KR102629338B1 - Analytical method for diagnosing obesity or diabetes and kit therefor - Google Patents

Analytical method for diagnosing obesity or diabetes and kit therefor Download PDF

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KR102629338B1
KR102629338B1 KR1020170000569A KR20170000569A KR102629338B1 KR 102629338 B1 KR102629338 B1 KR 102629338B1 KR 1020170000569 A KR1020170000569 A KR 1020170000569A KR 20170000569 A KR20170000569 A KR 20170000569A KR 102629338 B1 KR102629338 B1 KR 102629338B1
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백광현
최종호
임기환
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차의과학대학교 산학협력단
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
    • C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G01N33/6893—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids related to diseases not provided for elsewhere
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2800/00—Detection or diagnosis of diseases
    • G01N2800/04—Endocrine or metabolic disorders
    • G01N2800/042—Disorders of carbohydrate metabolism, e.g. diabetes, glucose metabolism
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2800/00—Detection or diagnosis of diseases
    • G01N2800/04—Endocrine or metabolic disorders
    • G01N2800/044—Hyperlipemia or hypolipemia, e.g. dyslipidaemia, obesity

Abstract

본 발명은 비만 또는 당뇨병 진단에 필요한 정보를 제공하기 위하여, 대상자의 시료 중 서열번호 1 내지 4의 단백질로 이루어진 군으로부터 선택된 단백질, 즉 USP19를 코딩하는 유전자의 발현량을 측정하는 단계를 포함하는 분석방법을 제공한다. 또한, 본 발명은 서열번호 1 내지 4의 단백질로 이루어진 군으로부터 선택된 단백질을 코딩하는 유전자의 발현량을 측정할 수 있는 분자를 포함하는 비만 또는 당뇨병 진단용 키트를 제공한다.The present invention is an analysis comprising measuring the expression level of a gene encoding USP19, a protein selected from the group consisting of proteins of SEQ ID NOs. 1 to 4 in a sample of a subject, in order to provide information necessary for diagnosing obesity or diabetes. Provides a method. Additionally, the present invention provides a kit for diagnosing obesity or diabetes, including a molecule capable of measuring the expression level of a gene encoding a protein selected from the group consisting of proteins of SEQ ID NOs: 1 to 4.

Description

비만 또는 당뇨병 진단을 위한 분석방법 및 키트{Analytical method for diagnosing obesity or diabetes and kit therefor}{Analytical method for diagnosing obesity or diabetes and kit therefor}

본 발명은 비만 또는 당뇨병 진단에 필요한 정보를 제공하기 위한 분석방법 및 비만 또는 당뇨병 진단용 키트에 관한 것이다.The present invention relates to an analysis method for providing information necessary for diagnosing obesity or diabetes, and a kit for diagnosing obesity or diabetes.

지방세포에 저장된 지방은 체내의 중요한 에너지원으로 사용된다. 그러나, 비만이 진행됨에 따라서 지방세포는 수적 증가가 일어날 뿐만 아니라 과다한 지방세포 분화에 의하여 지방세포의 크기증가를 포함한 형태적 변화와 여러 유전자 발현의 변화를 동반한다. 지방세포 분화는 인슐린이나 인슐린 성장인자-1(insulin like growth factor-1), 성장호르몬 등의 자극에 의하여 촉진되며, 이 과정에 PPAR-γ(peroxisome proliferator-activated receptor gamma) 등의 전사인자들의 증가가 관찰된다. 이들 전사인자들은 지방세포 조절인자와 더불어 지방세포의 분화를 촉진시키며 지방산 결합단백질인 aP2 또는 지방산 생합성효소(fatty acid synthase)과 같은 효소들의 발현량이 증가하게 된다. 비만이 진행되어 당뇨병으로 진행되는 제2형 당뇨병의 경우에 있어서 중성지방인 팔미테이트들이 점진적으로 인슐린 저항성을 유발하며 궁극적으로 췌장의 베타 세포를 파괴하여 당뇨를 유도한다(Shimabukuro M, et al. Proc. Natl. Acad. Sci. USA. 95, 2498-2502, 1998).Fat stored in fat cells is used as an important energy source in the body. However, as obesity progresses, not only do adipocytes increase in number, but excessive adipocyte differentiation is accompanied by morphological changes, including an increase in adipocyte size, and changes in the expression of several genes. Adipocyte differentiation is promoted by stimulation of insulin, insulin like growth factor-1 (insulin like growth factor-1), and growth hormones, and during this process, transcription factors such as PPAR-γ (peroxisome proliferator-activated receptor gamma) increase. is observed. These transcription factors, along with adipocyte regulatory factors, promote differentiation of adipocytes and increase the expression of enzymes such as fatty acid binding protein aP2 or fatty acid synthase. In the case of type 2 diabetes, in which obesity progresses to diabetes, palmitate, a neutral fat, gradually causes insulin resistance and ultimately destroys pancreatic beta cells, inducing diabetes (Shimabukuro M, et al. Proc . Natl. Acad. Sci. USA. 95, 2498-2502, 1998).

인슐린, TGF-beta 신호(transforming growth factor-beta), 섬유아세포 성장인자(fibroblast growth factor), 그리고 골 형성 단백질(bone morphogenetic protein)과 같은 다양한 자극 인자에 의해 촉진된 지방세포 분화 과정에 있어, 지방세포 조절 인자와 전사인자들은 유비퀴틴-프로테아솜 시스템 (Ubiquitin-proteasome system, UPS)에 의해 조절된다. 이를 통해, 유비퀴틴화 효소와 탈유비퀴틴화 효소가 다양한 유전자의 발현에 영향을 미치면서 항상성을 조절한다. 따라서, 이러한 항상성의 균형이 무너지면서 지방세포 분화와 관련된 유전자의 과발현 및 과활성이 되면 당뇨병이 유도되게 된다.In the process of adipocyte differentiation promoted by various stimulating factors such as insulin, TGF-beta signaling (transforming growth factor-beta), fibroblast growth factor, and bone morphogenetic protein, fat Cellular regulatory factors and transcription factors are regulated by the ubiquitin-proteasome system (UPS). Through this, ubiquitination and deubiquitination enzymes regulate homeostasis by affecting the expression of various genes. Therefore, when the balance of homeostasis is disrupted and genes related to adipocyte differentiation are overexpressed and overactivated, diabetes is induced.

한편, 이전 연구에서 NCoR 복합체(nuclear co-repressor complex)에 HDAC3, GPS2, TBL1, TBLR1, TAB2 및 CORO2A가 포함됨이 밝혀진 바 있다. NCoR은 THR(thyroid hormone receptor)과 RAR(retinoic acid receptor)과 같은 파트너들과의 상호작용을 통해서 핵의 수용체와 매개되는 유전자 억제에 중요한 기능을 한다. NCoR은 Akt와 연결된 인산화에 의해서 1450번째의 세린에서 인산화되어 단백질의 접힘(protein refolding)을 유도하며, 잘못 접혀지고 퇴화된 NCoR은 급성 골수성 백혈병과 연관된다(Nin DS, et al. PLoS One. 2013; 8:e70891). 그러나 이외의 다른 co-suppressor 단백질들의 번역 후 수정(post-translational modifications)의 역할은 거의 밝혀져 있지 않다. 몇몇의 연구들은 THR과 RAR이 NCoR 복합체를 동원하며, NcoR 복합체의 C-말단 도메인에 있는 corepressor-nuclear receptor(CoRNR) box 모티프가 RAR와 결합 시 중요한 역할을 한다고 보고된 바 있다(Park UH, et al. J Biol Chem . 2010; 285:34269-34278; Mengeling BJ, et al. J Biol Chem. 2011; 286:4236-4247; Hu X, et al. Nature. 1999; 402:93-96; Cohen RN, et al. Mol Endocrinol . 2001; 15:1049-1061; Webb P, et al. Mol Endocrinol . 2000; 14:1976-1985; Makowski A, et al. Mol Endocrinol. 2003; 17:273-286).Meanwhile, previous studies have shown that the NCoR complex (nuclear co-repressor complex) includes HDAC3, GPS2, TBL1, TBLR1, TAB2, and CORO2A. NCoR plays an important role in nuclear receptor-mediated gene repression through interactions with partners such as thyroid hormone receptor (THR) and retinoic acid receptor (RAR). NCoR is phosphorylated at serine at position 1450 by Akt-linked phosphorylation, leading to protein refolding, and misfolded and degenerated NCoR is associated with acute myeloid leukemia (Nin DS, et al. PLoS One . 2013 ;8:e70891). However, the role of post-translational modifications of other co-suppressor proteins is largely unknown. Several studies have reported that THR and RAR recruit the NCoR complex and that the corepressor-nuclear receptor (CoRNR) box motif in the C-terminal domain of the NcoR complex plays an important role in binding to RAR (Park UH, et al. al. J Biol. Chem . 2010; 285:34269-34278; Mengeling BJ, et al. J Biol Chem. 2011; 286:4236-4247; Hu X, et al. Nature . 1999; 402:93-96; Cohen RN, et al. Mol Endocrinol . 2001; 15:1049-1061; Webb P, et al. Mol Endocrinol . 2000; 14:1976-1985; Makowski A, et al. Mol Endocrinol . 2003; 17:273-286).

코로닌(Coronin)은 딕티오스텔리움 디스코이데움(Dictyostelium discoideum)의 수용성 액틴-결합 단백질로서 동정되었으며(de Hostos EL, et al. EMBO J. 1991; 10:4097-4104), 몇몇의 동형체(isoforms)가 존재한다. 코로닌의 동형체는 계통생리학적으로 3가지 유형으로 분류되며(Uetrecht AC, et al. Trends Cell Biol . 2006; 16:421-426), 가장 많이 발현되는 것은 CORO2A(코로닌 2A, coronin 2A)로서 IR10로도 지칭되고, CORO2A의 발현은 상대적으로 정소, 피질, 십이지장, 림프 노드, 난소 및 자궁에서 높게 나타난다(Cai L, et al. J Biol Chem . 2005; 280:31913-31923). 비록 CORO2A의 생물학적 기능은 잘 알려져 있지 않으나, 최근 연구에서 NCoR 복합체의 구성요소라는 것이 밝혀진 바 있다(Perissi V, et al. Nat Rev Genet. 2010; 11:109-123). 또한, NCoR과 SMRT는 PPAR-γ 유전자 전사를 억제시키며, NCoR은 지방세포 분화에 있어 PPAR-γ의 인산화와 연관되고, NCoR 복합체의 발현 억제(knock-down)는 지방생성(adipogenesis)을 촉진시키는 것으로 밝혀진 바 있다(Yu C, et al. J Biol Chem. 2005; 280:13600-13605). PPAR-γ는 당 대사에서 지방 조직의 필수 요소로서, 지방세포의 분화에 있어 중요한 역할을 한다(Jacobi D, et al. Adipocyte 2012; 1:4-12).Coronin (Dictyostelium discoideum)Dictyostelium discoideum) was identified as a soluble actin-binding protein (de Hostos EL, et al.EMBO J. 1991; 10:4097-4104), several isoforms exist. Isoforms of coronin are phylophysiologically classified into three types (Uetrecht AC, et al.Trends Cell Biol . 2006; 16:421-426), the most expressed one is CORO2A (coronin 2A), also referred to as IR10, and the expression of CORO2A is relatively high in testes, cortex, duodenum, lymph nodes, ovaries, and uterus (Cai L, et al.J Biol Chem . 2005; 280:31913-31923). Although the biological function of CORO2A is not well known, recent studies have shown that it is a component of the NCoR complex (Perissi V, et al.Nat Rev Genet.2010; 11:109-123). In addition, NCoR and SMRT inhibit PPAR-γ gene transcription, NCoR is associated with phosphorylation of PPAR-γ in adipocyte differentiation, and knock-down of the expression of the NCoR complex promotes adipogenesis. It has been found that (Yu C, et al.J Biol Chem. 2005; 280:13600-13605). PPAR-γ is an essential component of adipose tissue in glucose metabolism and plays an important role in the differentiation of adipocytes (Jacobi D, et al.Adipocyte 2012; 1:4-12).

따라서, NCoR 복합체의 구성요소 중 하나인 CORO2A의 결합물질(binding partner)를 밝혀내고, 또한 이들의 기능을 밝혀낼 경우, 지방생성(adipogenesis) 및 이와 밀접하게 연관된 질환, 예를 들어 비만 또는 당뇨병을 진단하는데 유용하게 사용될 수 있을 것으로 기대된다. Therefore, if the binding partner of CORO2A, one of the components of the NCoR complex, is discovered and its function is discovered, adipogenesis and diseases closely related to it, such as obesity or diabetes, can be prevented. It is expected that it will be useful for diagnosis.

본 발명자들은 CORO2A의 번역 후 수정(post-translational modifications)의 분자생물학적 기전을 규명하기 위하여 다양한 연구를 수행하였다. 본 발명자들은 탈유비퀴틴화 효소 중 하나인 USP19가 CORO2A의 탈유비퀴틴화를 통하여 CORO2A의 안정화를 유도함으로써 RAR의 전사를 억제한다는 것을 밝혀냈다. 따라서, USP19가 과발현될 경우 CORO2A를 활성화시켜 NCoR 복합체의 활성 억제로 이어지고, 이는 지방생성(adipogenesis)을 촉진시키게 된다. 즉, USP19의 과발현이 검출될 경우 비만 또는 당뇨병을 진단하는데 유용하게 사용될 수 있으며, USP19는 비만 또는 당뇨병 진단을 위한 바이오마커로서 사용될 수 있다.The present inventors conducted various studies to identify the molecular biological mechanism of post-translational modifications of CORO2A. The present inventors found that USP19, one of the deubiquitination enzymes, inhibits the transcription of RAR by inducing stabilization of CORO2A through deubiquitination of CORO2A. Therefore, when USP19 is overexpressed, it activates CORO2A, leading to inhibition of the activity of the NCoR complex, which promotes adipogenesis. That is, when overexpression of USP19 is detected, it can be usefully used to diagnose obesity or diabetes, and USP19 can be used as a biomarker for diagnosing obesity or diabetes.

따라서, 본 발명은 비만 또는 당뇨병 진단에 필요한 정보를 제공하기 위하여, USP19를 이용한 분석방법을 제공하는 것을 목적으로 한다.Therefore, the purpose of the present invention is to provide an analysis method using USP19 in order to provide information necessary for diagnosing obesity or diabetes.

또한, 본 발명은 상기 USP19를 코딩하는 유전자의 발현량을 측정할 수 있는 분자를 포함하는 비만 또는 당뇨병의 진단용 키트를 제공하는 것을 목적으로 한다.Additionally, the present invention aims to provide a diagnostic kit for obesity or diabetes that includes a molecule capable of measuring the expression level of the gene encoding USP19.

본 발명의 일 태양에 따라, 비만 또는 당뇨병 진단에 필요한 정보를 제공하기 위하여, 대상자의 시료 중 서열번호 1 내지 4의 단백질로 이루어진 군으로부터 선택된 단백질을 코딩하는 유전자의 발현량을 측정하는 단계를 포함하는 분석방법이 제공된다.According to one aspect of the present invention, in order to provide information necessary for diagnosing obesity or diabetes, it includes measuring the expression level of a gene encoding a protein selected from the group consisting of proteins of SEQ ID NOs: 1 to 4 in a sample of a subject. An analysis method is provided.

본 발명의 분석방법에 있어서, 상기 단백질을 코딩하는 유전자는 서열번호 4의 단백질을 코딩하는 유전자일 수 있다. 또한, 본 발명의 분석방법에 있어서, 상기 대상자의 시료는 혈액 또는 뇨일 수 있다. 상기 유전자의 발현량의 측정은 mRNA 또는 단백질의 양을 측정함으로써 수행될 수 있다. 일 구현예에서, 상기 단백질의 양의 측정은 웨스턴 블럿팅에 의하여 수행될 수 있다, 다른 구현예에서, 상기 mRNA 양의 측정은 RT-PCR 또는 실시간-PCR을 통하여 수행될 수 있다.In the analysis method of the present invention, the gene encoding the protein may be a gene encoding the protein of SEQ ID NO: 4. Additionally, in the analysis method of the present invention, the subject's sample may be blood or urine. Measurement of the expression level of the gene can be performed by measuring the amount of mRNA or protein. In one embodiment, the measurement of the amount of the protein may be performed by Western blotting. In another embodiment, the measurement of the amount of the mRNA may be performed by RT-PCR or real-time-PCR.

본 발명의 다른 태양에 따라, 서열번호 1 내지 4의 단백질로 이루어진 군으로부터 선택된 단백질을 코딩하는 유전자의 발현량을 측정할 수 있는 분자를 포함하는 비만 또는 당뇨병 진단용 키트로서, 상기 분자가 상기 단백질에 특이적으로 결합하는 항체, 기질, 리간드, 또는 보조인자(cofactor); 또는 상기 단백질을 코딩하는 유전자에 특이적인 상보적 서열을 갖는 프라이머인 비만 또는 당뇨병 진단용 키트가 제공된다.According to another aspect of the present invention, there is a kit for diagnosing obesity or diabetes, comprising a molecule capable of measuring the expression level of a gene encoding a protein selected from the group consisting of proteins of SEQ ID NOs: 1 to 4, wherein the molecule is attached to the protein. An antibody, substrate, ligand, or cofactor that specifically binds; Alternatively, a kit for diagnosing obesity or diabetes is provided, which is a primer having a specific complementary sequence to the gene encoding the protein.

본 발명의 진단용 키트에 있어서, 상기 단백질을 코딩하는 유전자는 서열번호 4의 단백질을 코딩하는 유전자일 수 있다. 일 구현예에서, 상기 분자는 검출가능한 표지로 표지될 수 있다. 다른 구현예에서, 상기 프라이머는 기판상에 고정화되어 있는 마이크로어레이 형태일 수 있다. 또한, 상기 프라이머는 서열번호 9 또는 10의 염기서열을 가질 수 있다.In the diagnostic kit of the present invention, the gene encoding the protein may be a gene encoding the protein of SEQ ID NO: 4. In one embodiment, the molecule can be labeled with a detectable label. In another embodiment, the primers may be in the form of a microarray immobilized on a substrate. Additionally, the primer may have the base sequence of SEQ ID NO: 9 or 10.

USP19가 CORO2A의 탈유비퀴틴화를 통하여 CORO2A의 안정화를 유도함으로써 RAR의 전사를 억제한다는 것이 본 발명에 의해 밝혀졌다. 즉, USP19가 과발현될 경우 CORO2A를 활성화시켜 NCoR 복합체의 활성 억제로 이어지고, 이는 지방생성(adipogenesis)을 촉진시키게 된다는 것이 본 발명에 의해 밝혀졌다. 따라서, 본 발명에 따른 분석방법 및 키트는 비만 또는 당뇨병을 진단하는데 유용하게 사용될 수 있다.The present invention has revealed that USP19 inhibits the transcription of RAR by inducing stabilization of CORO2A through deubiquitination of CORO2A. In other words, the present invention has revealed that when USP19 is overexpressed, it activates CORO2A, leading to inhibition of the activity of the NCoR complex, which promotes adipogenesis. Therefore, the analysis method and kit according to the present invention can be usefully used to diagnose obesity or diabetes.

도 1은 USP19이 CORO2A에 대하여 탈유비퀴틴화를 유도하여 안정성을 높임을 면역 침강법 및 웨스턴 블롯팅을 통해 나타낸 결과이다.
도 2는 면역 침강법 및 은 염색과 질량분석법을 통해 USP19과 결합하는 단백질을 웨스턴 블롯팅을 통해 결합을 확인한 결과이다.
도 3은 CORO2A를 과발현 시켰을 때 RAR(retinoic acid receptor) targeting elements의 전사 억제에 대한 효과를 웨스턴 블롯팅 및 루시퍼레이즈 분석 (luciferase assay)을 통해 나타낸 결과이다.
도 4는 USP19의 발현을 억제시켰을 때 나타나는 RAR의 활성을 웨스턴 블롯팅 및 루시퍼레이즈 분석을 통해 나타낸 결과이다.
도 5는 CORO2A의 발현과 USP19의 발현 억제 조건에서 지방세포 분화 시 마커인 Ets-1, HOXA1과 HOXA4의 전사 발현량을 비교한 결과이다.Figure 1 shows the results of immunoprecipitation and Western blotting showing that USP19 induces deubiquitination of CORO2A and increases its stability.
Figure 2 shows the results of confirming the binding of a protein that binds to USP19 through Western blotting using immunoprecipitation, silver staining, and mass spectrometry.
Figure 3 shows the results of Western blotting and luciferase assay showing the effect on transcriptional repression of RAR (retinoic acid receptor) targeting elements when CORO2A is overexpressed.
Figure 4 shows the results showing the activity of RAR when the expression of USP19 was suppressed through Western blotting and luciferase analysis.
Figure 5 shows the results of comparing the transcriptional expression levels of markers Ets-1, HOXA1, and HOXA4 during adipocyte differentiation under conditions of suppressing the expression of CORO2A and USP19.

본 발명자들은 지방생성(adipogenesis)에 대한 신규의 단백질 바이오마커를 면역 침강법 및 MALDLI TOF-MS, 및 단백질 서열분석 등을 수행하여 검색하였으며, 이 과정에서 탈유비퀴틴화 효소 중 하나인 USP19가 지방생성 과정에서 점차 감소하며 또한 NCoR 복합체의 구성요소 중 하나인 코로닌 2A(CORO2A)를 조절한다는 것을 밝혀냈다. USP19가 CORO2A를 탈유비퀴틴화시키며, 세포 내에서 NCoR 복합체를 안정화시킨다는 것을 확인하였다. USP19의 탈유비퀴틴화 활성을 통한 CORO2A의 조절은 RAR(retinoic acid receptor)의 전사 억제 활성에 영향을 주며, 이는 USP19가 RAR-매개 지방생성의 조절에 관여함을 나타낸다. 따라서, 대상자의 시료 중 USP19의 과발현을 검출할 경우 비만 또는 당뇨병을 진단하는데 유용하게 사용될 수 있으며, USP19는 비만 또는 당뇨병 진단을 위한 바이오마커로서 사용될 수 있다.The present inventors searched for novel protein biomarkers for adipogenesis by performing immunoprecipitation, MALDLI TOF-MS, and protein sequence analysis, and in this process, USP19, one of the deubiquitination enzymes, was used to induce adipogenesis. It was found that it gradually decreases in the process and also regulates coronin 2A (CORO2A), one of the components of the NCoR complex. It was confirmed that USP19 deubiquitinates CORO2A and stabilizes the NCoR complex within cells. Regulation of CORO2A through the deubiquitination activity of USP19 affects the transcriptional repression activity of retinoic acid receptor (RAR), indicating that USP19 is involved in the regulation of RAR-mediated lipogenesis. Therefore, detecting overexpression of USP19 in a subject's sample can be useful for diagnosing obesity or diabetes, and USP19 can be used as a biomarker for diagnosing obesity or diabetes.

탈유비퀴틴화(Deubiquitination)는 유비퀴틴화(ubiquitination)의 반대의 작용이며 탈유비퀴틴화 효소(deubiquitinating enzyme, DUB)에 의해 매개된다(Lim KH, et al. Cytokine Growth Factor Rev, 2013, 24, 427-431). 탈유비퀴틴화 효소는 유비퀴틴화된 기질들로부터 유비퀴틴 분자들을 떼어주는 능력을 가지며 이를 통해 기질의 활성과 안정화를 돕는다. 탈유비퀴틴화 효소 유전자는 인간 게놈에서 약 100여 개가 확인되어 있으며, USP19는 USP(Ubiquitin specific proteases) 중 하나이다. 몇몇의 연구를 통해 USP19는 세포 주기 조절, hypoxia에서의 반응, muscle atrophy를 포함하는 다양한 세포적 과정에서 역할을 하는 것으로 제안되어 왔으나(Lu Y, et al. PLoS One, 2011, 6, e15936; Altun M, et al. J Biol Chem, 2012, 287, 1962-1969; Combaret L, et al. Am J Physiol Endocrinol Metab, 2005, 288, E693-700), 지방생성지방생성(adipogenesis) 나아가 CORO2A에 의한 RAR-매개 지방생성의 조절에 대하여는 보고된 바 없다.Deubiquitination is the opposite of ubiquitination and is mediated by deubiquitinating enzyme (DUB) (Lim KH, et al. Cytokine Growth Factor Rev, 2013, 24, 427-431 ). Deubiquitination enzymes have the ability to remove ubiquitin molecules from ubiquitinated substrates, thereby helping to activate and stabilize the substrate. About 100 deubiquitinating enzyme genes have been identified in the human genome, and USP19 is one of USP (Ubiquitin specific proteases). Several studies have suggested that USP19 plays a role in various cellular processes, including cell cycle regulation, response to hypoxia, and muscle atrophy (Lu Y, et al. PLoS One , 2011, 6, e15936; Altun M, et al.J Biol Chem , 2012, 287, 1962-1969; Combaret L, et al. Am J Physiol Endocrinol Metab , 2005, 288, E693-700), adipogenesis. Furthermore, there has been no report on the regulation of RAR-mediated adipogenesis by CORO2A.

본 발명은 비만 또는 당뇨병 진단에 필요한 정보를 제공하기 위하여, 대상자의 시료 중 USP19 단백질을 코딩하는 유전자의 발현량을 측정하는 단계를 포함하는 분석방법을 제공한다.The present invention provides an analysis method that includes measuring the expression level of the gene encoding USP19 protein in a subject's sample in order to provide information necessary for diagnosing obesity or diabetes.

인간의 USP19는 4개의 동형체가 동정된 바 있으며, 각각 서열번호 1 내지 4의 단백질의 서열을 갖는다. 따라서, 본 명세서에서 달리 표시하지 않는 한, USP19는 서열번호 1 내지 4의 단백질로 이루어진 군으로부터 선택된 단백질을 포함하며, 바람직하게는 서열번호 4의 단백질을 포함한다. 일 구현예에서, 상기 USP19 단백질을 코딩하는 유전자는 서열번호 4의 단백질을 코딩하는 유전자이다.Four isoforms of human USP19 have been identified, each having the protein sequences of SEQ ID NOs: 1 to 4. Accordingly, unless otherwise indicated herein, USP19 includes a protein selected from the group consisting of proteins of SEQ ID NO: 1 to 4, and preferably includes the protein of SEQ ID NO: 4. In one embodiment, the gene encoding the USP19 protein is a gene encoding the protein of SEQ ID NO: 4.

본 발명의 분석방법에 있어서, 상기 대상자의 시료는 인체로부터 체외로 분리된 시료를 말하며, 예를 들어, 인체로부터 체외로 분리된 혈액, 뇨 등을 포함한다.In the analysis method of the present invention, the subject's sample refers to a sample separated outside the body from the human body, and includes, for example, blood and urine separated outside the body from the human body.

본 발명의 분석방법은 USP19, 즉 서열번호 1 내지 4의 단백질로 이루어진 군으로부터 선택된 단백질을 코딩하는 유전자의 발현량을 측정하는 단계를 포함한다. 상기 USP19를 코딩하는 유전자 서열은 유전자 은행 등에 공지되어 있는 공지의 유전자(예를 들어, 서열번호 5 내지 8의 염기서열로 이루어진 군으로부터 선택된 유전자)일 수 있다.The analysis method of the present invention includes the step of measuring the expression level of a gene encoding a protein selected from the group consisting of USP19, that is, proteins of SEQ ID NOs: 1 to 4. The gene sequence encoding USP19 may be a known gene (for example, a gene selected from the group consisting of base sequences of SEQ ID NOs: 5 to 8) known in gene banks, etc.

상기 유전자의 발현량의 측정은 해당 유전자의 mRNA 또는 단백질(즉, 서열번호 1 내지 4의 단백질로 이루어진 군으로부터 선택된 단백질)의 수준(level)을 생명공학 분야에서 통상적으로 사용하는 방법에 따라 측정함으로써 수행될 수 있다. USP19 단백질 양의 측정은 웨스턴 블럿팅 등의 방법에 의해 측정할 수 있다. 웨스턴 블럿팅 방법으로 단백질의 양을 측정하는 경우, 측정된 단백질 발현량이 정상인의 단백질 발현량에 비하여 유의성 있게 높은 경우 비만 또는 당뇨병 위험 환자로 판정할 수 있다. 또한, USP19 단백질을 코딩하는 유전자(예를 들어, 서열번호 5 내지 8의 염기서열로 이루어진 군으로부터 선택된 유전자)의 mRNA 양의 측정은 역전사-PCR(Reverse Transcription PCR) 또는 실시간-PCR(Real Time PCR) 등의 방법에 의해 측정할 수 있다. The expression level of the gene is measured by measuring the level of the mRNA or protein (i.e., a protein selected from the group consisting of proteins of SEQ ID NOs. 1 to 4) of the gene according to a method commonly used in the field of biotechnology. It can be done. The amount of USP19 protein can be measured by methods such as Western blotting. When measuring the amount of protein using the Western blotting method, if the measured protein expression level is significantly higher than that of normal people, the patient can be determined to be at risk of obesity or diabetes. In addition, the amount of mRNA of the gene encoding the USP19 protein (e.g., a gene selected from the group consisting of base sequences of SEQ ID NOs. 5 to 8) can be measured using reverse transcription PCR or real time PCR. ) can be measured by methods such as

본 발명은 또한 USP19 즉, 서열번호 1 내지 4의 단백질로 이루어진 군으로부터 선택된 단백질, 바람직하게는 서열번호 4의 단백질을 코딩하는 유전자의 발현량을 측정할 수 있는 분자를 포함하는 비만 또는 당뇨병 진단용 키트로서, 상기 분자가 상기 단백질에 특이적으로 결합하는 항체, 기질, 리간드, 또는 보조인자(cofactor); 또는 상기 단백질을 코딩하는 유전자에 특이적인 상보적 서열을 갖는 프라이머인 비만 또는 당뇨병 진단용 키트를 제공한다.The present invention also provides a kit for diagnosing obesity or diabetes, which includes a molecule capable of measuring the expression level of a gene encoding USP19, that is, a protein selected from the group consisting of proteins of SEQ ID NO: 1 to 4, preferably the protein of SEQ ID NO: 4. As, the molecule is an antibody, substrate, ligand, or cofactor that specifically binds to the protein; Alternatively, a kit for diagnosing obesity or diabetes is provided, which is a primer having a specific complementary sequence to the gene encoding the protein.

