KR20050115278A - 간엽계 간세포 검출용 마커 및 상기 마커를 이용한 간엽계간세포의 식별방법 - Google Patents
간엽계 간세포 검출용 마커 및 상기 마커를 이용한 간엽계간세포의 식별방법 Download PDFInfo
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Abstract
본 발명은, 간엽계 간세포를 검출, 분리·식별하기 위한 마커 및 상기 마커를 사용한 간엽계 간세포의 검출, 분리·식별방법을 제공하는 것이다. 배열표에 나타내는 유전자는 간엽계 간세포에서 특이적으로 발현한다. 상기 유전자를 간엽계 간세포 검출용의 마커로서 사용하는 것으로 이루어진다. 또, 본 발명은, 상기 간엽계 간세포 마커 유전자 검출용 프로브 및 상기 간엽계 간세포 유전자 마커의 검출시에, 피검세포의 상기 유전자를 증폭하기 위한 PCR용 프라이머를 포함하며, 또, 본 발명의 간엽계 간세포 마커 유전자가 발현하는 폴리펩티드로 이루어지는 간엽계 간세포 검출용 폴리펩티드 마커, 상기 폴리펩티드 마커를 검출하기 위한 상기 폴리펩티드 마커에 특이적으로 결합하는 항체, 또한 간엽계 간세포 마커 유전자 검출용 프로브, 폴리펩티드 마커에 특이적으로 결합하는 항체를 사용한 간엽계 간세포의 식별, 분리방법을 포함한다.
Description
본 발명은 간엽계(間葉系) 간세포의 검출, 분리·식별에 관한 것이며, 특히 간엽계 간세포를 검출, 분리·식별하는데에 사용하는 간엽계 간세포 검출용 유전자 마커 및 간엽계 간세포 검출용 폴리펩티드마커 및 상기 마커를 사용한 간엽계 간세포의 검출, 식별방법에 관한 것이다.
간엽계 간세포는 포유류의 골수 등에 존재하며, 지방세포, 연골세포, 골세포에 분화하는 다능성(多能性)의 간세포로서 알려져 있다. 간엽계 간세포는 그 분화다능성 때문에, 뼈, 연골, 힘줄, 금육, 지방, 치주조직 등, 많은 조직의 재생의료를 위한 이식(移植)재료로서 주목받고 있다(유전자 의학, Vol.4, No.2(2000) p58-61). 최근에는, 간엽계 간세포연구의 현황과 전망에 대한 총설이 발행되어 간엽계 간세포의 채취나 배양에 관한 연구가 보고되고 있다(실험의학, Vol.19, No.3(2월호)2001, p350-356). 또한, 지방조직에도 간엽계 간세포가 존재한다는 연구결과가 보고되었다. (Tissue Engineering, P.A. Zuk et al., Multilineage cells from human adipose tissue : implications for cell-based therapies. 7:211-228, 2001).
근래에, 간엽계 간세포의 배양, 분화 등에 관하여 몇개의 특허출원이 공개된바 있다. 예를 들면, 일본국 특표 평11-506610호 공보에는, 무혈청 환경하에서 인간의 간엽전구세포의 생존을 유지하는 조성물 및 방법에 대하여, 일본국 특표 평10-512756호 공보에는 간엽계 간세포의 분화를 유도하기 위하여, 프로스타그란딘, 아스코르빈산, 콜라겐세포외기질 등으로 이루어지는 골유도인자, 분화부수인자, 연골유도인자 등의 생물활성인자와 접촉시키는 것에 의하여 이루어지는 방법에 대하여, 일본국 특개2000-217576호 공보에는, 프로락틴 또는 그 동효물(同效物)의 공존하에서 다능성 간엽계 간세포를 배양하고, 간엽계 간세포를 지방세포로 분화시키는 방법에 대하여 각각 발명이 개시되어 있다.
본 발명자는, 앞서 기저막 세포외기질의 존재하에 있어서, 또는 섬유아(纖維芽)세포 증식인자(FGF) 등의 함유배지에서 간엽계 간세포를 배양하는 것에 의하여, 간엽계 간세포를 현저하게 빨리 증식시키고, 또한 그 분화능을 유지할 수 있다는 것을 발견하여, 종래의 배양방법과 비교하여 현저히 많은 간엽계 간세포를 얻는 배양방법을 개시한 바 있다(일본국 특개 2003-52360호 공보). 또, 간엽계 간세포의 채취 시에, 채취모체가 안전하고 또한 채취가 용이한 분리채취를 실시하기 위하여, 구강조직으로부터 간엽계 간세포를 분리채취하는 방법을 개시한 바 있다(일본국 특개 2003-52365호 공보).
최근, 재생의료의 발전과 함께, 간엽계 간세포는 그 분화다능성 때문에, 뼈, 연골, 힘줄, 근육, 지방, 치주조직 등, 많은 조직의 재생의료를 위한 이식재료로서 주목받고 있다. 간엽계 간세포를 조직의 재생의료에 이용하기 위해서는, 우선 이 간세포를 생체조직으로부터 채취하고, 그것을 증식시키고, 다시 그것을 분화증식하여 조직의 조제를 실시하는 일이 필요해진다. 간엽계 간세포는 골수나 골막에 존재하나, 조직재생의료에로의 실용화를 위해서는, 우선 이들 조직으로부터 간엽계 간세포를 안전하고 또한 용이하게 채취하는 방법을 개발하는 것이 필요하다. 그리고, 간엽계 간세포를 조직재생의료에 실용화하기 위해서는, 간엽계 간세포의 충분한 양을 확보하는 기술을 개발하는 것이 필요하다. 그렇게 하기 위해서는, 채취한 간엽계 간세포를 분화능력을 유지한 채로 배양증식시키는 기술을 개발하는 것이 중요하다.
본 발명자는, 상기한 바와 같이 간엽계 간세포의 채취에 있어서, 채취모체에 안전하고 또한 채취가 용이한 분리채취를 실시하기 위한 방법을 개발하였다. 또한 본 발명자는 기저막 세포외기질의 존재하에 있어서, 혹은 섬유아세포 증식인자(FGF) 등의 함유배지에서 간엽계 간세포를 배양하는 것에 의하여, 간엽계 간세포를 그 분화능을 유지한 채 폭발적으로 증식시키는 방법을 개발하였다. 그러나, 배양증식한 간엽계 간세포를 재생의료에 실용화하기 위해서는, 배양한 세포가 간엽계 간세포인 것을 확인할 필요가 있으며, 상기 간엽계 간세포를 검출, 식별하는 방법을 개발하는 것이 필요하다. 종래, 간엽계 간세포에 있어서, 그 세포를 특징짓는 마커유전자의 특정은 행해져 있지 않았다. 따라서, 간엽계 간세포를 특징짖는 마커유전자를 특정하고, 간엽계 간세포를 검출, 분리·식별할 수 있는 방법을 개발하는 것은, 간엽계 간세포를 재생의료에 실용화하는데 있어서 해결되어야 할 중요한 과제이다.
본 발명은, 간엽계 간세포를 검출, 식별하는 데에 사용하는 간엽계 간세포 검출용 유전자 마커 및 간엽계 간세포 검출용 폴리펩티드마커, 상기 유전자 마커를 검출하기 위한 PCR증폭용 프라이머, 및 상기 마커 및 프라이머를 사용한 간엽계 간세포의 검출과 식별방법을 제공하는 것에 있다.
본 발명자는, 간엽계 간세포 검출용의 마커가 되는 유전자를 찾아내기 위하여, 간엽계 간세포에 있어서의 여러가지 유전자의 발현에 대한 비교를 실시하고, 마커가 되는 유전자에 대하여 탐색한 결과, 배열표에 나타내는 13의 유전자에 관하여 간엽계 간세포와 섬유아세포와의 사이의 발현량에 차이가 발견되고, 간엽계 간세포에서 특이적으로 발현하고 있으며, 상기 유전자가 간엽계 간세포 검출용의 마커가 된다는 것을 발견하여 본 발명을 완성하는데 이르렀다. 본 발명은 상기 간엽계 간세포 마커유전자 검출용 프로브(probe), 및 상기 간엽계 간세포 유전자 마커의 검출에 있어서, 피검세포의 상기 유전자를 증폭하기 위한 PCR용 프라이머를 포함하는 것이다. 또, 본 발명은, 본 발명의 간엽계 간세포 마커유전자가 발현하는 폴리펩티드로 이루어지는 간엽계 간세포 검출용 폴리펩티드마커, 상기 폴리펩티드마커를 검출하기 위한 상기 폴리펩티드마커에 특이적으로 결합하는 항체를 포함하는 것이다. 또한, 본 발명은, 상기 간엽계 간세포 마커 유전자검출용 프로브, 폴리펩티드마커에 특이적으로 결합하는 항체를 사용한 간엽계 간세포의 식별방법 및 분리방법을 포함하는 것이다.
본원의 발명을 완성하기에 이른 경위에 대하여 설명하면, 각종의 간세포 중에서도 간엽계 간세포는, 뼈, 연골, 힘줄, 지방, 골격근, 심근, 혈관, 신경 등 많은 조직으로 분화하는 능력을 갖는 것이며, 재생용 세포로서의 기대도 크다. 그러나, 지금까지 간엽계 간세포를 특징짓는 마커유전자는 특정되어 있지 않았었다. 그래서, 본 발명자는, 간엽계 간세포를 특징짓는 마커유전자를 특정하기 위하여, 예의탐색을 실시하였다. 즉, 본 발명자는, 앞서 개발한 골수유래의 간엽계 간세포를 FGF등의 존재하에서 배양하는 것에 의하여, 그 분화능력을 유지한 채 폭발적으로 증식하는 방법을 사용하여 간엽계 간세포를 증식시키고, 이 방법에 의하여 얻은 간엽계 간세포와 인간섬유아세포를 FGF존재하에서 배양하여, 배양세포로부터 mRNA를 추출하고, 그 mRNA를 주형(鑄型)으로 하여 DNA칩(인사이트사 제품)을 프로브로 하여, DNA마이크로어레이법에 의하여 인간유래 간엽계 간세포와 섬유아세포에 발현하는 유전자를 비교하였다.
9400의 유전자 중 63유전자가 간엽계 간세포에 있어서 의미있게 발현이 높고, 141유전자가 의미있게 낮았다. 이 가운데, 발현량의 차가 3배 이상이었던 것은 87유전자(간엽계 간세포에서 높았던 유전자가 29유전자, 낮았던 유전자가 58유전자)였다. 3개체 유래의 간엽계 간세포와 3개체 유래의 섬유아세포로부터 전체 RNA를 추출하고, 이들을 주형으로 사용하여 상기 87유전자에 대하여 반정량적(半定量的) RT-PCR법에 의하여 그 발현량을 조사하였다. 그 결과, 발현량에 차이가 발견되지 않는 유전자가 대부분이었다. 또, 상이하는 개체의 간엽계 간세포, 또는 섬유아세포 사이에서도 발현량에 차이가 있고, 개체의 차이에 의하여 발현이 다르다고 생각되는 것도 많이 있었다.
RT-PCR에 의하여 간엽계 간세포에서 발현이 높은 것이 확인된 유전자는, 3유전자(tissue factor pathway inhibitor 2, major histocompatibility complex class2 DR beta 3, serine (or cysteine) proteinase inhibitor), 낮은 것이 확인된 유전자는 10종류(matrix metalloprotease 1, collagenase type XV alpha, CUG triplet repeat RNA binding protein, dermatopontin, protein tyrosine kinase 7, isocitrate dehydrogenase 2, Sam68-like phosphotyrosine protein, C-type lectin superfamily member 2, adrenomedullin, apolopoprotein D)였다. 이들 13유전자의 발현량을 측정하는 것에 의하여, 판별이 곤란했던 간엽계 간세포의 동정(同定)을 단시간에 또한 낮은 비용으로 실시할 수 있게 되었다. 또한, 본 발명에 있어서, 유전자의 탐색을 실시한 결과, 간엽계 간세포 또는 섬유아세포의 사이에서 발현량에 차이가 있는 많은 유전자를 확인하였다. 이들 유전자는 간엽계 간세포 검출용의 유전자 마커가 되는 것이며, 유전자 검출용 프로브를 구축하여 마이크로어레이 또는 DNA칩 등으로 하여, 간엽계 간세포의 검출에 사용할 수가 있는 것이다.
도1은, 본 발명의 실시예에 있어서, 반정량적 RT-PCR법에 의하여, 인간 간엽계 간세포 또는 인간 섬유아세포의 사이에서, 개체에 의한 불규칙성이 큰 유전자를 나타내는 도면.
도2는, 본 발명의 실시예에 있어서, 반정량적 RT-PCR에 의하여 MSC에서 발현이 높았던 유전자를 나타내는 도면.
도3은, 본 발명의 실시예에 있어서, 반정량적 RT-PCR에 의하여 MSC에서 발현이 낮았던 유전자를 나타내는 도면.
도4는, 간엽계 간세포에서 발현이 높음/낮음이 확인된 13유전자에 대하여, 7개체 유래의 간엽계 간세포와 4개체 유래의 섬유아세포로부터 추출한 전(全) RNA를 주형으로 한 반정량적 RT-PCR법에 의하여, 그 발현량의 확인을 실시한 결과를 나타내는 도면.
도5는, 본 발명의 실시예에 있어서, 간엽계 간세포와 섬유아세포에 있어서의 본 발명의 마커 유전자의 발현상황을 시험한 결과를 나타내는 도면.
도6은, 본 발명의 실시예에 있어서, 간엽계 간세포와 골아세포에 있어서의 각 마커 유전자의 디퍼렌셜한 유전자 발현의 상황을 나타내는 도면.
도7은, 본 발명의 실시예에 있어서, 간엽계 간세포와 골아세포에 있어서의 각 마커 유전자의 디퍼렌셜한 유전자 발현의 상황을 나타내는 도면.
도8은, 본 발명의 실시예에 있어서, 간엽계 간세포와 골아세포에 있어서의 각 마커 유전자의 디퍼렌셜한 유전자 발현의 상황을 나타내는 도면.
도9는, 본 발명의 실시예에 있어서, 각 마커 유전자에 대하여 간엽계 간세포와 인간섬유아세포에 있어서의 유전자 발현의 상황을 나타내는 도면.
도10은 본 발명의 실시예에 있어서, 각 마커 유전자에 대하여 간엽계 간세포와 인간섬유아세포에 있어서의 유전자 발현의 상황을 나타내는 도면.
도11은, 본 발명의 실시예에 있어서, 인간세포에 있어서의 간엽계 간세포 DR의 발현을 플로 사이토메트리(flow cytometry)를 사용하여 측정한 결과를 나타내는 도면.
도12는, 본 발명의 실시예에 있어서, 간엽계 간세포의 골아세포에로의 분화유도에 있어서의 각 유전자 발현의 상황을 측정한 결과를 나타내는 도면.
도13은, 본 발명의 실시예에 있어서, 각 마커 유전자에 대하여 골분화유도 4일~28일에서의 유전자 발현의 변화를 나타내는 도면.
도14는, 본 발명의 실시예에 있어서, 각 마커 유전자에 대하여 골분화유도 4일~28일에서의 유전자 발현의 변화를 나타내는 도면.
도15는, 본 발명의 실시예에 있어서, 각 마커 유전자에 대하여 골분화유도 4일~28일에서의 유전자 발현의 변화를 나타내는 도면.
도16은, 본 발명의 실시예에 있어서, 각 마커 유전자에 대하여 골분화유도 4일~28일에서의 유전자 발현의 변화를 나타내는 도면.
