selected from the sequence number 1 A variation in which G is replaced by A at base 1 to 25, a variation in which C is replaced by T at base 1 85 and a variation in which T is replaced by C at base 2874, or The procedure for determining the base of the polymorphic site of C at base 2875 with a T substitution. 7. —A method of judging the metabolic activity of a drug, which is characterized by containing (1) a step of preparing nucleic acid by an individual; this paper size applies the Chinese National Standard (CNS) A4 specification (210X297 mm) &quot; '&quot;- 74- (Please read the precautions on the back before filling this page). Order 200300798 A8 B8 C8 D8 __ VI. Patent application scope 2 (2) For the nucleic acid prepared in step (1), any one is selected from the sequence number A mutation in which G is replaced by A at base 125, a mutation in which C is replaced by T at base 1 85 and a mutation in which T is replaced by C at base 2874, or The step of determining the base of the variant polymorphic site where the C of base No. 2875 is substituted by T; and (3) predicting the metabolic activity of the individual based on the base information sequenced in step (2) step. 8. A gene characterized by having a base sequence according to any one of SEQ ID NOs: 2 to 5. 9. A protein characterized by having an amino acid sequence according to any one of SEQ ID NOs: 2 to 5. 10. A method for detecting a gene having a base sequence described in SEQ ID NO: 2 or SEQ ID NO: 5, which is characterized in that it contains a nucleic acid prepared by an individual and is the first in the base sequence described in SEQ ID NO: 1. A step of detecting whether there is a variation in which C is replaced by T at base 0, a variation in which C is replaced by T at base 2850, and a variation in which G is replaced by C at base 4180 is present. (Please read the notes on the back before filling out this page) Printed by the Consumer Cooperatives of the Intellectual Property Bureau of the Ministry of Economic Affairs This paper is sized for the Chinese National Standard (CNS) A4 (210 X 297 mm) 75-