USP19를 코딩하는 유전자의 발현량을 측정할 수 있는 분자로는, 상기 단백질에 특이적으로 결합하는 항체, 기질, 리간드, 또는 보조인자(cofactor); 또는 상기 단백질을 코딩하는 유전자에 특이적인 상보적 서열을 갖는 프라이머일 수 있다.Molecules that can measure the expression level of the gene encoding USP19 include antibodies, substrates, ligands, or cofactors that specifically bind to the protein; Alternatively, it may be a primer having a specific complementary sequence to the gene encoding the protein.

USP19 단백질은 생명공학 분야에서 통상적으로 사용되는 방법에 따라 폴리클론 항체 또는 단일클론 항체를 제조할 수 있으며, 해당 항체를 포함하는 진단용 키트를 제조할 수 있다. 또한, USP19는 탈유비퀴틴화 효소로서의 기능이 밝혀져 있으므로, 이에 대한 기질, 리간드, 또는 보조인자를 포함하도록 본 발명의 키트를 제작할 수도 있다. 또한, USP19를 코딩하는 유전자에 특이적인 상보적 서열을 갖는 프라이머를 생명공학 분야에서 통상적으로 사용되는 방법에 따라 제조할 수 있으며, 해당 프라이머를 포함한 진단용 키트를 제조할 수도 있다. 일 구현예에서, 상기 프라이머는 정방향 프라이머: 5'-TGT GGG CTA CTG CAA CCA-3' (서열번호 9) 및/또는 역방향 프라이머: 5'-GCT GAA TGG GGT CTC TCT-3' (서열번호 10)를 포함한다.USP19 protein can be used to produce polyclonal antibodies or monoclonal antibodies according to methods commonly used in the biotechnology field, and diagnostic kits containing the antibodies can be manufactured. In addition, since USP19 has been shown to function as a deubiquitination enzyme, the kit of the present invention can be prepared to include a substrate, ligand, or cofactor for it. In addition, primers having complementary sequences specific to the gene encoding USP19 can be prepared according to methods commonly used in the biotechnology field, and diagnostic kits containing the primers can also be manufactured. In one embodiment, the primers are forward primer: 5'-TGT GGG CTA CTG CAA CCA-3' (SEQ ID NO: 9) and/or reverse primer: 5'-GCT GAA TGG GGT CTC TCT-3' (SEQ ID NO: 10 ) includes.

본 발명의 진단용 키트에 있어서, USP19 단백질을 코딩하는 유전자의 발현량을 측정할 수 있는 분자는 검출가능한 표지(예를 들어, 발색단 등)로 표지될 수 있다. 또한, 본 발명의 진단용 키트는 상기 프라이머가 기판상에 고정화되어 있는 마이크로어레이 형태를 가짐으로써 DNA 칩 또는 단백질 칩 등의 칩(chip) 형태일 수도 있다.In the diagnostic kit of the present invention, the molecule capable of measuring the expression level of the gene encoding the USP19 protein may be labeled with a detectable label (eg, chromophore, etc.). Additionally, the diagnostic kit of the present invention has a microarray form in which the primers are immobilized on a substrate, so it may be in the form of a chip such as a DNA chip or protein chip.

이하, 본 발명을 실시예를 통하여 더욱 상세히 설명한다. 그러나, 하기 실시예는 본 발명을 예시하기 위한 것으로, 본 발명이 이들 실시예에 의해 제한되는 것은 아니다.Hereinafter, the present invention will be described in more detail through examples. However, the following examples are for illustrating the present invention, and the present invention is not limited by these examples.

실시예Example

1. 시험방법1. Test method

(1) (One) 바이오인포메틱스Bioinformatics (Bioinformatics) 및 LC-MS/MS 분석(Bioinformatics) and LC-MS/MS analysis

보존 도메인 예측(Conserved domain predictions)은 http://www.ncbi.nlm.nih.gov/Structure/cdd/wrpsb.cgi의 rpsblast 프로그램을 사용하여 분석하였다. Superfamily(http://supfam.mrc-lmb.cam.ac.uk/SUPERFAMILY/), SMART(http://smart.emblheidelberg.de/), Pfam(http://www.sanger.ac.uk/Software/Pfam/), InterPro(http://www.ebi.ac.uk/interpro/), 및 Prosite(http://us.expasy.org/prosite/)를 포함한 다른 프로그램을 또한 사용하였다. 정렬(Alignment)은 DNASTAR 프로그램 및 MultiAlign(http://prodes.toulouse.inra.fr/multalin/)을 사용하여 수행하였다. MALDI-TOF-MS 분석 후, 단백질은 단일동위원소 피크(monoisotopic peaks)를 사용하여 MASCOT(http://www.matrixscience.com) 및 MS-Fit(http://prospector.ucsf.edu)로 펩타이드 질량도(peptide mass maps)로부터 동정하였다. LC-MS/MS 분석을 위하여 이전에 보고된 방법(Cho YS, et al. J Immunoassay Immunochem. 2009; 30:291-304)에 따라 Agilent 1100 시리즈 나노-LC 및 LTQ-질량 스펙트로미터(Thermo Electron, Bremen, Germany)를 사용하였다. 모세관 컬럼은 Magic C18 고정상(stationary phase)(5 μm 입자, 100 Å 포어 사이즈)(Michrom Bioresources, Auburn, CA, USA)로 채웠다. LC 분리를 위한 이동상 A 및 B는 각각 탈이온수 또는 아세토니트릴 중의 0.1% 포름산을 사용하여 수행하였다. 크로마토그래피 구배 프로그램, 유속, 및 MS/MS 스캔은 이전에 보고된 방법(Cho YS, et al. J Immunoassay Immunochem. 2009; 30:291-304)에 따라 수행하였다. 펩타이드 서열의 동정은 SEQUEST 소프트웨어(Thermoquest, San Jose, CA, USA)를 사용하여 수행하였다.Conserved domain predictions were analyzed using the rpsblast program at http://www.ncbi.nlm.nih.gov/Structure/cdd/wrpsb.cgi. Superfamily (http://supfam.mrc-lmb.cam.ac.uk/SUPERFAMILY/), SMART (http://smart.emblheidelberg.de/), Pfam (http://www.sanger.ac.uk/ Other programs were also used, including Software/Pfam/), InterPro (http://www.ebi.ac.uk/interpro/), and Prosite (http://us.expasy.org/prosite/). Alignment was performed using the DNASTAR program and MultiAlign (http://prodes.toulouse.inra.fr/multalin/). After MALDI-TOF-MS analysis, proteins were analyzed using MASCOT (http://www.matrixscience.com) and MS-Fit (http://prospector.ucsf.edu) using monoisotopic peaks. It was identified from peptide mass maps. For LC-MS/MS analysis, Agilent 1100 Series Nano-LC and LTQ-Mass Spectrometer (Thermo Electron, Bremen, Germany) was used. The capillary column was filled with Magic C18 stationary phase (5 μm particles, 100 Å pore size) (Michrom Bioresources, Auburn, CA, USA). Mobile phases A and B for LC separation were performed using 0.1% formic acid in deionized water or acetonitrile, respectively. Chromatographic gradient programs, flow rates, and MS/MS scans were performed according to previously reported methods (Cho YS, et al. J Immunoassay Immunochem. 2009; 30:291-304). Identification of peptide sequences was performed using SEQUEST software (Thermoquest, San Jose, CA, USA).

(2) 플라스미드 및 (2) plasmid and siRNAsiRNA

인간 USP19(서열번호 4의 단백질)의 코딩 부위를 갖는 KIAA0891 클론은 일본의 카즈사 연구소(KAZUSA Institute)의 Nagase 박사에 의해 제공받았다. 인간 USP19의 코딩 부위를 pcDNA3-Myc(Invitrogen, Carlsbad, CA, USA)에 서브클로닝하였다. Myc-표지된 USP19 (C506S) 돌연변이는 부위-특이적 돌연변이(site-directed mutagenesis)에 의해 제조하였다. CORO2A의 코딩 부위를 갖는 클론(IMAGE clone: 3350035)을 사용하여, CORO2A를 pCS4-Flag 벡터(Invitrogen, Carlsbad, CA, USA)에 서브클로닝하였다. His-표지된 유비퀴틴은 이전에 보고된 방법(Park JJ, et al. Cell Death Dis . 2015; 6:e1654)에 따라 클로닝하였다. 상이한 농도의 USP19 siRNA(0.5, 1 및 1.5 nmol)을 Opti-MEM 및 RNAimax (Invitrogen, Carlsbad, CA, USA) 혼합물을 사용하여 제공자의 지침에 따라 세포에 형질감염시켰다. USP19 센스 siRNA 서열은 5'-GGA GGA GAU GGC AGU GGC A-3'이었으며, 안티센스 siRNA 서열은 5'-UGC CAC UGC CAU CUC CUC C-3'(UbiProtein Corp, Seongnam, Korea)이었다.The KIAA0891 clone, which has the coding region of human USP19 (protein of SEQ ID NO: 4), was provided by Dr. Nagase, KAZUSA Institute, Japan. The coding region of human USP19 was subcloned into pcDNA3-Myc (Invitrogen, Carlsbad, CA, USA). Myc-labeled USP19 (C506S) mutant was prepared by site-directed mutagenesis. Using a clone containing the coding region of CORO2A (IMAGE clone: 3350035), CORO2A was subcloned into the pCS4-Flag vector (Invitrogen, Carlsbad, CA, USA). His-labeled ubiquitin was cloned according to a previously reported method (Park JJ, et al. Cell Death Dis . 2015; 6:e1654). Different concentrations of USP19 siRNA (0.5, 1, and 1.5 nmol) were transfected into cells using a mixture of Opti-MEM and RNAimax (Invitrogen, Carlsbad, CA, USA) according to the provider's instructions. The USP19 sense siRNA sequence was 5'-GGA GGA GAU GGC AGU GGC A-3', and the antisense siRNA sequence was 5'-UGC CAC UGC CAU CUC CUC C-3' (UbiProtein Corp, Seongnam, Korea).

(3) 세포배양 및 형질감염(3) Cell culture and transfection

인간 배아 신장 293T 세포(Human embryonic kidney 293T cells), 3T3-L1 지방전구세포(3T3-L1 preadipocytes), 및 MCF7 유방암 세포는 10% 우태아혈청(GIBCO, Rockville, MD, USA) 및 1% 페니실린/스트렙토마이신(GIBCO Rockville, MD, USA)이 보충된 둘베코 변형 이글 배지(Dulbecco's modified Eagle's medium)(DMEM, GIBCO, Rockville, MD, USA)에서 배양하였다. 마우스 배아 섬유아세포(mouse embryo fibroblasts, MEFs)는 12.5일째의 마우스 배아로부터 얻었다(Baek KH, et al. PLoS One. 2012; 7:e44223). 3T3-L1 세포로부터 지방세포를 유도하기 위하여, 0.5 mM의 3-이소부틸-메틸산틴(3-isobutyl-1-methylxanthine, IBMX), 1 μM의 덱사메타손 및 10 μg/ml의 인슐린으로 3T3-L1 세포를 처리하였다. 세포는 5% CO2 인큐베이터에서 배양하였다. 293T 세포의 형질감염은 10 mM 폴리에틸렌이민(polyethlylenimine, PEI)(Polysciences, Warrington, PA, USA)를 사용하여 수행하였다.Human embryonic kidney 293T cells, 3T3-L1 preadipocytes, and MCF7 breast cancer cells were incubated with 10% fetal bovine serum (GIBCO, Rockville, MD, USA) and 1% penicillin. Cultured in Dulbecco's modified Eagle's medium (DMEM, GIBCO, Rockville, MD, USA) supplemented with streptomycin (GIBCO Rockville, MD, USA). Mouse embryo fibroblasts (MEFs) were obtained from day 12.5 mouse embryos (Baek KH, et al. PLoS One. 2012; 7:e44223). To induce adipocytes from 3T3-L1 cells, incubate 3T3-L1 cells with 0.5 mM 3-isobutyl-1-methylxanthine (IBMX), 1 μM dexamethasone, and 10 μg/ml insulin. was processed. Cells were cultured in a 5% CO 2 incubator. Transfection of 293T cells was performed using 10 mM polyethlylenimine (PEI) (Polysciences, Warrington, PA, USA).

(4) RNA 분리 및 정량적 실시간 (4) RNA isolation and quantitative real-time PCRPCR

제조사 프로토콜(Invitrogen, Carlsbad, CA, USA)에 따라, TRIzol을 사용하여 3T3-L1 및 MCF7 세포로부터 총 RNA를 분리하였다. RT-PCR 및 정량정 실시간 PCR은 SYBR green PCR 키트(Applied Biosystems, Foster City, CA, USA)를 사용하여 수행하였다.Total RNA was isolated from 3T3-L1 and MCF7 cells using TRIzol according to the manufacturer's protocol (Invitrogen, Carlsbad, CA, USA). RT-PCR and quantitative real-time PCR were performed using the SYBR green PCR kit (Applied Biosystems, Foster City, CA, USA).

(5) 은 염색, (5) silver dyeing, 면역침강법Immunoprecipitation , 면역 , immune 블랏팅Blotting , 및 , and 유비퀴틴화Ubiquitination 분석 analyze

은 염색(silver staining)을 위하여, pcDNA3-Myc 공벡터 또는 pcDNA3-Myc-USP19를 293T 세포에 형질감염시켰다. 48시간 후, 세포를 수확하고, 단백질 분해효소 저해제(protease inhibitor cocktail)(Roche Diagnostics, Mannheim, Germany) 및 페닐메틸술포닐 플루오라이드(phenylmethylsulfonyl fluoride, PMSF)(Sigma-Aldrich, St. Louis, MO, USA)를 함유하는 용해완충액(lysis buffer)(50 mM Tris-HCl [pH 7.8], 150 mM NaCl, 1% Triton X-100)을 사용하여 용해시켰다. 세포 용해물을 항-Myc 항체(9E10)로 4시간 동안 면역침강시켰다. 단백질 A/G 비드(Santa Cruz Biotechnology, Santa Cruz, CA, USA)와 함께 1시간 동안 인큐베이션한 후, 비드를 세척 완충액으로 3회 세척하였다. 은 염색은 silver stain plus 키트(Bio-Rad, Hercules, CA, USA)를 사용하여 제조사 지침에 따라 수행하였다. 즉, 메탄올 및 아세트산을 함유하는 고정 용액(fixing solution)으로 겔을 고정시켰다. 정제수로 2회 세척한 후, 겔을 염색 용액으로 염색하였다. 최종적으로 아세트산을 함유하는 정지 용액(stop solution)을 사용하였다. CORO2A와 USP19의 생체내(in vivo) 결합을 위하여 293T 세포를 pcDNA3-Myc-USP19 및/또는 pCS4-Flag-Coro2a 및/또는 pcDNA3-HA-Ubiquitin 으로 형질감염시켰다. 면역침강 분석을 위하여, 세포 용해물을 항체들과 함께 4℃에서 4시간 동안 배양한 후, 단백질 A/G PLUS 아가로즈 비드(Santa Cruz Biotechnology, Santa Cruz, CA, USA)를 첨가하고 1시간 동안 교반하였다. His-표지된 유비퀴틴은 이전에 보고된 방법(Baek KH, et al. PLoS One. 2012; 7:e44223)에 따라 정제하였다. 세포 용해물 유래의 단백질 및 면역침강물을 폴리아크릴아마이드 겔에 로딩한 다음, 폴리비닐리덴 플루오라이드(polyvinylidene fluoride, PVDF) 멤브레인(Millipore, Billerica, MA, USA)로 전사하였다. 항체로서, 항-Myc, 항-CORO2A(Santa Cruz Biotechnology, Santa Cruz, CA, USA), 항-HA(12CA5), 항-Flag(Sigma-Aldrich, St. Louis, MO, USA), 및 항-β-actin(Santa Cruz Biotechnology, Santa Cruz, CA, USA)를 사용하였다. 웨스턴 블롯팅 이미지는 Image J 프로그램을 사용하여 분석하였다.For silver staining, pcDNA3-Myc empty vector or pcDNA3-Myc- USP19 was transfected into 293T cells. After 48 hours, cells were harvested and incubated with protease inhibitor cocktail (Roche Diagnostics, Mannheim, Germany) and phenylmethylsulfonyl fluoride (PMSF) (Sigma-Aldrich, St. Louis, MO). USA) containing lysis buffer (50 mM Tris-HCl [pH 7.8], 150 mM NaCl, 1% Triton X-100). Cell lysates were immunoprecipitated with anti-Myc antibody (9E10) for 4 h. After incubation with protein A/G beads (Santa Cruz Biotechnology, Santa Cruz, CA, USA) for 1 hour, the beads were washed three times with washing buffer. Silver staining was performed using the silver stain plus kit (Bio-Rad, Hercules, CA, USA) according to the manufacturer's instructions. That is, the gel was fixed with a fixing solution containing methanol and acetic acid. After washing twice with purified water, the gel was stained with a staining solution. Finally, a stop solution containing acetic acid was used. For in vivo binding of CORO2A and USP19, 293T cells were transfected with pcDNA3-Myc- USP19 and/or pCS4-Flag- Coro2a and/or pcDNA3-HA- Ubiquitin . For immunoprecipitation analysis, cell lysates were incubated with antibodies at 4°C for 4 hours, then protein A/G PLUS agarose beads (Santa Cruz Biotechnology, Santa Cruz, CA, USA) were added and incubated for 1 hour. It was stirred. His-labeled ubiquitin was purified according to a previously reported method (Baek KH, et al. PLoS One. 2012; 7:e44223). Proteins and immunoprecipitates derived from cell lysates were loaded on a polyacrylamide gel and then transferred to a polyvinylidene fluoride (PVDF) membrane (Millipore, Billerica, MA, USA). Antibodies include anti-Myc, anti-CORO2A (Santa Cruz Biotechnology, Santa Cruz, CA, USA), anti-HA (12CA5), anti-Flag (Sigma-Aldrich, St. Louis, MO, USA), and anti- β-actin (Santa Cruz Biotechnology, Santa Cruz, CA, USA) was used. Western blotting images were analyzed using the Image J program.

(6) 항체 생산(6) Antibody production

USP19 C-말단 펩타이드(ASRIWQELEAEEEPVPEGSGP)(UbiProtein Corp, Seongnam, Korea)로 면역시킨 토끼에서 다클론 USP19 항혈청(antisera)을 생성시켰다. 토끼 다클론 항-USP19 항체는 rProtein A 아가로즈 컬럼(UbiProtein Corp, Seongnam, Korea)을 사용하여 정제(affinity-purified)하였고, 용래 완충액(1 M Tris [pH 9.0] 및 100 mM 구연산 [pH 3.0])으로 용리시켰다.Polyclonal USP19 antisera were generated in rabbits immunized with USP19 C-terminal peptide (ASRIWQELEAEEEPVPEGSGP) (UbiProtein Corp, Seongnam, Korea). The rabbit polyclonal anti-USP19 antibody was affinity-purified using an rProtein A agarose column (UbiProtein Corp, Seongnam, Korea) and eluted with elution buffer (1 M Tris [pH 9.0] and 100 mM citric acid [pH 3.0]). ) was eluted with.

(7) (7) 루시퍼레이즈Luciferase 분석( analyze( LuciferaseLuciferase assay) assay)

pcDNA3-Myc-USP19, RARE(RAR response elements)를 가지고 있는 USP19에 특이적인 siRNA, 및 PPAE 루시퍼레이즈 벡터(Cignal Report; Qiagen, Valencia, CA, USA)를 RARs를 발현하는 MCF7 세포에 형질감염시켰다. 48시간 후, 형질감염된 세포를 수동 용해 완충액(passive lysis buffer)을 사용하여 수확하고, 루시퍼레이즈 분석 완충액(Promega, Madison, WI, USA)과 함께 인큐베이션하였다. 상이한 조건하에서 각 세포 용해물의 루시퍼레이즈 활성을 루미노미터(luminometer)(Tecan, San Jose, CA, USA)를 사용하여 제조사 지침에 따라 검출하였다.pcDNA3-Myc- USP19 , siRNA specific for USP19 containing RAR response elements (RARE), and PPAE luciferase vector (Cignal Report; Qiagen, Valencia, CA, USA) were transfected into MCF7 cells expressing RARs. After 48 hours, transfected cells were harvested using passive lysis buffer and incubated with luciferase assay buffer (Promega, Madison, WI, USA). The luciferase activity of each cell lysate under different conditions was detected using a luminometer (Tecan, San Jose, CA, USA) according to the manufacturer's instructions.

2. 시험결과2. Test results

(1) (One) USP19의USP19 CORO2ACORO2A 유비퀴틴화Ubiquitination 조절 control

CORO2A의 유비퀴틴화는 아직 알려져 있지 않으나, 프로테아좀 저해제로서 MG132를 처리하여 CORO2A가 유비퀴틴-프로테아좀 경로를 통해 관여한다는 것을 확인하였다(도 1A, 레인 4). USP19의 과발현시 CORO2A의 유비퀴틴화 정도는 in vivo와 in vitro 조건에서 모두 유의성 있게 감소하였다(도 1B, 레인 4 및 도 1C, 레인 3). 그러나, USP19의 촉매작용이 억제된 돌연변이(C506S)는 CORO2A의 유비퀴틴화 정도를 감소시키지 못하였다(도 1C, 레인 4). 이러한 결과는 USP19가 프로테아좀을 통한 COO2A의 분해를 막는다는 것을 나타낸다. USP19는 CORO2A의 유비퀴틴화 정도를 감소시키기 때문에 USP19 발현량 증가에 따른 CORO2A의 발현량을 시험한 결과, CORO2A는 USP19 발현에 의해 점차로 증가하였다(도 1D). 그러나, 촉매작용이 억제된 돌연변이(C506S)는 CORO2A의 발현량에 어떠한 영향도 미치지 않았으며(도 1E), USP19 발현 억제 시 CORO2A의 발현량도 감소하였다(도 1F). 또한, USP19 발현 억제는 CORO2A의 유비퀴틴화 정도를 증가시켰다(도 1G). 따라서, 이 결과들은 CORO2A가 USP19의 결합 단백질이며 USP19의 탈유비퀴틴화 활성을 통하여 CORO2A를 안정화시킴을 나타낸다.Although the ubiquitination of CORO2A is not yet known, treatment with MG132 as a proteasome inhibitor confirmed that CORO2A is involved through the ubiquitin-proteasome pathway (Figure 1A, lane 4). Upon overexpression of USP19, the degree of CORO2A ubiquitination was significantly reduced both in vivo and in vitro (Figure 1B, lane 4 and Figure 1C, lane 3). However, a mutation (C506S) that inhibited the catalytic activity of USP19 did not reduce the degree of ubiquitination of CORO2A (Figure 1C, lane 4). These results indicate that USP19 prevents the degradation of COO2A through the proteasome. Because USP19 reduces the degree of ubiquitination of CORO2A, when the expression level of CORO2A was tested as the expression level of USP19 increased, CORO2A gradually increased due to the expression of USP19 (Figure 1D). However, the catalytic activity-inhibited mutation (C506S) did not have any effect on the expression level of CORO2A (Figure 1E), and the expression level of CORO2A was also reduced when USP19 expression was inhibited (Figure 1F). Additionally, inhibition of USP19 expression increased the degree of ubiquitination of CORO2A (Figure 1G). Therefore, these results indicate that CORO2A is a binding protein of USP19 and stabilizes CORO2A through the deubiquitination activity of USP19.

(2) (2) USP19의USP19 새로운 new 결합단백질로서의As a binding protein CORO2A의CORO2A's 확인 check

본 발명자들은 또한 면역침강법과 MALDI-TOF-MS 분석을 수행하여 USP19의 결합 단백질을 동정하였다. Myc-표지된 USP19가 과발현된 293T 세포로부터 결합 단백질들을 SDS-PAGE를 통해 분리한 다음, 은 염색 및 질량 분석(mass spectrometry)을 통해 다시 분리하였다(도 2A). 상이하게 나타난 단백질 밴드의 질량 분석 결과는 score value, 분자량, CORO2A의 부분적 아미노산 염기서열을 나타내었다(도 2B 및 도 2C). 이러한 결과는 CORO2A가 USP19의 결합 단백질임을 시사한다(도 2D 및 도 2C). 이어서 본 발명자들은 USP19과 CORO2A 사이의 연관성 및 USP19에 의한 CORO2A의 조절을 검증하였다. 293T 세포를 Flag-표지된 CORO2A 및 Myc-표지된 USP19로 형질감염시켰다. 면역침강 분석결과는 USP19와 CORO2A가 강하게 결합하고 있음을 나타내었다(도 2D 및 도 2E). 또한, 본 발명자들은 CORO2A와 USP19의 내인성 결합(endogenous binding)을 시험한 결과, 비-암세포(non-cancer cells)(293T와 3T3-L1 세포) 및 암세포(MCF7 세포)에서 USP19와 CORO2A가 모두 상호작용하고 있음을 확인하였다(도 2F-2H). 이들 결과는 USP19가 CORO2A의 안정성을 증가시키는 탈유비퀴틴화효소임을 나타낸다.We also performed immunoprecipitation and MALDI-TOF-MS analysis to identify the binding protein of USP19. Binding proteins were separated from 293T cells overexpressing Myc-tagged USP19 through SDS-PAGE and then again through silver staining and mass spectrometry (Figure 2A). The mass spectrometry results of the different protein bands showed the score value, molecular weight, and partial amino acid sequence of CORO2A (Figures 2B and 2C). These results suggest that CORO2A is a binding protein of USP19 (Figures 2D and 2C). Subsequently, the present inventors verified the association between USP19 and CORO2A and the regulation of CORO2A by USP19. 293T cells were transfected with Flag-tagged CORO2A and Myc-tagged USP19. Immunoprecipitation analysis results showed that USP19 and CORO2A were strongly bound (Figures 2D and 2E). In addition, the present inventors tested the endogenous binding of CORO2A and USP19 and found that USP19 and CORO2A interact with each other in non-cancer cells (293T and 3T3-L1 cells) and cancer cells (MCF7 cells). It was confirmed that it was working (Figures 2F-2H). These results indicate that USP19 is a deubiquitinating enzyme that increases the stability of CORO2A.

(3) (3) USP19와With USP19 CORO2A을CORO2A 통한 through RAR의RAR's 전사 억제 transcriptional repression

이전의 연구는 NCoR 복합체가 RAR에 대한 DNA-결합 부위를 통해 RARE(RAR response elements)의 전사를 조절한다고 보고한 바 있다(Germain P, et al. Pharmacol Rev. 2006; 58:712-725). 본 발명에 의해 CORO2A와 결합하는 것으로 밝혀진 USP19가 RARE의 전사 억제를 통해 RAR의 기능을 조절할 수 있는지 확인하였다. 본 발명자들은 루시퍼레이즈 분석을 수행하여 USP19 발현 시 RARE의 전사 수준을 특정한 결과, CORO2A가 MCF7 세포에서 발현할 때(도 3A), RARE의 루시퍼레이즈활성은 예상대로 CORO2A의 발현량이 점점 증가함에 따라 감소하였다(도 3B). 이러한 결과는 USP19의 발현이 RARE의 전사 수준에 또한 영향을 미친다는 것을 나타낸다. 흥미롭게도, USP19의 발현이 증가할수록 RARE의 루시퍼레이즈 활성은 약 20%까지 감소하였다(도 4A와 도 4B). 반면에, USP19에 특이적인 siRNA의 처리에 의해 매개된 USP19의 넉-다운은 RARE의 루시퍼레이즈 활성의 증가를 나타내었다(도 4C 및 도 4D). USP19 siRNA에 의한 오프-타겟 효과(off-target effect)를 확인하기 위해, USP19 siRNA 형질감염에 대한 발현을 구제하기 위하여 Myc-표지된 USP19를 사용하였으며, Myc-표지된 USP19의 발현은 USP19 결실 세포에서 RARE의 루시퍼레이즈 활성을 감소시켰다(도 4E). RAR 타켓 유전자들(Ets-1, Hoxa1, Hoxa4)은 CORO2A의 과발현에 의해 감소하였고, USP19의 발현 억제에 의해 증가하였다(도 5). CORO2A가 USP19-감염 조건에서 RARE 루시퍼레이즈 활성을 저해하는지 여부를 시험한 결과, USP siRNA에 의한 USP19 발현 억제와 CORO2A의 과발현은 USP19 siRNA 단독 존재 시에 비해서 RARE의 루시퍼레이즈 활성을 더 많이 증가시키지는 않았다(도 4F). 이러한 결과는 USP19는 CORO2A를 통해 RAR의 전사 억제를 조절함을 나타낸다.Previous studies have reported that the NCoR complex regulates the transcription of RAR response elements (RAREs) through the DNA-binding site for RAR (Germain P, et al. Pharmacol Rev. 2006; 58:712-725). It was confirmed whether USP19, which was found to bind to CORO2A according to the present invention, can regulate the function of RAR through transcriptional repression of RARE. The present inventors performed luciferase analysis to determine the transcription level of RARE upon USP19 expression. As a result, when CORO2A was expressed in MCF7 cells (Figure 3A), the luciferase activity of RARE decreased as the expression level of CORO2A gradually increased, as expected. (Figure 3B). These results indicate that expression of USP19 also affects the transcription level of RARE. Interestingly, as the expression of USP19 increased, the luciferase activity of RARE decreased by approximately 20% (Figures 4A and 4B). On the other hand, knock-down of USP19 mediated by treatment with siRNA specific for USP19 showed an increase in the luciferase activity of RARE (Figures 4C and 4D). To confirm the off-target effect caused by USP19 siRNA, Myc-labeled USP19 was used to rescue expression upon USP19 siRNA transfection, and expression of Myc-labeled USP19 was performed in USP19 deletion cells. reduced the luciferase activity of RARE (Figure 4E). RAR target genes (Ets-1, Hoxa1, Hoxa4) were decreased by overexpression of CORO2A and increased by inhibition of USP19 expression (Figure 5). As a result of testing whether CORO2A inhibits RARE luciferase activity under USP19-infected conditions, inhibition of USP19 expression by USP siRNA and overexpression of CORO2A did not increase the luciferase activity of RARE more than in the presence of USP19 siRNA alone. (Figure 4F). These results indicate that USP19 regulates transcriptional repression of RAR through CORO2A.