구체적으로 본 발명은, 배열표의 배열번호 1, 2, 4, 6, 7, 9, 10, 11, 12, 14, 15, 17, 19 또는 46에 나타내는 염기배열을 갖는 유전자인 것을 특징으로 하는 간엽계 간세포 검출용 유전자 마커(청구항1), 유전자 INTEGRIN, ALPHA 6 (ITGA6), MRNA (NM_000210) ; 유전자 SOLUTE CARRIER FAMILY 20 (PHOSPHATE TRANSPORTER), MEMBER 1 (NM_005415) ; 유전자 RIBONUCLEOTIDE REDUCTASE M2 POLYPEPTIDE (RRM2), MRNA (NM_001034) ; 유전자 FOLLISTATIN (FST), TRANSCRIPT VARIANT FST317, MRNA (NM_006350) ; 유전자 SPROUTY (DROSOPHILA) HOMOLOG 2 (SPRY2), MRNA(NM_005842) ; 유전자 RAB3B, MEMBER RAS ONCOGENE FAMILY (RAB3B), MRNA (NM_002867) ; 유전자 SOLUTE CARRIER FAMILY 2 (FACILITATED GLUCOSE TRANSPORTER) (NM_006516) ; 유전자 INTERLEUKIN 13 RECEPTOR, ALPHA 2 (IL13RA2), MRNA (NM_000640) ; 유전자 SERINE/THREONINE KINASE 12 (STK12), MRNA (NM_004217) ; 유전자 MINICHROMOSOME MAINTENANCE DEFICIENT (S. CEREVISIAE) 5 (CE)(NM_006739) ; 유전자 THYROID HORMONE RECEPTOR INTERACTOR 13 (TRIP13), MRNA(NM_004237) ; 유전자 KINESIN-LIKE 6 (MITOTIC CENTROMERE-ASSOCIATED KINESIN) (K)(NM_006845) ; 유전자 CYCLIN-DEPENDENT KINASE INHIBITOR 3 (CDK2-ASSOCIATED DUAL(NM_005192) ; 유전자 CHROMOSOME CONDENSATION PROTEIN G (HCAP-G),MRNA(NM_022346) ; 유전자 CDC28 PROTEIN KINASE 1 (CKS1), MRNA(NM_001826) ; 유전자 PROTEIN REGULATOR OF CYTOKINESIS 1 (PRC1), MRNA(NM_003981) ; 유전자 CELL DIVISION CYCLE 2, G1 TO S AND G2 TO M (CDC2), MRNA(NM_001786) ; 유전자 CDC20 (CELL DIVISION CYCLE 20, S. CEREVISIAE, HOMOLOG) (CD)(NM_001255) ; 유전자 LIKELY ORTHOLOG OF MATERNAL EMBRYONIC LEUCINE ZIPPER KINAS(NM_014791) ; 유전자 MINICHROMOSOME MAINTENANCE DEFICIENT (S. CEREVISIAE) 7 (M)(NM_005916) ; 유전자 CYCLIN A2 (CCNA2), MRNA (NM_001237) ; 유전자 THYMIDINE KINASE 1, SOLUBLE (TK1), MRNA (NM_003258) ; 또는 유전자 cyclin-dependent kinase inhibitor 2A (melanoma, p16, inhibits CDK4) (CDKN2A), mRNA (NM_000077)인 것을 특징으로 하는 간엽계 간세포 검출용 유전자 마커(청구항2), 유전자 EGF-containing fibulin-like extracellular matrix protein 1 (NM_018894) ; 유전자 Human insulin-like growth factor binding protein 5 (IGFBP5) mRNA (AU132011) ; 유전자 Homo sapiens clone 24775 mRNA sequence (AA402981) ; proteoglycan 1, secretory granule (AV734015) ; 유전자 insulin-like growth factor binding protein 5 (AA374325) ; 유전자 solute carrier family 21 (organic anion transporter), member 3 (AF085224) ; 유전 transglutaminase 2 (C polypeptide, protein-glutamine-gamma-glutamyl transferase) (AL552373) ; 유전자 coagulation factor II (thrombin) receptor (M62424) ; 유전자 plasminogen activator, urokinase (NM_002658) ; 유전자 tissue inhibitor of metalloproteinase 3 (Sorsby fundus dystrophy, pseudoinflammatory) (W96324) ; 유전자 matrix Gla protein (BF668572) ; 유전자 B-cell CLL/lymphoma 1 (Z23022) ; 유전자 CD74 antigen (invariant polypeptide of major histocompatibility complex, class II antigen-associated) (BG333618) ; 유전자 adducin 3 (gamma) (AL135243) ; 유전자 solute carrier family 16 (monocarboxylic acid transporters), member 4 (NM_004696) ; 유전자 protein S (alpha) (NM_000313) ; 유전자 phosphatidylinositol (4,5) bisphosphate 5-phosphatase homolog; phosphatidylinositol polyphosphate 5-phosphatase type IV (AF187891) ; 유전자 progesterone membrane binding protein (BE858855) ; 유전자 brain-derived neurotrophic factor (X60201) ; 또는 유전자 apolipoprotein E (BF967316)인 것을 특징으로 하는 간엽계 간세포 검출용 유전자 마커(청구항3), 유전자 HOMO SAPIENS WNT INHIBITORY FACTOR-1 (WIF-1), MRNA (NM_007191) ; 유전자 HOMO SAPIENS HYPOTHETICAL PROTEIN FLJ11175 (FLJ11175), MRNA (NM_018349) ; 유전자 HOMO SAPIENS RHO GDP DISSOCIATION INHIBITOR (GDI) BETA (ARHGDIB), MRNA (NM_001175) ; 유전자 HOMO SAPIENS POTASSIUM INTERMEDIATE/SMALL CONDUCTANCE CALCIUM-ACTIVATE (NM_021614) ; 유전자 HOMO SAPIENS NUCLEAR FACTOR (ERYTHROID-DERIVED 2)-LIKE 3 (NFE2L3), MRNA (NM_004289) ; 유전자 HOMO SAPIENS GS3955 PROTEIN (GS3955), MRNA (NM_021643) ; 유전자 HOMO SAPIENS CDNA 3' END SIMILAR TO SIMILAR TO GLYCOPROTEIN MUC18 (AA302605) ; 유전자 HOMO SAPIENS HYPOTHETICAL PROTEIN FLJ21841 (FLJ21841), MRNA (NM_024609) ; 유전자 HOMO SAPIENS ZINC FINGER PROTEIN 185 (LIM DOMAIN) (ZNF185), MRNA (NM_007150) ; 유전자 SC1=PUTATIVE TRANS-ACTING FACTOR INVOLVED IN CELL CYCLE CONTROL [HUMAN, MRNA, 24] (S53374) ; 유전자 HOMO SAPIENS NADH DEHYDROGENASE (UBIQUINONE) 1 ALPHA SUBCOMPLEX, 8 (19K) (NM_014222) ; 유전자 HOMO SAPIENS HOMOLOG OF YEAST MCM10; HYPOTHETICAL PROTEIN PRO2249 (PRO2) (NM_018518) ; 유전자 HOMO SAPIENS COLLAGEN, TYPE VII, ALPHA 1 (EPIDERMOLYSIS BULLOSA, DYSTRO(NM_000094) ; 유전자 HOMO SAPIENS AUTOCRINE MOTILITY FACTOR RECEPTOR (AMFR), MRNA(NM_001144) ; 유전자 HOMO SAPIENS CLONE 24421 MRNA SEQUENCE /CDS=UNKNOWN /GB=AF070641 /GI=3283914 /UG=(AF070641) ; 유전자 HOMO SAPIENS MRNA;유전자 CDNA DKFZP586E1621 (FROM CLONE DKFZP586E1621) /CDS=UNKNOWN /GB(AL080235)인 것을 특징으로 하는 간엽계 간세포 검출용 유전자 마커(청구항4), 유전자 HOMO SAPIENS RESERVED (KCNK12), MRNA (NM_022055) ; 유전자 HOMO SAPIENS UBIQUITIN-CONJUGATING ENZYME E2C (UBE2C), MRNA(NM_007019) ; 유전자 HOMO SAPIENS CDNA FLJ10517 FIS, CLONE NT2RP2000812 /CDS=(61,918) /GB=AK001379 /G(AK001379) ; 유전자 HOMO SAPIENS CARTILAGE LINKING PROTEIN 1 (CRTL1), MRNA(NM_001884) ; 유전자 HOMO SAPIENS MRNA; CDNA DKFZP434F2322 (FROM CLONE DKFZP434F2322) /CDS=UNKNOWN /GB(AL133105) ; 유전자 HOMO SAPIENS, CLONE MGC:9549 IMAGE:3857382, MRNA, COMPLETE CDS /CDS=(92,1285) /G (BC012453) ; 유전자 HOMO SAPIENS HYPOTHETICAL PROTEIN DKFZP586J1119 (DKFZP586J1119), MRNA(NM_032270) ; 유전자 HOMO SAPIENS GLIA MATURATION FACTOR, GAMMA (GMFG), MRNA (NM_004877) ; 유전자 HOMO SAPIENS CDNA: FLJ23095 FIS, CLONE LNG07413 /CDS=UNKNOWN /GB=AK026748 /GI=10(AK026748) ; 유전자 HOMO SAPIENS HYPOTHETICAL PROTEIN DKFZP762E1312 (DKFZP762E1312), MRNA (NM_018410) ; 유전자 HOMO SAPIENS IROQUOIS-CLASS HOMEODOMAIN PROTEIN (IRX-2A), MRNA (NM_005853) ; 또는 유전자 HOMO SAPIENS HYPOTHETICAL PROTEIN FLJ10540 (FLJ10540), MRNA(NM_018131)인 것을 특징으로 하는 간엽계 간세포 검출용 유전자 마커(청구항5), 유전자 HOMO SAPIENS MRNA; CDNA DKFZP434C1915 (FROM CLONE DKFZP434C1915); PARTIAL CDS /C(AL137698) ; 유전자 HOMO SAPIENS MINICHROMOSOME MAINTENANCE DEFICIENT (S. CEREVISIAE) 7 (M)(NM_005916) ; 유전자 HOMO SAPIENS DYNEIN, CYTOPLASMIC, INTERMEDIATE POLYPEPTIDE 1 (DNCI1), M (NM_004411) ; 유전자 HUMAN MRNA FOR PROTEIN GENE PRODUCT (PGP) 9.5. (X04741) ; 유전자 HOMO SAPIENS RAD51-INTERACTING PROTEIN (PIR51), MRNA (NM_006479) ; 유전자 HOMO SAPIENS BACULOVIRAL IAP REPEAT-CONTAINING 5 (SURVIVIN) (BIRC5), MR (NM_001168) ; 유전자 HOMO SAPIENS UDP-N-ACETYL-ALPHA-D-GALACTOSAMINE:POLYPEPTIDE N-ACETYLGAL (NM_004482) ; 유전자 HOMO SAPIENS CYCLIN B1 (CCNB1), MRNA (NM_031966) ; 유전자 HOMO SAPIENS FLAP STRUCTURE-SPECIFIC ENDONUCLEASE 1 (FEN1), MRNA( NM_004111) ; 유전자 HOMO SAPIENS SERINE/THREONINE KINASE 15 (STK15), MRNA (NM_003600) ; 유전자 HOMO SAPIENS MAD2 (MITOTIC ARREST DEFICIENT, YEAST, HOMOLOG)-LIKE 1 (MA) (NM_002358) ; 유전자 HOMO SAPIENS NUCLEOLAR PROTEIN ANKT (ANKT), MRNA (NM_018454) ; 유전자 HOMO SAPIENS HSPC150 PROTEIN SIMILAR TO UBIQUITIN-CONJUGATING ENZYME (H) (NM_014176) ; 유전자 HOMO SAPIENS CYTOCHROME P450 RETINOID METABOLIZING PROTEIN (P450RAI-2), (NM_019885) 유전자 HOMO SAPIENS NIMA (NEVER IN MITOSIS GENE A)-RELATED KINASE 2 (NEK2), MR（NM_002497）；유전자 HOMO SAPIENS BONE MORPHOGENETIC PROTEIN 4 (BMP4), MRNA（NM_001202）；유전자 HOMO SAPIENS CDNA FLJ10674 FIS, CLONE NT2RP2006436 /CDS=UNKNOWN /GB=AK001536 /GI（AK001536）；또는 유전자 HOMO SAPIENS CENTROMERE PROTEIN A (17KD) (CENPA), MRNA（NM_001809）인 것을 특징으로 하는 간엽계 간세포 검출용 유전자 마커(청구항6), 유전자 HOMO SAPIENS MRNA; CDNA DKFZP564P116 (FROM CLONE DKFZP564P116) /CDS=UNKNOWN /GB=A（AL049338）；유전자 HOMO SAPIENS TTK PROTEIN KINASE (TTK), MRNA（NM_003318）；유전자 HOMO SAPIENS KARYOPHERIN ALPHA 2 (RAG COHORT 1, IMPORTIN ALPHA 1) (KPNA（NM_002266）；유전자 HOMO SAPIENS POLYMERASE (DNA DIRECTED), THETA (POLQ), MRNA（NM_006596）；유전자 HOMO SAPIENS ETS VARIANT GENE 5 (ETS-RELATED MOLECULE) (ETV5), MRNA（NM_004454）；유전자 HOMO SAPIENS TRANSFORMING, ACIDIC COILED-COIL CONTAINING PROTEIN 3 (TAC)（NM_006342）；유전자 HOMO SAPIENS KINESIN-LIKE 1 (KNSL1), MRNA（NM_004523）；유전자 HOMO SAPIENS CENTROMERE PROTEIN E (312KD) (CENPE), MRNA（NM_001813）；유전자 HOMO SAPIENS CDNA, 3' END /CLONE=IMAGE:1651289 /CLONE_END=3' /GB=AI12（AI123815）；유전자 HOMO SAPIENS DISTAL-LESS HOMEO BOX 5 (DLX5), MRNA（NM_005221）；유전자 HOMO SAPIENS V-MYB AVIAN MYELOBLASTOSIS VIRAL ONCOGENE HOMOLOG-LIKE 2 （NM_002466）；또는 유전자 HOMO SAPIENS SERUM DEPRIVATION RESPONSE (PHOSPHATIDYLSERINE-BINDING PRO（NM_004657）인 것을 특징으로 하는 간엽계 간세포 검출용 유전자 마커(청구항7), 유전자 HOMO SAPIENS HYPOTHETICAL PROTEIN FLJ10604 (FLJ10604), MRNA（NM_018154）；유전자 HOMO SAPIENS RIBONUCLEASE HI, LARGE SUBUNIT (RNASEHI), MRNA（NM_006397）；유전자 HOMO SAPIENS ANILLIN (DROSOPHILA SCRAPS HOMOLOG), ACTIN BINDING PROTEIN（NM_018685）；유전자 HOMO SAPIENS H4 HISTONE FAMILY, MEMBER G (H4FG), MRNA（NM_003542）；유전자 HOMO SAPIENS G PROTEIN-COUPLED RECEPTOR 37 (ENDOTHELIN RECEPTOR TYPE B-（NM_005302）HOMO SAPIENS UBIQUITIN CARRIER PROTEIN (E2-EPF), MRNA（NM_014501）；유전자 HOMO SAPIENS, SIMILAR TO TUMOR DIFFERENTIALLY EXPRESSED 1, CLONE IMAGE:3639252, （BC007375）；유전자 HOMO SAPIENS HYPOTHETICAL PROTEIN MGC2577 (MGC2577), MRNA（NM_031299）；유전자 HOMO SAPIENS HYPOTHETICAL PROTEIN FLJ13912 (FLJ13912), MRNA（NM_022770）；유전자 HOMO SAPIENS ANNEXIN A3 (ANXA3), MRNA（NM_005139）；유전자 HOMO SAPIENS STATHMIN 1/ONCOPROTEIN 18 (STMN1), MRNA（NM_005563）HOMO SAPIENS, SIMILAR TO RIBOSOMAL PROTEIN L39, CLONE MGC:20168 IMAGE:4555759, M（BC012328）；유전자 HOMO SAPIENS POLO (DROSOPHIA)-LIKE KINASE (PLK), MRNA（NM_005030）인 것을 특징으로 하는 간엽계 간세포 검출용 유전자 마커(청구항8), 유전자 HOMO SAPIENS PROTEASOME (PROSOME, MACROPAIN) 26S SUBUNIT, ATPASE, 5 (PS)（NM_002805）；유전자 HOMO SAPIENS EUKARYOTIC TRANSLATION INITIATION FACTOR 4 GAMMA, 2 (EIF4G)（NM_001418）；유전자 HOMO SAPIENS DIHYDROPYRIMIDINASE-LIKE 3 (DPYSL3), MRNA）（NM_001387）；유전자 HOMO SAPIENS ADAPTOR-RELATED PROTEIN COMPLEX 1, SIGMA 2 SUBUNIT (AP1S2)（NM_003916）；유전자 HOMO SAPIENS PLECTIN 1, INTERMEDIATE FILAMENT BINDING PROTEIN, 500KD (P)（NM_000445）
；유전자 HOMO SAPIENS HETEROGENEOUS NUCLEAR RIBONUCLEOPROTEIN U (SCAFFOLD ATTAC（NM_031844）；유전자 HOMO SAPIENS HYPOTHETICAL PROTEIN MGC5306 (MGC5306), MRNA（NM_024116）；유전자 HOMO SAPIENS MHC CLASS I POLYPEPTIDE-RELATED SEQUENCE A (MICA), MRNA（NM_000247）；유전자 HOMO SAPIENS PROTEIN ASSOCIATED WITH PRK1 (AWP1), MRNA（NM_019006）；유전자 HOMO SAPIENS PROGESTERONE RECEPTOR MEMBRANE COMPONENT 2 (PGRMC2), MRNA（NM_006320）；유전자 HOMO SAPIENS POLYGLUTAMINE BINDING PROTEIN 1 (PQBP1), MRNA（NM_005710）；유전자 HOMO SAPIENS S-ADENOSYLMETHIONINE DECARBOXYLASE 1 (AMD1), MRNA（NM_001634）； HOMO SAPIENS INTERFERON-INDUCED, HEPATITIS C-ASSOCIATED MICROTUBULAR AG（NM_006417）；유전자 HOMO SAPIENS PROTEIN KINASE, AMP-ACTIVATED, GAMMA 1 NON-CATALYTIC SUBUN（NM_002733）；HOMO SAPIENS PROCOLLAGEN-PROLINE, 2-OXOGLUTARATE 4-DIOXYGENASE (PROLINE（NM_004199）；유전자 HOMO SAPIENS ENDOTHELIAL DIFFERENTIATION, LYSOPHOSPHATIDIC ACID G-PROTE（NM_012152）；또는 유전자 HOMO SAPIENS KIAA0127 GENE PRODUCT (KIAA0127), MRNA（NM_014755）인 것을 특징으로 하는 간엽계 간세포 검출용 유전자 마커(청구항8), 청구항1~9 중의 어느 한 항에 기재된 간엽계 간세포 검출용 마커 유전자와 스트린젠트한 조건(stringent condition) 하에서 하이브리다이즈(hybridige)하는 DNA배열을 갖는 간엽계 간세포 마커 유전자 검출용 프로브(청구항10), 청구항1~9 중의 어느 한 항에 기재된 염기배열의 안티센스 사슬의 전부 또는 일부로 이루어지는 청구항10에 기재된 간엽계 간세포 마커 유전자 검출용 프로브(청구항11), 청구항 10 또는 11에 기재된 DNA의 적어도 1개 이상을 고정화시킨 것을 특징으로 하는 간엽계 간세포 마커 유전자 검출용 마이크로어레이 또는 DNA칩(청구항12), 배열표의 배열번호 1, 2, 4, 6, 7, 9, 10, 11, 12, 14, 15, 17, 19 또는 46에 나타내는 염기배열을 갖는 유전자의 그룹, 및 청구항 2~9중 어느 한 항에 기재된 유전자 그룹의 2이상의 유전자를 검출하는 프로브를 고정화시킨 것을 특징으로 하는 청구항12 기재의 간엽계 간세포 마커 유전자 검출용 마이크로어레이 또는 DNA칩(청구항13), 배열표의 배열번호 3, 5, 8, 13, 16 또는 18에 나타내는 아미노산 배열을 갖는 폴리펩티드인 것을 특징으로 하는 간엽계 간세포 검출용 폴리펩티드 마커(청구항14), 청구항14에 기재된 폴리펩티드를 사용하여 유도되며, 상기 폴리펩티드에 특이적으로 결합하는 것을 특징으로 하는 항체(청구항15), 상기 항체가 모노클로널항체인 것을 특징으로 하는 청구항15에 기재된 항체(청구항16), 상기 항체가 폴리클로널항체인 것을 특징으로 하는 청구항15에 기재된 항체(청구항17)로 이루어진다.
또, 본 발명은 피검세포에 있어서의 청구항1~9 중의 어느 한 항에 기재된 간엽계 간세포 마커 유전자의 발현을 검출하는 것을 특징으로 하는 간엽계 간세포의 식별방법(청구항18), 간엽계 간세포 마커 유전자 발현의 검출을 노던 블로팅법을 사용하여 실시하는 것을 특징으로 하는 청구항18에 기재된 간엽계 간세포의 식별방법(청구항19), 간엽계 간세포 마커 유전자 발현의 검출을 청구항10 또는 11에 기재된 간엽계 간세포 마커 유전자 검출용 프로브를 사용하여 실시하는 것을 특징으로 하는 청구항18에 기재된 간엽계 간세포의 식별방법(청구항20), 간엽계 간세포 마커 유전자 발현의 검출을 청구항12 또는 13에 기재된 간엽계 간세포 마커 유전자 검출용 마이크로어레이 또는 DNA칩을 사용하여 실시하는 것을 특징으로 하는 청구항18에 기재된 간엽계 간세포의 식별방법(청구항21), 청구항18~21 중 어느 한 항에 기재된 간엽계 간세포 마커 유전자 발현의 검출이, 정량적 또는 반정량적 PCR의 사용을 포함하고 있는 것을 특징으로 하는 간엽계 간세포의 식별방법(청구항22), 정량적 또는 반정량적 PCR의 사용이 RT-PCR법인 것을 특징으로 하는 청구항22에 기재된 간엽계 간세포의 식별방법(청구항23)으로 이루어진다.
또한, 본 발명은, 배열표의 배열번호20에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호21에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호22에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호23에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호24에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호25에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호26에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호27에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호28에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호29에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호30에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호31에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호32에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호33에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호34에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호35에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호36에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호37에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호38에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호39에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호40에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호41에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호42에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호43에 나타내는 염기배열을 갖는 안티센스프라이머, 또는 배열표의 배열번호44에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호45에 나타내는 염기배열을 갖는 안티센스프라이머로 이루어지는 간엽계 간세포 마커 유전자 증폭을 위한 RT-PCR용 프라이머(청구항24), 배열표의 배열번호47에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호48에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호49에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호50에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호51에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호52에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호53에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호54에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호55에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호56에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호57에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호58에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호59에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호60에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호61에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호62에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호63에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호64에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호65에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호66에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호67에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호68에 나타내는 염기배열을 갖는 안티센스프라이머, 또는 배열표의 배열번호69에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호70에 나타내는 염기배열을 갖는 안티센스프라이머로 이루어지는 간엽계 간세포 마커 유전자 증폭을 위한 실시간 PCR용 프라이머(청구항25), 배열표의 배열번호71에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호72에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호73에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호74에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호75에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열표의 배열번호76에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호77에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호78에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호79에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호80에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호81에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호82에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호83에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호84에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호85에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호86에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호87에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호88에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호89에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호90에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호91에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호92에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호93에 나타내는 염기배열은 갖는 센스프라이머와 배열표의 배열번호94에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호95에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호96에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호97에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호98에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호99에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호 100에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호101에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호102에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호103에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호104에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호105에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호106에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호107에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호108에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호109에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호110에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호111에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호112에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호113에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호114에 나타내는 염기배열을 갖는 안티센스프라이머, 또는 배열표의 배열번호115에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호116에 나타내는 염기배열을 갖는 안티센스프라이머로 이루어지는 간엽계 간세포 마커 유전자 증폭을 위한 RT-PCR용 프라이머(청구항26)로 이루어진다.
또, 본 발명은 피검세포에 있어서의 유전자를, 청구항24~26 중의 어느 한 항에 기재된 센스프라이머 및 안티센스프라이머의 적어도 1쌍 이상의 프라이머를 사용하여 증폭하며, 증폭한 유전자를 청구항10 또는 11에 기재된 간엽계 간세포 마커 유전자 검출용 프로브를 사용하여 검출하는 것을 특징으로 하는 청구항18에 기재된 간엽계 간세포의 식별방법(청구항27), 청구항15~17 중의 어느 한 항에 기재된 항체를 사용하여 피검세포에 있어서의 간엽계 간세포 마커 폴리펩티드의 발현을 검출하는 것을 특징으로 하는 간엽계 간세포의 식별방법(청구항28), 피검세포에 있어서의 배열표의 배열번호 1, 11 및 19로 이루어지는 간엽계 간세포 마커 유전자 그룹의 1유전자 이상의 발현과, 배열표의 배열번호 2, 4, 6, 7, 9, 10, 12, 14, 15 및 17로 이루어지는 간엽계 간세포 마커 유전자 그룹의 1유전자 이상의 발현을 조합하여 평가하는 것을 특징으로 하는 간엽계 간세포의 식별방법(청구항29), 청구항10 또는 11에 기재된 간엽계 간세포 마커 유전자 검출용 프로브 및/또는 청구항15~17 중의 어느 한 항에 기재된 항체를 사용하여 피검세포에 있어서의 간엽계 간세포 검출용 마커 유전자 및/또는 간엽계 간세포 검출용 마커 폴리펩티드의 발현을 검출하며, 식별한 간엽계 간세포를 분리하는 것을 특징으로 하는 간엽계 간세포의 식별분리방법(청구항30), 청구항15~17 중의 어느 한 항에 기재된 항체를 사용한 형광항체법에 의하여 피검세포 중에 있어서의 간엽계 간세포를 표지화하고, 상기 표지화한 간엽계 간세포를 분리하는 것을 특징으로 하는 청구항30에 기재된 간엽계 간세포의 식별분리방법(청구항31), 청구항10 또는 11에 기재된 간엽계 간세포 마커 유전자 검출용 프로브, 상기 프로브를 고정한 청구항12 또는 13에 기재된 간엽계 간세포 마커 유전자 검출용 마이크로어레이 또는 DNA칩, 및 청구항15~17 중의 어느 한 항에 기재된 항체의 적어도 하나 이상을 장비하여 이루어지는 간엽계 간세포의 식별용 키트(청구항32)로 이루어진다.