3. 고찰3. Considerations

CORO2A는 NCoR 복합체의 구성요소 중 하나로 NCoR 조절을 통해 타켓 유전자들의 전사를 억제하는 역할을 한다고 알려져 있다(Huang W, et al. Nature, 2011, 470, 414-418). NCoR 억제제 복합체는 RAR, THR과 Sin3, HDAC1, HDAC3, TBL1/R1, GPS2, Kaiso, JMJD2A, CORO2A 등 다양한 분자들을 포함하여 핵 수용체를 통해 타켓 유전자들의 전사를 억제한다(Li J, et al. EMBO J, 2000, 19, 4342-4350; Yoon HG, et al. EMBO J, 2003, 22, 1336-1346; Yoon HG, et al. Mol Cell, 2003, 12, 723-734; Horlein AJ, et al. Nature, 1995, 377, 397-404; Nagy L, et al. Cell, 1997, 89, 373-380; Zhang D, et al. Mol Cell Biol, 2005, 25, 6404-6414; Zhang J, et al. Mol Cell, 2002, 9, 611-623]. RAR과 RXR(retinoid X receptor)은 RARE(RAR response elements) 또는 RXRE(RXR response elements)를 포함한 유전자들의 전사인자로서 작용한다(de Lera AR, et al. Nat Rev Drug Discov, 2007, 6, 811-820). 이 상호작용은 NCoR 복합체의 결합 단백질인 RAR의 전사 억제에 영향을 미친다(Atsumi A, et al. Biochem Biophys Res Commun, 2006, 345, 1471-1480). 예를 들어, RAR 타켓 유전자들의 전사 활성은 HDAC3의 억제에서는 증가하지만 RAR 발현 세포에서 NCoR의 억제에서는 증가하지 않는다(Atsumi A, et al. Biochem Biophys Res Commun, 2006, 345, 1471-1480). CORO2A is one of the components of the NCoR complex and is known to play a role in suppressing the transcription of target genes through NCoR regulation (Huang W, et al. Nature , 2011, 470, 414-418). The NCoR inhibitor complex inhibits the transcription of target genes through nuclear receptors, including RAR, THR, and various molecules such as Sin3, HDAC1, HDAC3, TBL1/R1, GPS2, Kaiso, JMJD2A, and CORO2A (Li J, et al. EMBO J , 2000, 19, 4342-4350; Yoon HG, et al. EMBO J , 2003, 22, 1336-1346; Yoon HG, et al. Mol Cell , 2003, 12, 723-734; Horlein AJ, et al. Nature , 1995, 377, 397-404; Nagy L, et al. Cell , 1997, 89, 373-380; Zhang D, et al. Mol Cell Biol , 2005, 25, 6404-6414; Zhang J, et al. Mol Cell , 2002, 9, 611-623]. RAR and RXR (retinoid . Nat Rev Drug Discov , 2007, 6, 811-820) This interaction affects transcriptional repression of RAR, a binding protein of the NCoR complex (Atsumi A, et al. Biochem Biophys Res Commun , 2006, 345, 1471-1480). For example, the transcriptional activity of RAR target genes increases upon inhibition of HDAC3 but does not increase upon inhibition of NCoR in RAR expressing cells (Atsumi A, et al. Biochem Biophys Res Commun , 2006, 345, 1471-1480).

NCoR 복합체 단백질인 CORO2A가 USP19 항체를 이용한 면역침강법을 통해 확인되었다. USP19는 CORO2A를 탈유비퀴틴화시키고(도 1), 이를 통해 USP19가 NCoR 복합체를 안정화시키는 것으로 확인되었다. USP19의 과발현은 RARE 전사를 억제하고 siRNA를 이용한 USP19의 발현 억제는 RARE의 전사를 증가시켰다 (도 3 및 도 4). 이를 통해 USP19는 탈유비퀴틴화 효소 활성을 통해 CORO2A의 안정성을 증가시키고 USP19와 CORO2A의 상호작용은 NCoR의 기능과 연관이 있는 것으로 보인다. 또한, 이러한 결과들은 지방생성(adipogenesis)에 관여하는 RAR의 조절에 있어서 USP19와 CORO2A의 상호작용의 중요성을 입증한다. 지금까지 지방생성(adipogenesis)과 관련된 탈유비퀴틴화 효소에 대한 연구가 없었고, 본 연구는 USP19가 지방세포 분화에 중요한 역할을 함을 증명하였다. USP19가 탈유비퀴틴화 효소 활성을 통해 CORO2A의 안정화를 유도하고 이로써 RAR의 전사가 억제됨을 확인하였고, 따라서 USP19는 비만 또는 당뇨병을 진단하는데 유용하게 사용될 수 있으며, USP19는 비만 또는 당뇨병 진단을 위한 바이오마커로서 사용될 수 있다.CORO2A, an NCoR complex protein, was identified through immunoprecipitation using USP19 antibody. USP19 deubiquitinates CORO2A (Figure 1), and it was confirmed that USP19 stabilizes the NCoR complex. Overexpression of USP19 inhibited RARE transcription, and inhibition of USP19 expression using siRNA increased transcription of RARE (Figures 3 and 4). Through this, USP19 increases the stability of CORO2A through deubiquitinating enzyme activity, and the interaction between USP19 and CORO2A appears to be related to the function of NCoR. Additionally, these results demonstrate the importance of the interaction between USP19 and CORO2A in the regulation of RAR involved in adipogenesis. Until now, there has been no research on deubiquitination enzymes related to adipogenesis, and this study demonstrated that USP19 plays an important role in adipocyte differentiation. It was confirmed that USP19 induces the stabilization of CORO2A through deubiquitinating enzyme activity, thereby suppressing the transcription of RAR. Therefore, USP19 can be usefully used to diagnose obesity or diabetes, and USP19 is a biomarker for diagnosing obesity or diabetes. It can be used as.

<110> College of Medicine Pochon CHA University Industry-Academic Cooperation Foundation <120> Analytical method for diagnosing obesity or diabetes and kit therefor <130> PN0798 <160> 10 <170> KopatentIn 2.0 <210> 1 <211> 1419 <212> PRT <213> Homo sapiens <400> 1 Met Ser Gly Gly Ala Ser Ala Thr Gly Pro Arg Arg Gly Pro Pro Gly 1 5 10 15 Leu Glu Asp Thr Thr Ser Lys Lys Lys Gln Lys Asp Arg Ala Asn Gln 20 25 30 Glu Ser Lys Asp Gly Asp Pro Arg Lys Glu Thr Gly Ser Arg Tyr Val 35 40 45 Ala Gln Ala Gly Leu Glu Pro Leu Ala Ser Gly Asp Pro Ser Ala Ser 50 55 60 Ala Ser His Ala Ala Gly Ile Thr Gly Ser Arg His Arg Thr Arg Leu 65 70 75 80 Phe Phe Pro Ser Ser Ser Gly Ser Ala Ser Thr Pro Gln Glu Glu Gln 85 90 95 Thr Lys Glu Gly Ala Cys Glu Asp Pro His Asp Leu Leu Ala Thr Pro 100 105 110 Thr Pro Glu Leu Leu Leu Asp Trp Arg Gln Ser Ala Glu Glu Val Ile 115 120 125 Val Lys Leu Arg Val Gly Val Gly Pro Leu Gln Leu Glu Asp Val Asp 130 135 140 Ala Ala Phe Thr Asp Thr Asp Cys Val Val Arg Phe Ala Gly Gly Gln 145 150 155 160 Gln Trp Gly Gly Val Phe Tyr Ala Glu Ile Lys Ser Ser Cys Ala Lys 165 170 175 Val Gln Thr Arg Lys Gly Ser Leu Leu His Leu Thr Leu Pro Lys Lys 180 185 190 Val Pro Met Leu Thr Trp Pro Ser Leu Leu Lys Lys Pro Leu Gly Thr 195 200 205 Gln Glu Leu Val Pro Gly Leu Arg Cys Gln Glu Asn Gly Gln Glu Leu 210 215 220 Ser Pro Ile Ala Leu Glu Pro Gly Pro Glu Pro His Arg Ala Lys Gln 225 230 235 240 Glu Ala Arg Asn Gln Lys Arg Ala Gln Gly Arg Gly Glu Val Gly Ala 245 250 255 Gly Ala Gly Pro Gly Ala Gln Ala Gly Pro Ser Ala Lys Arg Ala Val 260 265 270 His Leu Cys Arg Gly Pro Glu Gly Asp Gly Ser Arg Asp Asp Pro Gly 275 280 285 Pro Arg Gly Asp Ala Pro Pro Phe Val Ala Asp Pro Ala Thr Gln Val 290 295 300 Glu Ala Asp Glu Gln Leu Cys Ile Pro Pro Leu Asn Ser Gln Thr Cys 305 310 315 320 Leu Leu Gly Ser Glu Glu Asn Leu Ala Pro Leu Ala Gly Glu Lys Ala 325 330 335 Val Pro Pro Gly Asn Asp Pro Val Ser Pro Ala Met Val Arg Ser Arg 340 345 350 Asn Pro Gly Lys Asp Asp Cys Ala Lys Glu Glu Met Ala Val Ala Ala 355 360 365 Asp Ala Ala Thr Leu Val Asp Glu Pro Glu Ser Met Val Asn Leu Ala 370 375 380 Phe Val Lys Asn Asp Ser Tyr Glu Lys Gly Pro Asp Ser Val Val Val 385 390 395 400 His Val Tyr Val Lys Glu Ile Cys Arg Asp Thr Ser Arg Val Leu Phe 405 410 415 Arg Glu Gln Asp Phe Thr Leu Ile Phe Gln Thr Arg Asp Gly Asn Phe 420 425 430 Leu Arg Leu His Pro Gly Cys Gly Pro His Thr Thr Phe Arg Trp Gln 435 440 445 Val Lys Leu Arg Asn Leu Ile Glu Pro Glu Gln Cys Thr Phe Cys Phe 450 455 460 Thr Ala Ser Arg Ile Asp Ile Cys Leu Arg Lys Arg Gln Ser Gln Arg 465 470 475 480 Trp Gly Gly Leu Glu Ala Pro Ala Ala Arg Val Gly Gly Ala Lys Val 485 490 495 Ala Val Pro Thr Gly Pro Thr Pro Leu Asp Ser Thr Pro Pro Gly Gly 500 505 510 Ala Pro His Pro Leu Thr Gly Gln Glu Glu Ala Arg Ala Val Glu Lys 515 520 525 Asp Lys Ser Lys Ala Arg Ser Glu Asp Thr Gly Leu Asp Ser Val Ala 530 535 540 Thr Arg Thr Pro Met Glu His Val Thr Pro Lys Pro Glu Thr His Leu 545 550 555 560 Ala Ser Pro Lys Pro Thr Cys Met Val Pro Pro Met Pro His Ser Pro 565 570 575 Val Ser Gly Asp Ser Val Glu Glu Glu Glu Glu Glu Glu Lys Lys Val 580 585 590 Cys Leu Pro Gly Phe Thr Gly Leu Val Asn Leu Gly Asn Thr Cys Phe 595 600 605 Met Asn Ser Val Ile Gln Ser Leu Ser Asn Thr Arg Glu Leu Arg Asp 610 615 620 Phe Phe His Asp Arg Ser Phe Glu Ala Glu Ile Asn Tyr Asn Asn Pro 625 630 635 640 Leu Gly Thr Gly Gly Arg Leu Ala Ile Gly Phe Ala Val Leu Leu Arg 645 650 655 Ala Leu Trp Lys Gly Thr His His Ala Phe Gln Pro Ser Lys Leu Lys 660 665 670 Ala Ile Val Ala Ser Lys Ala Ser Gln Phe Thr Gly Tyr Ala Gln His 675 680 685 Asp Ala Gln Glu Phe Met Ala Phe Leu Leu Asp Gly Leu His Glu Asp 690 695 700 Leu Asn Arg Ile Gln Asn Lys Pro Tyr Thr Glu Thr Val Asp Ser Asp 705 710 715 720 Gly Arg Pro Asp Glu Val Val Ala Glu Glu Ala Trp Gln Arg His Lys 725 730 735 Met Arg Asn Asp Ser Phe Ile Val Asp Leu Phe Gln Gly Gln Tyr Lys 740 745 750 Ser Lys Leu Val Cys Pro Val Cys Ala Lys Val Ser Ile Thr Phe Asp 755 760 765 Pro Phe Leu Tyr Leu Pro Val Pro Leu Pro Gln Lys Gln Lys Val Leu 770 775 780 Pro Val Phe Tyr Phe Ala Arg Glu Pro His Ser Lys Pro Ile Lys Phe 785 790 795 800 Leu Val Ser Val Ser Lys Glu Asn Ser Thr Ala Ser Glu Val Leu Asp 805 810 815 Ser Leu Ser Gln Ser Val His Val Lys Pro Glu Asn Leu Arg Leu Ala 820 825 830 Glu Val Ile Lys Asn Arg Phe His Arg Val Phe Leu Pro Ser His Ser 835 840 845 Leu Asp Thr Val Ser Pro Ser Asp Thr Leu Leu Cys Phe Glu Leu Leu 850 855 860 Ser Ser Glu Leu Ala Lys Glu Arg Val Val Val Leu Glu Val Gln Gln 865 870 875 880 Arg Pro Gln Val Pro Ser Val Pro Ile Ser Lys Cys Ala Ala Cys Gln 885 890 895 Arg Lys Gln Gln Ser Glu Asp Glu Lys Leu Lys Arg Cys Thr Arg Cys 900 905 910 Tyr Arg Val Gly Tyr Cys Asn Gln Leu Cys Gln Lys Thr His Trp Pro 915 920 925 Asp His Lys Gly Leu Cys Arg Pro Glu Asn Ile Gly Tyr Pro Phe Leu 930 935 940 Val Ser Val Pro Ala Ser Arg Leu Thr Tyr Ala Arg Leu Ala Gln Leu 945 950 955 960 Leu Glu Gly Tyr Ala Arg Tyr Ser Val Ser Val Phe Gln Pro Pro Phe 965 970 975 Gln Pro Gly Arg Met Ala Leu Glu Ser Gln Ser Pro Gly Cys Thr Thr 980 985 990 Leu Leu Ser Thr Gly Ser Leu Glu Ala Gly Asp Ser Glu Arg Asp Pro 995 1000 1005 Ile Gln Pro Pro Glu Leu Gln Leu Val Thr Pro Met Ala Glu Gly Asp 1010 1015 1020 Thr Gly Leu Pro Arg Val Trp Ala Ala Pro Asp Arg Gly Pro Val Pro 1025 1030 1035 1040 Ser Thr Ser Gly Ile Ser Ser Glu Met Leu Ala Ser Gly Pro Ile Glu 1045 1050 1055 Val Gly Ser Leu Pro Ala Gly Glu Arg Val Ser Arg Pro Glu Ala Ala 1060 1065 1070 Val Pro Gly Tyr Gln His Pro Ser Glu Ala Met Asn Ala His Thr Pro 1075 1080 1085 Gln Phe Phe Ile Tyr Lys Ile Asp Ser Ser Asn Arg Glu Gln Arg Leu 1090 1095 1100 Glu Asp Lys Gly Asp Thr Pro Leu Glu Leu Gly Asp Asp Cys Ser Leu 1105 1110 1115 1120 Ala Leu Val Trp Arg Asn Asn Glu Arg Leu Gln Glu Phe Val Leu Val 1125 1130 1135 Ala Ser Lys Glu Leu Glu Cys Ala Glu Asp Pro Gly Ser Ala Gly Glu 1140 1145 1150 Ala Ala Arg Ala Gly His Phe Thr Leu Asp Gln Cys Leu Asn Leu Phe 1155 1160 1165 Thr Arg Pro Glu Val Leu Ala Pro Glu Glu Ala Trp Tyr Cys Pro Gln 1170 1175 1180 Cys Lys Gln His Arg Glu Ala Ser Lys Gln Leu Leu Leu Trp Arg Leu 1185 1190 1195 1200 Pro Asn Val Leu Ile Val Gln Leu Lys Arg Phe Ser Phe Arg Ser Phe 1205 1210 1215 Ile Trp Arg Asp Lys Ile Asn Asp Leu Val Glu Phe Pro Val Arg Asn 1220 1225 1230 Leu Asp Leu Ser Lys Phe Cys Ile Gly Gln Lys Glu Glu Gln Leu Pro 1235 1240 1245 Ser Tyr Asp Leu Tyr Ala Val Ile Asn His Tyr Gly Gly Met Ile Gly 1250 1255 1260 Gly His Tyr Thr Ala Cys Ala Arg Leu Pro Asn Asp Arg Ser Ser Gln 1265 1270 1275 1280 Arg Ser Asp Val Gly Trp Arg Leu Phe Asp Asp Ser Thr Val Thr Thr 1285 1290 1295 Val Asp Glu Ser Gln Val Val Thr Arg Tyr Ala Tyr Val Leu Phe Tyr 1300 1305 1310 Arg Arg Arg Asn Ser Pro Val Glu Arg Pro Pro Arg Ala Gly His Ser 1315 1320 1325 Glu His His Pro Asp Leu Gly Pro Ala Ala Glu Ala Ala Ala Ser Gln 1330 1335 1340 Ala Ser Arg Ile Trp Gln Glu Leu Glu Ala Glu Glu Glu Pro Val Pro 1345 1350 1355 1360 Glu Gly Ser Gly Pro Leu Gly Pro Trp Gly Pro Gln Asp Trp Val Gly 1365 1370 1375 Pro Leu Pro Arg Gly Pro Thr Thr Pro Asp Glu Gly Cys Leu Arg Tyr 1380 1385 1390 Phe Val Leu Gly Thr Val Ala Ala Leu Val Ala Leu Val Leu Asn Val 1395 1400 1405 Phe Tyr Pro Leu Val Ser Gln Ser Arg Trp Arg 1410 1415 <210> 2 <211> 1384 <212> PRT <213> Homo sapiens <400> 2 Met Ser Gly Gly Ala Ser Ala Thr Gly Pro Arg Arg Gly Pro Pro Gly 1 5 10 15 Leu Glu Asp Thr Thr Ser Lys Lys Lys Gln Lys Asp Arg Ala Asn Gln 20 25 30 Glu Ser Lys Asp Gly Asp Pro Arg Lys Glu Thr Gly Ser Arg Tyr Val 35 40 45 Ala Gln Ala Gly Leu Glu Pro Leu Ala Ser Gly Asp Pro Ser Ala Ser 50 55 60 Ala Ser His Ala Ala Gly Ile Thr Gly Ser Arg His Arg Thr Arg Leu 65 70 75 80 Phe Phe Pro Ser Ser Ser Gly Ser Ala Ser Thr Pro Gln Glu Glu Gln 85 90 95 Thr Lys Glu Gly Ala Cys Glu Asp Pro His Asp Leu Leu Ala Thr Pro 100 105 110 Thr Pro Glu Leu Leu Leu Asp Trp Arg Gln Ser Ala Glu Glu Val Ile 115 120 125 Val Lys Leu Arg Val Gly Val Gly Pro Leu Gln Leu Glu Asp Val Asp 130 135 140 Ala Ala Phe Thr Asp Thr Asp Cys Val Val Arg Phe Ala Gly Gly Gln 145 150 155 160 Gln Trp Gly Gly Val Phe Tyr Ala Glu Ile Lys Ser Ser Cys Ala Lys 165 170 175 Val Gln Thr Arg Lys Gly Ser Leu Leu His Leu Thr Leu Pro Lys Lys 180 185 190 Val Pro Met Leu Thr Trp Pro Ser Leu Leu Lys Lys Pro Leu Gly Thr 195 200 205 Gln Glu Leu Val Pro Gly Leu Arg Cys Gln Glu Asn Gly Gln Glu Leu 210 215 220 Ser Pro Ile Ala Leu Glu Pro Gly Pro Glu Pro His Arg Ala Lys Gln 225 230 235 240 Glu Ala Arg Asn Gln Lys Arg Ala Gln Gly Arg Gly Glu Val Gly Ala 245 250 255 Gly Ala Gly Pro Gly Ala Gln Ala Gly Pro Ser Ala Lys Arg Ala Val 260 265 270 His Leu Cys Arg Gly Pro Glu Gly Asp Gly Ser Arg Asp Asp Pro Gly 275 280 285 Pro Arg Gly Asp Ala Pro Pro Phe Val Ala Asp Pro Ala Thr Gln Val 290 295 300 Glu Ala Asp Glu Gln Leu Cys Ile Pro Pro Leu Asn Ser Gln Thr Cys 305 310 315 320 Leu Leu Gly Ser Glu Glu Asn Leu Ala Pro Leu Ala Gly Glu Lys Ala 325 330 335 Val Pro Pro Gly Asn Asp Pro Val Ser Pro Ala Met Val Arg Ser Arg 340 345 350 Asn Pro Gly Lys Asp Asp Cys Ala Lys Glu Glu Met Ala Val Ala Ala 355 360 365 Asp Ala Ala Thr Leu Val Asp Glu Pro Glu Ser Met Val Asn Leu Ala 370 375 380 Phe Val Lys Asn Asp Ser Tyr Glu Lys Gly Pro Asp Ser Val Val Val 385 390 395 400 His Val Tyr Val Lys Glu Ile Cys Arg Asp Thr Ser Arg Val Leu Phe 405 410 415 Arg Glu Gln Asp Phe Thr Leu Ile Phe Gln Thr Arg Asp Gly Asn Phe 420 425 430 Leu Arg Leu His Pro Gly Cys Gly Pro His Thr Thr Phe Arg Trp Gln 435 440 445 Val Lys Leu Arg Asn Leu Ile Glu Pro Glu Gln Cys Thr Phe Cys Phe 450 455 460 Thr Ala Ser Arg Ile Asp Ile Cys Leu Arg Lys Arg Gln Ser Gln Arg 465 470 475 480 Trp Gly Gly Leu Glu Ala Pro Ala Ala Arg Gly Ala Val Gly Gly Ala 485 490 495 Lys Val Ala Val Pro Thr Gly Pro Thr Pro Leu Asp Ser Thr Pro Pro 500 505 510 Gly Gly Ala Pro His Pro Leu Thr Gly Gln Glu Glu Ala Arg Ala Val 515 520 525 Glu Lys Asp Lys Ser Lys Ala Arg Ser Glu Asp Thr Gly Leu Asp Ser 530 535 540 Val Ala Thr Arg Thr Pro Met Glu His Val Thr Pro Lys Pro Glu Thr 545 550 555 560 His Leu Ala Ser Pro Lys Pro Thr Cys Met Val Pro Pro Met Pro His 565 570 575 Ser Pro Val Ser Gly Asp Ser Val Glu Glu Glu Glu Glu Glu Glu Lys 580 585 590 Lys Val Cys Leu Pro Gly Phe Thr Gly Leu Val Asn Leu Gly Asn Thr 595 600 605 Cys Phe Met Asn Ser Val Ile Gln Ser Leu Ser Asn Thr Arg Glu Leu 610 615 620 Arg Asp Phe Phe His Asp Arg Ser Phe Glu Ala Glu Ile Asn Tyr Asn 625 630 635 640 Asn Pro Leu Gly Thr Gly Gly Arg Leu Ala Ile Gly Phe Ala Val Leu 645 650 655 Leu Arg Ala Leu Trp Lys Gly Thr His His Ala Phe Gln Pro Ser Lys 660 665 670 Leu Lys Ala Ile Val Ala Ser Lys Ala Ser Gln Phe Thr Gly Tyr Ala 675 680 685 Gln His Asp Ala Gln Glu Phe Met Ala Phe Leu Leu Asp Gly Leu His 690 695 700 Glu Asp Leu Asn Arg Ile Gln Asn Lys Pro Tyr Thr Glu Thr Val Asp 705 710 715 720 Ser Asp Gly Arg Pro Asp Glu Val Val Ala Glu Glu Ala Trp Gln Arg 725 730 735 His Lys Met Arg Asn Asp Ser Phe Ile Val Asp Leu Phe Gln Gly Gln 740 745 750 Tyr Lys Ser Lys Leu Val Cys Pro Val Cys Ala Lys Val Ser Ile Thr 755 760 765 Phe Asp Pro Phe Leu Tyr Leu Pro Val Pro Leu Pro Gln Lys Gln Lys 770 775 780 Val Leu Pro Val Phe Tyr Phe Ala Arg Glu Pro His Ser Lys Pro Ile 785 790 795 800 Lys Phe Leu Val Ser Val Ser Lys Glu Asn Ser Thr Ala Ser Glu Val 805 810 815 Leu Asp Ser Leu Ser Gln Ser Val His Val Lys Pro Glu Asn Leu Arg 820 825 830 Leu Ala Glu Val Ile Lys Asn Arg Phe His Arg Val Phe Leu Pro Ser 835 840 845 His Ser Leu Asp Thr Val Ser Pro Ser Asp Thr Leu Leu Cys Phe Glu 850 855 860 Leu Leu Ser Ser Glu Leu Ala Lys Glu Arg Val Val Val Leu Glu Val 865 870 875 880 Gln Gln Arg Pro Gln Val Pro Ser Val Pro Ile Ser Lys Cys Ala Ala 885 890 895 Cys Gln Arg Lys Gln Gln Ser Glu Asp Glu Lys Leu Lys Arg Cys Thr 900 905 910 Arg Cys Tyr Arg Val Gly Tyr Cys Asn Gln Leu Cys Gln Lys Thr His 915 920 925 Trp Pro Asp His Lys Gly Leu Cys Arg Pro Glu Asn Ile Gly Tyr Pro 930 935 940 Phe Leu Val Ser Val Pro Ala Ser Arg Leu Thr Tyr Ala Arg Leu Ala 945 950 955 960 Gln Leu Leu Glu Gly Tyr Ala Arg Tyr Ser Val Ser Val Phe Gln Pro 965 970 975 Pro Phe Gln Pro Gly Arg Met Ala Leu Glu Ser Gln Ser Pro Gly Cys 980 985 990 Thr Thr Leu Leu Ser Thr Gly Ser Leu Glu Ala Gly Asp Ser Glu Arg 995 1000 1005 Asp Pro Ile Gln Pro Pro Glu Leu Gln Leu Val Thr Pro Met Ala Glu 1010 1015 1020 Gly Asp Thr Gly Leu Pro Arg Val Trp Ala Ala Pro Asp Arg Gly Pro 1025 1030 1035 1040 Val Pro Ser Thr Ser Gly Ile Ser Ser Glu Met Leu Ala Ser Gly Pro 1045 1050 1055 Ile Glu Val Gly Ser Leu Pro Ala Gly Glu Arg Val Ser Arg Pro Glu 1060 1065 1070 Ala Ala Val Pro Gly Tyr Gln His Pro Ser Glu Ala Met Asn Ala His 1075 1080 1085 Thr Pro Gln Phe Phe Ile Tyr Lys Ile Asp Ser Ser Asn Arg Glu Gln 1090 1095 1100 Arg Leu Glu Asp Lys Gly Asp Thr Pro Leu Glu Leu Gly Asp Asp Cys 1105 1110 1115 1120 Ser Leu Ala Leu Val Trp Arg Asn Asn Glu Arg Leu Gln Glu Phe Val 1125 1130 1135 Leu Val Ala Ser Lys Glu Leu Glu Cys Ala Glu Asp Pro Gly Ser Ala 1140 1145 1150 Gly Glu Ala Ala Arg Ala Gly His Phe Thr Leu Asp Gln Cys Leu Asn 1155 1160 1165 Leu Phe Thr Arg Pro Glu Val Leu Ala Pro Glu Glu Ala Trp Tyr Cys 1170 1175 1180 Pro Gln Cys Lys Gln His Arg Glu Ala Ser Lys Gln Leu Leu Leu Trp 1185 1190 1195 1200 Arg Leu Pro Asn Val Leu Ile Val Gln Leu Lys Arg Phe Ser Phe Arg 1205 1210 1215 Ser Phe Ile Trp Arg Asp Lys Ile Asn Asp Leu Val Glu Phe Pro Val 1220 1225 1230 Arg Asn Leu Asp Leu Ser Lys Phe Cys Ile Gly Gln Lys Glu Glu Gln 1235 1240 1245 Leu Pro Ser Tyr Asp Leu Tyr Ala Val Ile Asn His Tyr Gly Gly Met 1250 1255 1260 Ile Gly Gly His Tyr Thr Ala Cys Ala Arg Leu Pro Asn Asp Arg Ser 1265 1270 1275 1280 Ser Gln Arg Ser Asp Val Gly Trp Arg Leu Phe Asp Asp Ser Thr Val 1285 1290 1295 Thr Thr Val Asp Glu Ser Gln Val Val Thr Arg Tyr Ala Tyr Val Leu 1300 1305 1310 Phe Tyr Arg Arg Arg Asn Ser Pro Val Glu Arg Pro Pro Arg Ala Gly 1315 1320 1325 His Ser Glu His His Pro Asp Leu Gly Pro Ala Ala Glu Ala Ala Ala 1330 1335 1340 Ser Gln Gly Leu Gly Pro Gly Gln Ala Pro Glu Val Ala Pro Thr Arg 1345 1350 1355 1360 Thr Ala Pro Glu Arg Phe Ala Pro Pro Val Asp Arg Pro Ala Pro Thr 1365 1370 1375 Tyr Ser Asn Met Glu Glu Val Asp 1380 <210> 3 <211> 1372 <212> PRT <213> Homo sapiens <400> 3 Met Ser Gly Gly Ala Ser Ala Thr Gly Pro Arg Arg Gly Pro Pro Gly 1 5 10 15 Leu Glu Asp Thr Thr Ser Lys Lys Lys Gln Lys Asp Arg Ala Asn Gln 20 25 30 Glu Ser Lys Asp Gly Asp Pro Arg Lys Glu Thr Gly Ser Arg Tyr Val 35 40 45 Ala Gln Ala Gly Leu Glu Pro Leu Ala Ser Gly Asp Pro Ser Ala Ser 50 55 60 Ala Ser His Ala Ala Gly Ile Thr Gly Ser Arg His Arg Thr Arg Leu 65 70 75 80 Phe Phe Pro Ser Ser Ser Gly Ser Ala Ser Thr Pro Gln Glu Glu Gln 85 90 95 Thr Lys Glu Gly Ala Cys Glu Asp Pro His Asp Leu Leu Ala Thr Pro 100 105 110 Thr Pro Glu Leu Leu Leu Asp Trp Arg Gln Ser Ala Glu Glu Val Ile 115 120 125 Val Lys Leu Arg Val Gly Val Gly Pro Leu Gln Leu Glu Asp Val Asp 130 135 140 Ala Ala Phe Thr Asp Thr Asp Cys Val Val Arg Phe Ala Gly Gly Gln 145 150 155 160 Gln Trp Gly Gly Val Phe Tyr Ala Glu Ile Lys Ser Ser Cys Ala Lys 165 170 175 Val Gln Thr Arg Lys Gly Ser Leu Leu His Leu Thr Leu Pro Lys Lys 180 185 190 Lys Lys Pro Leu Gly Thr Gln Glu Leu Val Pro Gly Leu Arg Cys Gln 195 200 205 Glu Asn Gly Gln Glu Leu Ser Pro Ile Ala Leu Glu Pro Gly Pro Glu 210 215 220 Pro His Arg Ala Lys Gln Glu Ala Arg Asn Gln Lys Arg Ala Gln Gly 225 230 235 240 Arg Gly Glu Val Gly Ala Gly Ala Gly Pro Gly Ala Gln Ala Gly Pro 245 250 255 Ser Ala Lys Arg Ala Val His Leu Cys Arg Gly Pro Glu Gly Asp Gly 260 265 270 Ser Arg Asp Asp Pro Gly Pro Arg Gly Asp Ala Pro Pro Phe Val Ala 275 280 285 Asp Pro Ala Thr Gln Val Glu Ala Asp Glu Gln Leu Cys Ile Pro Pro 290 295 300 Leu Asn Ser Gln Thr Cys Leu Leu Gly Ser Glu Glu Asn Leu Ala Pro 305 310 315 320 Leu Ala Gly Glu Lys Ala Val Pro Pro Gly Asn Asp Pro Val Ser Pro 325 330 335 Ala Met Val Arg Ser Arg Asn Pro Gly Lys Asp Asp Cys Ala Lys Glu 340 345 350 Glu Met Ala Val Ala Ala Asp Ala Ala Thr Leu Val Asp Glu Pro Glu 355 360 365 Ser Met Val Asn Leu Ala Phe Val Lys Asn Asp Ser Tyr Glu Lys Gly 370 375 380 Pro Asp Ser Val Val Val His Val Tyr Val Lys Glu Ile Cys Arg Asp 385 390 395 400 Thr Ser Arg Val Leu Phe Arg Glu Gln Asp Phe Thr Leu Ile Phe Gln 405 410 415 Thr Arg Asp Gly Asn Phe Leu Arg Leu His Pro Gly Cys Gly Pro His 420 425 430 Thr Thr Phe Arg Trp Gln Val Lys Leu Arg Asn Leu Ile Glu Pro Glu 435 440 445 Gln Cys Thr Phe Cys Phe Thr Ala Ser Arg Ile Asp Ile Cys Leu Arg 450 455 460 Lys Arg Gln Ser Gln Arg Trp Gly Gly Leu Glu Ala Pro Ala Ala Arg 465 470 475 480 Val Gly Gly Ala Lys Val Ala Val Pro Thr Gly Pro Thr Pro Leu Asp 485 490 495 Ser Thr Pro Pro Gly Gly Ala Pro His Pro Leu Thr Gly Gln Glu Glu 500 505 510 Ala Arg Ala Val Glu Lys Asp Lys Ser Lys Ala Arg Ser Glu Asp Thr 515 520 525 Gly Leu Asp Ser Val Ala Thr Arg Thr Pro Met Glu His Val Thr Pro 530 535 540 Lys Pro Glu Thr His Leu Ala Ser Pro Lys Pro Thr Cys Met Val Pro 545 550 555 560 Pro Met Pro His Ser Pro Val Ser Gly Asp Ser Val Glu Glu Glu Glu 565 570 575 Glu Glu Glu Lys Lys Val Cys Leu Pro Gly Phe Thr Gly Leu Val Asn 580 585 590 Leu Gly Asn Thr Cys Phe Met Asn Ser Val Ile Gln Ser Leu Ser Asn 595 600 605 Thr Arg Glu Leu Arg Asp Phe Phe His Asp Arg Ser Phe Glu Ala Glu 610 615 620 Ile Asn Tyr Asn Asn Pro Leu Gly Thr Gly Gly Arg Leu Ala Ile Gly 625 630 635 640 Phe Ala Val Leu Leu Arg Ala Leu Trp Lys Gly Thr His His Ala Phe 645 650 655 Gln Pro Ser Lys Leu Lys Ala Ile Val Ala Ser Lys Ala Ser Gln Phe 660 665 670 Thr Gly Tyr Ala Gln His Asp Ala Gln Glu Phe Met Ala Phe Leu Leu 675 680 685 Asp Gly Leu His Glu Asp Leu Asn Arg Ile Gln Asn Lys Pro Tyr Thr 690 695 700 Glu Thr Val Asp Ser Asp Gly Arg Pro Asp Glu Val Val Ala Glu Glu 705 710 715 720 Ala Trp Gln Arg His Lys Met Arg Asn Asp Ser Phe Ile Val Asp Leu 725 730 735 Phe Gln Gly Gln Tyr Lys Ser Lys Leu Val Cys Pro Val Cys Ala Lys 740 745 750 Val Ser Ile Thr Phe Asp Pro Phe Leu Tyr Leu Pro Val Pro Leu Pro 755 760 765 Gln Lys Gln Lys Val Leu Pro Val Phe Tyr Phe Ala Arg Glu Pro His 770 775 780 Ser Lys Pro Ile Lys Phe Leu Val Ser Val Ser Lys Glu Asn Ser Thr 785 790 795 800 Ala Ser Glu Val Leu Asp Ser Leu Ser Gln Ser Val His Val Lys Pro 805 810 815 Glu Asn Leu Arg Leu Ala Glu Val Ile Lys Asn Arg Phe His Arg Val 820 825 830 Phe Leu Pro Ser His Ser Leu Asp Thr Val Ser Pro Ser Asp Thr Leu 835 840 845 Leu Cys Phe Glu Leu Leu Ser Ser Glu Leu Ala Lys Glu Arg Val Val 850 855 860 Val Leu Glu Val Gln Gln Arg Pro Gln Val Pro Ser Val Pro Ile Ser 865 870 875 880 Lys Cys Ala Ala Cys Gln Arg Lys Gln Gln Ser Glu Asp Glu Lys Leu 885 890 895 Lys Arg Cys Thr Arg Cys Tyr Arg Val Gly Tyr Cys Asn Gln Leu Cys 900 905 910 Gln Lys Thr His Trp Pro Asp His Lys Gly Leu Cys Arg Pro Glu Asn 915 920 925 Ile Gly Tyr Pro Phe Leu Val Ser Val Pro Ala Ser Arg Leu Thr Tyr 930 935 940 Ala Arg Leu Ala Gln Leu Leu Glu Gly Tyr Ala Arg Tyr Ser Val Ser 945 950 955 960 Val Phe Gln Pro Pro Phe Gln Pro Gly Arg Met Ala Leu Glu Ser Gln 965 970 975 Ser Pro Gly Cys Thr Thr Leu Leu Ser Thr Gly Ser Leu Glu Ala Gly 980 985 990 Asp Ser Glu Arg Asp Pro Ile Gln Pro Pro Glu Leu Gln Leu Val Thr 995 1000 1005 Pro Met Ala Glu Gly Asp Thr Gly Leu Pro Arg Val Trp Ala Ala Pro 1010 1015 1020 Asp Arg Gly Pro Val Pro Ser Thr Ser Gly Ile Ser Ser Glu Met Leu 1025 1030 1035 1040 Ala Ser Gly Pro Ile Glu Val Gly Ser Leu Pro Ala Gly Glu Arg Val 1045 1050 1055 Ser Arg Pro Glu Ala Ala Val Pro Gly Tyr Gln His Pro Ser Glu Ala 1060 1065 1070 Met Asn Ala His Thr Pro Gln Phe Phe Ile Tyr Lys Ile Asp Ser Ser 1075 1080 1085 Asn Arg Glu Gln Arg Leu Glu Asp Lys Gly Asp Thr Pro Leu Glu Leu 1090 1095 1100 Gly Asp Asp Cys Ser Leu Ala Leu Val Trp Arg Asn Asn Glu Arg Leu 1105 1110 1115 1120 Gln Glu Phe Val Leu Val Ala Ser Lys Glu Leu Glu Cys Ala Glu Asp 1125 1130 1135 Pro Gly Ser Ala Gly Glu Ala Ala Arg Ala Gly His Phe Thr Leu Asp 1140 1145 1150 Gln Cys Leu Asn Leu Phe Thr Arg Pro Glu Val Leu Ala Pro Glu Glu 1155 1160 1165 Ala Trp Tyr Cys Pro Gln Cys Lys Gln His Arg Glu Ala Ser Lys Gln 1170 1175 1180 Leu Leu Leu Trp Arg Leu Pro Asn Val Leu Ile Val Gln Leu Lys Arg 1185 1190 1195 1200 Phe Ser Phe Arg Ser Phe Ile Trp Arg Asp Lys Ile Asn Asp Leu Val 1205 1210 1215 Glu Phe Pro Val Arg Asn Leu Asp Leu Ser Lys Phe Cys Ile Gly Gln 1220 1225 1230 Lys Glu Glu Gln Leu Pro Ser Tyr Asp Leu Tyr Ala Val Ile Asn His 1235 1240 1245 Tyr Gly Gly Met Ile Gly Gly His Tyr Thr Ala Cys Ala Arg Leu Pro 1250 1255 1260 Asn Asp Arg Ser Ser Gln Arg Ser Asp Val Gly Trp Arg Leu Phe Asp 1265 1270 1275 1280 Asp Ser Thr Val Thr Thr Val Asp Glu Ser Gln Val Val Thr Arg Tyr 1285 1290 1295 Ala Tyr Val Leu Phe Tyr Arg Arg Arg Asn Ser Pro Val Glu Arg Pro 1300 1305 1310 Pro Arg Ala Gly His Ser Glu His His Pro Asp Leu Gly Pro Ala Ala 1315 1320 1325 Glu Ala Ala Ala Ser Gln Gly Leu Gly Pro Gly Gln Ala Pro Glu Val 1330 1335 1340 Ala Pro Thr Arg Thr Ala Pro Glu Arg Phe Ala Pro Pro Val Asp Arg 1345 1350 1355 1360 Pro Ala Pro Thr Tyr Ser Asn Met Glu Glu Val Asp 1365 1370 <210> 4 <211> 1318 <212> PRT <213> Homo sapiens <400> 4 Met Ser Gly Gly Ala Ser Ala Thr Gly Pro Arg Arg Gly Pro Pro Gly 1 5 10 15 Leu Glu Asp Thr Thr Ser Lys Lys Lys Gln Lys Asp Arg Ala Asn Gln 20 25 30 Glu Ser Lys Asp Gly Asp Pro Arg Lys Glu Thr Gly Ser Arg Tyr Val 35 40 45 Ala Gln Ala Gly Leu Glu Pro Leu Ala Ser Gly Asp Pro Ser Ala Ser 50 55 60 Ala Ser His Ala Ala Gly Ile Thr Gly Ser Arg His Arg Thr Arg Leu 65 70 75 80 Phe Phe Pro Ser Ser Ser Gly Ser Ala Ser Thr Pro Gln Glu Glu Gln 85 90 95 Thr Lys Glu Gly Ala Cys Glu Asp Pro His Asp Leu Leu Ala Thr Pro 100 105 110 Thr Pro Glu Leu Leu Leu Asp Trp Arg Gln Ser Ala Glu Glu Val Ile 115 120 125 Val Lys Leu Arg Val Gly Val Gly Pro Leu Gln Leu Glu Asp Val Asp 130 135 140 Ala Ala Phe Thr Asp Thr Asp Cys Val Val Arg Phe Ala Gly Gly Gln 145 150 155 160 Gln Trp Gly Gly Val Phe Tyr Ala Glu Ile Lys Ser Ser Cys Ala Lys 165 170 175 Val Gln Thr Arg Lys Gly Ser Leu Leu His Leu Thr Leu Pro Lys Lys 180 185 190 Val Pro Met Leu Thr Trp Pro Ser Leu Leu Val Glu Ala Asp Glu Gln 195 200 205 Leu Cys Ile Pro Pro Leu Asn Ser Gln Thr Cys Leu Leu Gly Ser Glu 210 215 220 Glu Asn Leu Ala Pro Leu Ala Gly Glu Lys Ala Val Pro Pro Gly Asn 225 230 235 240 Asp Pro Val Ser Pro Ala Met Val Arg Ser Arg Asn Pro Gly Lys Asp 245 250 255 Asp Cys Ala Lys Glu Glu Met Ala Val Ala Ala Asp Ala Ala Thr Leu 260 265 270 Val Asp Glu Pro Glu Ser Met Val Asn Leu Ala Phe Val Lys Asn Asp 275 280 285 Ser Tyr Glu Lys Gly Pro Asp Ser Val Val Val His Val Tyr Val Lys 290 295 300 Glu Ile Cys Arg Asp Thr Ser Arg Val Leu Phe Arg Glu Gln Asp Phe 305 310 315 320 Thr Leu Ile Phe Gln Thr Arg Asp Gly Asn Phe Leu Arg Leu His Pro 325 330 335 Gly Cys Gly Pro His Thr Thr Phe Arg Trp Gln Val Lys Leu Arg Asn 340 345 350 Leu Ile Glu Pro Glu Gln Cys Thr Phe Cys Phe Thr Ala Ser Arg Ile 355 360 365 Asp Ile Cys Leu Arg Lys Arg Gln Ser Gln Arg Trp Gly Gly Leu Glu 370 375 380 Ala Pro Ala Ala Arg Val Gly Gly Ala Lys Val Ala Val Pro Thr Gly 385 390 395 400 Pro Thr Pro Leu Asp Ser Thr Pro Pro Gly Gly Ala Pro His Pro Leu 405 410 415 Thr Gly Gln Glu Glu Ala Arg Ala Val Glu Lys Asp Lys Ser Lys Ala 420 425 430 Arg Ser Glu Asp Thr Gly Leu Asp Ser Val Ala Thr Arg Thr Pro Met 435 440 445 Glu His Val Thr Pro Lys Pro Glu Thr His Leu Ala Ser Pro Lys Pro 450 455 460 Thr Cys Met Val Pro Pro Met Pro His Ser Pro Val Ser Gly Asp Ser 465 470 475 480 Val Glu Glu Glu Glu Glu Glu Glu Lys Lys Val Cys Leu Pro Gly Phe 485 490 495 Thr Gly Leu Val Asn Leu Gly Asn Thr Cys Phe Met Asn Ser Val Ile 500 505 510 Gln Ser Leu Ser Asn Thr Arg Glu Leu Arg Asp Phe Phe His Asp Arg 515 520 525 Ser Phe Glu Ala Glu Ile Asn Tyr Asn Asn Pro Leu Gly Thr Gly Gly 530 535 540 Arg Leu Ala Ile Gly Phe Ala Val Leu Leu Arg Ala Leu Trp Lys Gly 545 550 555 560 Thr His His Ala Phe Gln Pro Ser Lys Leu Lys Ala Ile Val Ala Ser 565 570 575 Lys Ala Ser Gln Phe Thr Gly Tyr Ala Gln His Asp Ala Gln Glu Phe 580 585 590 Met Ala Phe Leu Leu Asp Gly Leu His Glu Asp Leu Asn Arg Ile Gln 595 600 605 Asn Lys Pro Tyr Thr Glu Thr Val Asp Ser Asp Gly Arg Pro Asp Glu 610 615 620 Val Val Ala Glu Glu Ala Trp Gln Arg His Lys Met Arg Asn Asp Ser 625 630 635 640 Phe Ile Val Asp Leu Phe Gln Gly Gln Tyr Lys Ser Lys Leu Val Cys 645 650 655 Pro Val Cys Ala Lys Val Ser Ile Thr Phe Asp Pro Phe Leu Tyr Leu 660 665 670 Pro Val Pro Leu Pro Gln Lys Gln Lys Val Leu Pro Val Phe Tyr Phe 675 680 685 Ala Arg Glu Pro His Ser Lys Pro Ile Lys Phe Leu Val Ser Val Ser 690 695 700 Lys Glu Asn Ser Thr Ala Ser Glu Val Leu Asp Ser Leu Ser Gln Ser 705 710 715 720 Val His Val Lys Pro Glu Asn Leu Arg Leu Ala Glu Val Ile Lys Asn 725 730 735 Arg Phe His Arg Val Phe Leu Pro Ser His Ser Leu Asp Thr Val Ser 740 745 750 Pro Ser Asp Thr Leu Leu Cys Phe Glu Leu Leu Ser Ser Glu Leu Ala 755 760 765 Lys Glu Arg Val Val Val Leu Glu Val Gln Gln Arg Pro Gln Val Pro 770 775 780 Ser Val Pro Ile Ser Lys Cys Ala Ala Cys Gln Arg Lys Gln Gln Ser 785 790 795 800 Glu Asp Glu Lys Leu Lys Arg Cys Thr Arg Cys Tyr Arg Val Gly Tyr 805 810 815 Cys Asn Gln Leu Cys Gln Lys Thr His Trp Pro Asp His Lys Gly Leu 820 825 830 Cys Arg Pro Glu Asn Ile Gly Tyr Pro Phe Leu Val Ser Val Pro Ala 835 840 845 Ser Arg Leu Thr Tyr Ala Arg Leu Ala Gln Leu Leu Glu Gly Tyr Ala 850 855 860 Arg Tyr Ser Val Ser Val Phe Gln Pro Pro Phe Gln Pro Gly Arg Met 865 870 875 880 Ala Leu Glu Ser Gln Ser Pro Gly Cys Thr Thr Leu Leu Ser Thr Gly 885 890 895 Ser Leu Glu Ala Gly Asp Ser Glu Arg Asp Pro Ile Gln Pro Pro Glu 900 905 910 Leu Gln Leu Val Thr Pro Met Ala Glu Gly Asp Thr Gly Leu Pro Arg 915 