본 발명은, 간엽계 간세포에서 특이적으로 발현하는 간엽계 간세포 검출용 마커 유전자 또는 간엽계 간세포 검출용 마커 폴리펩티드의 발현을 검출하여 간엽계 간세포의 식별을 실시하는 것에 의하여 이루어진다. 본 발명에 있어서, 간엽계 간세포 검출용 유전자 마커가 되는 유전자는, 배열표의 배열번호 1, 2, 4, 6, 7, 9, 10, 11, 12, 14, 15, 17 또는 19에 나타내는 염기배열을 갖는 유전자이다.
또한, 본 발명에 있어서, 간엽계 간세포 검출용의 폴리펩티드 마커가 되는 폴리펩티드는, 배열표의 배열번호 3, 5, 8, 13, 16 또는 18에 나타내는 아미노산 배열을 갖는 폴리펩티드이다.
배열표의 배열번호1에 나타나는 유전자는［serine (or cysteine) proteinase inhibitor］의 유전자이며, 배열표의 배열번호2에 나타내는 유전자는［adrenomedullin］의 유전자이며, 배열표의 배열번호4에 나타내는 유전자는［apolopoprotein D］의 유전자이며, 배열표의 배열번호6에 나타내는 유전자는［collagenase type XV alpha 1］의 유전자이며, 배열표의 배열번호7에 나타내는 유전자는［CUG triplet repeat RNA binding protein2］의 유전자이며, 배열표의 배열번호9에 나타내는 유전자는［dermatopontin］의 유전자이며, 배열표의 배열번호10에 나타내는 유전자는［isocitrate dehydrogenase 2］의 유전자이며, 배열표의 배열번호11에 나타내는 유전자는［major histocompatibility complex class2, DR beta 3］의 유전자이며, 배열표의 배열번호12에 나타내는 유전자는［protein tyrosine kinase 7］의 유전자이며, 배열표의 배열번호14에 나타내는 유전자는［Sam68-like phosphotyrosine protein］의 유전자이며, 배열표의 배열번호15에 나타내는 유전자는［C-type lectin superfamily member 2］의 유전자이며, 배열표의 배열번호17에 나타내는 유전자는［matrix metalloprotease 1］의 유전자이며, 또, 배열표의 배열번호19에 나타내는 유전자는［tissue factor pathway inhibitor 2］의 유전자이며, 배열표의 배열번호46에 나타내는 유전자는［major histocompatibility complex DR alpha］의 유전자이다.
본 발명의 간엽계 간세포 검출용 마커유전자의 DNA배열정보는, NCBI유전자 데이터 베이스에 있어서, 각각 액세션넘버:ＡＩ１３３６１３(배열번호1), ＮＭ＿００１１２４(배열번호2), ＮＭ＿００１６４７（배열번호４）、Ｌ０１６９７（배열번호６）、Ｕ６９５４６（배열번호７）、ＡＷ０１６４５１（배열번호９）、ＡＬ５４５９５３（배열번호１０）、ＢＦ７３２８２２（배열번호１１）、ＡＬ１５７４８６（배열번호１２）、ＡＡ１１２００１（배열번호１４）、ＸＭ＿００６６２６（배열번호１５）、ＮＭ＿００２４２１（배열번호１７）、ＡＬ５５０３５７（배열번호１９）、및 ＢＦ７９５９２９(배열번호４６)에 의하여 접근할 수가 있다.
또한 본 발명에 있어서 확인된, 섬유아세포에 비교하여, 간엽계 간세포에서 다량으로 발현하고 있는 유전자로서는, 이하의 것을 들 수 있다 :
유전자 INTEGRIN, ALPHA 6 (ITGA6), MRNA（NM_000210）；유전자 SOLUTE CARRIER FAMILY 20 (PHOSPHATE TRANSPORTER)、MEMBER 1（NM_005415）；유전자 RIBONUCLEOTIDE REDUCTASE M2 POLYPEPTIDE (RRM2), MRNA（NM_001034）；유전자 FOLLISTATIN (FST), TRANSCRIPT VARIANT FST317, MRNA（NM_006350）；유전자 SPROUTY (DROSOPHILA) HOMOLOG 2 (SPRY2), MRNA（NM_005842）；유전자 RAB3B, MEMBER RAS ONCOGENE FAMILY (RAB3B), MRNA（NM_002867）；유전자 SOLUTE CARRIER FAMILY 2 (FACILITATED GLUCOSE TRANSPORTER)（NM_006516）；유전자 INTERLEUKIN 13 RECEPTOR, ALPHA 2 (IL13RA2), MRNA（NM_000640）；유전자 SERINE/THREONINE KINASE 12 (STK12), MRNA（NM_004217）；유전자 MINICHROMOSOME MAINTENANCE DEFICIENT (S. CEREVISIAE) 5 (CE)（NM_006739）；유전자 THYROID HORMONE RECEPTOR INTERACTOR 13 (TRIP13), MRNA（NM_004237）；유전자 KINESIN-LIKE 6 (MITOTIC CENTROMERE-ASSOCIATED KINESIN) (K)（NM_006845）；유전자 CYCLIN-DEPENDENT KINASE INHIBITOR 3 (CDK2-ASSOCIATED DUAL（NM_005192）；유전자 CHROMOSOME CONDENSATION PROTEIN G (HCAP-G),MRNA（NM_022346）；유전자 CDC28 PROTEIN KINASE 1 (CKS1), MRNA（NM_001826）；유전자 PROTEIN REGULATOR OF CYTOKINESIS 1 (PRC1), MRNA（NM_003981）；유전자 CELL DIVISION CYCLE 2, G1 TO S AND G2 TO M (CDC2), MRNA（NM_001786）；유전자 CDC20 (CELL DIVISION CYCLE 20, S. CEREVISIAE, HOMOLOG) (CD)（NM_001255）；유전자 LIKELY ORTHOLOG OF MATERNAL EMBRYONIC LEUCINE ZIPPER KINAS（NM_014791）；유전자 MINICHROMOSOME MAINTENANCE DEFICIENT (S. CEREVISIAE) 7 (M)（NM_005916）；유전자 CYCLIN A2 (CCNA2), MRNA（NM_001237）；유전자 THYMIDINE KINASE 1, SOLUBLE (TK1), MRNA（NM_003258）；또는 유전자 cyclin-dependent kinase inhibitor 2A (melanoma, p16, inhibits CDK4) (CDKN2A), mRNA（NM_000077）。
유전자 EGF-containing fibulin-like extracellular matrix protein 1（NM_018894）；유전자 Human insulin-like growth factor binding protein 5 (IGFBP5) mRNA（AU132011）；유전자 Homo sapiens clone 24775 mRNA sequence（AA402981）；proteoglycan 1, secretory granule（AV734015）；유전자 insulin-like growth factor binding protein 5（AA374325）；유전자 solute carrier family 21 (organic anion transporter), member 3（AF085224）；유전 transglutaminase 2 (C polypeptide, protein-glutamine-gamma-glutamyl transferase)（AL552373）；유전자 coagulation factor II (thrombin) receptor（M62424）；유전자 plasminogen activator, urokinase（NM_002658）；유전자 tissue inhibitor of metalloproteinase 3 (Sorsby fundus dystrophy, pseudoinflammatory)（W96324）；유전자 matrix Gla protein（BF668572）；유전자 B-cell CLL/lymphoma 1（Z23022）；유전자 CD74 antigen (invariant polypeptide of major histocompatibility complex, class II antigen-associated)（BG333618）；유전자 adducin 3 (gamma)（AL135243）；유전자 solute carrier family 16 (monocarboxylic acid transporters), member 4（NM_004696）；유전자 protein S (alpha)（NM_000313）；유전자 phosphatidylinositol (4,5) bisphosphate 5-phosphatase homolog; phosphatidylinositol polyphosphate 5-phosphatase type IV（AF187891）；유전자 progesterone membrane binding protein（BE858855）；유전자 brain-derived neurotrophic factor（X60201）；또는 유전자 apolipoprotein E（BF967316）。
유전자 HOMO SAPIENS WNT INHIBITORY FACTOR-1 (WIF-1), MRNA（NM_007191）；유전자 HOMO SAPIENS HYPOTHETICAL PROTEIN FLJ11175 (FLJ11175), MRNA（NM_018349）；유전자 HOMO SAPIENS RHO GDP DISSOCIATION INHIBITOR (GDI) BETA (ARHGDIB), MRNA（NM_001175）；유전자 HOMO SAPIENS POTASSIUM INTERMEDIATE/SMALL CONDUCTANCE CALCIUM-ACTIVATE（NM_021614）；유전자 HOMO SAPIENS NUCLEAR FACTOR (ERYTHROID-DERIVED 2)-LIKE 3 (NFE2L3), MRNA（NM_004289）；유전자 HOMO SAPIENS GS3955 PROTEIN (GS3955), MRNA（NM_021643）；유전자 HOMO SAPIENS CDNA 3' END SIMILAR TO SIMILAR TO GLYCOPROTEIN MUC18（AA302605）；유전자 HOMO SAPIENS HYPOTHETICAL PROTEIN FLJ21841 (FLJ21841), MRNA（NM_024609）；유전자 HOMO SAPIENS ZINC FINGER PROTEIN 185 (LIM DOMAIN) (ZNF185), MRNA（NM_007150）；유전자 SC1=PUTATIVE TRANS-ACTING FACTOR INVOLVED IN CELL CYCLE CONTROL [HUMAN, MRNA, 24]（S53374）；유전자 HOMO SAPIENS NADH DEHYDROGENASE (UBIQUINONE) 1 ALPHA SUBCOMPLEX, 8 (19K)（NM_014222）；유전자 HOMO SAPIENS HOMOLOG OF YEAST MCM10; HYPOTHETICAL PROTEIN PRO2249 (PRO2)（NM_018518）；유전자 HOMO SAPIENS COLLAGEN, TYPE VII, ALPHA 1 (EPIDERMOLYSIS BULLOSA, DYSTRO（NM_000094）；유전자 HOMO SAPIENS AUTOCRINE MOTILITY FACTOR RECEPTOR (AMFR), MRNA（NM_001144）；유전자 HOMO SAPIENS CLONE 24421 MRNA SEQUENCE /CDS=UNKNOWN /GB=AF070641 /GI=3283914 /UG=（AF070641）；유전자 HOMO SAPIENS MRNA ; 유전자 CDNA DKFZP586E1621 (FROM CLONE DKFZP586E1621) /CDS=UNKNOWN /GB（AL080235）。
유전자 HOMO SAPIENS RESERVED (KCNK12), MRNA（NM_022055）；유전자 HOMO SAPIENS UBIQUITIN-CONJUGATING ENZYME E2C (UBE2C), MRNA（NM_007019）；유전자 HOMO SAPIENS CDNA FLJ10517 FIS, CLONE NT2RP2000812 /CDS=(61,918) /GB=AK001379 /G（AK001379）；유전자 HOMO SAPIENS CARTILAGE LINKING PROTEIN 1 (CRTL1), MRNA（NM_001884）；유전자 HOMO SAPIENS MRNA; CDNA DKFZP434F2322 (FROM CLONE DKFZP434F2322) /CDS=UNKNOWN /GB（AL133105）；유전자 HOMO SAPIENS, CLONE MGC:9549 IMAGE:3857382, MRNA, COMPLETE CDS /CDS=(92,1285) /G（BC012453）；유전자 HOMO SAPIENS HYPOTHETICAL PROTEIN DKFZP586J1119 (DKFZP586J1119), MRNA（NM_032270）；유전자 HOMO SAPIENS GLIA MATURATION FACTOR, GAMMA (GMFG), MRNA（NM_004877）；유전자 HOMO SAPIENS CDNA: FLJ23095 FIS, CLONE LNG07413 /CDS=UNKNOWN /GB=AK026748 /GI=10（AK026748）；유전자 HOMO SAPIENS HYPOTHETICAL PROTEIN DKFZP762E1312 (DKFZP762E1312), MRNA（NM_018410）；유전자 HOMO SAPIENS IROQUOIS-CLASS HOMEODOMAIN PROTEIN (IRX-2A), MRNA（NM_005853）；또는 유전자 HOMO SAPIENS HYPOTHETICAL PROTEIN FLJ10540 (FLJ10540), MRNA（NM_018131）。
유전자 HOMO SAPIENS MRNA; CDNA DKFZP434C1915 (FROM CLONE DKFZP434C1915); PARTIAL CDS /C（AL137698）；유전자 HOMO SAPIENS MINICHROMOSOME MAINTENANCE DEFICIENT (S. CEREVISIAE) 7 (M)（NM_005916）；유전자 HOMO SAPIENS DYNEIN, CYTOPLASMIC, INTERMEDIATE POLYPEPTIDE 1 (DNCI1), M（NM_004411）；유전자 HUMAN MRNA FOR PROTEIN GENE PRODUCT (PGP) 9.5.（X04741）；유전자 HOMO SAPIENS RAD51-INTERACTING PROTEIN (PIR51), MRNA（NM_006479）；유전자 HOMO SAPIENS BACULOVIRAL IAP REPEAT-CONTAINING 5 (SURVIVIN) (BIRC5), MR（NM_001168）；유전자 HOMO SAPIENS UDP-N-ACETYL-ALPHA-D-GALACTOSAMINE:POLYPEPTIDE N-ACETYLGAL（NM_004482）；유전자 HOMO SAPIENS CYCLIN B1 (CCNB1), MRNA（NM_031966）；유전자 HOMO SAPIENS FLAP STRUCTURE-SPECIFIC ENDONUCLEASE 1 (FEN1), MRNA（NM_004111）；유전자 HOMO SAPIENS SERINE/THREONINE KINASE 15 (STK15), MRNA（NM_003600）；유전자 HOMO SAPIENS MAD2 (MITOTIC ARREST DEFICIENT, YEAST, HOMOLOG)-LIKE 1 (MA)（NM_002358）；유전자 HOMO SAPIENS NUCLEOLAR PROTEIN ANKT (ANKT), MRNA（NM_018454）；유전자 HOMO SAPIENS HSPC150 PROTEIN SIMILAR TO UBIQUITIN-CONJUGATING ENZYME (H)（NM_014176）；유전자 HOMO SAPIENS CYTOCHROME P450 RETINOID METABOLIZING PROTEIN (P450RAI-2),（NM_019885）；유전자 HOMO SAPIENS NIMA (NEVER IN MITOSIS GENE A)-RELATED KINASE 2 (NEK2), MR（NM_002497）；유전자 HOMO SAPIENS BONE MORPHOGENETIC PROTEIN 4 (BMP4), MRNA（NM_001202）；유전자 HOMO SAPIENS CDNA FLJ10674 FIS, CLONE NT2RP2006436 /CDS=UNKNOWN /GB=AK001536 /GI（AK001536）；또는 유전자 HOMO SAPIENS CENTROMERE PROTEIN A (17KD) (CENPA), MRNA（NM_001809）。
유전자 HOMO SAPIENS MRNA; CDNA DKFZP564P116 (FROM CLONE DKFZP564P116) /CDS=UNKNOWN /GB=A（AL049338）；유전자 HOMO SAPIENS TTK PROTEIN KINASE (TTK), MRNA（ NM_003318）；유전자 HOMO SAPIENS KARYOPHERIN ALPHA 2 (RAG COHORT 1, IMPORTIN ALPHA 1) (KPNA（NM_002266）；유전자 HOMO SAPIENS POLYMERASE (DNA DIRECTED), THETA (POLQ), MRNA（NM_006596）；유전자 HOMO SAPIENS ETS VARIANT GENE 5 (ETS-RELATED MOLECULE) (ETV5), MRNA（NM_004454）；유전자 HOMO SAPIENS TRANSFORMING, ACIDIC COILED-COIL CONTAINING PROTEIN 3 (TAC)（NM_006342）；유전자 HOMO SAPIENS KINESIN-LIKE 1 (KNSL1), MRNA（NM_004523）；유전자 HOMO SAPIENS CENTROMERE PROTEIN E (312KD) (CENPE), MRNA（NM_001813）；유전자 HOMO SAPIENS CDNA, 3' END /CLONE=IMAGE:1651289 /CLONE_END=3' /GB=AI12（AI123815）；유전자 HOMO SAPIENS DISTAL-LESS HOMEO BOX 5 (DLX5), MRNA（ NM_005221）；유전자 HOMO SAPIENS V-MYB AVIAN MYELOBLASTOSIS VIRAL ONCOGENE HOMOLOG-LIKE 2 （ NM_002466）；또는 유전자 HOMO SAPIENS SERUM DEPRIVATION RESPONSE (PHOSPHATIDYLSERINE-BINDING PRO（NM_004657）。
유전자 HOMO SAPIENS HYPOTHETICAL PROTEIN FLJ10604 (FLJ10604), MRNA（NM_018154）；유전자 HOMO SAPIENS RIBONUCLEASE HI, LARGE SUBUNIT (RNASEHI), MRNA（NM_006397）；유전자 HOMO SAPIENS ANILLIN (DROSOPHILA SCRAPS HOMOLOG), ACTIN BINDING PROTEIN（NM_018685）；유전자 HOMO SAPIENS H4 HISTONE FAMILY, MEMBER G (H4FG), MRNA（NM_003542）；유전자 HOMO SAPIENS G PROTEIN-COUPLED RECEPTOR 37 (ENDOTHELIN RECEPTOR TYPE B-（NM_005302）HOMO SAPIENS UBIQUITIN CARRIER PROTEIN (E2-EPF), MRNA（NM_014501）；유전자 HOMO SAPIENS, SIMILAR TO TUMOR DIFFERENTIALLY EXPRESSED 1, CLONE IMAGE:3639252, （BC007375）；유전자 HOMO SAPIENS HYPOTHETICAL PROTEIN MGC2577 (MGC2577), MRNA（NM_031299）；유전자 HOMO SAPIENS HYPOTHETICAL PROTEIN FLJ13912 (FLJ13912), MRNA（NM_022770）；유전자 HOMO SAPIENS ANNEXIN A3 (ANXA3), MRNA（NM_005139）；유전자 HOMO SAPIENS STATHMIN 1/ONCOPROTEIN 18 (STMN1), MRNA（NM_005563）HOMO SAPIENS, SIMILAR TO RIBOSOMAL PROTEIN L39, CLONE MGC:20168 IMAGE:4555759, M（BC012328）；유전자 HOMO SAPIENS POLO (DROSOPHIA)-LIKE KINASE (PLK), MRNA（NM_005030）인 것을 특징으로 하는 간엽계 간세포 검출용 유전자 마커.
또, 본 발명에 있어서 확인된, 섬유아세포에 비교하여, 간엽계 간세포에서 발현하고 있지 않거나 또는 극히 소량만 발현하고 있는 유전자로서는 이하의 것을 들 수 있다：
유전자 HOMO SAPIENS PROTEASOME (PROSOME, MACROPAIN) 26S SUBUNIT, ATPASE, 5 (PS)（NM_002805）；유전자 HOMO SAPIENS EUKARYOTIC TRANSLATION INITIATION FACTOR 4 GAMMA, 2 (EIF4G)（NM_001418）；유전자 HOMO SAPIENS DIHYDROPYRIMIDINASE-LIKE 3 (DPYSL3), MRNA）（NM_001387）；유전자 HOMO SAPIENS ADAPTOR-RELATED PROTEIN COMPLEX 1, SIGMA 2 SUBUNIT (AP1S2)（NM_003916）；유전자 HOMO SAPIENS PLECTIN 1, INTERMEDIATE FILAMENT BINDING PROTEIN, 500KD (P)（ NM_000445）；유전자 HOMO SAPIENS HETEROGENEOUS NUCLEAR RIBONUCLEOPROTEIN U (SCAFFOLD ATTAC（NM_031844）；유전자 HOMO SAPIENS HYPOTHETICAL PROTEIN MGC5306 (MGC5306), MRNA（NM_024116）；유전자 HOMO SAPIENS MHC CLASS I POLYPEPTIDE-RELATED SEQUENCE A (MICA), MRNA（NM_000247）；유전자 HOMO SAPIENS PROTEIN ASSOCIATED WITH PRK1 (AWP1), MRNA（NM_019006）；유전자 HOMO SAPIENS PROGESTERONE RECEPTOR MEMBRANE COMPONENT 2 (PGRMC2), MRNA（NM_006320）；유전자 HOMO SAPIENS POLYGLUTAMINE BINDING PROTEIN 1 (PQBP1), MRNA（NM_005710）；유전자 HOMO SAPIENS S-ADENOSYLMETHIONINE DECARBOXYLASE 1 (AMD1), MRNA（NM_001634）； HOMO SAPIENS INTERFERON-INDUCED, HEPATITIS C-ASSOCIATED MICROTUBULAR AG（NM_006417）；유전자 HOMO SAPIENS PROTEIN KINASE, AMP-ACTIVATED, GAMMA 1 NON-CATALYTIC SUBUN（NM_002733）；HOMO SAPIENS PROCOLLAGEN-PROLINE, 2-OXOGLUTARATE 4-DIOXYGENASE (PROLINE（NM_004199）；유전자 HOMO SAPIENS ENDOTHELIAL DIFFERENTIATION, LYSOPHOSPHATIDIC ACID G-PROTE（NM_012152）；또는 유전자 HOMO SAPIENS KIAA0127 GENE PRODUCT (KIAA0127), MRNA（NM_014755）인 것을 특징으로 하는 간엽계 간세포 검출용 유전자 마커.