920 925 Val Trp Ala Ala Pro Asp Arg Gly Pro Val Pro Ser Thr Ser Gly Ile 930 935 940 Ser Ser Glu Met Leu Ala Ser Gly Pro Ile Glu Val Gly Ser Leu Pro 945 950 955 960 Ala Gly Glu Arg Val Ser Arg Pro Glu Ala Ala Val Pro Gly Tyr Gln 965 970 975 His Pro Ser Glu Ala Met Asn Ala His Thr Pro Gln Phe Phe Ile Tyr 980 985 990 Lys Ile Asp Ser Ser Asn Arg Glu Gln Arg Leu Glu Asp Lys Gly Asp 995 1000 1005 Thr Pro Leu Glu Leu Gly Asp Asp Cys Ser Leu Ala Leu Val Trp Arg 1010 1015 1020 Asn Asn Glu Arg Leu Gln Glu Phe Val Leu Val Ala Ser Lys Glu Leu 1025 1030 1035 1040 Glu Cys Ala Glu Asp Pro Gly Ser Ala Gly Glu Ala Ala Arg Ala Gly 1045 1050 1055 His Phe Thr Leu Asp Gln Cys Leu Asn Leu Phe Thr Arg Pro Glu Val 1060 1065 1070 Leu Ala Pro Glu Glu Ala Trp Tyr Cys Pro Gln Cys Lys Gln His Arg 1075 1080 1085 Glu Ala Ser Lys Gln Leu Leu Leu Trp Arg Leu Pro Asn Val Leu Ile 1090 1095 1100 Val Gln Leu Lys Arg Phe Ser Phe Arg Ser Phe Ile Trp Arg Asp Lys 1105 1110 1115 1120 Ile Asn Asp Leu Val Glu Phe Pro Val Arg Asn Leu Asp Leu Ser Lys 1125 1130 1135 Phe Cys Ile Gly Gln Lys Glu Glu Gln Leu Pro Ser Tyr Asp Leu Tyr 1140 1145 1150 Ala Val Ile Asn His Tyr Gly Gly Met Ile Gly Gly His Tyr Thr Ala 1155 1160 1165 Cys Ala Arg Leu Pro Asn Asp Arg Ser Ser Gln Arg Ser Asp Val Gly 1170 1175 1180 Trp Arg Leu Phe Asp Asp Ser Thr Val Thr Thr Val Asp Glu Ser Gln 1185 1190 1195 1200 Val Val Thr Arg Tyr Ala Tyr Val Leu Phe Tyr Arg Arg Arg Asn Ser 1205 1210 1215 Pro Val Glu Arg Pro Pro Arg Ala Gly His Ser Glu His His Pro Asp 1220 1225 1230 Leu Gly Pro Ala Ala Glu Ala Ala Ala Ser Gln Ala Ser Arg Ile Trp 1235 1240 1245 Gln Glu Leu Glu Ala Glu Glu Glu Pro Val Pro Glu Gly Ser Gly Pro 1250 1255 1260 Leu Gly Pro Trp Gly Pro Gln Asp Trp Val Gly Pro Leu Pro Arg Gly 1265 1270 1275 1280 Pro Thr Thr Pro Asp Glu Gly Cys Leu Arg Tyr Phe Val Leu Gly Thr 1285 1290 1295 Val Ala Ala Leu Val Ala Leu Val Leu Asn Val Phe Tyr Pro Leu Val 1300 1305 1310 Ser Gln Ser Arg Trp Arg 1315 <210> 5 <211> 4878 <212> DNA <213> Homo sapiens <400> 5 agcttggcgc tccgcctccc tcggccttag ctagaagtcg aaacaaaaca agccgctaag 60 gcggtggcgg cggcgccggg acgggggagg ggcgcgccgg aaccggaacc gacctgcgcc 120 ggaaccggaa cggagagcgg gttgccaggg cccgaagagg gctggctgcg gcggtctcgc 180 tcggctgtcc gttccttgct ggagaatttg gccacaaaga gttgccaaga tagctgggcc 240 aggaagaaag cgccgcagcc ctgacccaga cgctgttgcc gaccccgggg cactctggct 300 gtcgaccaag cggctcaaga tgtctggcgg ggccagtgcc acaggcccaa ggagagggcc 360 cccaggactg gaggacacca ctagtaagaa gaagcagaag gatcgagcaa accaggagag 420 caaggatgga gatcctagga aagagacagg gtctcgatat gttgcccagg ctggtcttga 480 acctctggcc tcaggtgatc cttctgcctc agcctcccat gcagctggga tcacaggctc 540 acgccaccgt acccggctgt tctttccttc atcgtcaggg tcagcatcca ctcctcaaga 600 ggagcagacc aaagagggag cttgtgaaga ccctcatgat ctcttggcta ctcccactcc 660 agagttgttg ctcgattgga ggcagagtgc agaagaggtg attgtcaagc ttcgtgtggg 720 agtaggtccc ctgcagctgg aggatgtaga tgctgctttc acagatacag actgtgtggt 780 gcggtttgca ggtggtcagc agtggggtgg tgtcttctat gctgagataa aaagctcttg 840 tgctaaagtg caaacccgca agggcagtct cctgcacctg acactgccca aaaaggtgcc 900 tatgctcacg tggccctccc tcctgaagaa acctctaggg acccaggagc tggtgccggg 960 gctgcggtgc caggagaatg ggcaggaact gtctcccatt gccctggagc caggccctga 1020 gccccaccgg gctaagcagg aggcccggaa ccagaagcgg gcccagggcc gtggtgaggt 1080 aggcgcaggg gctggccccg gggcccaggc agggcccagc gccaagaggg ctgtgcatct 1140 ctgcagaggg ccagaggggg acgggtccag ggatgaccct ggaccccggg gtgatgcccc 1200 acccttcgtg gctgacccag ccacccaggt tgaggctgat gaacagcttt gcataccacc 1260 gctgaactcc caaacctgcc tcctgggctc agaggagaat ttagcccctt tggcaggaga 1320 gaaagcagtg cctcccggga atgacccagt ctctccagcc atggtccgga gcagaaaccc 1380 tgggaaagat gactgtgcca aggaggagat ggcagtggca gcagatgctg caaccttggt 1440 ggatgagccc gagtcgatgg tgaacctggc gtttgtcaag aatgactcgt atgagaaggg 1500 cccggattca gtggtggtgc acgtgtacgt gaaggagatc tgcagggaca cctcaagagt 1560 acttttccgt gagcaggact tcacgctcat cttccagacc agggatggaa acttcctgag 1620 gctgcacccg ggctgtgggc cccacaccac cttccgttgg caggtgaagc tcaggaatct 1680 gattgagcca gagcagtgca ccttctgttt cacggcttct cgcatcgaca tctgccttcg 1740 taagaggcag agtcagcgct gggggggcct ggaggccccg gctgcacgag tgggtggtgc 1800 aaaggttgcc gtgccgacag gtccaacccc tctggattca accccaccag gaggtgctcc 1860 ccaccccctg acaggccagg aggaggcccg ggctgtggag aaggataaat ccaaggcacg 1920 atctgaggac acagggctag acagtgtggc aacccgcaca cccatggagc atgtaacccc 1980 aaagccagag acacacctgg cctcgcccaa gcctacatgc atggtgcctc ccatgcccca 2040 cagcccagtt agtggagaca gcgtggagga ggaggaagag gaagagaaga aggtgtgtct 2100 gccaggcttc actggccttg tcaatttagg caacacctgc ttcatgaaca gcgtcattca 2160 gtctctgtcc aacactcggg aactccggga cttcttccat gaccgctcct ttgaggctga 2220 gatcaactac aacaacccac tagggactgg tgggcgtctg gccattggct ttgccgtgct 2280 gcttcgggcg ctgtggaagg gcacccacca tgccttccag ccttccaagt tgaaggccat 2340 tgtggcgagt aaggccagcc agttcacagg ctatgcacag catgatgccc aggagttcat 2400 ggctttcctg ctggatgggc tgcacgagga cctgaatcgc attcagaaca agccctacac 2460 agagaccgtg gattcagatg ggcggcccga tgaggtggta gctgaggaag catggcagcg 2520 gcacaagatg aggaatgact ctttcatcgt ggacctattt caggggcagt acaagtcgaa 2580 gctggtgtgc cctgtgtgtg ccaaggtctc catcactttt gacccgtttc tttatctgcc 2640 ggtgcccttg ccacaaaagc aaaaggttct ccctgtcttt tattttgccc gagagcccca 2700 cagcaagccc atcaagttcc tggtgagcgt cagcaaggag aactccactg cgagcgaagt 2760 attggactcc ctctctcaga gtgttcatgt gaagcctgag aacctgcgtt tggcggaggt 2820 aattaagaat cgttttcatc gtgtgttcct accctcccac tcactggaca ctgtgtcccc 2880 atctgatacg ctcctctgct ttgagctgct atcctcagag ttggctaagg agcgggtagt 2940 ggtgctagag gtgcaacagc gcccccaggt gcccagcgtc cccatctcca agtgtgcagc 3000 ctgccagcgg aagcaacagt cggaggatga aaagctgaag cgctgtaccc ggtgctaccg 3060 tgtgggctac tgcaaccagc tctgccagaa aacccactgg cctgaccaca agggcctctg 3120 ccgacctgag aacattggct accccttcct ggtcagtgta cctgcctcac gcctcactta 3180 tgcccgcctc gctcagttgc tagagggcta tgcccggtac tctgtgagtg tattccagcc 3240 accctttcag ccaggccgca tggccttgga gtctcagagc cctggctgca ccacactgct 3300 ctccacaggt tccctggagg ctggggacag cgagagagac cccattcagc cacctgagct 3360 ccagctggtg acccctatgg ctgaggggga cacagggctt ccccgggtgt gggcagcccc 3420 tgaccggggt cctgtgccca gcaccagtgg aatttcttct gagatgctgg ccagtgggcc 3480 cattgaggtt ggctccttgc cagctggcga gagggtgtcc cgacccgaag ctgctgtgcc 3540 tgggtaccag catccaagtg aagctatgaa tgcccacaca ccccagttct tcatctataa 3600 aattgattca tccaaccgag agcagcggct agaggacaaa ggagacaccc cactggagct 3660 gggtgacgac tgtagcctgg ctctcgtctg gcggaacaat gagcgcttgc aggagtttgt 3720 gttggtagcc tccaaggagc tggaatgtgc tgaggatcca ggctctgccg gtgaggctgc 3780 ccgggccggc cacttcaccc tggaccagtg cctcaacctc ttcacacggc ctgaggtgct 3840 ggcacccgag gaggcctggt actgcccaca gtgcaaacag caccgtgagg cctccaagca 3900 gctgttgcta tggcgcctgc caaatgttct catcgtgcag ctcaagcgct tctcctttcg 3960 tagttttatc tggcgtgaca agatcaatga cttggtggag ttccctgtta ggaacctgga 4020 cctgagcaag ttctgcattg gtcagaaaga ggagcagctg cccagctacg atctatatgc 4080 tgtcatcaac cactatggag gcatgattgg tggccactac actgcctgtg cacgcctgcc 4140 caatgatcgt agcagtcagc gcagtgacgt gggctggcgc ttgtttgatg acagcacagt 4200 gacaacggta gacgagagcc aggttgtgac gcgttatgcc tatgtactct tctaccgccg 4260 gcggaactct cctgtggaga ggccccccag ggcaggtcac tctgagcacc acccagacct 4320 aggccctgca gctgaggctg ctgccagcca ggcttcccgg atttggcagg agctggaggc 4380 tgaggaggag ccggtgcctg aggggtctgg gcccctgggt ccctgggggc cccaagactg 4440 ggtgggcccc ctaccacgtg gccctaccac accagatgag ggctgcctcc ggtactttgt 4500 cctgggcacc gtggcggctt tggtggccct cgtgctcaac gtgttctatc ctctggtatc 4560 ccagagtcgc tggagatgag ctcgcctgca ggcagctgct gtgagctggc ctacctgcct 4620 gccccaggcc atgcctgcct ttgttgtggg gaacacctct gggctttggg cctcagctta 4680 tgcatctggt gggagagggt ggggaggttg tggcccctgc aggggcagag tatcctaggg 4740 tgtgtatcca tctggctgtc tgtccattca tcctgctgct ctgacccttg gcctcaggct 4800 tggccctgcc caagctactt cctgtactta aaagtgttaa taaaaccaga ctattcaggc 4860 ccaaaaaaaa aaaaaaaa 4878 <210> 6 <211> 4883 <212> DNA <213> Homo sapiens <400> 6 agcttggcgc tccgcctccc tcggccttag ctagaagtcg aaacaaaaca agccgctaag 60 gcggtggcgg cggcgccggg acgggggagg ggcgcgccgg aaccggaacc gacctgcgcc 120 ggaaccggaa cggagagcgg gttgccaggg cccgaagagg gctggctgcg gcggtctcgc 180 tcggctgtcc gttccttgct ggagaatttg gccacaaaga gttgccaaga tagctgggcc 240 aggaagaaag cgccgcagcc ctgacccaga cgctgttgcc gaccccgggg cactctggct 300 gtcgaccaag cggctcaaga tgtctggcgg ggccagtgcc acaggcccaa ggagagggcc 360 cccaggactg gaggacacca ctagtaagaa gaagcagaag gatcgagcaa accaggagag 420 caaggatgga gatcctagga aagagacagg gtctcgatat gttgcccagg ctggtcttga 480 acctctggcc tcaggtgatc cttctgcctc agcctcccat gcagctggga tcacaggctc 540 acgccaccgt acccggctgt tctttccttc atcgtcaggg tcagcatcca ctcctcaaga 600 ggagcagacc aaagagggag cttgtgaaga ccctcatgat ctcttggcta ctcccactcc 660 agagttgttg ctcgattgga ggcagagtgc agaagaggtg attgtcaagc ttcgtgtggg 720 agtaggtccc ctgcagctgg aggatgtaga tgctgctttc acagatacag actgtgtggt 780 gcggtttgca ggtggtcagc agtggggtgg tgtcttctat gctgagataa aaagctcttg 840 tgctaaagtg caaacccgca agggcagtct cctgcacctg acactgccca aaaaggtgcc 900 tatgctcacg tggccctccc tcctgaagaa acctctaggg acccaggagc tggtgccggg 960 gctgcggtgc caggagaatg ggcaggaact gtctcccatt gccctggagc caggccctga 1020 gccccaccgg gctaagcagg aggcccggaa ccagaagcgg gcccagggcc gtggtgaggt 1080 aggcgcaggg gctggccccg gggcccaggc agggcccagc gccaagaggg ctgtgcatct 1140 ctgcagaggg ccagaggggg acgggtccag ggatgaccct ggaccccggg gtgatgcccc 1200 acccttcgtg gctgacccag ccacccaggt tgaggctgat gaacagcttt gcataccacc 1260 gctgaactcc caaacctgcc tcctgggctc agaggagaat ttagcccctt tggcaggaga 1320 gaaagcagtg cctcccggga atgacccagt ctctccagcc atggtccgga gcagaaaccc 1380 tgggaaagat gactgtgcca aggaggagat ggcagtggca gcagatgctg caaccttggt 1440 ggatgagccc gagtcgatgg tgaacctggc gtttgtcaag aatgactcgt atgagaaggg 1500 cccggattca gtggtggtgc acgtgtacgt gaaggagatc tgcagggaca cctcaagagt 1560 acttttccgt gagcaggact tcacgctcat cttccagacc agggatggaa acttcctgag 1620 gctgcacccg ggctgtgggc cccacaccac cttccgttgg caggtgaagc tcaggaatct 1680 gattgagcca gagcagtgca ccttctgttt cacggcttct cgcatcgaca tctgccttcg 1740 taagaggcag agtcagcgct gggggggcct ggaggccccg gctgcacgag gtgcagtggg 1800 tggtgcaaag gttgccgtgc cgacaggtcc aacccctctg gattcaaccc caccaggagg 1860 tgctccccac cccctgacag gccaggagga ggcccgggct gtggagaagg ataaatccaa 1920 ggcacgatct gaggacacag ggctagacag tgtggcaacc cgcacaccca tggagcatgt 1980 aaccccaaag ccagagacac acctggcctc gcccaagcct acatgcatgg tgcctcccat 2040 gccccacagc ccagttagtg gagacagcgt ggaggaggag gaagaggaag agaagaaggt 2100 gtgtctgcca ggcttcactg gccttgtcaa tttaggcaac acctgcttca tgaacagcgt 2160 cattcagtct ctgtccaaca ctcgggaact ccgggacttc ttccatgacc gctcctttga 2220 ggctgagatc aactacaaca acccactagg gactggtggg cgtctggcca ttggctttgc 2280 cgtgctgctt cgggcgctgt ggaagggcac ccaccatgcc ttccagcctt ccaagttgaa 2340 ggccattgtg gcgagtaagg ccagccagtt cacaggctat gcacagcatg atgcccagga 2400 gttcatggct ttcctgctgg atgggctgca cgaggacctg aatcgcattc agaacaagcc 2460 ctacacagag accgtggatt cagatgggcg gcccgatgag gtggtagctg aggaagcatg 2520 gcagcggcac aagatgagga atgactcttt catcgtggac ctatttcagg ggcagtacaa 2580 gtcgaagctg gtgtgccctg tgtgtgccaa ggtctccatc acttttgacc cgtttcttta 2640 tctgccggtg cccttgccac aaaagcaaaa ggttctccct gtcttttatt ttgcccgaga 2700 gccccacagc aagcccatca agttcctggt gagcgtcagc aaggagaact ccactgcgag 2760 cgaagtattg gactccctct ctcagagtgt tcatgtgaag cctgagaacc tgcgtttggc 2820 ggaggtaatt aagaatcgtt ttcatcgtgt gttcctaccc tcccactcac tggacactgt 2880 gtccccatct gatacgctcc tctgctttga gctgctatcc tcagagttgg ctaaggagcg 2940 ggtagtggtg ctagaggtgc aacagcgccc ccaggtgccc agcgtcccca tctccaagtg 3000 tgcagcctgc cagcggaagc aacagtcgga ggatgaaaag ctgaagcgct gtacccggtg 3060 ctaccgtgtg ggctactgca accagctctg ccagaaaacc cactggcctg accacaaggg 3120 cctctgccga cctgagaaca ttggctaccc cttcctggtc agtgtacctg cctcacgcct 3180 cacttatgcc cgcctcgctc agttgctaga gggctatgcc cggtactctg tgagtgtatt 3240 ccagccaccc tttcagccag gccgcatggc cttggagtct cagagccctg gctgcaccac 3300 actgctctcc acaggttccc tggaggctgg ggacagcgag agagacccca ttcagccacc 3360 tgagctccag ctggtgaccc ctatggctga gggggacaca gggcttcccc gggtgtgggc 3420 agcccctgac cggggtcctg tgcccagcac cagtggaatt tcttctgaga tgctggccag 3480 tgggcccatt gaggttggct ccttgccagc tggcgagagg gtgtcccgac ccgaagctgc 3540 tgtgcctggg taccagcatc caagtgaagc tatgaatgcc cacacacccc agttcttcat 3600 ctataaaatt gattcatcca accgagagca gcggctagag gacaaaggag acaccccact 3660 ggagctgggt gacgactgta gcctggctct cgtctggcgg aacaatgagc gcttgcagga 3720 gtttgtgttg gtagcctcca aggagctgga atgtgctgag gatccaggct ctgccggtga 3780 ggctgcccgg gccggccact tcaccctgga ccagtgcctc aacctcttca cacggcctga 3840 ggtgctggca cccgaggagg cctggtactg cccacagtgc aaacagcacc gtgaggcctc 3900 caagcagctg ttgctatggc gcctgccaaa tgttctcatc gtgcagctca agcgcttctc 3960 ctttcgtagt tttatctggc gtgacaagat caatgacttg gtggagttcc ctgttaggaa 4020 cctggacctg agcaagttct gcattggtca gaaagaggag cagctgccca gctacgatct 4080 atatgctgtc atcaaccact atggaggcat gattggtggc cactacactg cctgtgcacg 4140 cctgcccaat gatcgtagca gtcagcgcag tgacgtgggc tggcgcttgt ttgatgacag 4200 cacagtgaca acggtagacg agagccaggt tgtgacgcgt tatgcctatg tactcttcta 4260 ccgccggcgg aactctcctg tggagaggcc ccccagggca ggtcactctg agcaccaccc 4320 agacctaggc cctgcagctg aggctgctgc cagccaggga ctaggccctg gccaggcccc 4380 cgaggtggcc cccacgcgga cagcccctga acgcttcgcc ccccctgtgg atcggccagc 4440 ccccacctac agcaacatgg aggaggtgga ttagcaggtc cctggctgat gggggggact 4500 gggtttggga cacccacaca gagggccagc tccttgccgc ttctccttct ctaacccaga 4560 ggacactggc tctgtcaatg ggaagctgag gggtatgatt tgggtgtgga gacctctcag 4620 gttgggactt cttgtcagct tggacccctg accagtgggc tttggcttct ccagccgcct 4680 ccagtgctgc gtgatttgat tctgttgtac cttcaattct tctgacccgc attataaaca 4740 ttataatttt attctaaaaa ttgtaatttt ttttgcattt tggaagtgac tgctgctgtt 4800 taaatatatt ttaaaaataa ataataagta agtagactta aaaaaaaaaa aaaaaaaaaa 4860 aaaaaaaaaa aaaaaaaaaa aaa 4883 <210> 7 <211> 4847 <212> DNA <213> Homo sapiens <400> 7 agcttggcgc tccgcctccc tcggccttag ctagaagtcg aaacaaaaca agccgctaag 60 gcggtggcgg cggcgccggg acgggggagg ggcgcgccgg aaccggaacc gacctgcgcc 120 ggaaccggaa cggagagcgg gttgccaggg cccgaagagg gctggctgcg gcggtctcgc 180 tcggctgtcc gttccttgct ggagaatttg gccacaaaga gttgccaaga tagctgggcc 240 aggaagaaag cgccgcagcc ctgacccaga cgctgttgcc gaccccgggg cactctggct 300 gtcgaccaag cggctcaaga tgtctggcgg ggccagtgcc acaggcccaa ggagagggcc 360 cccaggactg gaggacacca ctagtaagaa gaagcagaag gatcgagcaa accaggagag 420 caaggatgga gatcctagga aagagacagg gtctcgatat gttgcccagg ctggtcttga 480 acctctggcc tcaggtgatc cttctgcctc agcctcccat gcagctggga tcacaggctc 540 acgccaccgt acccggctgt tctttccttc atcgtcaggg tcagcatcca ctcctcaaga 600 ggagcagacc aaagagggag cttgtgaaga ccctcatgat ctcttggcta ctcccactcc 660 agagttgttg ctcgattgga ggcagagtgc agaagaggtg attgtcaagc ttcgtgtggg 720 agtaggtccc ctgcagctgg aggatgtaga tgctgctttc acagatacag actgtgtggt 780 gcggtttgca ggtggtcagc agtggggtgg tgtcttctat gctgagataa aaagctcttg 840 tgctaaagtg caaacccgca agggcagtct cctgcacctg acactgccca aaaagaagaa 900 acctctaggg acccaggagc tggtgccggg gctgcggtgc caggagaatg ggcaggaact 960 gtctcccatt gccctggagc caggccctga gccccaccgg gctaagcagg aggcccggaa 1020 ccagaagcgg gcccagggcc gtggtgaggt aggcgcaggg gctggccccg gggcccaggc 1080 agggcccagc gccaagaggg ctgtgcatct ctgcagaggg ccagaggggg acgggtccag 1140 ggatgaccct ggaccccggg gtgatgcccc acccttcgtg gctgacccag ccacccaggt 1200 tgaggctgat gaacagcttt gcataccacc gctgaactcc caaacctgcc tcctgggctc 1260 agaggagaat ttagcccctt tggcaggaga gaaagcagtg cctcccggga atgacccagt 1320 ctctccagcc atggtccgga gcagaaaccc tgggaaagat gactgtgcca aggaggagat 1380 ggcagtggca gcagatgctg caaccttggt ggatgagccc gagtcgatgg tgaacctggc 1440 gtttgtcaag aatgactcgt atgagaaggg cccggattca gtggtggtgc acgtgtacgt 1500 gaaggagatc tgcagggaca cctcaagagt acttttccgt gagcaggact tcacgctcat 1560 cttccagacc agggatggaa acttcctgag gctgcacccg ggctgtgggc cccacaccac 1620 cttccgttgg caggtgaagc tcaggaatct gattgagcca gagcagtgca ccttctgttt 1680 cacggcttct cgcatcgaca tctgccttcg taagaggcag agtcagcgct gggggggcct 1740 ggaggccccg gctgcacgag tgggtggtgc aaaggttgcc gtgccgacag gtccaacccc 1800 tctggattca accccaccag gaggtgctcc ccaccccctg acaggccagg aggaggcccg 1860 ggctgtggag aaggataaat ccaaggcacg atctgaggac acagggctag acagtgtggc 1920 aacccgcaca cccatggagc atgtaacccc aaagccagag acacacctgg cctcgcccaa 1980 gcctacatgc atggtgcctc ccatgcccca cagcccagtt agtggagaca gcgtggagga 2040 ggaggaagag gaagagaaga aggtgtgtct gccaggcttc actggccttg tcaatttagg 2100 caacacctgc ttcatgaaca gcgtcattca gtctctgtcc aacactcggg aactccggga 2160 cttcttccat gaccgctcct ttgaggctga gatcaactac aacaacccac tagggactgg 2220 tgggcgtctg gccattggct ttgccgtgct gcttcgggcg ctgtggaagg gcacccacca 2280 tgccttccag ccttccaagt tgaaggccat tgtggcgagt aaggccagcc agttcacagg 2340 ctatgcacag catgatgccc aggagttcat ggctttcctg ctggatgggc tgcacgagga 2400 cctgaatcgc attcagaaca agccctacac agagaccgtg gattcagatg ggcggcccga 2460 tgaggtggta gctgaggaag catggcagcg gcacaagatg aggaatgact ctttcatcgt 2520 ggacctattt caggggcagt acaagtcgaa gctggtgtgc cctgtgtgtg ccaaggtctc 2580 catcactttt gacccgtttc tttatctgcc ggtgcccttg ccacaaaagc aaaaggttct 2640 ccctgtcttt tattttgccc gagagcccca cagcaagccc atcaagttcc tggtgagcgt 2700 cagcaaggag aactccactg cgagcgaagt attggactcc ctctctcaga gtgttcatgt 2760 gaagcctgag aacctgcgtt tggcggaggt aattaagaat cgttttcatc gtgtgttcct 2820 accctcccac tcactggaca ctgtgtcccc atctgatacg ctcctctgct ttgagctgct 2880 atcctcagag ttggctaagg agcgggtagt ggtgctagag gtgcaacagc gcccccaggt 2940 gcccagcgtc cccatctcca agtgtgcagc ctgccagcgg aagcaacagt cggaggatga 3000 aaagctgaag cgctgtaccc ggtgctaccg tgtgggctac tgcaaccagc tctgccagaa 3060 aacccactgg cctgaccaca agggcctctg ccgacctgag aacattggct accccttcct 3120 ggtcagtgta cctgcctcac gcctcactta tgcccgcctc gctcagttgc tagagggcta 3180 tgcccggtac tctgtgagtg tattccagcc accctttcag ccaggccgca tggccttgga 3240 gtctcagagc cctggctgca ccacactgct ctccacaggt tccctggagg ctggggacag 3300 cgagagagac cccattcagc cacctgagct ccagctggtg acccctatgg ctgaggggga 3360 cacagggctt ccccgggtgt gggcagcccc tgaccggggt cctgtgccca gcaccagtgg 3420 aatttcttct gagatgctgg ccagtgggcc cattgaggtt ggctccttgc cagctggcga 3480 gagggtgtcc cgacccgaag ctgctgtgcc tgggtaccag catccaagtg aagctatgaa 3540 tgcccacaca ccccagttct tcatctataa aattgattca tccaaccgag agcagcggct 3600 agaggacaaa ggagacaccc cactggagct gggtgacgac tgtagcctgg ctctcgtctg 3660 gcggaacaat gagcgcttgc aggagtttgt gttggtagcc tccaaggagc tggaatgtgc 3720 tgaggatcca ggctctgccg gtgaggctgc ccgggccggc cacttcaccc tggaccagtg 3780 cctcaacctc ttcacacggc ctgaggtgct ggcacccgag gaggcctggt actgcccaca 3840 gtgcaaacag caccgtgagg cctccaagca gctgttgcta tggcgcctgc caaatgttct 3900 catcgtgcag ctcaagcgct tctcctttcg tagttttatc tggcgtgaca agatcaatga 3960 cttggtggag ttccctgtta ggaacctgga cctgagcaag ttctgcattg gtcagaaaga 4020 ggagcagctg cccagctacg atctatatgc tgtcatcaac cactatggag gcatgattgg 4080 tggccactac actgcctgtg cacgcctgcc caatgatcgt agcagtcagc gcagtgacgt 4140 gggctggcgc ttgtttgatg acagcacagt gacaacggta gacgagagcc aggttgtgac 4200 gcgttatgcc tatgtactct tctaccgccg gcggaactct cctgtggaga ggccccccag 4260 ggcaggtcac tctgagcacc acccagacct aggccctgca gctgaggctg ctgccagcca 4320 gggactaggc cctggccagg cccccgaggt ggcccccacg cggacagccc ctgaacgctt 4380 cgccccccct gtggatcggc cagcccccac ctacagcaac atggaggagg tggattagca 4440 ggtccctggc tgatgggggg gactgggttt gggacaccca cacagagggc cagctccttg 4500 ccgcttctcc ttctctaacc cagaggacac tggctctgtc aatgggaagc tgaggggtat 4560 gatttgggtg tggagacctc tcaggttggg acttcttgtc agcttggacc cctgaccagt 4620 gggctttggc ttctccagcc gcctccagtg ctgcgtgatt tgattctgtt gtaccttcaa 4680 ttcttctgac ccgcattata aacattataa ttttattcta aaaattgtaa ttttttttgc 4740 attttggaag tgactgctgc tgtttaaata tattttaaaa ataaataata agtaagtaga 4800 cttaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaa 4847 <210> 8 <211> 4575 <212> DNA <213> Homo sapiens <400> 8 agcttggcgc tccgcctccc tcggccttag ctagaagtcg aaacaaaaca agccgctaag 60 gcggtggcgg cggcgccggg acgggggagg ggcgcgccgg aaccggaacc gacctgcgcc 120 ggaaccggaa cggagagcgg gttgccaggg cccgaagagg gctggctgcg gcggtctcgc 180 tcggctgtcc gttccttgct ggagaatttg gccacaaaga gttgccaaga tagctgggcc 240 aggaagaaag cgccgcagcc ctgacccaga cgctgttgcc gaccccgggg cactctggct 300 gtcgaccaag cggctcaaga tgtctggcgg ggccagtgcc acaggcccaa ggagagggcc 360 cccaggactg gaggacacca ctagtaagaa gaagcagaag gatcgagcaa accaggagag 420 caaggatgga gatcctagga aagagacagg gtctcgatat gttgcccagg ctggtcttga 480 acctctggcc tcaggtgatc cttctgcctc agcctcccat gcagctggga tcacaggctc 540 acgccaccgt acccggctgt tctttccttc atcgtcaggg tcagcatcca ctcctcaaga 600 ggagcagacc aaagagggag cttgtgaaga ccctcatgat ctcttggcta ctcccactcc 660 agagttgttg ctcgattgga ggcagagtgc agaagaggtg attgtcaagc ttcgtgtggg 720 agtaggtccc ctgcagctgg aggatgtaga tgctgctttc acagatacag actgtgtggt 780 gcggtttgca ggtggtcagc agtggggtgg tgtcttctat gctgagataa aaagctcttg 840 tgctaaagtg caaacccgca agggcagtct cctgcacctg acactgccca aaaaggtgcc 900 tatgctcacg tggccctccc tcctggttga ggctgatgaa cagctttgca taccaccgct 960 gaactcccaa acctgcctcc tgggctcaga ggagaattta gcccctttgg caggagagaa 1020 agcagtgcct cccgggaatg acccagtctc tccagccatg gtccggagca gaaaccctgg 1080 gaaagatgac tgtgccaagg aggagatggc agtggcagca gatgctgcaa ccttggtgga 1140 tgagcccgag tcgatggtga acctggcgtt tgtcaagaat gactcgtatg agaagggccc 1200 ggattcagtg gtggtgcacg tgtacgtgaa ggagatctgc agggacacct caagagtact 1260 tttccgtgag caggacttca cgctcatctt ccagaccagg gatggaaact tcctgaggct 1320 gcacccgggc tgtgggcccc acaccacctt ccgttggcag gtgaagctca ggaatctgat 1380 tgagccagag cagtgcacct tctgtttcac ggcttctcgc atcgacatct gccttcgtaa 1440 gaggcagagt cagcgctggg ggggcctgga ggccccggct gcacgagtgg gtggtgcaaa 1500 ggttgccgtg ccgacaggtc caacccctct ggattcaacc ccaccaggag gtgctcccca 1560 ccccctgaca ggccaggagg aggcccgggc tgtggagaag gataaatcca aggcacgatc 1620 tgaggacaca gggctagaca gtgtggcaac ccgcacaccc atggagcatg taaccccaaa 1680 gccagagaca cacctggcct cgcccaagcc tacatgcatg gtgcctccca tgccccacag 1740 cccagttagt ggagacagcg tggaggagga ggaagaggaa gagaagaagg tgtgtctgcc 1800 aggcttcact ggccttgtca atttaggcaa cacctgcttc atgaacagcg tcattcagtc 1860 tctgtccaac actcgggaac tccgggactt cttccatgac cgctcctttg aggctgagat 1920 caactacaac aacccactag ggactggtgg gcgtctggcc attggctttg ccgtgctgct 1980 tcgggcgctg tggaagggca cccaccatgc cttccagcct tccaagttga aggccattgt 2040 ggcgagtaag gccagccagt tcacaggcta tgcacagcat gatgcccagg agttcatggc 2100 tttcctgctg gatgggctgc acgaggacct gaatcgcatt cagaacaagc cctacacaga 2160 gaccgtggat tcagatgggc ggcccgatga ggtggtagct gaggaagcat ggcagcggca 2220 caagatgagg aatgactctt tcatcgtgga cctatttcag gggcagtaca agtcgaagct 2280 ggtgtgccct gtgtgtgcca aggtctccat cacttttgac ccgtttcttt atctgccggt 2340 gcccttgcca caaaagcaaa aggttctccc tgtcttttat tttgcccgag agccccacag 2400 caagcccatc aagttcctgg tgagcgtcag caaggagaac tccactgcga gcgaagtatt 2460 ggactccctc tctcagagtg ttcatgtgaa gcctgagaac ctgcgtttgg cggaggtaat 2520 taagaatcgt tttcatcgtg tgttcctacc ctcccactca ctggacactg tgtccccatc 2580 tgatacgctc ctctgctttg agctgctatc ctcagagttg gctaaggagc gggtagtggt 2640 gctagaggtg caacagcgcc cccaggtgcc cagcgtcccc atctccaagt gtgcagcctg 2700 ccagcggaag caacagtcgg aggatgaaaa gctgaagcgc tgtacccggt gctaccgtgt 2760 gggctactgc aaccagctct gccagaaaac ccactggcct gaccacaagg gcctctgccg 2820 acctgagaac attggctacc ccttcctggt cagtgtacct gcctcacgcc tcacttatgc 2880 ccgcctcgct cagttgctag agggctatgc ccggtactct gtgagtgtat tccagccacc 2940 ctttcagcca ggccgcatgg ccttggagtc tcagagccct ggctgcacca cactgctctc 3000 cacaggttcc ctggaggctg gggacagcga gagagacccc attcagccac ctgagctcca 3060 gctggtgacc cctatggctg agggggacac agggcttccc cgggtgtggg cagcccctga 3120 ccggggtcct gtgcccagca ccagtggaat ttcttctgag atgctggcca gtgggcccat 3180 tgaggttggc tccttgccag ctggcgagag ggtgtcccga cccgaagctg ctgtgcctgg 3240 gtaccagcat ccaagtgaag ctatgaatgc ccacacaccc cagttcttca tctataaaat 3300 tgattcatcc aaccgagagc agcggctaga ggacaaagga gacaccccac tggagctggg 3360 tgacgactgt agcctggctc tcgtctggcg gaacaatgag cgcttgcagg agtttgtgtt 3420 ggtagcctcc aaggagctgg aatgtgctga ggatccaggc tctgccggtg aggctgcccg 3480 ggccggccac ttcaccctgg accagtgcct caacctcttc acacggcctg aggtgctggc 3540 acccgaggag gcctggtact gcccacagtg caaacagcac cgtgaggcct ccaagcagct 3600 gttgctatgg cgcctgccaa atgttctcat cgtgcagctc aagcgcttct cctttcgtag 3660 ttttatctgg cgtgacaaga tcaatgactt ggtggagttc cctgttagga acctggacct 3720 gagcaagttc tgcattggtc agaaagagga gcagctgccc agctacgatc tatatgctgt 3780 catcaaccac tatggaggca tgattggtgg ccactacact gcctgtgcac gcctgcccaa 3840 tgatcgtagc agtcagcgca gtgacgtggg ctggcgcttg tttgatgaca gcacagtgac 3900 aacggtagac gagagccagg ttgtgacgcg ttatgcctat gtactcttct accgccggcg 3960 gaactctcct gtggagaggc cccccagggc aggtcactct gagcaccacc cagacctagg 4020 ccctgcagct gaggctgctg ccagccaggc ttcccggatt tggcaggagc tggaggctga 4080 ggaggagccg gtgcctgagg ggtctgggcc cctgggtccc tgggggcccc aagactgggt 4140 gggcccccta ccacgtggcc ctaccacacc agatgagggc tgcctccggt actttgtcct 4200 gggcaccgtg gcggctttgg tggccctcgt gctcaacgtg ttctatcctc tggtatccca 4260 gagtcgctgg agatgagctc gcctgcaggc agctgctgtg agctggccta cctgcctgcc 4320 ccaggccatg cctgcctttg ttgtggggaa cacctctggg ctttgggcct cagcttatgc 4380 atctggtggg agagggtggg gaggttgtgg cccctgcagg ggcagagtat cctagggtgt 4440 gtatccatct ggctgtctgt ccattcatcc tgctgctctg acccttggcc tcaggcttgg 4500 ccctgcccaa gctacttcct gtacttaaaa gtgttaataa aaccagacta ttcaggccca 4560 aaaaaaaaaa aaaaa 4575 <210> 9 <211> 18 <212> DNA <213> Artificial Sequence <220> <223> Primer <400> 9 tgtgggctac tgcaacca 18 <210> 10 <211> 18 <212> DNA <213> Artificial Sequence <220> <223> Primer <400> 10 gctgaatggg gtctctct 18 <110> College of Medicine Pochon CHA University Industry-Academic Cooperation Foundation <120> Analytical method for diagnosing obesity or diabetes and kit therefor <130> PN0798 <160> 10 <170> KopatentIn 2.0 <210> 1 <211> 1419 <212 > PRT <213> Homo sapiens <400> 1 Met Ser Gly Gly Ala Ser Ala Thr Gly Pro Arg Arg Gly Pro Pro Gly 1 5 10 15 Leu Glu Asp Thr Thr Ser Lys Lys Lys Gln Lys Asp Arg Ala Asn Gln 20 25 30 Glu Ser Lys Asp Gly Asp Pro Arg Lys Glu Thr Gly Ser Arg Tyr Val 35 40 45 Ala Gln Ala Gly Leu Glu Pro Leu Ala Ser Gly Asp Pro Ser Ala Ser 50 55 60 Ala Ser His Ala Ala Gly Ile Thr Gly Ser Arg His Arg Thr Arg Leu 65 70 75 80 Phe Phe Pro Ser Ser Ser Gly Ser Ala Ser Thr Pro Gln Glu Glu Gln 85 90 95 Thr Lys Glu Gly Ala Cys Glu Asp Pro His Asp Leu Leu Ala Thr Pro 100 105 110 Thr Pro Glu Leu Leu Leu Asp Trp Arg Gln Ser Ala Glu Glu Val Ile 115 120 125 Val Lys Leu Arg Val Gly Val Gly Pro Leu Gln Leu Glu Asp Val Asp 130 135 140 Ala Ala Phe Thr Asp Thr Asp Cys Val Val Arg Phe Ala Gly Gly Gln 145 150 155 160 Gln Trp Gly Gly Val Phe Tyr Ala Glu Ile Lys Ser Ser Cys Ala Lys 165 170 175 Val Gln Thr Arg Lys Gly Ser Leu Leu His Leu Thr Leu Pro Lys Lys 180 185 190 Val Pro Met Leu Thr Trp Pro Ser Leu Leu Lys Lys Pro Leu Gly Thr 195 200 205 Gln Glu Leu Val Pro Gly Leu Arg Cys Gln Glu Asn Gly Gln Glu Leu 210 215 220 Ser Pro Ile Ala Leu Glu Pro Gly Pro Glu Pro His Arg Ala Lys Gln 225 230 235 240 Glu Ala Arg Asn Gln Lys Arg Ala Gln Gly Arg Gly Glu Val Gly Ala 245 250 255 Gly Ala Gly Pro Gly Ala Gln Ala Gly Pro Ser Ala Lys Arg Ala Val 260 265 270 His Leu Cys Arg Gly Pro Glu Gly Asp Gly Ser Arg Asp Asp Pro Gly 275 280 285 Pro Arg Gly Asp Ala Pro Pro Phe Val Ala Asp Pro Ala Thr Gln Val 290 295 300 Glu Ala Asp Glu Gln Leu Cys Ile Pro Pro Leu Asn Ser Gln Thr Cys 305 310 315 320 Leu Leu Gly Ser Glu Glu Asn Leu Ala Pro Leu Ala Gly Glu Lys Ala 325 330 335 Val Pro Pro Gly Asn Asp Pro Val Ser Pro Ala Met Val Arg Ser Arg 340 345 350 Asn Pro Gly Lys Asp Asp Cys Ala Lys Glu Glu Met Ala Val Ala Ala 355 360 365 Asp Ala Ala Thr Leu Val Asp Glu Pro Glu Ser Met Val Asn Leu Ala 370 375 380 Phe Val Lys Asn Asp Ser Tyr Glu Lys Gly Pro Asp Ser Val Val Val 385 390 395 400 His Val Tyr Val Lys Glu Ile Cys Arg Asp Thr Ser Arg Val Leu Phe 405 410 415 Arg Glu Gln Asp Phe Thr Leu Ile Phe Gln Thr Arg Asp Gly Asn Phe 420 425 430 Leu Arg Leu His Pro Gly Cys Gly Pro His Thr Thr Phe Arg Trp Gln 435 440 445 Val Lys Leu Arg Asn Leu Ile Glu Pro Glu Gln Cys Thr Phe Cys Phe 450 455 460 Thr Ala Ser Arg Ile Asp Ile Cys Leu Arg Lys Arg Gln Ser Gln Arg 465 470 475 480 Trp Gly Gly Leu Glu Ala Pro Ala Ala Arg Val Gly Gly Ala Lys Val 485 490 495 Ala Val Pro Thr Gly Pro Thr Pro Leu Asp Ser Thr Pro Pro Gly Gly 500 505 510 Ala Pro His Pro Leu Thr Gly Gln Glu Glu Ala Arg Ala Val Glu Lys 515 520 525 Asp Lys Ser Lys Ala Arg Ser Glu Asp Thr Gly Leu Asp Ser Val Ala 530 535 540 Thr Arg Thr Pro Met Glu His Val Thr Pro Lys Pro Glu Thr His Leu 545 550 555 560 Ala Ser Pro Lys Pro Thr Cys Met Val Pro Pro Met Pro His Ser Pro 565 570 575 Val Ser Gly Asp Ser Val Glu Glu Glu Glu Glu Glu Glu Lys Lys Val 580 585 590 Cys Leu Pro Gly Phe Thr Gly Leu Val Asn Leu Gly Asn Thr Cys Phe 595 600 605 Met Asn Ser Val Ile Gln Ser Leu Ser Asn Thr Arg Glu Leu Arg Asp 610 615 620 Phe Phe His Asp Arg Ser Phe Glu Ala Glu Ile Asn Tyr Asn Asn Pro 625 630 635 640 Leu Gly Thr Gly Gly Arg Leu Ala Ile Gly Phe Ala Val Leu Leu Arg 645 650 655 Ala Leu Trp Lys Gly Thr His His Ala Phe Gln Pro Ser Lys Leu Lys 660 665 670 Ala Ile Val Ala Ser Lys Ala Ser Gln Phe Thr Gly Tyr Ala Gln His 675 680 685 Asp Ala Gln Glu Phe Met Ala Phe Leu Leu Asp Gly Leu His Glu Asp 690 695 700 Leu Asn Arg Ile Gln Asn Lys Pro Tyr Thr Glu Thr Val Asp Ser Asp 705 710 715 720 Gly Arg Pro Asp Glu Val Val Ala Glu Glu Ala Trp Gln Arg His Lys 725 730 735 Met Arg Asn Asp Ser Phe Ile Val Asp Leu Phe Gln Gly Gln Tyr Lys 740 745 750 Ser Lys Leu Val Cys Pro Val Cys Ala Lys Val Ser Ile Thr Phe Asp 755 760 765 Pro Phe Leu Tyr Leu Pro Val Pro Leu Pro Gln Lys Gln Lys Val Leu 770 775 780 Pro Val Phe Tyr Phe Ala Arg Glu Pro His Ser Lys Pro Ile Lys Phe 785 790 795 800 Leu Val Ser Val Ser Lys Glu Asn Ser Thr Ala Ser Glu Val Leu Asp 805 810 815 Ser Leu Ser Gln Ser Val His Val Lys Pro Glu Asn Leu Arg Leu Ala 820 825 830 Glu Val Ile Lys Asn Arg Phe His Arg Val Phe Leu Pro Ser His Ser 835 840 845 Leu Asp Thr Val Ser Pro Ser Asp Thr Leu Leu Cys Phe Glu Leu Leu 850 855 860 Ser Ser Glu Leu Ala Lys Glu Arg Val Val Val Leu Glu Val Gln Gln 865 870 875 880 Arg Pro Gln Val Pro Ser Val Pro Ile Ser Lys Cys Ala Ala Cys Gln 885 890 895 Arg Lys Gln Gln Ser Glu Asp Glu Lys Leu Lys Arg Cys Thr Arg Cys 900 905 910 Tyr Arg Val Gly Tyr Cys Asn Gln Leu Cys Gln Lys Thr His Trp Pro 915 920 925 Asp His Lys Gly Leu Cys Arg Pro Glu Asn Ile Gly Tyr Pro Phe Leu 930 935 940 Val Ser Val Pro Ala Ser Arg Leu Thr Tyr Ala Arg Leu Ala Gln Leu 945 950 955 960 Leu Glu Gly Tyr Ala Arg Tyr Ser Val Ser Val Phe Gln Pro Pro Phe 965 970 975 Gln Pro Gly Arg Met Ala Leu Glu Ser Gln Ser Pro Gly Cys Thr 980 985 990 Leu Leu Ser Thr Gly Ser Leu Glu Ala Gly Asp Ser Glu Arg Asp Pro 995 1000 1005 Ile Gln Pro Pro Glu Leu Gln Leu Val Thr Pro Met Ala Glu Gly Asp 1010 1015 1020 Thr Gly Leu Pro Arg Val Trp Ala Ala Pro Asp Arg Gly Pro Val Pro 1025 1030 1035 1040 Ser Thr Ser Gly Ile Ser Ser Glu Met Leu Ala Ser Gly Pro Ile Glu 1045 1050 1055 Val Gly Ser Leu Pro Ala Gly Glu Arg Val Ser Arg Pro Glu Ala Ala 1060 1065 1070 Val Pro Gly Tyr Gln His Pro Ser Glu Ala Met Asn Ala His Thr Pro 1075 1080 1085 Gln Phe Phe Ile Tyr Lys Ile Asp Ser Ser Asn Arg Glu Gln Arg Leu 1090 1095 1100 Glu Asp Lys Gly Asp Thr Pro Leu Glu Leu Gly Asp Asp Cys Ser Leu 1105 1110 1115 1120 Ala Leu Val Trp Arg Asn Asn Glu Arg Leu Gln Glu Phe Val Leu Val 1125 1130 1135 Ala Ser Lys Glu Leu Glu Cys Ala Glu Asp Pro Gly Ser Ala Gly Glu 1140 1145 1150 Ala Ala Arg Ala Gly His Phe Thr Leu Asp Gln Cys Leu Asn Leu Phe 1155 1160 1165 Thr Arg Pro Glu Val Leu Ala Pro Glu Glu Ala Trp Tyr Cys Pro Gln 1170 1175 1180 Cys Lys Gln His Arg Glu Ala Ser Lys Gln Leu Leu Leu Trp Arg Leu 1185 1190 1195 1200 Pro Asn Val Leu Ile Val Gln Leu Lys Arg Phe Ser Phe Arg Ser Phe 1205 1210 1215 Ile Trp Arg Asp Lys Ile Asn Asp Leu Val Glu Phe Pro Val Arg Asn 1220 1225 1230 Leu Asp Leu Ser Lys Phe Cys Ile Gly Gln Lys Glu Glu Gln Leu Pro 1235 1240 1245 Ser Tyr Asp Leu Tyr Ala Val Ile Asn His Tyr Gly Gly Met Ile Gly 1250 1255 1260 Gly His Tyr Thr Ala Cys Ala Arg Leu Pro Asn Asp Arg Ser Ser Gln 1265 1270 1275 1280 Arg Ser Asp Val Gly Trp Arg Leu Phe Asp Asp Ser Thr Val Thr Thr 1285 1290 1295 Val Asp Glu Ser Gln Val Val Thr Arg Tyr Ala Tyr Val Leu Phe Tyr 1300 1305 1310 Arg Arg Arg Asn Ser Pro Val Glu Arg Pro Pro Arg Ala Gly His Ser 1315 1320 1325 Glu His His Pro Asp Leu Gly Pro Ala Ala Glu Ala Ala Ala Ser Gln 1330 1335 1340 Ala Ser Arg Ile Trp Gln Glu Leu Glu Ala Glu Glu Glu Pro Val Pro 1345 1350 1355 1360 Glu Gly Ser Gly Pro Leu Gly Pro Trp Gly Pro Gln Asp Trp Val Gly 1365 1370 1375 Pro Leu Pro Arg Gly Pro Thr Thr Pro Asp Glu Gly Cys Leu Arg Tyr 1380 1385 1390 Phe Val Leu Gly Thr Val Ala Ala Leu Val Ala Leu Val Leu Asn Val 1395 1400 1405 Phe Tyr Pro Leu Val Ser Gln Ser Arg Trp Arg 1410 1415 <210> 2 <211> 1384 <212> PRT <213> Homo sapiens <400> 2 Met Ser Gly Gly Ala Ser Ala Thr Gly Pro Arg Arg Gly Pro Pro Gly 1 5 10 15 Leu Glu Asp Thr Thr Ser Lys Lys Lys Gln Lys Asp Arg Ala Asn Gln 20 25 30 Glu Ser Lys Asp Gly Asp Pro Arg Lys Glu Thr Gly Ser Arg Tyr Val 35 40 45 Ala Gln Ala Gly Leu Glu Pro Leu Ala Ser Gly Asp Pro Ser Ala Ser 50 55 60 Ala Ser His Ala Ala Gly Ile Thr Gly Ser Arg His Arg Thr Arg Leu 65 70 75 80 Phe Phe Pro Ser Ser Ser Gly Ser Ala Ser Thr Pro Gln Glu Glu Gln 85 90 95 Thr Lys Glu Gly Ala Cys Glu Asp Pro His Asp Leu Leu Ala Thr Pro 100 105 110 Thr Pro Glu Leu Leu Leu Asp Trp Arg Gln Ser Ala Glu Glu Val Ile 115 120 125 Val Lys Leu Arg Val Gly Val Gly Pro Leu Gln Leu Glu Asp Val Asp 130 135 140 Ala Ala Phe Thr Asp Thr Asp Cys Val Val Arg Phe Ala Gly Gly Gln 145 150 155 160 Gln Trp Gly Gly Val Phe Tyr Ala Glu Ile Lys Ser Ser Cys Ala Lys 165 170 175 Val Gln Thr Arg Lys Gly Ser Leu Leu His Leu Thr Leu Pro Lys Lys 180 185 190 Val Pro Met Leu Thr Trp Pro Ser Leu Leu Lys Lys Pro Leu Gly Thr 195 200 205 Gln Glu Leu Val Pro Gly Leu Arg Cys Gln Glu Asn Gly Gln Glu Leu 210 215 220 Ser Pro Ile Ala Leu Glu Pro Gly Pro Glu Pro His Arg Ala Lys Gln 225 230 235 240 Glu Ala Arg Asn Gln Lys Arg Ala Gln Gly Arg Gly Glu Val Gly Ala 245 250 255 Gly Ala Gly Pro Gly Ala Gln Ala Gly Pro Ser Ala Lys Arg Ala Val 260 265 270 His Leu Cys Arg Gly Pro Glu Gly Asp Gly Ser Arg Asp Asp Pro Gly 275 280 285 Pro Arg Gly Asp Ala Pro Pro Phe Val Ala Asp Pro Ala Thr Gln Val 290 295 300 Glu Ala Asp Glu Gln Leu Cys Ile Pro Pro Leu Asn Ser Gln Thr Cys 305 310 315 320 Leu Leu Gly Ser Glu Glu Asn Leu Ala Pro Leu Ala Gly Glu Lys Ala 325 330 335 Val Pro Pro Gly Asn Asp Pro Val Ser Pro Ala Met Val Arg Ser Arg 340 345 350 Asn Pro Gly Lys Asp Asp Cys Ala Lys Glu Glu Met Ala Val Ala Ala 355 360 365 Asp Ala Ala Thr Leu Val Asp Glu Pro Glu Ser Met Val Asn Leu Ala 370 375 380 Phe Val Lys Asn Asp Ser Tyr Glu Lys Gly Pro Asp Ser Val Val Val 385 390 395 400 His Val Tyr Val Lys Glu Ile Cys Arg Asp Thr Ser Arg Val Leu Phe 405 410 415 Arg Glu Gln Asp Phe Thr Leu Ile Phe Gln Thr Arg Asp Gly Asn Phe 420 425 430 Leu Arg Leu His Pro Gly Cys Gly Pro His Thr Thr Phe Arg Trp Gln 435 440 445 Val Lys Leu Arg Asn Leu Ile Glu Pro Glu Gln Cys Thr Phe Cys Phe 450 455 460 Thr Ala Ser Arg Ile Asp Ile Cys Leu Arg Lys Arg Gln Ser Gln Arg 465 470 475 480 Trp Gly Gly Leu Glu Ala Pro Ala Ala Arg Gly Ala Val Gly Gly Ala 485 490 495 Lys Val Ala Val Pro Thr Gly Pro Thr Pro Leu Asp Ser Thr Pro Pro 500 505 510 Gly Gly Ala Pro His Pro Leu Thr Gly Gln Glu Glu Ala Arg Ala Val 515 520 525 Glu Lys Asp Lys Ser Lys Ala Arg Ser Glu Asp Thr Gly Leu Asp Ser 530 535 540 Val Ala Thr Arg Thr Pro Met Glu His Val Thr Pro Lys Pro Glu Thr 545 550 555 560 His Leu Ala Ser Pro Lys Pro Thr Cys Met Val Pro Pro Met Pro His 565 570 575 Ser Pro Val Ser Gly Asp Ser Val Glu Glu Glu Glu Glu Glu Glu Lys 580 585 590 Lys Val Cys Leu Pro Gly Phe Thr Gly Leu Val Asn Leu Gly Asn Thr 595 600 605 Cys Phe Met Asn Ser Val Ile Gln Ser Leu Ser Asn Thr Arg Glu Leu 610 615 620 Arg Asp Phe Phe His Asp Arg Ser Phe Glu Ala Glu Ile Asn Tyr Asn 625 630 635 640 Asn Pro Leu Gly Thr Gly Gly Arg Leu Ala Ile Gly Phe Ala Val Leu 645 650 655 Leu Arg Ala Leu Trp Lys Gly Thr His His Ala Phe Gln Pro Ser Lys 660 665 670 Leu Lys Ala Ile Val Ala Ser Lys Ala Ser Gln Phe Thr Gly Tyr Ala 675 680 685 Gln His Asp Ala Gln Glu Phe Met Ala Phe Leu Leu Asp Gly Leu His 690 695 700 Glu Asp Leu Asn Arg Ile Gln Asn Lys Pro Tyr Thr Glu Thr Val Asp 705 710 715 720 Ser Asp Gly Arg Pro Asp Glu Val Val Ala Glu Glu Ala Trp Gln Arg 725 730 735 His Lys Met Arg Asn Asp Ser Phe Ile Val Asp Leu Phe Gln Gly Gln 740 745 750 Tyr Lys Ser Lys Leu Val Cys Pro Val Cys Ala Lys Val Ser Ile Thr 755 760 765 Phe Asp Pro Phe Leu Tyr Leu Pro Val Pro Leu Pro Gln Lys Gln Lys 770 775 780 Val Leu Pro Val Phe Tyr Phe Ala Arg Glu Pro His Ser Lys Pro Ile 785 790 795 800 Lys Phe Leu Val Ser Val Ser Lys Glu Asn Ser Thr Ala Ser Glu Val 805 810 815 Leu Asp Ser Leu Ser Gln Ser Val His Val Lys Pro Glu Asn Leu Arg 820 825 830 Leu Ala Glu Val Ile Lys Asn Arg Phe His Arg Val Phe Leu Pro Ser 835 840 845 His Ser Leu Asp Thr Val Ser Pro Ser Asp Thr Leu Leu Cys Phe Glu 850 855 860 Leu Leu Ser Ser Glu Leu Ala Lys Glu Arg Val Val Val Leu Glu Val 865 870 875 880 Gln Gln Arg Pro Gln Val Pro Ser Val Pro Ile Ser Lys Cys Ala Ala 885 890 895 Cys Gln Arg Lys Gln Gln Ser Glu Asp Glu Lys Leu Lys Arg Cys Thr 900 905 910 Arg Cys Tyr Arg Val Gly Tyr Cys Asn Gln Leu Cys Gln Lys Thr His 915 920 925 Trp Pro Asp His Lys Gly Leu Cys Arg Pro Glu Asn Ile Gly Tyr Pro 930 935 940 Phe Leu Val Ser Val Pro Ala Ser Arg Leu Thr Tyr Ala Arg Leu Ala 945 950 955 960 Gln Leu Leu Glu Gly Tyr Ala Arg Tyr Ser Val Ser Val Phe Gln Pro 965 970 975 Pro Phe Gln Pro Gly Arg Met Ala Leu Glu Ser Gln Ser Pro Gly Cys 980 985 990 Thr Thr Leu Leu Ser Thr Gly Ser Leu Glu Ala Gly Asp Ser Glu Arg 995 1000 1005 Asp Pro Ile Gln Pro Pro Glu Leu Gln Leu Val Thr Pro Met Ala Glu 1010 1015 1020 Gly Asp Thr Gly Leu Pro Arg Val Trp Ala Ala Pro Asp Arg Gly Pro 1025 1030 1035 1040 Val Pro Ser Thr Ser Gly Ile Ser Ser Glu Met Leu Ala Ser Gly Pro 1045 1050 1055 Ile Glu Val Gly Ser Leu Pro Ala Gly Glu Arg Val Ser Arg Pro Glu 1060 1065 1070 Ala Ala Val Pro Gly Tyr Gln His Pro Ser Glu Ala Met Asn Ala His 1075 1080 1085 Thr Pro Gln Phe Phe Ile Tyr Lys Ile Asp Ser Ser Asn Arg Glu Gln 1090 1095 1100 Arg Leu Glu Asp Lys Gly Asp Thr Pro Leu Glu Leu Gly Asp Asp Cys 1105 1110 1115 1120 Ser Leu Ala Leu Val Trp Arg Asn Asn Glu Arg Leu Gln Glu Phe Val 1125 1130 1135 Leu Val Ala Ser Lys Glu Leu Glu Cys Ala Glu Asp Pro Gly Ser Ala 1140 1145 1150 Gly Glu Ala Ala Arg Ala Gly His Phe Thr Leu Asp Gln Cys Leu Asn 1155 1160 1165 Leu Phe Thr Arg Pro Glu Val Leu Ala Pro Glu Glu Ala Trp Tyr Cys 1170 1175 1180 Pro Gln Cys Lys Gln His Arg Glu Ala Ser Lys Gln Leu Leu Leu Trp 1185 1190 1195 1200 Arg Leu Pro Asn Val Leu Ile Val Gln Leu Lys Arg Phe Ser Phe Arg 1205 1210 1215 Ser Phe Ile Trp Arg Asp Lys Ile Asn Asp Leu Val Glu Phe Pro Val 1220 1225 1230 Arg Asn Leu Asp Leu Ser Lys Phe Cys Ile Gly Gln Lys Glu Glu Gln 1235 1240 1245 Leu Pro Ser Tyr Asp Leu Tyr Ala Val Ile Asn His Tyr Gly Gly Met 1250 1255 1260 Ile Gly Gly His Tyr Thr Ala Cys Ala Arg Leu Pro Asn Asp Arg Ser 1265 1270 1275 1280 Ser Gln Arg Ser Asp Val Gly Trp Arg Leu Phe Asp Asp Ser Thr Val 1285 1290 1295 Thr Thr Val Asp Glu Ser Gln Val Val Thr Arg Tyr Ala Tyr Val Leu 1300 1305 1310 Phe Tyr Arg Arg Arg Asn Ser Pro Val Glu Arg Pro Pro Arg Ala Gly 1315 1320 1325 His Ser Glu His His Pro Asp Leu Gly Pro Ala Ala Glu Ala Ala Ala 1330 1335 1340 Ser Gln Gly Leu Gly Pro Gly Gln Ala Pro Glu Val Ala Pro Thr Arg 1345 1350 1355 1360 Thr Ala Pro Glu Arg Phe Ala Pro Pro Val Asp Arg Pro Ala Pro Thr 1365 1370 1375 Tyr Ser Asn Met Glu Glu Val Asp 1380 <210> 3 <211> 1372 <212> PRT <213> Homo sapiens <400> 3 Met Ser Gly Gly Ala Ser Ala Thr Gly Pro Arg Arg Gly Pro Pro Gly 1 5 10 15 Leu Glu Asp Thr Thr Ser Lys Lys Lys Gln Lys Asp Arg Ala Asn Gln 20 25 30 Glu Ser Lys Asp Gly Asp Pro Arg Lys Glu Thr Gly Ser Arg Tyr Val 35 40 45 Ala Gln Ala Gly Leu Glu Pro Leu Ala Ser Gly Asp Pro Ser Ala Ser 50 55 60 Ala Ser His Ala Ala Gly Ile Thr Gly Ser Arg His Arg Thr Arg Leu 65 70 75 80 Phe Phe Pro Ser Ser Ser Gly Ser Ala Ser Thr Pro Gln Glu Glu Gln 85 90 95 Thr Lys Glu Gly Ala Cys Glu Asp Pro His Asp Leu Leu Ala Thr Pro 100 105 110 Thr Pro Glu Leu Leu Leu Asp Trp Arg Gln Ser Ala Glu Glu Val Ile 115 120 125 Val Lys Leu Arg Val Gly Val Gly Pro Leu Gln Leu Glu Asp Val Asp 130 135 140 Ala Ala Phe Thr Asp Thr Asp Cys Val Val Arg Phe Ala Gly Gly Gln 145 150 155 160 Gln Trp Gly Gly Val Phe Tyr Ala Glu Ile Lys Ser Ser Cys Ala Lys 165 170 175 Val Gln Thr Arg Lys Gly Ser Leu Leu His Leu Thr Leu Pro Lys Lys 180 185 190 Lys Lys Pro Leu Gly Thr Gln Glu Leu Val Pro Gly Leu Arg Cys Gln 195 200 205 Glu Asn Gly Gln Glu Leu Ser Pro Ile Ala Leu Glu Pro Gly Pro Glu 210 215 220 Pro His Arg Ala Lys Gln Glu Ala Arg Asn Gln Lys Arg Ala Gln Gly 225 230 235 240 Arg Gly Glu Val Gly Ala Gly Ala Gly Pro Gly Ala Gln Ala Gly Pro 245 250 255 Ser Ala Lys Arg Ala Val His Leu Cys Arg Gly Pro Glu Gly Asp Gly 260 265 270 Ser Arg Asp Asp Pro Gly Pro Arg Gly Asp Ala Pro Pro Phe Val Ala 275 280 285 Asp Pro Ala Thr Gln Val Glu Ala Asp Glu Gln Leu Cys Ile Pro Pro 290 295 300 Leu Asn Ser Gln Thr Cys Leu Leu Gly Ser Glu Glu Asn Leu Ala Pro 305 310 315 320 Leu Ala Gly Glu Lys Ala Val Pro Pro Gly Asn Asp Pro Val Ser Pro 325 330 335 Ala Met Val Arg Ser Arg Asn Pro Gly Lys Asp Asp Cys Ala Lys Glu 340 345 350 Glu Met Ala Val Ala Ala Asp Ala Ala Thr Leu Val Asp Glu Pro Glu 355 360 365 Ser Met Val Asn Leu Ala Phe Val Lys Asn Asp Ser Tyr Glu Lys Gly 370 375 380 Pro Asp Ser Val Val Val His Val Tyr Val Lys Glu Ile Cys Arg Asp 385 390 395 400 Thr Ser Arg Val Leu Phe Arg Glu Gln Asp Phe Thr Leu Ile Phe Gln 405 410 415 Thr Arg Asp Gly Asn Phe Leu Arg Leu His Pro Gly Cys Gly Pro His 420 425 430 Thr Thr Phe Arg Trp Gln Val Lys Leu Arg Asn Leu Ile Glu Pro Glu 435 440 445 Gln Cys Thr Phe Cys Phe Thr Ala Ser Arg Ile Asp Ile Cys Leu Arg 450 455 460 Lys Arg Gln Ser Gln Arg Trp Gly Gly Leu Glu Ala Pro Ala Ala Arg 465 470 475 480 Val Gly Gly Ala Lys Val Ala Val Pro Thr Gly Pro Thr Pro Leu Asp 485 490 495 Ser Thr Pro Pro Gly Gly Ala Pro His Pro Leu Thr Gly Gln Glu Glu 500 505 510 Ala Arg Ala Val Glu Lys Asp Lys Ser Lys Ala Arg Ser Glu Asp Thr 515 520 525 Gly Leu Asp Ser Val Ala Thr Arg Thr Pro Met Glu His Val Thr Pro 530 535 540 Lys Pro Glu Thr His Leu Ala Ser Pro Lys Pro Thr Cys Met Val Pro 545 550 555 560 Pro Met Pro His Ser Pro Val Ser Gly Asp Ser Val Glu Glu Glu Glu 565 570 575 Glu Glu Glu Lys Lys Val Cys Leu Pro Gly Phe Thr Gly Leu Val Asn 580 585 590 Leu Gly Asn Thr Cys Phe Met Asn Ser Val Ile Gln Ser Leu Ser Asn 595 600 605 Thr Arg Glu Leu Arg Asp Phe Phe His Asp Arg Ser Phe Glu Ala Glu 610 615 620 Ile Asn Tyr Asn Asn Pro Leu Gly Thr Gly Gly Arg Leu Ala Ile Gly 625 630 635 640 Phe Ala Val Leu Leu Arg Ala Leu Trp Lys Gly Thr His His Ala Phe 645 650 655 Gln Pro Ser Lys Leu Lys Ala Ile Val Ala Ser Lys Ala Ser Gln Phe 660 665 670 Thr Gly Tyr Ala Gln His Asp Ala Gln Glu Phe Met Ala Phe Leu Leu 675 680 685 Asp Gly Leu His Glu Asp Leu Asn Arg Ile Gln Asn Lys Pro Tyr Thr 690 695 700 Glu Thr Val Asp Ser Asp Gly Arg Pro Asp Glu Val Val Ala Glu Glu 705 710 715 720 Ala Trp Gln Arg His Lys Met Arg Asn Asp Ser Phe Ile Val Asp Leu 725 730 735 Phe Gln Gly Gln Tyr Lys Ser Lys Leu Val Cys Pro Val Cys Ala Lys 740 745 750 Val Ser Ile Thr Phe Asp Pro Phe Leu Tyr Leu Pro Val Pro Leu Pro 755 760 765 Gln Lys Gln Lys Val Leu Pro Val Phe Tyr Phe Ala Arg Glu Pro His 770 775 780 Ser Lys Pro Ile Lys Phe Leu Val Ser Val Ser Lys Glu Asn Ser Thr 785 790 795 800 Ala Ser Glu Val Leu Asp Ser Leu Ser Gln Ser Val His Val Lys Pro 805 810 815 Glu Asn Leu Arg Leu Ala Glu Val Ile Lys Asn Arg Phe His Arg Val 820 825 830 Phe Leu Pro Ser His Ser Leu Asp Thr Val Ser Pro Ser Asp Thr Leu 835 840 845 Leu Cys Phe Glu Leu Leu Ser Ser Glu Leu Ala Lys Glu Arg Val Val 850 855 860 Val Leu Glu Val Gln Gln Arg Pro Gln Val Pro Ser Val Pro Ile Ser 865 870 875 880 Lys Cys Ala Ala Cys Gln Arg Lys Gln Gln Ser Glu Asp Glu Lys Leu 885 890 895 Lys Arg Cys Thr Arg Cys Tyr Arg Val Gly Tyr Cys Asn Gln Leu Cys 900 905 910 Gln Lys Thr His Trp Pro Asp His Lys Gly Leu Cys Arg Pro Glu Asn 915 920 925 Ile Gly Tyr Pro Phe Leu Val Ser Val Pro Ala Ser Arg Leu Thr Tyr 930 935 940 Ala Arg Leu Ala Gln Leu Leu Glu Gly Tyr Ala Arg Tyr Ser Val Ser 945 950 955 960 Val Phe Gln Pro Pro Phe Gln Pro Gly Arg Met Ala Leu Glu Ser Gln 965 970 975 Ser Pro Gly Cys Thr Thr Leu Leu Ser Thr Gly Ser Leu Glu Ala Gly 980 985 990 Asp Ser Glu Arg Asp Pro Ile Gln Pro Pro Glu Leu Gln Leu Val Thr 995 1000 1005 Pro Met Ala Glu Gly Asp Thr Gly Leu Pro Arg Val Trp Ala Ala Pro 1010 1015 1020 Asp Arg Gly Pro Val Pro Ser Thr Ser Gly Ile Ser Ser Glu Met Leu 1025 1030 1035 1040 Ala Ser Gly Pro Ile Glu Val Gly Ser Leu Pro Ala Gly Glu Arg Val 1045 1050 1055 Ser Arg Pro Glu Ala Ala Val Pro Gly Tyr Gln His Pro Ser Glu Ala 1060 1065 1070 Met Asn Ala His Thr Pro Gln Phe Phe Ile Tyr Lys Ile Asp Ser Ser 1075 1080 1085 Asn Arg Glu Gln Arg Leu Glu Asp Lys Gly Asp Thr Pro Leu Glu Leu 1090 1095 1100 Gly Asp Asp Cys Ser Leu Ala Leu Val Trp Arg Asn Asn Glu Arg Leu 1105 1110 1115 1120 Gln Glu Phe Val Leu Val Ala Ser Lys Glu Leu Glu Cys Ala Glu Asp 1125 1130 1135 Pro Gly Ser Ala Gly Glu Ala Ala Arg Ala Gly His Phe Thr Leu Asp 1140 1145 1150 Gln Cys Leu Asn Leu Phe Thr Arg Pro Glu Val Leu Ala Pro Glu Glu 1155 1160 1165 Ala Trp Tyr Cys Pro Gln Cys Lys Gln His Arg Glu Ala Ser Lys Gln 1170 1175 1180 Leu Leu Leu Trp Arg Leu Pro Asn Val Leu Ile Val Gln Leu Lys Arg 1185 1190 1195 1200 Phe Ser Phe Arg Ser Phe Ile Trp Arg Asp Lys Ile Asn Asp Leu Val 1205 1210 1215 Glu Phe Pro Val Arg Asn Leu Asp Leu Ser Lys Phe Cys Ile Gly Gln 1220 1225 1230 Lys Glu Glu Gln Leu Pro Ser Tyr Asp Leu Tyr Ala Val Ile Asn His 1235 1240 1245 Tyr Gly Gly Met Ile Gly Gly His Tyr Thr Ala Cys Ala Arg Leu Pro 1250 1255 1260 Asn Asp Arg Ser Ser Gln Arg Ser Asp Val Gly Trp Arg Leu Phe Asp 1265 1270 1275 1280 Asp Ser Thr Val Thr Thr Val Asp Glu Ser Gln Val Val Thr Arg Tyr 1285 1290 1295 Ala Tyr Val Leu Phe Tyr Arg Arg Arg Asn Ser Pro Val Glu Arg Pro 1300 1305 1310 Pro Arg Ala Gly His Ser Glu His His Pro Asp Leu Gly Pro Ala Ala 1315 1320 1325 Glu Ala Ala Ala Ser Gln Gly Leu Gly Pro Gly Gln Ala Pro Glu Val 1330 1335 1340 Ala Pro Thr Arg Thr Ala Pro Glu Arg Phe Ala Pro Pro Pro Val Asp Arg 1345 1350 1355 1360 Pro Ala Pro Thr Tyr Ser Asn Met Glu Glu Val Asp 1365 1370 <210> 4 <211> 1318 <212> PRT <213> Homo sapiens <400> 4 Met Ser Gly Gly Ala Ser Ala Thr Gly Pro Arg Arg Gly Pro Pro Gly 1 5 10 15 Leu Glu Asp Thr Thr Ser Lys Lys Lys Gln Lys Asp Arg Ala Asn Gln 20 25 30 Glu Ser Lys Asp Gly Asp Pro Arg Lys Glu Thr Gly Ser Arg Tyr Val 35 40 45 Ala Gln Ala Gly Leu Glu Pro Leu Ala Ser Gly Asp Pro Ser Ala Ser 50 55 60 Ala Ser His Ala Ala Gly Ile Thr Gly Ser Arg His Arg Thr Arg Leu 65 70 75 80 Phe Phe Pro Ser Ser Ser Gly Ser Ala Ser Thr Pro Gln Glu Glu Gln 85 90 95 Thr Lys Glu Gly Ala Cys Glu Asp Pro His Asp Leu Leu Ala Thr Pro 100 105 110 Thr Pro Glu Leu Leu Leu Asp Trp Arg Gln Ser Ala Glu Glu Val Ile 115 120 125 Val Lys Leu Arg Val Gly Val Gly Pro Leu Gln Leu Glu Asp Val Asp 130 135 140 Ala Ala Phe Thr Asp Thr Asp Cys Val Val Arg Phe Ala Gly Gly Gln 145 150 155 160 Gln Trp Gly Gly Val Phe Tyr Ala Glu Ile Lys Ser Ser Cys Ala Lys 165 170 175 Val Gln Thr Arg Lys Gly Ser Leu Leu His Leu Thr Leu Pro Lys Lys 180 185 190 Val Pro Met Leu Thr Trp Pro Ser Leu Leu Val Glu Ala Asp Glu Gln 195 200 205 Leu Cys Ile Pro Pro Leu Asn Ser Gln Thr Cys Leu Leu Gly Ser Glu 210 215 220 Glu Asn Leu Ala Pro Leu Ala Gly Glu Lys Ala Val Pro Pro Gly Asn 225 230 235 240 Asp Pro Val Ser Pro Ala Met Val Arg Ser Arg Asn Pro Gly Lys Asp 245 250 255 Asp Cys Ala Lys Glu Glu Met Ala Val Ala Ala Asp Ala Ala Thr Leu 260 265 270 Val Asp Glu Pro Glu Ser Met Val Asn Leu Ala Phe Val Lys Asn Asp 275 280 285 Ser Tyr Glu Lys Gly Pro Asp Ser Val Val Val His Val Tyr Val Lys 290 295 300 Glu Ile Cys Arg Asp Thr Ser Arg Val Leu Phe Arg Glu Gln Asp Phe 305 310 315 320 Thr Leu Ile Phe Gln Thr Arg Asp Gly Asn Phe Leu Arg Leu His Pro 325 330 335 Gly Cys Gly Pro His Thr Thr Phe Arg Trp Gln Val Lys Leu Arg Asn 340 345 350 Leu Ile Glu Pro Glu Gln Cys Thr Phe Cys Phe Thr Ala Ser Arg Ile 355 360 365 Asp Ile Cys Leu Arg Lys Arg Gln Ser Gln Arg Trp Gly Gly Leu Glu 370 375 380 Ala Pro Ala Ala Arg Val Gly Gly Ala Lys Val Ala Val Pro Thr Gly 385 390 395 400 Pro Thr Pro Leu Asp Ser Thr Pro Pro Gly Gly Ala Pro His Pro Leu 405 410 415 Thr Gly Gln Glu Glu Ala Arg Ala Val Glu Lys Asp Lys Ser Lys Ala 420 425 430 Arg Ser Glu Asp Thr Gly Leu Asp Ser Val Ala Thr Arg Thr Pro Met 435 440 445 Glu His Val Thr Pro Lys Pro Glu Thr His Leu Ala Ser Pro Lys Pro 450 455 460 Thr Cys Met Val Pro Pro Met Pro His Ser Pro Val Ser Gly Asp Ser 465 470 475 480 Val Glu Glu Glu Glu Glu Glu Glu Lys Lys Val Cys Leu Pro Gly Phe 485 490 495 Thr Gly Leu Val Asn Leu Gly Asn Thr Cys Phe Met Asn Ser Val Ile 500 505 510 Gln Ser Leu Ser Asn Thr Arg Glu Leu Arg Asp Phe Phe His Asp Arg 515 520 525 Ser Phe Glu Ala Glu Ile Asn Tyr Asn Asn Pro Leu Gly Thr Gly Gly 530 535 540 Arg Leu Ala Ile Gly Phe Ala Val Leu Leu Arg Ala Leu Trp Lys Gly 545 550 555 560 Thr His His Ala Phe Gln Pro Ser Lys Leu Lys Ala Ile Val Ala Ser 565 570 575 Lys Ala Ser Gln Phe Thr Gly Tyr Ala Gln His Asp Ala Gln Glu Phe 580 585 590 Met Ala Phe Leu Leu Asp Gly Leu His Glu Asp Leu Asn Arg Ile Gln 595 600 605 Asn Lys Pro Tyr Thr Glu Thr Val Asp Ser Asp Gly Arg Pro Asp Glu 610 615 620 Val Val Ala Glu Glu Ala Trp Gln Arg His Lys Met Arg Asn Asp Ser 625 630 635 640 Phe Ile Val Asp Leu Phe Gln Gly Gln Tyr Lys Ser Lys Leu Val Cys 645 650 655 Pro Val Cys Ala Lys Val Ser Ile Thr Phe Asp Pro Phe Leu Tyr Leu 660 665 670 Pro Val Pro Leu Pro Gln Lys Gln Lys Val Leu Pro Val Phe Tyr Phe 675 680 685 Ala Arg Glu Pro His Ser Lys Pro Ile Lys Phe Leu Val Ser Val Ser 690 695 700 Lys Glu Asn Ser Thr Ala Ser Glu Val Leu Asp Ser Leu Ser Gln Ser 705 710 715 720 Val His Val Lys Pro Glu Asn Leu Arg Leu Ala Glu Val Ile Lys Asn 725 730 735 Arg Phe His Arg Val Phe Leu Pro Ser His Ser Leu Asp Thr Val Ser 740 745 750 Pro Ser Asp Thr Leu Leu Cys Phe Glu Leu Leu Ser Ser Glu Leu Ala 755 760 765 Lys Glu Arg Val Val Val Leu Glu Val Gln Gln Arg Pro Gln Val Pro 770 775 780 Ser Val Pro Ile Ser Lys Cys Ala Ala Cys Gln Arg Lys Gln Gln Ser 785 790 795 800 Glu Asp Glu Lys Leu Lys Arg Cys Thr Arg Cys Tyr Arg Val Gly Tyr 805 810 815 Cys Asn Gln Leu Cys Gln Lys Thr His Trp Pro Asp His Lys Gly Leu 820 825 830 Cys Arg Pro Glu Asn Ile Gly Tyr Pro Phe Leu Val Ser Val Pro Ala 835 840 845 Ser Arg Leu Thr Tyr Ala Arg Leu Ala Gln Leu Leu Glu Gly Tyr Ala 850 855 860 Arg Tyr Ser Val Ser Val Phe Gln Pro Pro Phe Gln Pro Gly Arg Met 865 870 875 880 Ala Leu Glu Ser Gln Ser Pro Gly Cys Thr Thr Leu Leu Ser Thr Gly 885 890 895 Ser Leu Glu Ala Gly Asp Ser Glu Arg Asp Pro Ile Gln Pro Pro Glu 900 905 910 Leu Gln Leu Val Thr Pro Met Ala Glu Gly Asp Thr Gly Leu Pro Arg 915 920 925 Val Trp Ala Ala Pro Asp Arg Gly