본 발명에 있어서, 본 발명의 간엽계 간세포 마커 유전자의 발현을 검출하기 위해서는 공지의 유전자 발현의 검출방법을 사용할 수가 있다. 예를 들면, 본 발명의 간엽계 간세포 마커 유전자의 발현을 검출하기 위하여, 노던 블로팅법을 사용할 수가 있다. 또, 본 발명의 유전자 마커에 의하여, 간엽계 간세포를 검출, 식별하기 위하여, 본 발명의 간엽계 간세포 유전자 마커의 DNA배열과 스트린젠트한 조건하에서 하이브리다이즈하는 DNA배열을 갖는 프로브를 사용할 수가 있다. 상기 프로브를 사용하여 간엽계 간세포를 검출하기 위해서는, 공지의 방법을 사용하여 적절히 실시할 수가 있다. 예를 들면, 배열표에 나타낸 유전자 마커의 DNA배열로부터 적당한 길이의 DNA프로브를 제작하고, 적당한 형광표지 등의 표지를 부여해 놓고, 이것을 피검체와 하이브리다이즈하는 것에 의하여, 간엽계 간세포의 검출을 실시한다. 상기 DNA프로브로서는 배열표에 나타낸 본 발명의 유전자 마커의 염기배열의 안티센스사슬의 전부 또는 일부로 이루어지는 간엽계 간세포 마커 유전자 검출용 프로브를 사용할 수가 있다. 또, 상기 프로브를, 적어도 1개 이상을 고정화시킨 마커 유전자 검출용의 마이크로어레이 또는 DNA칩의 형태로 사용할 수도 있다.
또한, 상기 DNA프로브의 제작시에, 본 발명의 염기배열에 있어서,「간엽계 간세포 검출용 마커 유전자의 DNA배열과 스트린젠트한 조건하에서 하이브리다이즈하는」조건으로서는, 예를 들면, 42℃에서의 하이브리다이제이션, 및 1×SSC(0.15M NaCl, 0.015M 쿠엔산 나트륨), 0.1%의 SDS(Sodium dodecyl sulfate)를 포함하는 완충액에 의한 42℃에서의 세정처리를 들 수 있으며, 65℃에서의 하이브리다이제이션 및 0.1×SSC, 0.1%의 SDS를 포함하는 완충액에 의한 65℃에서의 세정처리를 보다 바람직하게 들 수 있다. 또한, 하이브리다이제이션의 스트린젠시에 영향을 미치는 요소로서는 상기 온도조건 이외에 여러가지의 요소가 있으며, 당업자라면 여러가지의 요소를 조합하여 상기 예시한 하이브리다이제이션의 스트린젠시와 동등한 스트린젠시를 실현할 수가 있다.
또, 본 발명에 있어서는, 본 발명의 간엽계 간세포 검출용 폴리펩티드 마커의 검출시에, 상기 단백질의 폴리펩티드에 의하여 유도되며, 상기 폴리펩티드에 특이적으로 결합하는 항체를 사용할 수가 있다. 상기 항체로서는 모노클로널항체 및 폴리클로널항체를 들 수 있다. 상기 항체의 제작은, 본 발명의 폴리펩티드 마커를 항원으로 하여 상법에 의하여 제작할 수가 있다. 본 발명의 항체를 사용하여 피검세포에 있어서의 간엽계 간세포 검출용 마커 폴리펩티드의 발현을 검출하기 위해서는, 공지의 항체를 사용한 면역학적 측정법을 사용하여 실시할 수가 있다. 상기 면역학적 측정법으로서는, 예를 들면, RIA법, ELISA법, 형광항체법 등의 공지의 면역학적 측정법을 들 수가 있다.
본 발명에 있어서는, 본 발명의 간엽계 간세포 검출용 프로브 및/또는 본 발명의 간엽계 간세포 검출용 항체를 사용하여, 피검세포에 있어서의 간엽계 간세포 검출용 마커 유전자 및/또는 간엽계 간세포 검출용 마커 폴리펩티드의 발현을 검출하고, 식별하여 간엽계 간세포를 분리하여 취득할 수가 있다. 피검세포에 있어서의 간엽계 간세포 검출용 마커 유전자의 검출시에는, 피검세포의 유전자를 증폭시키기 위하여 정량적 또는 반정량적 PCR을 사용할 수 있다. 상기 PCR로서는 RT-PCR(역전사(逆轉寫)PCR)을 사용할 수가 있다. 상기 PCR을 실시할 때에는, 본 발명의 간엽계 간세포 검출용 마커 유전자를 증폭시키기 위한 센스프라이머 및 안티센스프라이머로 이루어지는 프라이머를 사용한다.
본 발명의 간엽계 간세포 검출용 마커 유전자를 증폭시키기 위한 프라이머를 열거하면: 배열표의 배열번호20에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호21에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호22에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호23에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호24에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호25에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호26에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호27에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호28에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호29에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호30에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호31에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호32에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호33에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호34에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호35에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호36에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호37에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호38에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호39에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호40에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호41에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호42에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호43에 나타내는 염기배열을 갖는 안티센스프라이머, 또는 배열표의 배열번호44에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호45에 나타내는 염기배열을 갖는 안티센스프라이머를 들 수가 있다.
또, 본 발명의 간엽계 간세포 검출용 마커 유전자를 증폭하기 위한 프라이머를 열거하면 : 배열표의 배열번호47에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호48에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호49에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호50에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호51에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호52에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호53에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호54에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호55에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호56에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호57에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호58에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호59에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호60에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호61에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호62에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호63에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호64에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호65에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호66에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호67에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호68에 나타내는 염기배열을 갖는 안티센스프라이머, 또는 배열표의 배열번호69에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호70에 나타내는 염기배열을 갖는 안티센스프라이머로 이루어지는 간엽계 간세포 마커 유전자 증폭을 위한 실시간 PCR용 프라이머를 들 수가 있다.
또, 본 발명의 간엽계 간세포 검출용 마커 유전자를 증폭하기 위한 프라이머를 열거하면 : 배열표의 배열번호71에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호72에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호73에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호74에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호75에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호76에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호77에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호78에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호79에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호80에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호81에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호82에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호83에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호84에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호85에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호86에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호87에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호88에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호89에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호90에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호91에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호92에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호93에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호94에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호95에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호96에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호97에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호98에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호99에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호100에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호101에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호102에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호103에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호104에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호105에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호106에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호107에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호108에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호109에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호110에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호111에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호112에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호113에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호114에 나타내는 염기배열을 갖는 안티센스프라이머, 또는 배열표의 배열번호115에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호116에 나타내는 염기배열을 갖는 안티센스프라이머로 이루어지는 간엽계 간세포 마커 유전자 증폭을 위한 RT-PCR용 프라이머를 들 수가 있다.
본 발명에 있어서, 간엽계 간세포의 식별을 실시하기 위해서는, 피검세포에 있어서의 유전자를 상기의 센스프라이머 및 안티센스프라이머의 적어도 1쌍 이상의 프라이머를 사용하여 증폭하고, 증폭한 유전자를 본 발명의 간엽계 간세포 마커 유전자 검출용 프로브를 사용하여 검출한다.
본 발명의 간엽계 간세포 검출용 마커 유전자는, 간엽계 간세포에 있어서 특이적으로 높게 발현하는 유전자와, 특이적으로 낮게 발현하는 유전자가 있으며, 그들 유전자의 발현을 조합하여 검출하는 것에 의하여, 간엽계 간세포의 검출정밀도를 높일수가 있다. 덧붙여 말하면, 간엽계 간세포에 있어서 특이적으로 높게 발현하는 유전자로서는, 배열표의 배열번호 1, 11 및 19로 이루어지는 간엽계 간세포 마커 유전자를 들 수 있으며, 특이적으로 낮게 발현하는 유전자로서는, 배열표의 배열번호 2, 4, 6, 7, 9, 10, 12, 14, 15, 17 및 46으로 이루어지는 간엽계 간세포 마커 유전자를 들 수 있다. 또, 본 발명에서 확인된 간엽계 간세포 마커 유전자를 들 수가 있다. 따라서, 각각의 유전자 그룹의 1유전자 이상의 발현을 조합하는 것에 의하여, 보다 검출정밀도가 높은 간엽계 간세포의 식별을 실시할 수가 있다.
본 발명의 항체를 사용하여 형광항체법을 따라서, 피검세포에 있어서의 간엽계 간세포 마커 폴리펩티드의 발현을 검출하고, 간엽계 간세포를 식별, 분리하기 위해서는, 공지의 방법에 의하여 간엽계 간세포를 표지화하여, 분리할 수가 있다. 상기 표지화 방법으로서는, 예를 들면, 형광항체법을 들 수가 있다. 형광항체법에 의하여 미분화 조혈세포를 표지화하기 위해서는, 본 발명의 간엽계 간세포 검출용 폴리펩티드 마커에 특이적으로 결합하는 항체를 형광표지하고, 이를 항원을 발현하고 있는 간엽계 간세포에 결합시켜서 간엽계 간세포를 표지화하거나(직접 형광항체법), 또는 항원을 발현하고 있는 간엽계 간세포에, 미표지의 본 발명의 특이항체를 결합시킨 후에, 표지화한 2차항체(항면역 글로블린항체)를 결합시켜서 간엽계 간세포를 표지화하여(간접 형광항체법), 상기 표지화한 간엽계 간세포를 분리, 채취한다.
본 발명의 간엽계 간세포의 검출, 식별에 사용하는 간엽계 간세포 마커 유전자 검출용 프로브, 상기 프로브를 고정한 간엽계 간세포 마커 유전자 검출용 마이크로 어레이 또는 DNA칩, 및 본 발명의 간엽계 간세포 검출용의 항체는, 그들을 장비한 간엽계 간세포의 식별용 키트로서 제품화해서 놓을 수가 있다.
이하, 실시예에 따라서 본 발명을 보다 구체적으로 설명한다. 단, 본 발명의 기술적 범위는 이들 예시에 한정되는 것이 아니다.
실시예1 [재료와 방법]
(세포)
인간유래 간엽계 간세포(human mesenchymal stem cell）, 및 인간섬유아세포(human fibroblast）는 Dalbecco's modified Eagle's medium（low glucose）（Sigma사 제품）에 10％ fetal calf serum、1ng／ml의 bFGF를 첨가한 배지를 사용하여 37℃에서 배양하였다.
(DNA마이크로어레이)
10㎝ 접시 1장의 각 세포로부터 TRIZOL reagent（Invitrogen사 제품)를 사용하여 전(全) RNA를 추출한 후, micro poly (A) purist（Ambion사 제품)를 사용하여 mRNA를 정제하고, DNA microarray（Incyte Genomics사：Kurabo Life Array analysis servis；Lot. No. KL01081）를 실시하였다. 또한, DNA 마이크로어레이 해석은 kurabo사에 위탁하였다.
(반정량적 RT-PCR)
각 세포로부터 TRIZOL reagent（Invitrogen사 제품)를 사용하여 전체 RNA를 추출한 후, 각 전체 RNA 1㎎을 주형으로 하여 SuperScript first-strand synthesis system for RT-PCR（Invitrogen사 제품）을 사용하여 42℃, 50분간의 역전사 반응을 실시하여 cDNA를 합성하였다. 합성된 cDNA를 주형으로 하여 Advantage 2 PCR enzyme system（Clontech사 제품）을 사용하여 각 유전자를 증폭시켰다. 94℃ 30초간에서 변성(變性), 68℃ 1분간에서 어닐링을 30사이클 실시하였다. 증폭에 사용한 각 유전자의 프라이머배열을 표1에 정리하였다. 증폭한 유전자를 1% 아가로스겔을 사용한 전기영동으로 분리하고, 에티듐 브로마이드 염색에 의하여 검출하였다.
< 실시예2[검출결과] >
(DNA마이크로어레이)
DNA마이크로어레이의 결과, 9400유전자 중, 63유전자가 섬유아세포와 비교하여 간엽계 간세포에 있어서 의미있게 발현이 높고, 141유전자가 낮다는 것이 명백해졌다. 또, 그 이외의 유전자에서는 우위의 차이가 확인되지 않았다. 이 중, 3배 이상의 차이가 확인된 87유전자 중, 간엽계 간세포 쪽이 발현이 높았던 유전자가 29유전자, 낮았던 유전자가 58유전자였다.
(반정량적 RT-PCR)
3개체 유래의 간엽계 간세포와 3개체 유래의 섬유아세포로부터 전체 RNA를 추출하고, 이들을 주형으로 사용하여 상기 87유전자에 대하여 반정량적 RT-PCR법에 의하여 그 발현량을 조사하였다. 그 결과, 발현량에 차이가 나타나지 않는 유전자가 대부분이었다. 또, 상이한 개체의 간엽계 간세포, 또는 섬유아세포 사이에서도 발현량에 차이가 있으며, 개체차에 따라서 발현이 다르다고 생각되는 것도 많았다(도1).
RT-PCR에 의하여 간엽계 간세포에서 발현이 높다는 것이 확인된 유전자는, serine (or cysteine) proteinase inhibitor(배열표의 배열번호1)、major histocompatibility complex class2 DR beta 3（배열번호11）、및 tissue factor pathway inhibitor 2(배열번호19)의 3유전자였다(도2).
또, 낮다는 것이 확인된 유전자는, adrenomedullin（배열번호2）、apolipoprotein D（배열번호４）、collagenase type XV alpha 1（배열번호６）、CUG triplet repeat RNA binding protein（배열번호7）、dermatopontin protein（배열번호９）、isocitrate dehydrogenase 2（배열번호10）、tyrosine kinase 7（배열번호12）、Sam68-like phosphotyrosine protein（배열번호14）、C-type lectin superfamily member 2（배열번호15)、및 matrix metalloprotease 1(배열번호17)의 10유전자(도3)였다. 또, 이들 13유전자에 대하여 7개체 유래의 간엽계 간세포와 4개체 유래의 섬유아세포로부터 추출한 전체 RNA를 주형으로 한 반정량적 RT-PCR법에 의하여, 그 발현량의 확인을 실시하였다(도4).
실시예3[간엽계 간세포에 있어서의 각 유전자 발현의 측정 I]
실시예1과 동일하게 하여, 본 발명의 유전자 마커의 인간유래 간엽계 간세포 및 인간 섬유아세포에 있어서의 발현을 측정하였다. 사용한 RT-PCR 및 실시간 PCR의 프라이머와 실시간 PCR용의 프로브를 표2에 나타낸다.
간엽계 간세포와 인간섬유아세포에 있어서의 mRNA 레벨의 발현의 상황을 도5에 나타낸다. 또, 간엽계 간세포와 인간섬유아세포에 있어서의 상대적인 mRNA레벨의 발현 상황을 도6, 7, 8에 나타낸다.
< 실시예4 [간엽계 간세포에 있어서의 각 유전자 발현의 측정 II] >
실시예1과 동일하게 하여, 본 발명의 각 유전자 마커의 인간유래 간엽계 간세포 및 인간섬유아세포에 있어서의 발현을 측정하였다. 사용한 RT-PCR의 센스 및 안티센스프라이머는 배열표의 배열번호71~116에 나타낸다. 간엽계 간세포와 인간섬유아세포에 있어서의 유전자 발현의 상황을 도9 및 도10에 나타낸다. 또, 간엽계 간세포/섬유아세포의 발현의 비율을 표3에 나타낸다.
< 실시예5 [간엽계 간세포에 있어서의 각 유전자 발현의 측정 III] >
실시예1과 동일하게 하여, 본 발명의 각 유전자 마커의 인간유래 간엽계 간세포 및 인간섬유아세포에 있어서의 발현을 측정하였다. 간엽계 간세포/섬유아세포의 발현의 비율을 표4에 나타낸다.