Pro Val Pro Ser Thr Ser Gly Ile 930 935 940 Ser Ser Glu Met Leu Ala Ser Gly Pro Ile Glu Val Gly Ser Leu Pro 945 950 955 960 Ala Gly Glu Arg Val Ser Arg Pro Glu Ala Ala Val Pro Gly Tyr Gln 965 970 975 His Pro Ser Glu Ala Met Asn Ala His Thr Pro Gln Phe Phe Ile Tyr 980 985 990 Lys Ile Asp Ser Ser Asn Arg Glu Gln Arg Leu Glu Asp Lys Gly Asp 995 1000 1005 Thr Pro Leu Glu Leu Gly Asp Asp Cys Ser Leu Ala Leu Val Trp Arg 1010 1015 1020 Asn Asn Glu Arg Leu Gln Glu Phe Val Leu Val Ala Ser Lys Glu Leu 1025 1030 1035 1040 Glu Cys Ala Glu Asp Pro Gly Ser Ala Gly Glu Ala Ala Arg Ala Gly 1045 1050 1055 His Phe Thr Leu Asp Gln Cys Leu Asn Leu Phe Thr Arg Pro Glu Val 1060 1065 1070 Leu Ala Pro Glu Glu Ala Trp Tyr Cys Pro Gln Cys Lys Gln His Arg 1075 1080 1085 Glu Ala Ser Lys Gln Leu Leu Leu Trp Arg Leu Pro Asn Val Leu Ile 1090 1095 1100 Val Gln Leu Lys Arg Phe Ser Phe Arg Ser Phe Ile Trp Arg Asp Lys 1105 1110 1115 1120 Ile Asn Asp Leu Val Glu Phe Pro Val Arg Asn Leu Asp Leu Ser Lys 1125 1130 1135 Phe Cys Ile Gly Gln Lys Glu Glu Gln Leu Pro Ser Tyr Asp Leu Tyr 1140 1145 1150 Ala Val Ile Asn His Tyr Gly Gly Met Ile Gly Gly His Tyr Thr Ala 1155 1160 1165 Cys Ala Arg Leu Pro Asn Asp Arg Ser Ser Gln Arg Ser Asp Val Gly 1170 1175 1180 Trp Arg Leu Phe Asp Asp Ser Thr Val Thr Thr Val Asp Glu Ser Gln 1185 1190 1195 1200 Val Val Thr Arg Tyr Ala Tyr Val Leu Phe Tyr Arg Arg Arg Asn Ser 1205 1210 1215 Pro Val Glu Arg Pro Pro Arg Ala Gly His Ser Glu His His Pro Asp 1220 1225 1230 Leu Gly Pro Ala Ala Glu Ala Ala Ala Ser Gln Ala Ser Arg Ile Trp 1235 1240 1245 Gln Glu Leu Glu Ala Glu Glu Glu Pro Val Pro Glu Gly Ser Gly Pro 1250 1255 1260 Leu Gly Pro Trp Gly Pro Gln Asp Trp Val Gly Pro Leu Pro Arg Gly 1265 1270 1275 1280 Pro Thr Thr Pro Asp Glu Gly Cys Leu Arg Tyr Phe Val Leu Gly Thr 1285 1290 1295 Val Ala Ala Leu Val Ala Leu Val Leu Asn Val Phe Tyr Pro Leu Val 1300 1305 1310 Ser Gln Ser Arg Trp Arg 1315 <210> 5 <211> 4878 <212> DNA <213> Homo sapiens <400> 5 agcttggcgc tccgcctccc tcggccttag ctagaagtcg aaaaaaaca agccgctaag 60 gcggtggcgg cggcgccggg acgggggagg ggcgcgccgg aaccggaacc gacctgcgcc 120 ggaaccggaa cggagagcgg gttgccaggg cccgaagagg gctggctgcg gcggtctcgc 180 tcggctgtcc gttccttgct ggagaattt g gccacaaaga gttgccaaga tagctgggcc 240 aggaagaaag cgccgcagcc ctgacccaga cgctgttgcc gaccccgggg cactctggct 300 gtcgaccaag cggctcaaga tgtctggcgg ggccagtgcc acaggcccaa ggagagggcc 360 cccaggactg gaggacacca ctagtaagaa gaagcaga ag gatcgagcaa accaggagag 420 caaggatgga gatcctagga aagagacagg gtctcgatat gttgcccagg ctggtcttga 480 acctctggcc tcaggtgatc cttctgcctc agcctcccat gcagctggga tcacaggctc 540 acgccaccgt acccggctgt tctttccttc atcgtcaggg tcagcatcca ctcctcaaga 600 ggagcagacc aaagagggag cttgtgaaga ccctcatga t ctcttggcta ctcccactcc 660 agagttgttg ctcgattgga ggcagagtgc agaagaggtg attgtcaagc ttcgtgtggg 720 agtaggtccc ctgcagctgg aggatgtaga tgctgctttc acagatacag actgtgtggt 780 gcggtttgca ggtgg tcagc agtggggtgg tgtcttctat gctgagataa aaagctcttg 840 tgctaaagtg caaacccgca agggcagtct cctgcacctg acactgccca aaaaggtgcc 900 tatgctcacg tggccctccc tcctgaagaa acctctaggg acccaggagc tggtgccggg 960 gctgcggtgc caggagaatg ggcaggaact gtctcccatt gccctggagc caggccctga 1020 gccccaccgg gctaagcagg aggccccggaa ccagaagcgg gcccagggcc gtggtgaggt 1080 aggcgcaggg gctggccccg gggcccaggc agggcccagc gccaagaggg ctgtgcatct 1140 ctgcagaggg ccagaggggg acgggtccag ggatgaccct ggaccccggg gtgatgcccc 1200 acccttcgtg gctgacccag ccacccaggt tgaggctgat gaacagctt t gcataccacc 1260 gctgaactcc caaacctgcc tcctgggctc agaggagaat ttagcccctt tggcaggaga 1320 gaaagcagtg cctcccggga atgacccagt ctctccagcc atggtccgga gcagaaaccc 1380 tgggaaagat gactgtgcca aggaggagat ggcagtggca gcagatgctg caaccttggt 1440 ggatgagccc gagtcgatgg tgaacctggc gtttgtcaag aatgactcgt atgaga aggg 1500 cccggattca gtggtggtgc acgtgtacgt gaaggagatc tgcagggaca cctcaagagt 1560 acttttccgt gagcaggact tcacgctcat cttccagacc agggatggaa acttcctgag 1620 gctgcacccg ggctgtgggc cccacaccac cttccgttgg caggtga agc tcaggaatct 1680 gattgagcca gagcagtgca ccttctgttt cacggcttct cgcatcgaca tctgccttcg 1740 taagaggcag agtcagcgct gggggggcct ggaggccccg gctgcacgag tgggtggtgc 1800 aaaggttgcc gtgccgacag gtccaacccc tctggattca accccaccag gaggtgctcc 1860 ccaccccctg acaggccagg aggaggcccg ggctgtggag aaggataaat ccaaggcacg 192 0 atctgaggac acagggctag acagtgtggc aacccgcaca cccatggagc atgtaacccc 1980 aaagccagag acacacctgg cctcgcccaa gcctacatgc atggtgcctc ccatgcccca 2040 cagcccagtt agtggagaca gcgtggagga ggaggaagag gaagagaaga aggtgtgtct 2100 gccaggcttc actggccttg tcaatttagg caacacctgc ttcatgaaca gcgtcattca 2160 gtctctgtcc aacactcggg aactccggga cttcttccat gaccgctcct ttgaggctga 2220 gatcaactac aacaacccac tagggactgg tgggcgtctg gccattggct ttgccgtgct 2280 gcttcgggcg ctgtggaagg gcacccacca tgccttccag ccttccaagt tgaaggccat 2340 tgtggcgagt aaggccagcc agttcacagg ctatgcacag catgatgccc aggagttcat 2400 ggctttcctg ctggatgggc tgcacgagga cctgaatcgc attcagaaca agccctacac 2460 agagaccgtg gattcagatg ggcggcccga tgaggtggta gctgaggaag catggcagcg 252 0 gcacaagatg aggaatgact ctttcatcgt ggacctattt caggggcagt acaagtcgaa 2580 gctggtgtgc cctgtgtgtg ccaaggtctc catcactttt gacccgtttc tttatctgcc 2640 ggtgcccttg ccacaaaagc aaaaggttct ccctgtcttt tattttgccc gagagcccca 2700 cagcaagccc atcaagttcc tggtgagcgt cagcaaggag aactccactg cgagcgaagt 2760 attggactcc ctctctca ga gtgttcatgt gaagcctgag aacctgcgtt tggcggaggt 2820 aattaagaat cgttttcatc gtgtgttcct accctcccac tcactggaca ctgtgtcccc 2880 atctgatacg ctcctctgct ttgagctgct atcctcagag ttggctaagg agcgggtagt 294 0 ggtgctagag gtgcaacagc gcccccaggt gcccagcgtc cccatctcca agtgtgcagc 3000 ctgccagcgg aagcaacagt cggaggatga aaagctgaag cgctgtaccc ggtgctaccg 3060 tgtgggctac tgcaaccagc tctgccagaa aacccactgg cctgaccaca agggcctctg 3120 ccgacctgag aacattggct accccttcct ggtcagtgta cctgcctcac gcctcactta 3180 tgcccgcctc gctcagttgc tagag ggcta tgcccggtac tctgtgagtg tattccagcc 3240 accctttcag ccaggccgca tggccttgga gtctcagagc cctggctgca ccacactgct 3300 ctccacaggt tccctggagg ctggggacag cgagagagac cccattcagc cacctgagct 3360 ccagctggtg accccta tgg ctgaggggga cacagggctt ccccgggtgt gggcagcccc 3420 tgaccggggt cctgtgccca gcaccagtgg aatttcttct gagatgctgg ccagtgggcc 3480 cattgaggtt ggctccttgc cagctggcga gagggtgtcc cgacccgaag ctgctgtgcc 3540 tgggtaccag catccaagtg aagctatgaa tgcccacaca ccccagttct tcatctataa 3600 aattgattca tccaaccgag agcagcggct agaggacaaa gg agacacc cactggagct 3660 gggtgacgac tgtagcctgg ctctcgtctg gcggaacaat gagcgcttgc aggagtttgt 3720 gttggtagcc tccaaggagc tggaatgtgc tgaggatcca ggctctgccg gtgaggctgc 3780 ccgggccggc cacttcaccc tggaccagtg cctcaacctc ttcacacggc ctgaggtgct 3840 ggcacccgag gaggcctggt actgcccaca gtgcaaacag caccgtgagg cctccaagca 3900 gctgttgcta tggcgcctgc caaatgttct catcgtgcag ctcaagcgct tctcctttcg 3960 tagttttatc tggcgtgaca agatcaatga cttggtggag ttccctgtta ggaacctgga 4020 cctgagcaag ttctgcattg gtcagaaaga ggagcagctg cccagctacg atctatatgc 4080 tgtcatcaac cactatggag gcatgattgg tggccactac actgcctgtg cacgcctgcc 4140 caatgatcgt agcagtcagc gcagtgacgt gggctggcgc ttgtttgatg acagcacagt 4200 gacaacggta gacgagagcc aggttg tgac gcgttatgcc tatgtactct tctaccgccg 4260 gcggaactct cctgtggaga ggccccccag ggcaggtcac tctgagcacc acccagacct 4320 aggccctgca gctgaggctg ctgccagcca ggcttcccgg atttggcagg agctggaggc 4380 tgaggaggag ccggtgcctg aggggtctgg gcccctgggt ccctggggggc cccaagactg 4440 ggtgggcccc ctaccacgtg gccctaccac accagatgag ggctgcctcc ggt actttgt 4500 cctgggcacc gtggcggctt tggtggccct cgtgctcaac gtgttctatc ctctggtatc 4560 ccagagtcgc tggagatgag ctcgcctgca ggcagctgct gtgagctggc ctacctgcct 4620 gccccaggcc atgcctgcct ttgttgtgg g gaacacctct gggctttggg cctcagctta 4680 tgcatctggt gggagagggt ggggaggttg tggcccctgc aggggcagag tatcctaggg 4740 tgtgtatcca tctggctgtc tgtccattca tcctgctgct ctgacccttg gcctcaggct 4800 tggccctgcc caagctactt cctgtactta aaagtgttaa taaaaccaga ctattcaggc 4860 ccaaaaaaaa aaaaaaaaa 4878 <210> 6 <211> 4883 <212> DNA <213> Homo sap iens <400> 6 agcttggcgc tccgcctccc tcggccttag ctagaagtcg aaaaaaaca agccgctaag 60 gcggtggcgg cggcgccggg acgggggagg ggcgcgccgg aaccggaacc gacctgcgcc 120 ggaaccggaa cggagagcgg gttgccaggg cccgaagagg gctggctgcg gcggtctcgc 180 tcggctgtcc gttccttgct ggagaatttg gccacaaaga gttgccaaga tagctgggcc 240 aggaagaaag cgccgcagcc ctgacccaga cgctgttgcc gaccccgggg cact ctggct 300 gtcgaccaag cggctcaaga tgtctggcgg ggccagtgcc acaggcccaa ggagagggcc 360 cccaggactg gaggacacca ctagtaagaa gaagcagaag gatcgagcaa accaggagag 420 caaggatgga gatcctagga aagagacagg gtctcgatat gttgcccagg ctggtcttga 48 0 acctctggcc tcaggtgatc cttctgcctc agcctcccat gcagctggga tcacaggctc 540 acgccaccgt acccggctgt tctttccttc atcgtcaggg tcagcatcca ctcctcaaga 600 ggagcagacc aaagagggag cttgtgaaga ccctcatgat ctcttggcta ctcccactcc 660 agagttgttg ctcgattgga ggcagagtgc agaagaggtg attgtcaag c ttcgtgtggg 720 agtaggtccc ctgcagctgg aggatgtaga tgctgctttc acagatacag actgtgtggt 780 gcggtttgca ggtggtcagc agtggggtgg tgtcttctat gctgagataa aaagctcttg 840 tgctaaagtg caaacccgca agggcagt ct cctgcacctg acactgccca aaaaggtgcc 900 tatgctcacg tggccctccc tcctgaagaa acctctaggg acccaggagc tggtgccggg 960 gctgcggtgc caggagaatg ggcaggaact gtctcccatt gccctggagc caggccctga 1020 gccccaccgg gctaagcagg aggcccggaa ccagaagcgg gcccagggcc gtggtgaggt 1080 aggcgcaggg gctggccccg gggcccaggc agggcccagc gccaagaggg ctgtgcatct 1140 ctgcaga ggg ccagaggggg acgggtccag ggatgaccct ggaccccggg gtgatgcccc 1200 acccttcgtg gctgacccag ccacccaggt tgaggctgat gaacagcttt gcataccacc 1260 gctgaactcc caaacctgcc tcctgggctc agaggagaat ttagcccctt tggcaggaga 1320 gaaagca gtg cctccccggga atgacccagt ctctccagcc atggtccgga gcagaaaccc 1380 tgggaaagat gactgtgcca aggaggat ggcagtggca gcagatgctg caaccttggt 1440 ggatgagccc gagtcgatgg tgaacctggc gtttgtcaag aatgactcgt atgagaaggg 1500 cccggattca gtggtggtgc acgtgtacgt gaaggagatc tgcagggaca cctcaagagt 1560 acttt tccgt gagcaggact tcacgctcat cttccagacc agggatggaa acttcctgag 1620 gctgcacccg ggctgtgggc cccacaccac cttccgttgg caggtgaagc tcaggaatct 1680 gattgagcca gagcagtgca ccttctgttt cacggcttct cgcatcgaca tctgccttcg 17 40 taagaggcag agtcagcgct gggggggcct ggaggccccg gctgcacgag gtgcagtggg 1800 tggtgcaaag gttgccgtgc cgacaggtcc aacccctctg gattcaaccc caccaggagg 1860 tgctccccac cccctgacag gccaggagga ggcccgggct gtggagaagg ataaatccaa 1920 ggcacgatct gaggacacag ggctagacag tgtggcaacc cgcacaccca tggagcatgt 1980 aaccccaaag ccagagacac acctggcctc g cccaagcct acatgcatgg tgcctcccat 2040 gccccacagc ccagttagtg gagacagcgt ggaggaggag gaagaggaag agaagaaggt 2100 gtgtctgcca ggcttcactg gccttgtcaa tttaggcaac acctgcttca tgaacagcgt 2160 cattcagtct ctgtccaaca ct cgggaact ccgggacttc ttccatgacc gctcctttga 2220 ggctgagatc aactacaaca acccactagg gactggtggg cgtctggcca ttggctttgc 2280 cgtgctgctt cgggcgctgt ggaagggcac ccaccatgcc ttccagcctt ccaagttgaa 2340 ggccattgtg gcgagtaagg ccagccagtt cacaggctat gcacagcatg atgcccagga 2400 gttcatggct ttcctgctgg at gggctgca cgaggacctg aatcgcattc agaacaagcc 2460 ctacacagag accgtggatt cagatgggcg gcccgatgag gtggtagctg aggaagcatg 2520 gcagcggcac aagatgagga atgactcttt catcgtggac ctatttcagg ggcagtacaa 2580 gtcgaagctg gtg tgccctg tgtgtgccaa ggtctccatc acttttgacc cgtttcttta 2640 tctgccggtg cccttgccac aaaagcaaaa ggttctccct gtcttttatt ttgcccgaga 2700 gccccacagc aagcccatca agttcctggt gagcgtcagc aaggagaact ccactgcgag 2760 cgaagtattg gactccctct ctcagagtgt tcatgtgaag cctgagaacc tgcgtttggc 2820 ggaggtaatt aagaatcgtt ttcatcgtgt gttcc taccc tcccactcac tggacactgt 2880 gtccccatct gatacgctcc tctgctttga gctgctatcc tcagagttgg ctaaggagcg 2940 ggtagtggtg ctagaggtgc aacagcgccc ccaggtgccc agcgtcccca tctccaagtg 3000 tgcagcctgc cagcgga agc aacagtcgga ggatgaaaag ctgaagcgct gtacccggtg 3060 ctaccgtgtg ggctactgca accagctctg ccagaaaacc cactggcctg accacaaggg 3120 cctctgccga cctgagaaca ttggctaccc cttcctggtc agtgtacctg cctcacgcct 3180 cacttatgcc cgcctcgctc agttgctaga gggctatgcc cggtactctg tgagtgtatt 3240 ccagccaccc tttcagccag gccgcatggc cttggagtct caga gccctg gctgcaccac 3300 actgctctcc acaggttccc tggaggctgg ggacagcgag agagaccca ttcagccacc 3360 tgagctccag ctggtgaccc ctatggctga gggggacaca gggcttcccc gggtgtgggc 3420 agcccctgac cggggtcctg tgcccagcac cag tggaatt tcttctgaga tgctggccag 3480 tgggcccatt gaggttggct ccttgccagc tggcgagagg gtgtcccgac ccgaagctgc 3540 tgtgcctggg taccagcatc caagtgaagc tatgaatgcc cacacacccc agttcttcat 3600 ctataaaatt gattcatcca accgagagca gcggctagag gacaaaggag acaccccact 3660 ggagctgggt gacgactgta gcctggctct cgtctggcgg aacaatgagc gcttgcagga 372 0 gtttgtgttg gtagcctcca aggagctgga atgtgctgag gatccaggct ctgccggtga 3780 ggctgcccgg gccggccact tcaccctgga ccagtgcctc aacctcttca cacggcctga 3840 ggtgctggca cccgaggagg cctggtactg cccacagtgc aaacagcacc gt gaggcctc 3900 caagcagctg ttgctatggc gcctgccaaa tgttctcatc gtgcagctca agcgcttctc 3960 ctttcgtagt tttatctggc gtgacaagat caatgacttg gtggagttcc ctgttaggaa 4020 cctggacctg agcaagttct gcattggtca gaaagaggag cagctgccca gctacgatct 4080 atatgctgtc atcaaccact atggaggcat gattggtggc cactacactg cctgtgcacg 4140 cctgcccaat gatcgtagca gtcagcgcag tgacgtgggc tggcgcttgt ttgatgacag 4200 cacagtgaca acggtagacg agagccaggt tgtgacgcgt tatgcctatg tactcttcta 4260 ccgccggcgg aactctcctg tggagaggcc ccccagggca ggtcactctg agcaccaccc 4320 agacctaggc cctgcagctg aggctgctgc cagccaggga ctaggccctg gccaggcccc 4380 cgaggtggcc cccacgcgga cagcccctga acgcttcgcc ccccctgtgg atcggccagc 4440 ccccacctac agcaacatgg aggaggtgga ttagcaggtc cctggctgat gggggggact 4500 gggtttggga cacccacaca gagggccagc tccttgccgc ttctccttct ctaacccaga 4560 ggacactggc tctg tcaatg ggaagctgag gggtatgatt tgggtgtgga gacctctcag 4620 gttgggactt cttgtcagct tggacccctg accagtgggc tttggcttct ccagccgcct 4680 ccagtgctgc gtgatttgat tctgttgtac cttcaattct tctgacccgc attataaaca 4 740 ttataatttt attctaaaaa ttgtaatttt ttttgcattt tggaagtgac tgctgctgtt 4800 taaatatatt ttaaaaataa ataataagta agtagactta aaaaaaaaa aaaaaaaaaa 4860 aaaaaaaaaaa aaaaaaaaaaa aaa 4883 <210> 7 <211> 4847 <212> DNA <213> Homo sapiens <400> 7 agcttggcgc tccgcctccc tcggccttag ctagaagtcg aaacaaaaaca agccgctaag 6 0 gcggtggcgg cggcgccggg acgggggagg ggcgcgccgg aaccggaacc gacctgcgcc 120 ggaaccggaa cggagagcgg gttgccaggg cccgaagagg gctggctgcg gcggtctcgc 180 tcggctgtcc gttccttgct ggagaatttg gccacaaaga gttgccaaga tagctgggcc 240 aggaagaaag cgccgcagcc ctgacccaga cgctgttgcc gaccccgggg cactctggct 300 gtcgaccaag cggctcaaga tgtctggcgg ggccagtgcc acaggcccaa ggagagggcc 36 0 cccaggactg gaggacacca ctagtaagaa gaagcagaag gatcgagcaa accaggagag 420 caaggatgga gatcctagga aagagacagg gtctcgatat gttgcccagg ctggtcttga 480 acctctggcc tcaggtgatc cttctgcctc agcctcccat gcagctggga tcacaggctc 540 acg ccaccgt acccggctgt tctttccttc atcgtcaggg tcagcatcca ctcctcaaga 600 ggagcagacc aaagagggag cttgtgaaga ccctcatgat ctcttggcta ctcccactcc 660 agagttgttg ctcgattgga ggcagagtgc agaagaggtg attgtcaagc ttcgtgtggg 720 agtaggtccc ctgcagctgg aggatgtaga tgctgctttc acagatacag actgtg tggt 780 gcggtttgca ggtggtcagc agtggggtgg tgtcttctat gctgagataa aaagctcttg 840 tgctaaagtg caaacccgca agggcagtct cctgcacctg acactgccca aaaagaagaa 900 acctctaggg acccaggagc tggtgccggg gctgcggtgc cagg agaatg ggcaggaact 960 gtctcccatt gccctggagc caggccctga gccccaccgg gctaagcagg aggcccggaa 1020 ccagaagcgg gcccagggcc gtggtgaggt aggcgcaggg gctggccccg gggcccaggc 1080 agggcccagc gccaagaggg ctgtgcatct ctgcagaggg ccagaggggg acgggtccag 1140 ggatgaccct ggaccccggg gtgatgcccc acccttcgtg gctgacccag ccacccaggt 1200 tgaggct gat gaacagcttt gcataccacc gctgaactcc caaacctgcc tcctgggctc 1260 agaggagaat ttagcccctt tggcaggaga gaaagcagtg cctcccggga atgacccagt 1320 ctctccagcc atggtccgga gcagaaaccc tgggaaagat gactgtgcca aggaggagat 1380 ggcagtgg ca gcagatgctg caaccttggt ggatgagccc gagtcgatgg tgaacctggc 1440 gtttgtcaag aatgactcgt atgagaaggg cccggattca gtggtggtgc acgtgtacgt 1500 gaaggagatc tgcagggaca cctcaagagt acttttccgt gagcaggact tcacgctcat 1560 cttccagacc agggatggaa acttcctgag gctgcacccg ggctgtgggc cccacaccac 1620 cttccgttgg caggtgaag c tcaggaatct gattgagcca gagcagtgca ccttctgttt 1680 cacggcttct cgcatcgaca tctgccttcg taagaggcag agtcagcgct gggggggcct 1740 ggaggccccg gctgcacgag tgggtggtgc aaaggttgcc gtgccgacag gtccaacccc 1800 tctgg attca accccaccag gaggtgctcc ccaccccctg acaggccagg aggaggcccg 1860 ggctgtggag aaggataaat ccaaggcacg atctgaggac acagggctag acagtgtggc 1920 aacccgcaca cccatggagc atgtaacccc aaagccagag acacacctgg cctcgcccaa 1980 gcctacatgc atggtgcctc ccatgcccca cagcccagtt agtggagaca gcgtggag ga 2040 ggaggaagag gaagagaaga aggtgtgtct gccaggcttc actggccttg tcaatttagg 2100 caacacctgc ttcatgaaca gcgtcattca gtctctgtcc aacactcggg aactccggga 2160 cttcttccat gaccgctcct ttgaggctga gatcaactac aacaacccac tagggactgg 2220 tgggcgtctg gccattggct ttgccgtgct gcttcgggcg ctgtggaagg gcacccacca 2280 tgccttccag ccttccaagt tgaaggccat tgtggcgagt aaggccagcc agttcacagg 2340 ctatgcacag catgatgccc aggagttcat ggctttcctg ctggatgggc tgcacgagga 2400 cctgaatcgc attcagaaca agccctacac agagaccgtg gattcagatg ggcggcccga 2460 tgaggtggta gctgaggaag catggcagcg gcacaagatg aggaatgact ctttcatcgt 2520 ggacctattt caggggcagt acaagtcgaa gctggtgtgc cctgtgtgtg ccaaggtctc 2580 catcactttt gacccgtttc tttatctgcc ggtgcccttg c cacaaaagc aaaaggttct 2640 ccctgtcttt tattttgccc gagagcccca cagcaagccc atcaagttcc tggtgagcgt 2700 cagcaaggag aactccactg cgagcgaagt attggactcc ctctctcaga gtgttcatgt 2760 gaagcctgag aacctgcgtt tggcggaggt aattaagaat cgttttcatc gtgtgttcct 2820 accctcccac tcactggaca ctgtgtcccc atctgatacg ctcctctgct ttgagctgct 2880 atcctcagag ttggctaagg agcgggtagt ggtgctagag gtgcaacagc gcccccaggt 2940 gcccagcgtc cccatctcca agtgtgcagc ctgccagcgg aagcaacagt cggagggatga 3000 aaagctgaag cgctgtaccc ggtgctaccg tgtgggctac tgcaaccagc tctgccagaa 3060 aacccactgg cctgaccaca agggcctctg ccgacctgag aacattggct accccttcct 3120 ggtcagtgta cctgcctcac gcctcactta tgcccgcctc gctcagttgc tagagggcta 3180 tgcccggtac tctgtgagtg tattccagcc accctttcag ccaggccgca tggccttgga 3240 gtctcagagc cctggctgca ccacactgct ctccacaggt tccctggagg ctggggacag 3300 cgagagagac cccattca gc cacctgagct ccagctggtg acccctatgg ctgaggggga 3360 cacagggctt ccccgggtgt gggcagcccc tgaccggggt cctgtgccca gcaccagtgg 3420 aatttcttct gagatgctgg ccagtgggcc cattgaggtt ggctccttgc cagctggcga 3480 gagggtgtcc cgacccgaag ctgctgtgcc tgggtaccag catccaagtg aagctatgaa 3540 tgcccacaca ccccagttct tcatctataa aattgattca tccaaccgag agcagcggct 3600 agaggacaaa ggagacaccc cactggagct gggtgacgac tgtagcctgg ctctcgtctg 3660 gcggaacaat gagcgcttgc aggagtttgt gttggtagcc tccaaggagc tggaatgtgc 3720 tgaggatcca ggctctgccg gtgagg ctgc ccgggccggc cacttcaccc tggaccagtg 3780 cctcaacctc ttcacacggc ctgaggtgct ggcacccgag gaggcctggt actgcccaca 3840 gtgcaaacag caccgtgagg cctccaagca gctgttgcta tggcgcctgc caaatgttct 3900 catcgtgcag ctcaagcgct tctcctttcg tagttttatc tggcgtgaca agatcaatga 3960 cttggtggag ttccctgtta ggaacctgga cctgagcaag ttctgcattg gtcagaaaga 4020 ggagcagctg cccagctacg atctatatgc tgtcatcaac cactatggag gcatgattgg 4080 tggccactac actgcctgtg cacgcctgcc caatgatcgt agcagtcagc gcagtgacgt 4140 gggctggcgc ttgtttgatg acagcacagt gaca acggta gacgagagcc aggttgtgac 4200 gcgttatgcc tatgtactct tctaccgccg gcggaactct cctgtggaga ggccccccag 4260 ggcaggtcac tctgagcacc acccagacct aggccctgca gctgaggctg ctgccagcca 4320 gggactaggc cctggccagg cccccgagg t ggcccccacg cggacagccc ctgaacgctt 4380 cgccccccct gtggatcggc cagcccccac ctacagcaac atggaggagg tggattagca 4440 ggtccctggc tgatgggggg gactgggttt gggacaccca cacagagggc cagctccttg 4500 ccgcttctcc ttctctaacc cagaggacac tggctctgtc aatgggaagc tgaggggtat 4560 gatttgggtg tggagacctc tcaggttggg acttcttgtc agcttggac c cctgaccagt 4620 gggctttggc ttctccagcc gcctccagtg ctgcgtgatt tgattctgtt gtaccttcaa 4680 ttcttctgac ccgcattata aacattataa ttttattcta aaaattgtaa ttttttttgc 4740 attttggaag tgactgctgc tgtttaaata tattttaaaa ataaataata agtaagtaga 4800 cttaaaaaaa aaaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaa 4847 < 210> 8 <211> 4575 <212> DNA <213> Homo sapiens <400> 8 agcttggcgc tccgcctccc tcggccttag ctagaagtcg aaacaaaaca agccgctaag 60 gcggtggcgg cggcgccggg acgggggagg ggcgcgccgg aaccggaacc gacctgcg cc 120 ggaaccggaa cggagagcgg gttgccaggg cccgaagagg gctggctgcg gcggtctcgc 180 tcggctgtcc gttccttgct ggagaatttg gccacaaaga gttgccaaga tagctgggcc 240 aggaagaaag cgccgcagcc ctgacccaga cgctgttgcc gaccccgggg cactctggct 300 gtcgaccaag cggctcaaga tgtctggcgg ggccagtgcc acaggcccaa ggagagggcc 360 cccaggactg gaggacacca ctagtaagaa gaagcagaag gatcgagcaa accaggagag 420 caaggatgga gatcctagga aagagacagg g tctcgatat gttgcccagg ctggtcttga 480 acctctggcc tcaggtgatc cttctgcctc agcctcccat gcagctggga tcacaggctc 540 acgccaccgt acccggctgt tctttccttc atcgtcaggg tcagcatcca ctcctcaaga 600 ggagcagacc aaagagggag cttgtgaaga ccctcatgat ctcttggcta ctcccactcc 660 agagttgttg ctcgatgga ggcagagtgc agaagaggtg attgtcaagc ttcgtgtggg 720 agtaggtccc ctgcagctgg aggatgtaga tgctgctttc acagatacag actgtgtggt 780 gcggtttgca ggtggtcagc agtggggtgg tgtcttctat gctgagataa aaagctcttg 840 tgctaaagt g caaacccgca agggcagtct cctgcacctg acactgccca aaaaggtgcc 900 tatgctcacg tggccctccc tcctggttga ggctgatgaa cagctttgca taccaccgct 960 gaactcccaa acctgcctcc tgggctcaga ggagaattta gcccctttgg caggagagaa 1020 agca gtgcct cccgggaatg acccagtctc tccagccatg gtccggagca gaaaccctgg 1080 gaaagatgac tgtgccaagg aggagatggc agtggcagca gatgctgcaa ccttggtgga 1140 tgagcccgag tcgatggtga acctggcgtt tgtcaagaat gactcgtatg agaagggccc 1200 ggattcagtg gtggtgcacg tgtacgtgaa ggagatctgc agggacacct caagagtact 1260 tttccgtgag caggacttca cgctcatctt ccagaccagg gatggaaact tcctgaggct 1320 gcacccgggc tgtgggcccc acaccacctt ccgttggcag gtgaagctca ggaatctgat 1380 tgagccagag cagtgcacct tctgtttcac ggcttctcgc atcgacatct gccttcgtaa 1440 gaggcagagt cagcgctggg ggggcctgga ggccccggct gcacgagtgg gtggtgcaaa 1500 ggttgccgtg ccgacaggtc caacccctct ggattcaacc ccaccaggag gtgctcccca 1560 ccccctgaca ggccaggagg aggcccgggc tgtggagaag gataaatcca aggcacgatc 1620 tgaggacaca gggctagaca gtgtggcaac ccgcacaccc atggagcatg taaccccaaa 1680 gccagagaca cacctggcct cgcccaagcc tacatgcatg gtgcctccca tgccccacag 1740 cccagttagt ggagacagcg tggaggagga ggaagaggaa gagaagaagg tgtgtctgcc 1800 aggcttcact ggccttgtca atttaggcaa cacctgcttc atgaacagcg tcattcagtc 1860 tctgtccaac actcgggaac tccgggactt cttccatgac cgct cctttg aggctgagat 1920 caactacaac aacccactag ggactggtgg gcgtctggcc attggctttg ccgtgctgct 1980 tcgggcgctg tggaagggca cccaccatgc cttccagcct tccaagttga aggccattgt 2040 ggcgagtaag gccagccagt tcacaggcta tgcacagcat gatgcccagg agttcatggc 2100 tttcctgctg gatgggctgc acgaggacct gaatcgcatt cagaac aagc cctacacaga 2160 gaccgtggat tcagatgggc ggcccgatga ggtggtagct gaggaagcat ggcagcggca 2220 caagatgagg aatgactctt tcatcgtgga cctatttcag gggcagtaca agtcgaagct 2280 ggtgtgccct gtgtgtgcca aggtctccat cacttt tgac ccgtttcttt atctgccggt 2340 gcccttgcca caaaagcaaa aggttctccc tgtcttttat tttgcccgag agccccacag 2400 caagcccatc aagttcctgg tgagcgtcag caaggagaac tccactgcga gcgaagtatt 2460 ggactccctc tctcagagtg ttcatgtgaa gcctgagaac ctgcgtttgg cggaggtaat 2520 taagaatcgt tttcatcgtg tgttcctacc ctcccactca ctggacactg tgt ccccatc 2580 tgatacgctc ctctgctttg agctgctatc ctcagagttg gctaaggagc gggtagtggt 2640 gctagaggtg caacagcgcc cccaggtgcc cagcgtcccc atctccaagt gtgcagcctg 2700 ccagcggaag caacagtcgg aggatgaaaa gctgaagc gc tgtacccggt gctaccgtgt 2760 gggctactgc aaccagctct gccagaaaac ccactggcct gaccacaagg gcctctgccg 2820 acctgagaac attggctacc ccttcctggt cagtgtacct gcctcacgcc tcacttatgc 2880 ccgcctcgct cagttgctag agggctatgc ccggtactct gtgagtgtat tccagccacc 2940 ctttcagcca ggccgcatgg ccttggagtc tcagagccct ggctgcacca cactgctct c 3000 cacaggttcc ctggaggctg gggacagcga gagagacccc attcagccac ctgagctcca 3060 gctggtgacc cctatggctg agggggacac agggcttccc cgggtgtggg cagcccctga 3120 ccggggtcct gtgcccagca ccagtggaat ttcttctgag atgctggcca gtgggcccat 3180 tgaggttggc tccttgccag ctggcgagag ggtgtcccga cccgaagctg ctgtgcctgg 3240 gtaccagcat ccaagtgaag ctatgaatgc ccacacaccc cagttcttca tctataaaat 3300 tgattcatcc aaccgagagc agcggctaga ggacaaagga gacaccccac tggagctggg 3360 tgacgactgt agcctggctc tcgtctggcg gaacaatgag cgcttgcagg agtttgtgtt 3420 gg tagcctcc aaggagctgg aatgtgctga ggatccaggc tctgccggtg aggctgcccg 3480 ggccggccac ttcaccctgg accagtgcct caacctcttc acacggcctg aggtgctggc 3540 acccgaggag gcctggtact gcccacagtg caaacagcac cgtgaggcct ccaagcagct 3600 gttgctatgg cgcctgccaa atgttctcat cgtgcagctc aagcgcttct cctttcgtag 3660 ttttatctgg cgtgacaaga tcaatgactt ggtggagttc cctgttagga acctggacct 3720 gagcaagttc tgcattggtc agaaagagga gcagctgccc agctacgatc tatatgctgt 3780 catcaaccac tatggaggca tgattggtgg ccactacact gcctgtgcac gcctgcccaa 3840 tgatcgtagc agtcagc gca gtgacgtggg ctggcgcttg tttgatgaca gcacagtgac 3900 aacggtagac gagagccagg ttgtgacgcg ttatgcctat gtactcttct accgccggcg 3960 gaactctcct gtggagaggc cccccagggc aggtcactct gagcaccacc cagacctagg 4020 ccctgcag ct gaggctgctg ccagccaggc ttcccggatt tggcaggagc tggaggctga 4080 ggaggagccg gtgcctgagg ggtctgggcc cctgggtccc tgggggcccc aagactgggt 4140 gggccccta ccacgtggcc ctaccacacc agatgagggc tgcctccggt actttgtcct 4200 gggcaccgtg gcggctttgg tggccctcgt gctcaacgtg ttctatcctc tggtatccca 4260 gagtcgctgg agatgagctc gcctg caggc agctgctgtg agctggccta cctgcctgcc 4320 ccaggccatg cctgcctttg ttgtggggaa cacctctggg ctttgggcct cagcttatgc 4380 atctggtggg agagggtggg gaggttgtgg cccctgcagg ggcagagtat cctagggtgt 4440 gtatc catct ggctgtctgt ccattcatcc tgctgctctg acccttggcc tcaggcttgg 4500 ccctgcccaa gctacttcct gtacttaaaa gtgttaataa aaccagacta ttcaggccca 4560 aaaaaaaaaaa aaaaa 4575 <210> 9 <211> 18 <212> DNA <213> Artificial Sequence <220> <223> Primer <400> 9 tgtgggctac tgcaacca 18 <210> 10 <211> 18 <212> DNA <213 > Artificial Sequence <220> <223> Primer<400> 10 gctgaatggg gtctctct 18