| (간엽계 간세포/섬유아세포) 유 전 자 명 |
| 8.9 EGF-containing fibulin-like extracellular matrix protein 1 |
| 8.4 Human insulin-like growth factor binding protein 5 (IGFBP5) mRNA |
| 7.7 Homo sapiens clone 24775 mRNA sequence |
| 7 "proteoglycan 1, secretory granule" |
| 6.8 insulin-like growth factor binding protein 5 |
| 6.7 "solute carrier family 21 (organic anion transporter), member 3" |
| 6.5 "transglutaminase 2 (C polypeptide,proteinglutamine-gamma-glutamyl transferase)" |
| 5.8 coagulation factor II (thrombin) receptor |
| 5.6 "plasminogen activator, urokinase" |
| 5.5 "tissue inhibitor of metalloproteinase 3 (Sorsby fundus dystrophy, pseudoinflammatory)" |
| 5.1 matrix Gla protein |
| 4.6 B-cell CLL/lymphoma 1 |
| 4.6 "CD74 antigen (invariant polypeptide of major histocompatibility complex, class II antigen-associated)" |
| 4.2 adducin 3 (gamma) |
| 4.2 "solute carrier family 16 (monocarboxylic acid transporters), member 4" |
| 3.4 protein S (alpha) |
| 3.3 "phosphatidylinositol (4,5) bisphosphate 5-phosphatase homolog; phosphatidylinositol polyphosphate 5-phosphatase type IV" |
| 3.2 progesterone membrane binding protein |
| 3.2 brain-derived neurotrophic factor |
| 3 apolipoprotein E |
| 38.5 "GI|6005949|REF|NM_007191.1| HOMO SAPIENS WNT INHIBITORY FACTOR-1 (WIF-1), MRNA" |
| 12.7 "GI|8922916|REF|NM_018349.1| HOMO SAPIENS HYPOTHETICAL PROTEIN FLJ11175 (FLJ11175), MRNA" |
| 11.2 "GI|10835001|REF|NM_001175.1| HOMO SAPIENS RHO GDP DISSOCIATION INHIBITOR (GDI) BETA (ARHGDIB), MRNA" |
| 10.3 "GI|11055973|REF|NM_021614.1| HOMO SAPIENS POTASSIUM INTERMEDIATE/SMALL CONDUCTANCE CALCIUM-ACTIVATE" |
| 8.7 "GI|5731346|REF|NM_004289.3| HOMO SAPIENS NUCLEAR FACTOR (ERYTHROID-DERIVED 2)-LIKE 3 (NFE2L3), MRNA" |
| 7.6 "GI|11056053|REF|NM_021643.1| HOMO SAPIENS GS3955 PROTEIN (GS3955), MRNA" |
| 7.5 "EST10324 ADIPOSE TISSUE, WHITE I HOMO SAPIENS CDNA 3' END SIMILAR TO SIMILAR TO GLYCOPROTEIN MUC18," |
| 6.6 "GI|13375818|REF|NM_024609.1| HOMO SAPIENS HYPOTHETICAL PROTEIN FLJ21841 (FLJ21841), MRNA" |
| 6.5 "GI|6005971|REF|NM_007150.1| HOMO SAPIENS ZINC FINGER PROTEIN 185 (LIM DOMAIN) (ZNF185), MRNA" |
| 6.5 "GNL|UG|HS#S2334464 SC1=PUTATIVE TRANS-ACTING FACTOR INVOLVED IN CELL CYCLE CONTROL [HUMAN, MRNA, 24" |
| 6.4 "GI|7657368|REF|NM_014222.1| HOMO SAPIENS NADH DEHYDROGENASE (UBIQUINONE) 1 ALPHA SUBCOMPLEX, 8 (19K" |
| 6.3 "GI|8924142|REF|NM_018518.1| HOMO SAPIENS HOMOLOG OF YEAST MCM10; HYPOTHETICAL PROTEIN PRO2249 (PRO2" |
| 6.2 "GI|4502960|REF|NM_000094.1| HOMO SAPIENS COLLAGEN, TYPE VII, ALPHA 1 (EPIDERMOLYSIS BULLOSA, DYSTRO" |
| 6.2 "GI|4502074|REF|NM_001144.1| HOMO SAPIENS AUTOCRINE MOTILITY FACTOR RECEPTOR (AMFR), MRNA" |
| 6.2 GNL|UG|HS#S1090551 HOMO SAPIENS CLONE 24421 MRNA SEQUENCE /CDS=UNKNOWN /GB=AF070641 /GI=3283914 /UG= |
| 6.2 GNL|UG|HS#S1570341 HOMO SAPIENS MRNA; CDNA DKFZP586E1621 (FROM CLONE DKFZP586E1621) /CDS=UNKNOWN /GB |
| 6.1 "GI|11545760|REF|NM_022055.1| HOMO SAPIENS RESERVED (KCNK12), MRNA" |
| 6.0 "GI|5902145|REF|NM_007019.1| HOMO SAPIENS UBIQUITIN-CONJUGATING ENZYME E2C (UBE2C), MRNA" |
| 5.9 "GNL|UG|HS#S1970614 HOMO SAPIENS CDNA FLJ10517 FIS, CLONE NT2RP2000812 /CDS=(61,918) /GB=AK001379 /G" |
| 5.8 "GI|4503052|REF|NM_001884.1| HOMO SAPIENS CARTILAGE LINKING PROTEIN 1 (CRTL1), MRNA" |
| 5.8 GNL|UG|HS#S1972520 HOMO SAPIENS MRNA; CDNA DKFZP434F2322 (FROM CLONE DKFZP434F2322) /CDS=UNKNOWN /GB |
| 5.7 "GNL|UG|HS#S3837475 HOMO SAPIENS, CLONE MGC:9549 IMAGE:3857382, MRNA, COMPLETE CDS /CDS=(92,1285) /G" |
| 5.4 "GI|14150008|REF|NM_032270.1| HOMO SAPIENS HYPOTHETICAL PROTEIN DKFZP586J1119 (DKFZP586J1119), MRNA" |
| 5.4 "GI|4758439|REF|NM_004877.1| HOMO SAPIENS GLIA MATURATION FACTOR, GAMMA (GMFG), MRNA" |
| 5.4 "GNL|UG|HS#S2654530 HOMO SAPIENS CDNA: FLJ23095 FIS, CLONE LNG07413 /CDS=UNKNOWN /GB=AK026748 /GI=10" |
| 5.3 "GI|8922180|REF|NM_018410.1| HOMO SAPIENS HYPOTHETICAL PROTEIN DKFZP762E1312 (DKFZP762E1312), MRNA" |
| 5.3 "GI|5031806|REF|NM_005853.1| HOMO SAPIENS IROQUOIS-CLASS HOMEODOMAIN PROTEIN (IRX-2A), MRNA" |
| 5.2 "GI|8922501|REF|NM_018131.1| HOMO SAPIENS HYPOTHETICAL PROTEIN FLJ10540 (FLJ10540), MRNA" |
| 5.1 "GNL|UG|HS#S1972896 HOMO SAPIENS MRNA; CDNA DKFZP434C1915 (FROM CLONE DKFZP434C1915); PARTIAL CDS /C" |
| 5.1 "GI|14149622|REF|NM_005916.1| HOMO SAPIENS MINICHROMOSOME MAINTENANCE DEFICIENT (S. CEREVISIAE) 7 (M" |
| 5.0 "GI|4758177|REF|NM_004411.1| HOMO SAPIENS DYNEIN, CYTOPLASMIC, INTERMEDIATE POLYPEPTIDE 1 (DNCI1), M" |
| 5.0 HUMAN MRNA FOR PROTEIN GENE PRODUCT (PGP) 9.5. |
| 5.0 "GI|5453903|REF|NM_006479.1| HOMO SAPIENS RAD51-INTERACTING PROTEIN (PIR51), MRNA" |
| 4.9 "GI|4502144|REF|NM_001168.1| HOMO SAPIENS BACULOVIRAL IAP REPEAT-CONTAINING 5 (SURVIVIN) (BIRC5), MR" |
| 4.9 "GI|9945386|REF|NM_004482.2| HOMO SAPIENS UDP-N-ACETYL-ALPHA -D-GALACTOSAMINE:POLYPEPTIDE N-ACETYLGAL" |
| 4.9 "GI|14327895|REF|NM_031966.1| HOMO SAPIENS CYCLIN B1 (CCNB1), MRNA" |
| 4.8 "GI|6325465|REF|NM_004111.3| HOMO SAPIENS FLAP STRUCTURE-SPECIFIC ENDONUCLEASE 1 (FEN1), MRNA" |
| 4.8 "GI|4507274|REF|NM_003600.1| HOMO SAPIENS SERINE/THREONINE KINASE 15 (STK15), MRNA" |
| 4.8 "GI|6466452|REF|NM_002358.2| HOMO SAPIENS MAD2 (MITOTIC ARREST DEFICIENT, YEAST, HOMOLOG)-LIKE 1 (MA" |
| 4.7 "GI|15421859|REF|NM_018454.2| HOMO SAPIENS NUCLEOLAR PROTEIN ANKT (ANKT), MRNA" |
| 4.7 "GI|7661807|REF|NM_014176.1| HOMO SAPIENS HSPC150 PROTEIN SIMILAR TO UBIQUITIN-CONJUGATING ENZYME (H" |
| 4.7 "GI|9845284|REF|NM_019885.1| HOMO SAPIENS CYTOCHROME P450 RETINOID METABOLIZING PROTEIN (P450RAI-2)," |
| 4.7 "GI|4505372|REF|NM_002497.1| HOMO SAPIENS NIMA (NEVER IN MITOSIS GENE A)-RELATED KINASE 2 (NEK2), MR" |
| 4.7 "GI|4502422|REF|NM_001202.1| HOMO SAPIENS BONE MORPHOGENETIC PROTEIN 4 (BMP4), MRNA" |
| 4.7 "GNL|UG|HS#S1970775 HOMO SAPIENS CDNA FLJ10674 FIS, CLONE NT2RP2006436 /CDS=UNKNOWN /GB=AK001536 /GI" |
| 4.7 "GI|4585861|REF|NM_001809.2| HOMO SAPIENS CENTROMERE PROTEIN A (17KD) (CENPA), MRNA" |
| 4.6 GNL|UG|HS#S1368141 HOMO SAPIENS MRNA; CDNA DKFZP564P116 (FROM CLONE DKFZP564P116) /CDS=UNKNOWN /GB=A |
| 4.6 "GI|4507718|REF|NM_003318.1| HOMO SAPIENS TTK PROTEIN KINASE (TTK), MRNA" |
| 4.6 "GI|4504896|REF|NM_002266.1| HOMO SAPIENS KARYOPHERIN ALPHA 2 (RAG COHORT 1, IMPORTIN ALPHA 1) (KPNA" |
| 4.6 "GI|5729983|REF|NM_006596.1| HOMO SAPIENS POLYMERASE (DNA DIRECTED), THETA (POLQ), MRNA" |
| 4.5 "GI|4758315|REF|NM_004454.1| HOMO SAPIENS ETS VARIANT GENE 5 (ETS-RELATED MOLECULE) (ETV5), MRNA" |
| 4.5 "HOMO SAPIENS TRANSFORMING, ACIDIC COILED-COIL CONTAINING PROTEIN 3 (TAC" |
| 4.5 "HOMO SAPIENS KINESIN-LIKE 1 (KNSL1), MRNA" |
| 4.5 "HOMO SAPIENS CENTROMERE PROTEIN E (312KD) (CENPE), MRNA" |
| 4.5 " HOMO SAPIENS CDNA, 3' END /CLONE=IMAGE:1651289 /CLONE_END=3' /GB=AI12" |
| 4.4 "HOMO SAPIENS DISTAL-LESS HOMEO BOX 5 (DLX5), MRNA" |
| 4.4 "HOMO SAPIENS V-MYB AVIAN MYELOBLASTOSIS VIRAL ONCOGENE HOMOLOG-LIKE 2 " |
| 4.4 "HOMO SAPIENS SERUM DEPRIVATION RESPONSE (PHOSPHATIDYLSERINE -BINDING PRO" |
| 4.4 "HOMO SAPIENS HYPOTHETICAL PROTEIN FLJ10604 (FLJ10604), MRNA" |
| 4.4 "HOMO SAPIENS RIBONUCLEASE HI, LARGE SUBUNIT (RNASEHI), MRNA |
| 4.4 "HOMO SAPIENS ANILLIN (DROSOPHILA SCRAPS HOMOLOG), ACTIN BINDING PROTEIN" |
| 4.4 "HOMO SAPIENS H4 HISTONE FAMILY, MEMBER G (H4FG), MRNA" |
| 4.3 "HOMO SAPIENS G PROTEIN-COUPLED RECEPTOR 37 (ENDOTHELIN RECEPTOR TYPE B-" |
| 4.3 "HOMO SAPIENS UBIQUITIN CARRIER PROTEIN (E2-EPF), MRNA" |
| 4.3 "HOMO SAPIENS, SIMILAR TO TUMOR DIFFERENTIALLY EXPRESSED 1, CLONE IMAGE:3639252, " |
| 4.2 "HOMO SAPIENS HYPOTHETICAL PROTEIN MGC2577 (MGC2577), MRNA" |
| 4.2 "HOMO SAPIENS HYPOTHETICAL PROTEIN FLJ13912 (FLJ13912), MRNA" |
| 4.2 "HOMO SAPIENS ANNEXIN A3 (ANXA3), MRNA" |
| 4.2 "HOMO SAPIENS STATHMIN 1/ONCOPROTEIN 18 (STMN1), MRNA" |
| 4.2 "GNL|UG|HS#S3837587 HOMO SAPIENS, SIMILAR TO RIBOSOMAL PROTEIN L39, CLONE MGC:20168 IMAGE:4555759, M" |
| 4.2 "HOMO SAPIENS POLO (DROSOPHIA)-LIKE KINASE (PLK), MRNA" |
< 실시예6 [간엽계 간세포에 있어서의 각 유전자 발현의 측정 VI] >
실시예1과 동일하게 하여, 본 발명의 각 유전자 마커의 인간유래 간엽계 간세포 및 인간섬유아세포에 있어서의 발현을 측정하였다. 간엽계 간세포/섬유아세포의 발현의 비율을 표5에 나타낸다. 표 중의 - 은 간엽계 간세포에 있어서, 유전자가 발현하고 있지 않거나, 또는 극히 소량의 발현인 것을 나타낸다.
| (간엽계 간세포/섬유아세포) 유 전 자 명 |
| -53.576 "HOMO SAPIENS PROTEASOME (PROSOME, MACROPAIN) 26S SUBUNIT, ATPASE, 5 (PS" |
| -45.692 "HOMO SAPIENS EUKARYOTIC TRANSLATION INITIATION FACTOR 4 GAMMA, 2 (EIF4G" |
| -27.6354 "HOMO SAPIENS DIHYDROPYRIMIDINASE-LIKE 3 (DPYSL3), MRNA" |
| -20.5446 "HOMO SAPIENS ADAPTOR-RELATED PROTEIN COMPLEX 1, SIGMA 2 SUBUNIT (AP1S2)" |
| -19.5809 "HOMO SAPIENS PLECTIN 1, INTERMEDIATE FILAMENT BINDING PROTEIN, 500KD P" |
| -16.9025 "HOMO SAPIENS HETEROGENEOUS NUCLEAR RIBONUCLEOPROTEIN U (SCAFFOLD ATTAC" |
| -14.97 "HOMO SAPIENS HYPOTHETICAL PROTEIN MGC5306 (MGC5306), MRNA" |
| -14.6106 "HOMO SAPIENS MHC CLASS I POLYPEPTIDE-RELATED SEQUENCE A (MICA), MRNA" |
| -13.8886 "HOMO SAPIENS PROTEIN ASSOCIATED WITH PRK1 (AWP1), MRNA" |
| -13.6578 "HOMO SAPIENS PROGESTERONE RECEPTOR MEMBRANE COMPONENT 2 (PGRMC2), MRNA" |
| -11.4783 "HOMO SAPIENS POLYGLUTAMINE BINDING PROTEIN 1 (PQBP1), MRNA" |
| -10.2392 "HOMO SAPIENS S-ADENOSYLMETHIONINE DECARBOXYLASE 1 (AMD1), MRNA" |
| -9.9506 "HOMO SAPIENS INTERFERON-INDUCED, HEPATITIS C-ASSOCIATED MICROTUBULAR AG" |
| -9.2594 "HOMO SAPIENS PROTEIN KINASE, AMP-ACTIVATED, GAMMA 1 NON-CATALYTIC SUBUN" |
| -7.518 "HOMO SAPIENS PROCOLLAGEN-PROLINE, 2-OXOGLUTARATE 4-DIOXYGENASE (PROLINE" |
| -6.0365 "HOMO SAPIENS ENDOTHELIAL DIFFERENTIATION, LYSOPHOSPHATIDIC ACID G-PROTE" |
| -5.4442 "HOMO SAPIENS KIAA0127 GENE PRODUCT (KIAA0127), MRNA""" |
< 실시예7 [인간세포에 있어서의 간엽계 간세포 DR의 발현의 측정] >
인간세포에 있어서의 간엽계 간세포 DR의 발현을 플로 사이토메트리를 사용하여 측정하였다. 결과를 도11에 나타낸다.
< 실시예8[간엽계 간세포의 분화유도에 있어서의 각 유전자 발현의 변화]>
실시예1과 동일하게 RT-PCR을 사용하여 간엽계 간세포의 분화유도에 있어서의 각 유전자 발현의 변화를 측정하였다. 골아세포의 분화유도에 있어서의 각 유전자의 발현을 도12~16에 나타낸다.
최근, 재생의료의 진전과 함께, 간엽계 간세포가 많은 조직의 재생의료를 위한 이식재료로서 주목받고 있다. 간엽계 간세포를 조직의 재생의료에 이용하기 위해서는, 간엽계 간세포의 충분한 양을 확보하는 기술을 개발하는 것이 필요함과 동시에, 배양한 세포가 간엽계 간세포인 것을 확인할 필요가 있으며, 상기 간엽계 간세포를 검출, 식별하는 방법이 중요해진다. 또, 항체 등을 사용하여 다종류의 세포의 집단으로부터 간엽계 간세포를 분리하는 것에도 도움이 된다. 본 발명에 의하여, 간엽계 간세포를 특징짓는 마커 유전자를 특정하는 것에 의하여, 용이하고 또한 확실하게 간엽계 간세포를 검출, 식별하는 것이 가능해지며, 간엽계 간세포를 재생의료에 실용화하는 데에 크게 공헌할 것을 기대할 수 있다.
<110> Japan Science and Technology Agency
KATO, Yukio
Two Cells Co. Ltd
<120> MARKER FOR DETECTING MESENCHYMAL STEM CELL AND METHOD OF
DISTINGUISHING MESENCHYMAL STEM CELL USING THE MARKER
<150> JP2003-63077
<151> 2000-03-10
<160> 116
<170> KopatentIn 1.71
<210> 1
<211> 652
<212> DNA
<213> Homo sapiens
<400> 1
gaattgccaa ttccttgttt gtgcaaaatg gatttcatgt caatgaggag tttttgcaaa 60
tgatgaaaaa atattttaat gcagcagtaa atcatgtgga cttcagtcaa aatgtagccg 120
tggccaacta catcaataag tgggtggaga ataacacaaa caatctggtg aaagatttgg 180
tatccccaag ggattttgat gctgccactt atctggccct cattaatgct gtctatttca 240
aggggaactg gaagtcgcag tttaggcctg aaaatactag aaccttttct ttcactaaag 300
atgatgaaag tgaagtccaa attccaatga tgtatcagca aggagaattt tattatgggg 360
aatttagtga tggctccaat gaagctggtg gtatctacca agtcctagaa ataccatatg 420
aaggagatga aataagcatg atgctggtgc tgtccagaca ggaagttcct cttgctactc 480
tggagccatt agtcaaagca cagctggttg aagaatgggc aaactctgtg aagaagcaaa 540
aagtagaagt atacctgccc aggttcacag tggaacagga aattgattta aaagatgttt 600
tgaaggctct tggaataact gaaaattttc atcaaagatg caaattttga ca 652
<210> 2
<211> 1450
<212> DNA
<213> Homo sapiens
<400> 2
ctggatagaa cagctcaagc cttgccactt cgggcttctc actgcagctg ggcttggact 60
tcggagtttt gccattgcca gtgggacgtc tgagactttc tccttcaagt acttggcaga 120
tcactctctt agcagggtct gcgcttcgca gccgggatga agctggtttc cgtcgccctg 180
atgtacctgg gttcgctcgc cttcctaggc gctgacaccg ctcggttgga tgtcgcgtcg 240
gagtttcgaa agaagtggaa taagtgggct ctgagtcgtg ggaagaggga actgcggatg 300
tccagcagct accccaccgg gctcgctgac gtgaaggccg ggcctgccca gacccttatt 360
cggccccagg acatgaaggg tgcctctcga agccccgaag acagcagtcc ggatgccgcc 420
cgcatccgag tcaagcgcta ccgccagagc atgaacaact tccagggcct ccggagcttt 480
ggctgccgct tcgggacgtg cacggtgcag aagctggcac accagatcta ccagttcaca 540
gataaggaca aggacaacgt cgcccccagg agcaagatca gcccccaggg ctacggccgc 600
cggcgccggc gctccctgcc cgaggccggc ccgggtcgga ctctggtgtc ttctaagcca 660
caagcacacg gggctccagc ccccccgagt ggaagtgctc cccactttct ttaggattta 720
ggcgcccatg gtacaaggaa tagtcgcgca agcatcccgc tggtgcctcc cgggacgaag 780
gacttcccga gcggtgtggg gaccgggctc tgacagccct gcggagaccc tgagtccggg 840
aggcaccgtc cggcggcgag ctctggcttt gcaagggccc ctccttctgg gggcttcgct 900
tccttagcct tgctcaggtg caagtgcccc agggggcggg gtgcagaaga atccgagtgt 960
ttgccaggct taaggagagg agaaactgag aaatgaatgc tgagaccccc ggagcagggg 1020
tctgagccac agccgtgctc gcccacaaac tgatttctca cggcgtgtca ccccaccagg 1080
gcgcaagcct cactattact tgaactttcc aaaacctaaa gaggaaaagt gcaatgcgtg 1140
ttgtacatac agaggtaact atcaatattt aagtttgttg ctgtcaagat tttttttgta 1200
acttcaaata tagagatatt tttgtacgtt atatattgta ttaagggcat tttaaaagca 1260
attatattgt cctcccccta ttttaagacg tgaatgtctc agcgaggtgt aaagttgttc 1320
gccgcgtgga atgtgagtgt gtttgtgtgc atgaaagaga aagactgatt acctcctgtg 1380
tggaagaagg aaacaccgag tctctgtata atctatttac ataaaatggg tgatatgcga 1440
acagcaaacc 1450
<210> 3
<211> 185
<212> PRT
<213> Homo sapiens
<400> 3
Met Lys Leu Val Ser Val Ala Leu Met Tyr Leu Gly Ser Leu Ala Phe
1 5 10 15
Leu Gly Ala Asp Thr Ala Arg Leu Asp Val Ala Ser Glu Phe Arg Lys
20 25 30
Lys Trp Asn Lys Trp Ala Leu Ser Arg Gly Lys Arg Glu Leu Arg Met
35 40 45
Ser Ser Ser Tyr Pro Thr Gly Leu Ala Asp Val Lys Ala Gly Pro Ala
50 55 60
Gln Thr Leu Ile Arg Pro Gln Asp Met Lys Gly Ala Ser Arg Ser Pro
65 70 75 80
Glu Asp Ser Ser Pro Asp Ala Ala Arg Ile Arg Val Lys Arg Tyr Arg
85 90 95
Gln Ser Met Asn Asn Phe Gln Gly Leu Arg Ser Phe Gly Cys Arg Phe
100 105 110
Gly Thr Cys Thr Val Gln Lys Leu Ala His Gln Ile Tyr Gln Phe Thr
115 120 125
Asp Lys Asp Lys Asp Asn Val Ala Pro Arg Ser Lys Ile Ser Pro Gln
130 135 140
Gly Tyr Gly Arg Arg Arg Arg Arg Ser Leu Pro Glu Ala Gly Pro Gly
145 150 155 160
Arg Thr Leu Val Ser Ser Lys Pro Gln Ala His Gly Ala Pro Ala Pro
165 170 175
Pro Ser Gly Ser Ala Pro His Phe Leu
180 185
<210> 4
<211> 809
<212> DNA
<213> Homo sapiens
<400> 4
atgcctgtct tcatcttgaa agaaaagctc caggtccctt ctccagccac ccagccccaa 60
gatggtgatg ctgctgctgc tgctttccgc actggctggc ctcttcggtg cggcagaggg 120
acaagcattt catcttggga agtgccccaa tcctccggtg caggagaatt ttgacgtgaa 180
taagtatctc ggaagatggt acgaaattga gaagatccca acaacctttg agaatggacg 240
ctgcatccag gccaactact cactaatgga aaacggaaag atcaaagtgt taaaccagga 300
gttgagagct gatggaactg tgaatcaaat cgaaggtgaa gccaccccag ttaacctcac 360
agagcctgcc aagctggaag ttaagttttc ctggtttatg ccatcggcac cgtactggat 420
cctggccacc gactatgaga actatgccct cgtgtattcc tgtacctgca tcatccaact 480
ttttcacgtg gattttgctt ggatcttggc aagaaaccct aatctccctc cagaaacagt 540
ggactctcta aaaaatatcc tgacttctaa taacattgat gtcaagaaaa tgacggtcac 600
agaccaggtg aactgcccca agctctcgta accaggttct acagggaggc tgcacccact 660
ccatgttact tctgcttcgc tttcccctac cccacccccc cccataaaga caaaccaatc 720
aaccacgaca aaggaagttg acctaaacat gtaaccatgc cctaccctgt taccttgcta 780
gctgcaaaat aaacttgttg ctgacctgc 809
<210> 5
<211> 189
<212> PRT
<213> Homo sapiens
<400> 5
Met Val Met Leu Leu Leu Leu Leu Ser Ala Leu Ala Gly Leu Phe Gly
1 5 10 15
Ala Ala Glu Gly Gln Ala Phe His Leu Gly Lys Cys Pro Asn Pro Pro
20 25 30
Val Gln Glu Asn Phe Asp Val Asn Lys Tyr Leu Gly Arg Trp Tyr Glu
35 40 45
Ile Glu Lys Ile Pro Thr Thr Phe Glu Asn Gly Arg Cys Ile Gln Ala
50 55 60
Asn Tyr Ser Leu Met Glu Asn Gly Lys Ile Lys Val Leu Asn Gln Glu
65 70 75 80
Leu Arg Ala Asp Gly Thr Val Asn Gln Ile Glu Gly Glu Ala Thr Pro
85 90 95
Val Asn Leu Thr Glu Pro Ala Lys Leu Glu Val Lys Phe Ser Trp Phe
100 105 110
Met Pro Ser Ala Pro Tyr Trp Ile Leu Ala Thr Asp Tyr Glu Asn Tyr
115 120 125
Ala Leu Val Tyr Ser Cys Thr Cys Ile Ile Gln Leu Phe His Val Asp
130 135 140
Phe Ala Trp Ile Leu Ala Arg Asn Pro Asn Leu Pro Pro Glu Thr Val
145 150 155 160
Asp Ser Leu Lys Asn Ile Leu Thr Ser Asn Asn Ile Asp Val Lys Lys
165 170 175
Met Thr Val Thr Asp Gln Val Asn Cys Pro Lys Leu Ser
180 185
<210> 6
<211> 2127
<212> DNA
<213> Homo sapiens
<400> 6
gaaagatgga tcactccagc tcaaagagaa catgtgggaa tgaaaggaca ggctgggccc 60
aaaggagaaa agggtgatgc tggggaggag cttcctggcc ctcctgaacc ttctgggcct 120
gttggaccca cggcaggagc agaagcagag ggctctggcc taggctgggg ctcggacgtc 180
ggctctggct ctggtgacct ggtgggcagt gagcagctgc tgagaggtcc tccaggaccc 240
ccagggccac ctggcttacc tgggattcca ggaaaaccag gaactgatgt tttcatggga 300
ccccctggat ctcctggaga ggatggacct gctggtgaac ctgggccccc gggccctgag 360
ggacagcctg gagttgatgg agccaccggc cttcccggga tgaaagggga gaagggagca 420
agagggccta atggctcagt tggtgaaaag ggtgaccctg gcaacagagg cttacctgga 480
cccccgggga aaaagggaca agctggccct cctggggtca tgggaccccc agggcctcct 540
ggaccccctg ggcccccagg ccctggatgc acaatgggac ttggattcga ggataccgaa 600
ggctctggaa gcacccagct attgaatgaa cccaaactct ccagaccaac ggctgcaatt 660
ggtctcaaag gagagaaagg agaccgggga cccaagggag aaagggggat ggatggagcc 720
agtattgtgg gaccccctgg gccgagaggg ccacctgggc acatcaaggt cttgtctaat 780
tccttgatca atatcaccca tggattcatg aatttctcgg acattcctga gctggtgggg 840
cctccggggc cggacgggtt gcctgggctg ccaggatttc caggtcctag aggaccaaaa 900
ggtgacactg gtttacctgg ctttccagga ctaaaaggag aacagggcga gaagggagag 960
ccgggtgcca tcctgacaga ggacattcct ctggaaaggc tgatggggaa aaagggtgaa 1020
cctggaatgc atggagcccc aggaccaatg gggcccaaag gaccaccagg acataaagga 1080
gaatttggcc ttcccgggcg acctggtcgc ccaggactga atggcctcaa gggtaccaaa 1140
ggagatccag gggtcattat gcagggccca cctggcttac ctggccctcc aggcccccct 1200
gggccacctg gagctgtgat taacatcaaa ggagccattt tcccaatacc cgtccgacca 1260
cactgcaaaa tgccagttga tactgctcat cctgggagtc cagagctcat cacttttcac 1320
ggtgttaaag gagagaaagg atcctggggt cttcctggct caaagggaga aaaaggcgac 1380
cagggagccc agggaccacc aggtcctcca cttgatctag cttacctgag acactttctg 1440
aacaacttga agggggagaa tggagacaag gggttcaaag gtgaaaaagg agaaaaagga 1500
gacattaatg gcagcttcct tatgtctggg cctccaggcc tgcccggaaa tccaggcccg 1560
gctggccaaa aaggggagac agtcgttggg ccccaaggac ccccaggtgc tcctggtctg 1620
cctgggccac ctggctttgg aagacctggt gatcctgggc caccggggcc cccggggcca 1680
ccaggacctc cagctatcct gggagcagct gtggcccttc caggtccccc tggccctcca 1740
ggacagccag ggcttcccgg atccagaaac ctggtcacag cattcagcaa catggatgac 1800
atgctgcaga aagcgcattt ggttatagaa ggaacattca tctacctgag ggacagcact 1860
gagtttttca ttcgtgttag agatggctgg aaaaaattac agctgggaga actgatcccc 1920
attcctgccg acagccctcc accccctgcg ctttccagca acccacatca gcttctgcct 1980
ccaccaaacc ctatttcaag tgccaattat gagaagcctg ctctgcattt ggctgctctg 2040
aacatgccat tttctgggga cattcgagct gattttcagt gcttcaagca ggccagagct 2100
gcaggactgt tgtccaccta ccgagca 2127
<210> 7
<211> 2384
<212> DNA
<213> Homo sapiens
<400> 7
cctgctcgtt cctgacagaa ttaacggcac agccaacaag atgaacggag ctttggatca 60
ctcagaccaa ccagacccag atgccattaa gatgtttgtc ggacagatcc cccggtcatg 120
gtcggaaaag gagctgaaag aactttttga gccttacgga gccgtctacc agatcaacgt 180
cctccgggac cggagtcaga accctccgca gagtaaaggt tgttgtttcg taacatttta 240
tacaagaaaa gctgcacttg aggcccagaa tgcactgcac aatattaaaa ctttacctgg 300
gatgcatcat cccattcaga tgaaacctgc agatagtgaa aagtccaacg ctgtggaaga 360
cagaaaattg ttcataggaa tggtttcgaa gaaatgtaat gagaacgaca tcagggtgat 420
gttctctcca tttggccaga tagaagaatg ccggatcctc cggggacctg atgggctgag 480
tcgaggctgt gcgtttgtca cattttctac aagggcaatg gcacagaatg caatcaaagc 540
catgcatcag tctcagacca tggagggctg ctcttcacct atcgtggtga agtttgctga 600
cactcagaag gacaaagagc aaaggcgcct ccagcagcag ctcgctcagc agatgcagca 660
gctcaacact gccacctggg ggaacctgac agggctgggc ggactgaccc cacagtatct 720
ggcgctcctg cagcaggcca cctcctccag caacctgggt gcgttcagcg gcattcaaca 780
aatggcaggc atgaatgctt tacagttgca gaacctggcg acgctggctg ctgctgcagc 840
tgcggcccag acctcagcca ccagcaccaa tgcaaaccct ctctctacca cgagcagcgc 900
cctgggagcc ctcacgagtc ccgtggctgc ttcaaccccc aactccactg ctggtgcagc 960
catgaactcc ttgacctctc tcgggactct gcaaggactg gctggagcca ctgttggact 1020
gaataatatt aatgcactag cagttgctca aatgctctca ggtatggcgg ctctgaatgg 1080
aggacttggc gccacaggct tgacgaatgg cacggctggc accatggacg ccctcaccca 1140
ggcctactca ggaattcaac agtacgcagc cgccgcgctg cccactctgt acagccagag 1200
cctgctgcag cagcagagcg ctgcaggcag ccagaaggaa ggtccagagg gggcaaacct 1260
ctttatttac caccttccac aggaatttgg agaccagcac attctgcaga tgttcatgcc 1320
ttttggaaat gttatctctg ctaaagtctt cattgacaaa cagaccaatc tgagcaagtg 1380
ctttggtttt gttagctacg acaatccagt ctctgcacaa gctgctatcc aagctatgaa 1440
tggctttcag atcggcatga aacgcttgaa ggtgcagctg aagcgttcca aaaacgacag 1500
caaaccttac tgatcctaac cccagaggct ccctgctctc attttagctt tcttaggaca 1560
tcttcatgcc cgttagttca tcgtttgcct agcatgtccc tgtggcgtct caaaaaaaag 1620
tttcatcgtc ccgtcattgt ttctgatgtc tttctgacct cacatcatat ttggttctcc 1680
tactgacctt tgatctagtt tgacctttga aatttgcatg tgacctcatc tagctatgaa 1740
ttctgggaag tcaatgtgaa aaacattgct gcattcatgc aagactgaaa tttattatta 1800
gacaaattca ttatagaaaa aacctgtggc aaaaacgttt ctttcttatt ttttttcttt 1860
tcctaaaaca gacttgaaag tattatacag ggattggcat tcttcccggt cactggtaac 1920
aatagcaata tgtgtccagg gacacagaat gttggtttct aacagactac ttccaaaaac 1980
agtttgagaa aaaaactgtc tgattttaag tctctagagg tctgtaatag tttttacatt 2040
tttcaggcag tgtaaagttt tttgataagg ccattttagg tggctcactt tctcattaag 2100
atatatatat agaaccactt tttgtagatt agtataagaa aaatatttac cctgttttgg 2160
ggcaaatgct acctatttgt gtcacctttt gctgaactca cagttagaca atccatggtt 2220
taatgcacat gaaattacct atattttata ctgtttcaat gtacaggaga aaggttactg 2280
taaactgtgt tatgttggtg cttctgtgaa ttaagttgtg gtttcatcat gagtcttaat 2340
gttctttgtt gataagacaa gtttagaatt ggtttactta atac 2384
<210> 8
<211> 490
<212> PRT
<213> Homo sapiens
<400> 8
Met Asn Gly Ala Leu Asp His Ser Asp Gln Pro Asp Pro Asp Ala Ile
1 5 10 15
Lys Met Phe Val Gly Gln Ile Pro Arg Ser Trp Ser Glu Lys Glu Leu
20 25 30
Lys Glu Leu Phe Glu Pro Tyr Gly Ala Val Tyr Gln Ile Asn Val Leu
35 40 45
Arg Asp Arg Ser Gln Asn Pro Pro Gln Ser Lys Gly Cys Cys Phe Val
50 55 60
Thr Phe Tyr Thr Arg Lys Ala Ala Leu Glu Ala Gln Asn Ala Leu His
65 70 75 80
Asn Ile Lys Thr Leu Pro Gly Met His His Pro Ile Gln Met Lys Pro
85 90 95
Ala Asp Ser Glu Lys Ser Asn Ala Val Glu Asp Arg Lys Leu Phe Ile
100 105 110
Gly Met Val Ser Lys Lys Cys Asn Glu Asn Asp Ile Arg Val Met Phe
115 120 125
Ser Pro Phe Gly Gln Ile Glu Glu Cys Arg Ile Leu Arg Gly Pro Asp
130 135 140
Gly Leu Ser Arg Gly Cys Ala Phe Val Thr Phe Ser Thr Arg Ala Met
145 150 155 160
Ala Gln Asn Ala Ile Lys Ala Met His Gln Ser Gln Thr Met Glu Gly
165 170 175
Cys Ser Ser Pro Ile Val Val Lys Phe Ala Asp Thr Gln Lys Asp Lys
180 185 190
Glu Gln Arg Arg Leu Gln Gln Gln Leu Ala Gln Gln Met Gln Gln Leu
195 200 205
Asn Thr Ala Thr Trp Gly Asn Leu Thr Gly Leu Gly Gly Leu Thr Pro
210 215 220
Gln Tyr Leu Ala Leu Leu Gln Gln Ala Thr Ser Ser Ser Asn Leu Gly
225 230 235 240
Ala Phe Ser Gly Ile Gln Gln Met Ala Gly Met Asn Ala Leu Gln Leu
245 250 255
Gln Asn Leu Ala Thr Leu Ala Ala Ala Ala Ala Ala Ala Gln Thr Ser
260 265 270
Ala Thr Ser Thr Asn Ala Asn Pro Leu Ser Thr Thr Ser Ser Ala Leu
275 280 285
Gly Ala Leu Thr Ser Pro Val Ala Ala Ser Thr Pro Asn Ser Thr Ala
290 295 300
Gly Ala Ala Met Asn Ser Leu Thr Ser Leu Gly Thr Leu Gln Gly Leu
305 310 315 320
Ala Gly Ala Thr Val Gly Leu Asn Asn Ile Asn Ala Leu Ala Val Ala
325 330 335
Gln Met Leu Ser Gly Met Ala Ala Leu Asn Gly Gly Leu Gly Ala Thr
340 345 350
Gly Leu Thr Asn Gly Thr Ala Gly Thr Met Asp Ala Leu Thr Gln Ala
355 360 365
Tyr Ser Gly Ile Gln Gln Tyr Ala Ala Ala Ala Leu Pro Thr Leu Tyr
370 375 380
Ser Gln Ser Leu Leu Gln Gln Gln Ser Ala Ala Gly Ser Gln Lys Glu
385 390 395 400
Gly Pro Glu Gly Ala Asn Leu Phe Ile Tyr His Leu Pro Gln Glu Phe
405 410 415
Gly Asp Gln His Ile Leu Gln Met Phe Met Pro Phe Gly Asn Val Ile
420 425 430
Ser Ala Lys Val Phe Ile Asp Lys Gln Thr Asn Leu Ser Lys Cys Phe
435 440 445
Gly Phe Val Ser Tyr Asp Asn Pro Val Ser Ala Gln Ala Ala Ile Gln
450 455 460
Ala Met Asn Gly Phe Gln Ile Gly Met Lys Arg Leu Lys Val Gln Leu
465 470 475 480
Lys Arg Ser Lys Asn Asp Ser Lys Pro Tyr
485 490
<210> 9
<211> 419
<212> DNA
<213> Homo sapiens
<400> 9
tttttttttt tttttttcag catagtgaac gttaatattt ttatttcttt tgtaatgaag 60
agtacaaatg gttgggagca ggacatcaca ggaggaggaa agatagcgcc atctctgcag 120
aagaactcct gagccacaca cagaaggaaa gttgatcccc agggcagcct ttcccaccaa 180
aaaaatcagg cccaatccag gagagtttgc cagtagctcc ccagggttcc agggtgtctg 240
ccagccttcc taggaatcgt gggcaggctt ctaggtgcca gtgactcaaa ctcctttttc 300
cacttcccag ttcaacctgg tcactctcat ccccacaagt tcccaatctg aatcccattc 360
tctgaccatt ctctgcttcc ttgtttttaa tctcatttga gagtgatcct cacgggttc 419
<210> 10
<211> 762
<212> DNA
<213> Homo sapiens
<400> 10
gccgggagga gcggcgcgcg ctcggacctc tcccgccctg ctcgttcgct ctccagcttg 60
ggatggccgg ctacctgcgg gtcgtgcgct cgctctgcag agcctcaggc tcgcggccgg 120
cctgggcgcc ggcggccctg acagccccca cctcgcaaga gcagccgcgg cgccactatg 180
ccgacaaaag gatcaaggtg gcgaagccgt ggtggagatg gatggtgatg agatgacccg 240
tattatctgg cagttcatca aggagaagct catcctgccc cacgtggaca tccagctaaa 300
gtattttgac ctcgggctcc caaaccgtga ccagactgat gaccaggtca ccattgactc 360
tgcactggcc acccagaagt acagtgtggc tgtcaagtgt gccaccatca cccctgatga 420
ggcccgtgtg gaagagttca agctgaagaa gatgtggaaa agtcccaatg gaactatccg 480
gaacatcctg ggggggactg tcttccggga gcccatcatc tgcaaaaaca tcccacgcct 540
agtccctggc tggaccaagc ccatcaccat tggcaggcac gcccatggcg accagtacaa 600
ggccacagac tttgtggcag accgggccgg cactttcaaa atggtcttca ccccaaaaga 660
tggcagtggt gtcaaggagt gggaagtgta caacttcccc gcaggcgcgc gtgggcatgg 720
gcatgtacma caccgacgag tccatctcag gttttgcgca ca 762
<210> 11
<211> 689
<212> DNA
<213> Homo sapiens
<400> 11
aactccatct ttgaaaaaca tttaataatg taatgtgttt gtggtacagg gtgagtacag 60
atgcacagga ggccataggg tttaggcaaa ggggagcaca aaagttgaag atgaggcgct 120
gccatcaatg ctgggacttc aggccaaggg caggaactga ggaagccaca agggaggaca 180
ttttctgcag ttgctgaacc agtagcaact aggtcctgag aaagccctct ctcgtggaag 240
aataacagcc aggcgggaaa gcttttcatc ctgcaaagct ggggaagaag attcttcctt 300
aaattgtcat ctgcacttca gctcaggaat cctgttggct gaagtccaga gtgtcctttc 360
tgattcctga agtagatgaa cagcccggcc ccaaggaaga gcaggcccag cacaaagccc 420
ccgactccac tcagcatctt gctctgtgca gattcagacc gtgcactcca ttccactgtg 480
agagggctcg ttacgcttgg gtgctccact tggcaggtgt aaacctctcc actccgagga 540
actgtttcta gcatcaccag ggtctggaat gtccagtctc cattctggat caggcgcgtg 600
gacaccaccc cagccttctc ttcctgcccg ttccggaacc agctgacttc aatgctgcct 660
ggatagaaac cactcacaga gcagaccag 689
<210> 12
<211> 1959
<212> DNA
<213> Homo sapiens
<400> 12
gggagcacgc gggagtccgg ctgcccaccc atggcagggt ctaccagaag ggccacgagc 60
tggtgttggc caatattgct gaaagtgatg ctggtgtcta cacctgccac gcggccaacc 120
tggctggtca gcggagacag gatgtcaaca tcactgtggc cactgtgccc tcctggctga 180
agaagcccca agacagccag ctggaggagg gcaaacccgg ctacttggat tgcctgaccc 240
aggccacacc aaaacctaca gttgtctggt acagaaacca gatgctcatc tcagaggact 300
cacggttcga ggtgttcctg gcaaaggctc agggcttgga ggagggagtg gcagagaccc 360
tggtacttgt gaagagcctg cagagcaagg atgagcagca gcagctggac ttccggaggg 420
agttggagat gtttgggaag ctgaaccacg ccaacgtggt gcggctcctg gggctgtgcc 480
gggaggctga gccccactac atggtgctgg aatatgtgga tctgggagac ctcaagcagt 540
tcctgaggat ttccaagagc aaggatgaaa aattgaagtc acagcccctc agcaccaagc 600
agaaggtggc cctatgcacc caggtagccc tgggcatgga gcacctgtcc aacaaccgct 660
ttgtgcataa ggacttggct gcgcgtaact gcctggtcag tgcccagaga caagtgaagg 720
tgtctgccct gggcctcagc aaggatgtgt acaacagtga gtactaccac ttccgccagg 780
cctgggtgcc gctgcgctgg atgtcccccg aggccatcct ggagggtgac ttctctacca 840
agtctgatgt ctgggccttc ggtgtgctga tgtgggaagt gtttacacat ggagagatgc 900
cccatggtgg gcaggcagat gatgaagtac tggcagattt gcaggctggg aaggctagac 960
ttcctcagcc cgagggctgc ccttccaaac tctatcggct gatgcagcgc tgctgggccc 1020
tcagccccaa ggaccggccc tccttcagtg agattgccag cgccctggga gacagcaccg 1080
tggacagcaa gccgtgagga gggagcccgc tcaggatggc ctgggcaggg gaggacatct 1140
ctagagggaa gctcacagca tgatgggcaa gatccctgtc ctcctgggcc ctgaggcccc 1200
tgccctagtg caacaggcat tgctgaggtc tgagcagggc ctggcctttc ctcctcttcc 1260
tcaccctcat cctttgggag gctgacttgg acccaaactg ggcgactagg gctttgagct 1320
gggcagtttt ccctgccacc tcttcctcta tcagggacag tgtgggtgcc acaggtaacc 1380
ccaatttctg gccttcaact tctccccttg accgggtcca actctgccac tcatctgcca 1440
actttgcctg gggagggcta ggcttgggat gagctgggtt tgtggggagt tccttaatat 1500
tctcaagttc tgggcacaca gggttaatga gtctcttggc ccactggtcc cacttggggg 1560
tctagaccag gattatagag gacacagcaa gtgagtcctc cccactctgg gcttgtgcac 1620
actgacccag acccacgtct tccccaccct tctctccttt cctcatccta agtgcctggc 1680
agatgaagga gttttcagga gcttttgaca ctatataaac cgcccttttt gtatgcacca 1740
cgggcggctt ttatatgtaa ttgcagcgtg gggtgggtgg gcatgggagg taggggtggg 1800
ccctggagat gaggagggtg ggccatcctt accccacact tttattgttg tcgttttttg 1860