Claims (11)

비만 진단에 필요한 정보를 제공하기 위하여, 대상자의 시료 중 서열번호 1 내지 4의 단백질로 이루어진 군으로부터 선택된 단백질을 코딩하는 유전자의 발현량을 측정하는 단계를 포함하는 분석방법으로서,
상기 대상자의 시료 중의 서열번호 1 내지 4의 단백질로 이루어진 군으로부터 선택된 단백질의 발현량 또는 이를 코딩하는 유전자의 발현량이 정상인의 시료 중의 서열번호 1 내지 4의 단백질로 이루어진 군으로부터 선택된 단백질의 발현량 또는 이를 코딩하는 유전자의 발현량에 비하여 높을 경우, 비만 위험 환자로 분류하는 분석방법.In order to provide information necessary for diagnosing obesity, an analysis method comprising measuring the expression level of a gene encoding a protein selected from the group consisting of proteins of SEQ ID NOs: 1 to 4 in a sample of a subject,
The expression level of a protein selected from the group consisting of proteins of SEQ ID NOs. 1 to 4 in a sample of the subject or the expression level of a gene encoding the same, the expression level of a protein selected from the group consisting of proteins of SEQ ID NOs. 1 to 4 in a sample of a normal person, or An analysis method that classifies patients at risk of obesity when it is higher than the expression level of the gene that codes for it. 제1항에 있어서, 상기 단백질을 코딩하는 유전자가 서열번호 4의 단백질을 코딩하는 유전자인 것을 특징으로 하는 분석방법.The analysis method according to claim 1, wherein the gene encoding the protein is a gene encoding the protein of SEQ ID NO: 4. 제1항에 있어서, 상기 대상자의 시료가 혈액 또는 뇨인 것을 특징으로 하는 분석방법.The analysis method according to claim 1, wherein the subject's sample is blood or urine. 제1항 내지 제3항 중 어느 한에 있어서, 상기 유전자의 발현량의 측정이 mRNA 또는 단백질의 양을 측정하는 것을 특징으로 하는 분석방법.The analysis method according to any one of claims 1 to 3, wherein the expression level of the gene is measured by measuring the amount of mRNA or protein. 제4항에 있어서, 상기 단백질의 양의 측정이 웨스턴 블럿팅에 의하여 수행되는 것을 특징으로 하는 분석방법.The analysis method according to claim 4, wherein the amount of the protein is measured by Western blotting. 제4항에 있어서, 상기 mRNA 양의 측정이 RT-PCR 또는 실시간-PCR을 수행하여 측정하는 것을 특징으로 하는 분석방법.The analysis method according to claim 4, wherein the amount of mRNA is measured by performing RT-PCR or real-time PCR. 서열번호 1 내지 4의 단백질로 이루어진 군으로부터 선택된 단백질을 코딩하는 유전자의 발현량을 측정할 수 있는 분자를 포함하는 비만 진단용 키트로서,
상기 분자가 상기 단백질에 특이적으로 결합하는 항체, 기질, 리간드, 또는 보조인자(cofactor); 또는 상기 단백질을 코딩하는 유전자에 특이적인 상보적 서열을 갖는 프라이머인 비만 진단용 키트.A kit for diagnosing obesity comprising a molecule capable of measuring the expression level of a gene encoding a protein selected from the group consisting of proteins of SEQ ID NOs: 1 to 4,
An antibody, substrate, ligand, or cofactor that specifically binds the molecule to the protein; Or a kit for diagnosing obesity, which is a primer having a specific complementary sequence to the gene encoding the protein. 제7항에 있어서, 상기 단백질을 코딩하는 유전자가 서열번호 4의 단백질을 코딩하는 유전자인 것을 특징으로 하는 비만 진단용 키트.The kit for diagnosing obesity according to claim 7, wherein the gene encoding the protein is a gene encoding the protein of SEQ ID NO: 4. 제7항에 있어서, 상기 분자가 검출가능한 표지로 표지된 것임을 특징으로 하는 비만 진단용 키트.The kit for diagnosing obesity according to claim 7, wherein the molecule is labeled with a detectable label. 제7항에 있어서, 상기 프라이머가 기판상에 고정화되어 있는 마이크로어레이 형태인 것을 특징으로 하는 비만 진단용 키트.The kit for diagnosing obesity according to claim 7, wherein the primers are in the form of a microarray immobilized on a substrate. 제7항에 있어서, 상기 프라이머가 서열번호 9 또는 10의 염기서열을 갖는 것을 특징으로 하는 비만 진단용 키트.The kit for diagnosing obesity according to claim 7, wherein the primer has a base sequence of SEQ ID NO: 9 or 10.
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US20110003296A1 (en) 1999-01-06 2011-01-06 Choong-Chin Liew Method for the detection of gene transcripts in blood and uses thereof
KR100794706B1 (en) * 2006-07-14 2008-01-14 (주) 차바이오텍 Probes or primers for diagnosing obesity or diabetes derived from genes coding deubiquitinating enzymes and methods for diagnosing obesity or diabetes using the same

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