tttgttttgt ttttttgttt ttgtttttgt ttttacactc gctgctctca ataaataagc 1920
cttttttaca aaaaaaaaaa aaaaaaaaaa aaaaaaaaa 1959
<210> 13
<211> 364
<212> PRT
<213> Homo sapiens
<400> 13
Glu His Ala Gly Val Arg Leu Pro Thr His Gly Arg Val Tyr Gln Lys
1 5 10 15
Gly His Glu Leu Val Leu Ala Asn Ile Ala Glu Ser Asp Ala Gly Val
20 25 30
Tyr Thr Cys His Ala Ala Asn Leu Ala Gly Gln Arg Arg Gln Asp Val
35 40 45
Asn Ile Thr Val Ala Thr Val Pro Ser Trp Leu Lys Lys Pro Gln Asp
50 55 60
Ser Gln Leu Glu Glu Gly Lys Pro Gly Tyr Leu Asp Cys Leu Thr Gln
65 70 75 80
Ala Thr Pro Lys Pro Thr Val Val Trp Tyr Arg Asn Gln Met Leu Ile
85 90 95
Ser Glu Asp Ser Arg Phe Glu Val Phe Leu Ala Lys Ala Gln Gly Leu
100 105 110
Glu Glu Gly Val Ala Glu Thr Leu Val Leu Val Lys Ser Leu Gln Ser
115 120 125
Lys Asp Glu Gln Gln Gln Leu Asp Phe Arg Arg Glu Leu Glu Met Phe
130 135 140
Gly Lys Leu Asn His Ala Asn Val Val Arg Leu Leu Gly Leu Cys Arg
145 150 155 160
Glu Ala Glu Pro His Tyr Met Val Leu Glu Tyr Val Asp Leu Gly Asp
165 170 175
Leu Lys Gln Phe Leu Arg Ile Ser Lys Ser Lys Asp Glu Lys Leu Lys
180 185 190
Ser Gln Pro Leu Ser Thr Lys Gln Lys Val Ala Leu Cys Thr Gln Val
195 200 205
Ala Leu Gly Met Glu His Leu Ser Asn Asn Arg Phe Val His Lys Asp
210 215 220
Leu Ala Ala Arg Asn Cys Leu Val Ser Ala Gln Arg Gln Val Lys Val
225 230 235 240
Ser Ala Leu Gly Leu Ser Lys Asp Val Tyr Asn Ser Glu Tyr Tyr His
245 250 255
Phe Arg Gln Ala Trp Val Pro Leu Arg Trp Met Ser Pro Glu Ala Ile
260 265 270
Leu Glu Gly Asp Phe Ser Thr Lys Ser Asp Val Trp Ala Phe Gly Val
275 280 285
Leu Met Trp Glu Val Phe Thr His Gly Glu Met Pro His Gly Gly Gln
290 295 300
Ala Asp Asp Glu Val Leu Ala Asp Leu Gln Ala Gly Lys Ala Arg Leu
305 310 315 320
Pro Gln Pro Glu Gly Cys Pro Ser Lys Leu Tyr Arg Leu Met Gln Arg
325 330 335
Cys Trp Ala Leu Ser Pro Lys Asp Arg Pro Ser Phe Ser Glu Ile Ala
340 345 350
Ser Ala Leu Gly Asp Ser Thr Val Asp Ser Lys Pro
355 360
<210> 14
<211> 430
<212> DNA
<213> Homo sapiens
<400> 14
ggttgatgtt tcaacaaagt agttttaagt tccatccatt cagattttaa gcattttgat 60
ttatttaaaa agggattgtt catagaaaaa aattactttg aacagtatac aggactggtg 120
gagattggct atttcacaac atcagacagt acaatcagta tctgccatat ggctggtctc 180
tgtagacgcc ctttgctgtc ctcgctgaag gtgccttgtg tcttgagtta gtccactctt 240
cttgcccgta ggaatcataa gtctcctcac tgagtccatg tccgtaatca tagtaatcag 300
caccactttg ggctggggtg ctatagctgt tatcatagga atcataactc tgttcatcat 360
aagcagtgcc atatccatca tcatagtcat attctccata agtctcttgg tgtcgggggt 420
ggcggtggaa 430
<210> 15
<211> 616
<212> DNA
<213> Homo sapiens
<400> 15
ggtatgatga ccaaacataa aaagtgtttt ataattgttg gtgttttaat aacaactaat 60
attattactc tgatagttaa actaactcga gattctcaga gtttatgccc ctatgattgg 120
attggtttcc aaaacaaatg ctattatttc tctaaagaag aaggagattg gaattcaagt 180
aaatacaact gttccactca acatgccgac ctaactataa ttgacaacat agaagaaatg 240
aattttctta ggcggtataa atgcagttct gatcactgga ttggactgaa gatggcaaaa 300
aatcgaacag gacaatgggt agatggagct acatttacca aatcgtttgg catgagaggg 360
agtgaaggat gtgcctacct cagcgatgat ggtgcagcaa cagctagatg ttacaccgaa 420
agaaaatgga tttgcaggaa aagaatacac taagttaatg tctaagataa tggggaaaat 480
agaaaataac attattaagt gtaaaaccag caaagtactt ttttaattaa acaaagttcg 540
agttttgtac ctgtctggtt aattctgctt acgtgtcagg ctacacataa aagccacttc 600
aaagattggc aaaaaa 616
<210> 16
<211> 149
<212> PRT
<213> Homo sapiens
<400> 16
Met Met Thr Lys His Lys Lys Cys Phe Ile Ile Val Gly Val Leu Ile
1 5 10 15
Thr Thr Asn Ile Ile Thr Leu Ile Val Lys Leu Thr Arg Asp Ser Gln
20 25 30
Ser Leu Cys Pro Tyr Asp Trp Ile Gly Phe Gln Asn Lys Cys Tyr Tyr
35 40 45
Phe Ser Lys Glu Glu Gly Asp Trp Asn Ser Ser Lys Tyr Asn Cys Ser
50 55 60
Thr Gln His Ala Asp Leu Thr Ile Ile Asp Asn Ile Glu Glu Met Asn
65 70 75 80
Phe Leu Arg Arg Tyr Lys Cys Ser Ser Asp His Trp Ile Gly Leu Lys
85 90 95
Met Ala Lys Asn Arg Thr Gly Gln Trp Val Asp Gly Ala Thr Phe Thr
100 105 110
Lys Ser Phe Gly Met Arg Gly Ser Glu Gly Cys Ala Tyr Leu Ser Asp
115 120 125
Asp Gly Ala Ala Thr Ala Arg Cys Tyr Thr Glu Arg Lys Trp Ile Cys
130 135 140
Arg Lys Arg Ile His
145
<210> 17
<211> 1973
<212> DNA
<213> Homo sapiens
<400> 17
gggatattgg agtagcaaga ggctgggaag ccatcactta ccttgcactg agaaagaaga 60
caaaggccag tatgcacagc tttcctccac tgctgctgct gctgttctgg ggtgtggtgt 120
ctcacagctt cccagcgact ctagaaacac aagagcaaga tgtggactta gtccagaaat 180
acctggaaaa atactacaac ctgaagaatg atgggaggca agttgaaaag cggagaaata 240
gtggcccagt ggttgaaaaa ttgaagcaaa tgcaggaatt ctttgggctg aaagtgactg 300
ggaaaccaga tgctgaaacc ctgaaggtga tgaagcagcc cagatgtgga gtgcctgatg 360
tggctcagtt tgtcctcact gaggggaacc ctcgctggga gcaaacacat ctgacctaca 420
ggattgaaaa ttacacgcca gatttgccaa gagcagatgt ggaccatgcc attgagaaag 480
ccttccaact ctggagtaat gtcacacctc tgacattcac caaggtctct gagggtcaag 540
cagacatcat gatatctttt gtcaggggag atcatcggga caactctcct tttgatggac 600
ctggaggaaa tcttgctcat gcttttcaac caggcccagg tattggaggg gatgctcatt 660
ttgatgaaga tgaaaggtgg accaacaatt tcagagagta caacttacat cgtgttgcgg 720
ctcatgaact cggccattct cttggactct cccattctac tgatatcggg gctttgatgt 780
accctagcta caccttcagt ggtgatgttc agctagctca ggatgacatt gatggcatcc 840
aagccatata tggacgttcc caaaatcctg tccagcccat cggcccacaa accccaaaag 900
cgtgtgacag taagctaacc tttgatgcta taactacgat tcggggagaa gtgatgttct 960
ttaaagacag attctacatg cgcacaaatc ccttctaccc ggaagttgag ctcaatttca 1020
tttctgtttt ctggccacaa ctgccaaatg ggcttgaagc tgcttacgaa tttgccgaca 1080
gagatgaagt ccggtttttc aaagggaata agtactgggc tgttcaggga cagaatgtgc 1140
tacacggata ccccaaggac atctacagct cctttggctt ccctagaact gtgaagcata 1200
tcgatgctgc tctttctgag gaaaacactg gaaaaaccta cttctttgtt gctaacaaat 1260
actggaggta tgatgaatat aaacgatcta tggatccagg ttatcccaaa atgatagcac 1320
atgactttcc tggaattggc cacaaagttg atgcagtttt catgaaagat ggatttttct 1380
atttctttca tggaacaaga caatacaaat ttgatcctaa aacgaagaga attttgactc 1440
tccagaaagc taatagctgg ttcaactgca ggaaaaattg aacattacta atttgaatgg 1500
aaaacacatg gtgtgagtcc aaagaaggtg ttttcctgaa gaactgtcta ttttctcagt 1560
catttttaac ctctagagtc actgatacac agaatataat cttatttata cctcagtttg 1620
catatttttt tactatttag aatgtagccc tttttgtact gatataattt agttccacaa 1680
atggtgggta caaaaagtca agtttgtggc ttatggattc atataggcca gagttgcaaa 1740
gatcttttcc agagtatgca actctgacgt tgatcccaga gagcagcttc agtgacaaac 1800
atatcctttc aagacagaaa gagacaggag acatgagtct ttgccggagg aaaagcagct 1860
caagaacaca tgtgcagtca ctggtgtcac cctggatagg caagggataa ctcttctaac 1920
acaaaataag tgttttatgt ttggaataaa gtcaaccttg tttctactgt ttt 1973
<210> 18
<211> 469
<212> PRT
<213> Homo sapiens
<400> 18
Met His Ser Phe Pro Pro Leu Leu Leu Leu Leu Phe Trp Gly Val Val
1 5 10 15
Ser His Ser Phe Pro Ala Thr Leu Glu Thr Gln Glu Gln Asp Val Asp
20 25 30
Leu Val Gln Lys Tyr Leu Glu Lys Tyr Tyr Asn Leu Lys Asn Asp Gly
35 40 45
Arg Gln Val Glu Lys Arg Arg Asn Ser Gly Pro Val Val Glu Lys Leu
50 55 60
Lys Gln Met Gln Glu Phe Phe Gly Leu Lys Val Thr Gly Lys Pro Asp
65 70 75 80
Ala Glu Thr Leu Lys Val Met Lys Gln Pro Arg Cys Gly Val Pro Asp
85 90 95
Val Ala Gln Phe Val Leu Thr Glu Gly Asn Pro Arg Trp Glu Gln Thr
100 105 110
His Leu Thr Tyr Arg Ile Glu Asn Tyr Thr Pro Asp Leu Pro Arg Ala
115 120 125
Asp Val Asp His Ala Ile Glu Lys Ala Phe Gln Leu Trp Ser Asn Val
130 135 140
Thr Pro Leu Thr Phe Thr Lys Val Ser Glu Gly Gln Ala Asp Ile Met
145 150 155 160
Ile Ser Phe Val Arg Gly Asp His Arg Asp Asn Ser Pro Phe Asp Gly
165 170 175
Pro Gly Gly Asn Leu Ala His Ala Phe Gln Pro Gly Pro Gly Ile Gly
180 185 190
Gly Asp Ala His Phe Asp Glu Asp Glu Arg Trp Thr Asn Asn Phe Arg
195 200 205
Glu Tyr Asn Leu His Arg Val Ala Ala His Glu Leu Gly His Ser Leu
210 215 220
Gly Leu Ser His Ser Thr Asp Ile Gly Ala Leu Met Tyr Pro Ser Tyr
225 230 235 240
Thr Phe Ser Gly Asp Val Gln Leu Ala Gln Asp Asp Ile Asp Gly Ile
245 250 255
Gln Ala Ile Tyr Gly Arg Ser Gln Asn Pro Val Gln Pro Ile Gly Pro
260 265 270
Gln Thr Pro Lys Ala Cys Asp Ser Lys Leu Thr Phe Asp Ala Ile Thr
275 280 285
Thr Ile Arg Gly Glu Val Met Phe Phe Lys Asp Arg Phe Tyr Met Arg
290 295 300
Thr Asn Pro Phe Tyr Pro Glu Val Glu Leu Asn Phe Ile Ser Val Phe
305 310 315 320
Trp Pro Gln Leu Pro Asn Gly Leu Glu Ala Ala Tyr Glu Phe Ala Asp
325 330 335
Arg Asp Glu Val Arg Phe Phe Lys Gly Asn Lys Tyr Trp Ala Val Gln
340 345 350
Gly Gln Asn Val Leu His Gly Tyr Pro Lys Asp Ile Tyr Ser Ser Phe
355 360 365
Gly Phe Pro Arg Thr Val Lys His Ile Asp Ala Ala Leu Ser Glu Glu
370 375 380
Asn Thr Gly Lys Thr Tyr Phe Phe Val Ala Asn Lys Tyr Trp Arg Tyr
385 390 395 400
Asp Glu Tyr Lys Arg Ser Met Asp Pro Gly Tyr Pro Lys Met Ile Ala
405 410 415
His Asp Phe Pro Gly Ile Gly His Lys Val Asp Ala Val Phe Met Lys
420 425 430
Asp Gly Phe Phe Tyr Phe Phe His Gly Thr Arg Gln Tyr Lys Phe Asp
435 440 445
Pro Lys Thr Lys Arg Ile Leu Thr Leu Gln Lys Ala Asn Ser Trp Phe
450 455 460
Asn Cys Arg Lys Asn
465
<210> 19
<211> 1141
<212> DNA
<213> Homo sapiens
<400> 19
ccgccaggcg ctttctcgga cgccttgccc agcgggccgc ccgaccccct gcaccatgga 60
ccccgctcgc cccctggggc tgtcgattct gctgcttttc ctgacggagg ctgcactggg 120
cgatgctgct caggagccaa caggaaataa cgcggagatc tgtctcctgc ccctagacta 180
cggaccctgc cgggccctac ttctccgtta ctactacgac aggtacacgc agagctgccg 240
ccagttcctg tacgggggct gcgagggcaa cgccaacaat ttctacacct gggaggcttg 300
cgacgatgct tgctggagga tagaaaaagt tcccaaagtt tgccggctgc aagtgagtgt 360
ggacgaccag tgtgaggggt ccacagaaaa gtatttcttt aatctaagtt ccatgacatg 420
tgaaaaattc ttttccggtg ggtgtcaccg gaaccggatt gagaacaggt ttccagatga 480
agctacttgt atgggcttct gcgcaccaaa gaaaattcca tcattttgct acagtccaaa 540
agatgaggga ctgtgctctg ccaatgtgac tcgctattat tttaatccaa gatacagaac 600
ctgtgatgct ttcacctata ctggctgtgg agggaatgac aataactttg ttagcaggga 660
ggattgcaaa cgtgcatgtg caaaagcttt gaaaaagaaa aagaagatgc caaagcttcg 720
ctttgccagt agaatccgga aaattcggaa gaagcaattt taaacattct taatatgtca 780
tcttgtttgt ctttatggct tatttgcctt tatggttgta tctgaagaat aatatgacag 840
catgaggaaa caaatcattg gtgatttatt caccagtttt tattaataca agtcactttt 900
taaaaaattt ggattttttt atatataact agctgctatt caaatgtgag tctaccattt 960
ttaatttatg gttcaactgt ttgtgagact gaattcttgc aatgcataag atataaaagc 1020
aaatatgact cactcatttc ttggggtcgt attcctgatt tcagaagagg atcataactg 1080
aaacaacata agacaatata atcatgtgct tttaacatat ttgagaataa aaaggactag 1140
c 1141
<210> 20
<211> 22
<212> DNA
<213> Artificial Sequence
<220>
<223> Sequence:serine (or cysteine) proteinase inhibitor sense primer
<400> 20
ccaaactttg aaagccaagg tg 22
<210> 21
<211> 21
<212> DNA
<213> Artificial Sequence
<220>
<223> serine (or cysteine) proteinase inhibitor antisense primer
<400> 21
tgcacttcaa agaccagcag g 21
<210> 22
<211> 21
<212> DNA
<213> Artificial Sequence
<220>
<223> adrenomedullin sense primer
<400> 22
aggaatagtc gcgcaagcat c 21
<210> 23
<211> 22
<212> DNA
<213> Artificial Sequence
<220>
<223> adrenomedullin antisense primer
<400> 23
cacgcattgc acttttcctc tt 22
<210> 24
<211> 21
<212> DNA
<213> Artificial Sequence
<220>
<223> apolipoprotein D sense primer
<400> 24
tgaagccacc ccagttaacc t 21
<210> 25
<211> 23
<212> DNA
<213> Artificial Sequence
<220>
<223> apolipoprotein D antisense primer
<400> 25
ggtcaacttc ctttgtgcgt ggt 23
<210> 26
<211> 22
<212> DNA
<213> Artificial Sequence
<220>
<223> collagen type 15 alpha 1 sense primer
<400> 26
accaaaccct atttcaagtg cc 22
<210> 27
<211> 21
<212> DNA
<213> Artificial Sequence
<220>
<223> collagen type 15 alpha 1 antisense primer
<400> 27
tagttatcca caaggcggac g 21
<210> 28
<211> 21
<212> DNA
<213> Artificial Sequence
<220>
<223> CUG triplet repeat RNA-binding protein 2 sense primer
<400> 28
atgtttgtcg gacagatccc c 21
<210> 29
<211> 21
<212> DNA
<213> Artificial Sequence
<220>
<223> CUG triplet repeat RNA-binding protein 2 antisense primer
<400> 29
atgtgacaaa cgcacagcct c 21
<210> 30
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223> dermatopontin sense primer
<400> 30
atttgaaccg gcaaggcttc 20
<210> 31
<211> 21
<212> DNA
<213> Artificial Sequence
<220>
<223> dermatopontinanti sense primer
<400> 31
aagtggttgt tgctcctcgg a 21
<210> 32
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223> isocitrate dehydrogenase sense primer
<400> 32
tgccatccag aagaaatggc 20
<210> 33
<211> 19
<212> DNA
<213> Artificial Sequence
<220>
<223> isocitrate dehydrogenase antisense primer
<400> 33
aggcaaagat gctggcgat 19
<210> 34
<211> 22
<212> DNA
<213> Artificial Sequence
<220>
<223> MHC Class 2 DR beta3 sense primer
<400> 34
tagctgtgga caaagccaac ct 22
<210> 35
<211> 22
<212> DNA
<213> Artificial Sequence
<220>
<223> MHC Class 2 DR beta3 antisense primer
<400> 35
ggcacacacc acgttctctg ta 22
<210> 36
<211> 22
<212> DNA
<213> Artificial Sequence
<220>
<223> protein tyrosine kinase 7 sense primer
<400> 36
tggaaccaga gcgtacgact gt 22
<210> 37
<211> 19
<212> DNA
<213> Artificial Sequence
<220>
<223> protein tyrosine kinase 7 antisense primer
<400> 37
tggctttgca gcgcttctt 19
<210> 38
<211> 22
<212> DNA
<213> Artificial Sequence
<220>
<223> Sam-68 like phosphotyrosine protein, T-STAR sense primer
<400> 38
aggagaaggc aaggatgaag aa 22
<210> 39
<211> 22
<212> DNA
<213> Artificial Sequence
<220>
<223> Sam-68 like phosphotyrosine protein, T-STAR antisense primer
<400> 39
tgttaactcc tggagctgtg ct 22
<210> 40
<211> 22
<212> DNA
<213> Artificial Sequence
<220>
<223> C-type lectin superfamily member 2 sense primer
<400> 40
gcccctatga ttggattggt tt 22
<210> 41
<211> 22
<212> DNA
<213> Artificial Sequence
<220>
<223> C-type lectin superfamily member 2 antisense primer
<400> 41
catccttcac tccctctcat gc 22
<210> 42
<211> 25
<212> DNA
<213> Artificial Sequence
<220>
<223> MMP (martix metalloproteinase) 1 sense primer
<400> 42
cgactctaga aacacaagag caaga 25
<210> 43
<211> 25
<212> DNA
<213> Artificial Sequence
<220>
<223> MMP (martix metalloproteinase) 1 antisense primer
<400> 43
aaggttagct tactgtcaca cgctt 25
<210> 44
<211> 17
<212> DNA
<213> Artificial Sequence
<220>
<223> TFPI (tissue factor pathway inhibitor) 2 sense primer
<400> 44
gtcgattctg cttttcc 17
<210> 45
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223> TFPI (tissue factor pathway inhibitor) 2 antisense primer
<400> 45
atggaatttt ctttggtgcg 20
<210> 46
<211> 1045
<212> DNA
<213> Homo sapiens
<400> 46
ctcccgagct ctactgactc ccaacagagc gcccaagcaa gaaaatggcc ataagtggag 60
tccctgtgct aggatttttc atcatagctg tgctgatgag cgctcaggaa tcatgggcta 120
tcaaagaaga acatgtgatc atccaggccg agttctatct gaatcctgac caatcaggcg 180
agtttatgtt tgactttgat ggtgatgaga ttttccatgt ggatatggca aagaaggaga 240
cggtctggcg gcttgaagaa tttggacgat ttgccagctt tgaggctcaa ggtgcattgg 300
ccaacatagc tgtggacaaa gccaacctgg aaatcatgac aaagcgctcc aactatactc 360
cgatcaccaa tgtacctcca gaggtaactg tgctcacgaa cagccctgtg gaactgagag 420
agcccaacgt cctcatctgt ttcatcgaca agttcacccc accagtggtc aatgtcacgt 480
ggcttcgaaa tggaaaacct gtcaccacag gagtgtcaga gacagtcttc ctgcccaggg 540
aagaccacct tttccgcaag ttccactatc tccccttcct gccctcaact gaggacgttt 600
acgactgcag ggtggagcac tggggcttgg actgagcctc ttctcaagca ctgggaggtt 660
gatgctccaa gcccgtctcc agagacttac gagacgtggt gtgtgccctg gggctgactg 720
tgggtctggt cgggcttcct ttgggaccct ctttcatcat ccacggcgag tgcgccaaaa 780
gcattgccca gaaacccagg gggccctctg taaggcctgt gcgggggcag aacggtccct 840
taaaaagaag aacctcgtgg gcaaaacctg gcgcaagacg tcatacaaag ggggggcata 900
taacacgcgt gccccagaac gaccgcactc agggctcgcc gcctataaca ccccggtgtg 960
gtacccccaa ggcgcgctaa aactaaacag cgcgcacata gtagctcagg cgatacacca 1020
ctcacatata taatagcacc gctaa 1045
<210> 47
<211> 18
<212> DNA
<213> Artificial Sequence
<220>
<223> adrenomedullin primer for RT-PCR
<400> 47
ggtttccgtc gccctgat 18
<210> 48
<211> 24
<212> DNA
<213> Artificial Sequence
<220>
<223> adrenomedullin primer for RT-PCR
<400> 48
gagcccactt attccacttc tttc 24
<210> 49
<211> 22
<212> DNA
<213> Artificial Sequence
<220>
<223> matrix metaroproteinase 1 primer for RT-PCR
<400> 49
gatggacctg gaggaaatct tg 22
<210> 50
<211> 24
<212> DNA
<213> Artificial Sequence
<220>
<223> matrix metaroproteinase 1 primer for RT-PCR
<400> 50
ccgcaacacg atgtaagttg tact 24
<210> 51
<211> 21
<212> DNA
<213> Artificial Sequence
<220>
<223> tissue factor pathway inhibitor 2 primer for RT-PCR
<400> 51
ggcaacgcca acaatttcta c 21
<210> 52
<211> 26
<212> DNA
<213> Artificial Sequence
<220>
<223> tissue factor pathway inhibitor 2 primer for RT-PCR
<400> 52
caaactttgg gaactttttc tatcct 26
<210> 53
<211> 23
<212> DNA
<213> Artificial Sequence
<220>
<223> serine (or cysteine) proteinase inhibitor primer for PT-PCR
<400> 53
tgggtggaga ataacacaaa caa 23
<210> 54
<211> 22
<212> DNA
<213> Artificial Sequence
<220>
<223> serine (or cysteine) proteinase inhibitor primer for RT-PCR
<400> 54
ccagataagt ggcagcatca aa 22
<210> 55
<211> 28
<212> DNA
<213> Artificial Sequence
<220>
<223> protein tyrosine kinase 7 primer for RT-PCR
<400> 55
gggagttcct taatattctc aagttctg 28
<210> 56
<211> 27
<212> DNA
<213> Artificial Sequence
<220>
<223> protein tyrosine kinase 7
<400> 56
gctgtgtcct ctataatcct ggtctag 27
<210> 57
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223> collagen, type XV, alpha 1 primer for RT-PCR
<400> 57
ccagcaaccc acatcagctt 20
<210> 58
<211> 23
<212> DNA
<213> Artificial Sequence
<220>
<223> collagen, type XV, alpha 1 primer for RT-PCR
<400> 58
atgcagagca ggcttctcat aat 23
<210> 59
<211> 29
<212> DNA
<213> Artificial Sequence
<220>
<223> Sam68-like phosphotyrosine protein primer for RT-PCR
<400> 59
tcacaacatc agacagtaca atcagtatc 29
<210> 60
<211> 22
<212> DNA
<213> Artificial Sequence
<220>
<223> Sam68-like phosphotyrosine protein primer for RT-PCR
<400> 60
acgggcaaga agagtggact aa 22
<210> 61
<211> 23
<212> DNA
<213> Artificial Sequence
<220>
<223> apolipoprotein D primer for RT-PCR
<400> 61
tgagaagatc ccaacaacct ttg 23
<210> 62
<211> 24
<212> DNA
<213> Artificial Sequence
<220>
<223> apolipoprotein D primer for PT-PCR
<400> 62
tgatctttcc gttttccatt agtg 24
<210> 63
<211> 22
<212> DNA
<213> Artificial Sequence
<220>
<223> major histocompatibility complex DR beta
<400> 63
ggctgaagtc cagagtgtcc tt 22
<210> 64
<211> 18
<212> DNA
<213> Artificial Sequence
<220>
<223> major histocompatibility complex DR beta primer for RT-PCR
<400> 64
gctgggcctg ctcttcct 18
<210> 65
<211> 19
<212> DNA
<213> Artificial Sequence
<220>
<223> major histocompatibility complex DR alpha primer for RT-PCR
<400> 65
gcccagggaa gaccacctt 19
<210> 66
<211> 23
<212> DNA
<213> Artificial Sequence
<220>
<223> major histocompatibility complex DR alpha primer for RT-PCR
<400> 66
cagtcgtaaa cgtcctcagt tga 23
<210> 67
<211> 27
<212> DNA
<213> Artificial Sequence
<220>
<223> C-type lectin primer for RT-PCR
<400> 67
atccattttc tttcggtgta acatcta 27
<210> 68
<211> 23
<212> DNA
<213> Artificial Sequence
<220>
<223> C-type lectin primer for RT-PCR
<400> 68
catgagaggg agtgaaggat gtg 23
<210> 69
<211> 24
<212> DNA
<213> Artificial Sequence
<220>
<223> CUG triplet repeat RNA-binding protein 2 primer for RT-PCR
<400> 69
catgaatgct ttacagttgc agaa 24
<210> 70
<211> 19
<212> DNA
<213> Artificial Sequence
<220>
<223> CUG triplet repeat RNA-binding protein 2 primer for RT-PCR
<400> 70
gcgctgctcg tggtagaga 19
<210> 71
<211> 22
<212> DNA
<213> Artificial Sequence
<220>
<223> INTEGRIN, ALPHA 6 (ITGA6), MRNA
<400> 71
gatgacagtg ttccccgata cc 22
<210> 72
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223> INTEGRIN, ALPHA 6 (ITGA6), MRNA
<400> 72
tgtaagtcag ccacgccaaa 20
<210> 73
<211> 21
<212> DNA
<213> Artificial Sequence
<220>
<223> SOLUTE CARRIER FAMILY 20 (PHOSPHATE TRANSPORTER), MEMBER 1
<400> 73
aatggagaag ctgacatggc c 21
<210> 74
<211> 21
<212> DNA
<213> Artificial Sequence
<220>
<223> SOLUTE CARRIER FAMILY 20 (PHOSPHATE TRANSPORTER), MEMBER 1
<400> 74
ccaaacccac agaccaacac a 21
<210> 75
<211> 21
<212> DNA
<213> Artificial Sequence
<220>
<223> RIBONUCLEOTIDE REDUCTASE M2 POLYPEPTIDE (RRM2)
<400> 75
tccaaggaca ttcagcactg g 21
<210> 76
<211> 19
<212> DNA
<213> Artificial Sequence
<220>
<223> RIBONUCLEOTIDE REDUCTASE M2 POLYPEPTIDE (RRM2), MRNA
<400> 76
atcgacgcaa aagaaccgg 19
<210> 77
<211> 22
<212> DNA
<213> Artificial Sequence
<220>
<223> FOLLISTATIN (FST), TRANSCRIPT VARIANT FST317, MRNA
<400> 77
ggaagtccag taccaaggca ga 22
<210> 78
<211> 19
<212> DNA
<213> Artificial Sequence
<220>
<223> FOLLISTATIN (FST), TRANSCRIPT VARIANT FST317, MRNA
<400> 78
tggcattgtc actggcaca 19
<210> 79
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223> SPROUTY (DROSOPHILA) HOMOLOG 2 (SPRY2), MRNA
<400> 79
gccatccgaa acaccaatga 20
<210> 80
<211> 22
<212> DNA
<213> Artificial Sequence
<220>
<223> SPROUTY (DROSOPHILA) HOMOLOG 2 (SPRY2), MRNA
<400> 80
tgccacagtc ctcacacctg ta 22
<210> 81
<211> 21
<212> DNA
<213> Artificial Sequence
<220>
<223> RAB3B, MEMBER RAS ONCOGENE FAMILY (RAB3B), MRNA
<400> 81
atgctgatga cacgttcacc c 21
<210> 82
<211> 22
<212> DNA
<213> Artificial Sequence
<220>
<223> RAB3B, MEMBER RAS ONCOGENE FAMILY (RAB3B), MRNA
<400> 82
ggcctgcctt acactgatgt tc 22
<210> 83
<211> 21
<212> DNA
<213> Artificial Sequence
<220>
<223> SOLUTE CARRIER FAMILY 2 (FACILITATED GLUCOSE TRANSPORTER)
<400> 83
gccttttcgt taaccgcttt g 21
<210> 84
<211> 19
<212> DNA
<213> Artificial Sequence
<220>
<223> SOLUTE CARRIER FAMILY 2 (FACILITATED GLUCOSE TRANSPORTER)
<400> 84
tcctcgttgc ggttgatga 19
<210> 85
<211> 21
<212> DNA
<213> Artificial Sequence
<220>
<223> INTERLEUKIN 13 RECEPTOR, ALPHA 2 (IL13RA2), MRNA
<400> 85
tgaaggctat ttgaagtcgc c 21
<210> 86
<211> 19
<212> DNA
<213> Artificial Sequence
<220>
<223> INTERLEUKIN 13 RECEPTOR, ALPHA 2 (IL13RA2), MRNA
<400> 86
cttcgcttca atgcccttg 19
<210> 87
<211> 21
<212> DNA
<213> Artificial Sequence
<220>
<223> SERINE/THREONINE KINASE 12 (STK12), MRNA
<400> 87
tggaaacgtg tacttggctc g 21
<210> 88
<211> 21
<212> DNA
<213> Artificial Sequence
<220>
<223> SERINE/THREONINE KINASE 12 (STK12), MRNA
<400> 88
accagccgaa gtcagcaatc t 21
<210> 89
<211> 22
<212> DNA
<213> Artificial Sequence
<220>
<223> MINICHROMOSOME MAINTENANCE DEFICIENT (S. CEREVISIAE) 5
<400> 89
aagtttggcc tgactaccag ca 22
<210> 90
<211> 22
<212> DNA
<213> Artificial Sequence
<220>
<223> MINICHROMOSOME MAINTENANCE DEFICIENT (S. CEREVISIAE)
<400> 90
ccagacgtgt ataccccaat gg 22
<210> 91
<211> 22
<212> DNA
<213> Artificial Sequence
<220>
<223> THYROID HORMONE RECEPTOR INTERACTOR 13 (TRIP13), MRNA
<400> 91
cctgtgtaaa gcgttagccc ag 22
<210> 92
<211> 22
<212> DNA
<213> Artificial Sequence
<220>
<223> THYROID HORMONE RECEPTOR INTERACTOR 13 (TRIP13), MRNA
<400> 92
gtggcccaat gtactgcttg at 22
<210> 93
<211> 22
<212> DNA
<213> Artificial Sequence
<220>
<223> KINESIN-LIKE 6 (MITOTIC CENTROMERE-ASSOCIATED KINESIN)
<400> 93
ccagctttaa cgaagccatg ac 22
<210> 94
<211> 22
<212> DNA
<213> Artificial Sequence
<220>
<223> KINESIN-LIKE 6 (MITOTIC CENTROMERE-ASSOCIATED KINESIN)
<400> 94
gcatttacca gaacctgtcc ca 22
<210> 95
<211> 22
<212> DNA
<213> Artificial Sequence
<220>
<223> CYCLIN-DEPENDENT KINASE INHIBITOR 3
<400> 95
tcggtttatg tgctcttcca gg 22
<210> 96
<211> 22
<212> DNA
<213> Artificial Sequence
<220>
<223> CYCLIN-DEPENDENT KINASE INHIBITOR 3
<400> 96
atcctcttag gtctcgcagg ct 22
<210> 97
<211> 22
<212> DNA
<213> Artificial Sequence
<220>
<223> CHROMOSOME CONDENSATION PROTEIN G (HCAP-G), MRNA
<400> 97
gctcgccaga aattcgagtc ta 22
<210> 98
<211> 22
<212> DNA
<213> Artificial Sequence
<220>
<223> CHROMOSOME CONDENSATION PROTEIN G (HCAP-G), MRNA
<400> 98
catgatcact ttcagagtcg gc 22
<210> 99
<211> 21
<212> DNA
<213> Artificial Sequence
<220>
<223> CDC28 PROTEIN KINASE 1 (CKS1), MRNA
<400> 99
gcccactacc caagaaacca a 21
<210> 100
<211> 22
<212> DNA
<213> Artificial Sequence
<220>
<223> CDC28 PROTEIN KINASE 1 (CKS1), MRNA
<400> 100
cataaatccg caagtcacca ca 22
<210> 101
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223> PROTEIN REGULATOR OF CYTOKINESIS 1 (PRC1), MRNA
<400> 101
cccaagtgga attgatgcga 20
<210> 102
<211> 22
<212> DNA
<213> Artificial Sequence
<220>
<223> PROTEIN REGULATOR OF CYTOKINESIS 1 (PRC1), MRNA
<400> 102
cccctcacac actgcttcat tt 22
<210> 103
<211> 21
<212> DNA
<213> Artificial Sequence
<220>
<223> CELL DIVISION CYCLE 2, G1 TO S AND G2 TO M (CDC2), MRNA
<400> 103
ctgattttgg ccttgccaga g 21
<210> 104
<211> 22
<212> DNA
<213> Artificial Sequence
<220>
<223> CELL DIVISION CYCLE 2, G1 TO S AND G2 TO M (CDC2), MRNA
<400> 104
ggatgattca gtgccatttt gc 22
<210> 105
<211> 21
<212> DNA
<213> Artificial Sequence
<220>
<223> CDC20 (CELL DIVISION CYCLE 20, S. CEREVISIAE, HOMOLOG)
<400> 105
acagttcgcg ttcgagagtg a 21
<210> 106
<211> 21
<212> DNA
<213> Artificial Sequence
<220>
<223> CDC20 (CELL DIVISION CYCLE 20, S. CEREVISIAE, HOMOLOG)
<400> 106
ttccactgag ccgaaggatc t 21
<210> 107
<211> 22
<212> DNA
<213> Artificial Sequence
<220>
<223> LIKELY ORTHOLOG OF MATERNAL EMBRYONIC LEUCINE ZIPPER KINAS
<400> 107
tggctctctc ccagtagcat tc 22
<210> 108
<211> 21
<212> DNA
<213> Artificial Sequence
<220>
<223> LIKELY ORTHOLOG OF MATERNAL EMBRYONIC LEUCINE ZIPPER KINAS
<400> 108
tcacttgcgg tcacatcttc c 21
<210> 109
<211> 22
<212> DNA
<213> Artificial Sequence
<220>
<223> MINICHROMOSOME MAINTENANCE DEFICIENT (S. CEREVISIAE) 7
<400> 109
agagaacaca aggattgccc ag 22
<210> 110
<211> 22
<212> DNA
<213> Artificial Sequence
<220>
<223> MINICHROMOSOME MAINTENANCE DEFICIENT (S. CEREVISIAE) 7
<400> 110
atgacaggag ctgagacttg gc 22
<210> 111
<211> 22
<212> DNA
<213> Artificial Sequence
<220>
<223> CYCLIN A2 (CCNA2), MRNA
<400> 111
ggcactgctg ctatgctgtt ag 22
<210> 112
<211> 21
<212> DNA
<213> Artificial Sequence
<220>
<223> CYCLIN A2 (CCNA2), MRNA
<400> 112
tgaaggtcca tgagacaagg c 21
<210> 113
<211> 19
<212> DNA
<213> Artificial Sequence
<220>
<223> THYMIDINE KINASE 1, SOLUBLE (TK1), MRNA
<400> 113
ggcgcaatga gctgcatta 19
<210> 114
<211> 22
<212> DNA
<213> Artificial Sequence
<220>
<223> THYMIDINE KINASE 1, SOLUBLE (TK1), MRNA
<400> 114
aaatggcttc ctctggaagg tc 22
<210> 115
<211> 21
<212> DNA
<213> Artificial Sequence
<220>
<223> cyclin-dependent kinase inhibitor 2A (melanoma, p16, inhibits
CDK4) (CDKN2A), mRNA
<400> 115
tccccgattg aaagaaccag a 21
<210> 116
<211> 22
<212> DNA
<213> Artificial Sequence
<220>
<223> cyclin-dependent kinase inhibitor 2A (melanoma, p16, inhibits
CDK4) (CDKN2A), mRNA
<400> 116
aagagaagcc agtaaccccc ct 22
Claims (32)
- 배열표의 배열번호 1, 2, 4, 6, 7, 9, 10, 11, 12, 14, 15, 17, 19 또는 46에 나타내는 염기배열을 갖는 유전자인 것을 특징으로 하는 간엽계 간세포 검출용 유전자 마커.
- 유전자 INTEGRIN, ALPHA 6 (ITGA6), MRNA（NM_000210）；유전자 SOLUTE CARRIER FAMILY 20 (PHOSPHATE TRANSPORTER)、MEMBER 1（NM_005415）；유전자RIBONUCLEOTIDE REDUCTASE M2 POLYPEPTIDE (RRM2), MRNA（NM_001034）；유전자FOLLISTATIN (FST), TRANSCRIPT VARIANT FST317, MRNA（NM_006350）；유전자 SPROUTY (DROSOPHILA) HOMOLOG 2 (SPRY2), MRNA（NM_005842）；유전자 RAB3B, MEMBER RAS ONCOGENE FAMILY (RAB3B), MRNA（NM_002867）；유전자 SOLUTE CARRIER FAMILY 2 (FACILITATED GLUCOSE TRANSPORTER)（NM_006516）；유전자 INTERLEUKIN 13 RECEPTOR, ALPHA 2 (IL13RA2), MRNA（NM_000640）；유전자 SERINE/THREONINE KINASE 12 (STK12), MRNA（NM_004217）；유전자 MINICHROMOSOME MAINTENANCE DEFICIENT (S. CEREVISIAE) 5 (CE)（NM_006739）；유전자 THYROID HORMONE RECEPTOR INTERACTOR 13 (TRIP13), MRNA（NM_004237）；유전자 KINESIN-LIKE 6 (MITOTIC CENTROMERE-ASSOCIATED KINESIN) (K)（NM_006845）；유전자 CYCLIN-DEPENDENT KINASE INHIBITOR 3 (CDK2-ASSOCIATED DUAL（NM_005192）；유전자 CHROMOSOME CONDENSATION PROTEIN G (HCAP-G),MRNA（NM_022346）；유전자 CDC28 PROTEIN KINASE 1 (CKS1), MRNA（NM_001826）；유전자 PROTEIN REGULATOR OF CYTOKINESIS 1 (PRC1), MRNA（NM_003981）；유전자 CELL DIVISION CYCLE 2, G1 TO S AND G2 TO M (CDC2), MRNA（NM_001786）；유전자 CDC20 (CELL DIVISION CYCLE 20, S. CEREVISIAE, HOMOLOG) (CD)（NM_001255）；유전자 LIKELY ORTHOLOG OF MATERNAL EMBRYONIC LEUCINE ZIPPER KINAS（NM_014791）；유전자 MINICHROMOSOME MAINTENANCE DEFICIENT (S. CEREVISIAE) 7 (M)（NM_005916）；유전자 CYCLIN A2 (CCNA2), MRNA（NM_001237）；유전자 THYMIDINE KINASE 1, SOLUBLE (TK1), MRNA（NM_003258）；또는 유전자 cyclin-dependent kinase inhibitor 2A (melanoma, p16, inhibits CDK4) (CDKN2A), mRNA（NM_000077）인 것을 특징으로 하는 간엽계 간세포 검출용 유전자 마커.
- 유전자 EGF-containing fibulin-like extracellular matrix protein 1（NM_018894）；유전자 Human insulin-like growth factor binding protein 5 (IGFBP5) mRNA（AU132011）；유전자 Homo sapiens clone 24775 mRNA sequence（AA402981）；proteoglycan 1, secretory granule（AV734015）；유전자 insulin-like growth factor binding protein 5（AA374325）；유전자 solute carrier family 21 (organic anion transporter), member 3（AF085224）；유전 transglutaminase 2 (C polypeptide, protein-glutamine-gamma-glutamyl transferase)（AL552373）；유전자 coagulation factor II (thrombin) receptor（M62424）；유전자 plasminogen activator, urokinase（NM_002658）；유전자 tissue inhibitor of metalloproteinase 3 (Sorsby fundus dystrophy, pseudoinflammatory)（W96324）；유전자 matrix Gla protein（BF668572）；유전자 B-cell CLL/lymphoma 1（Z23022）；유전자 CD74 antigen (invariant polypeptide of major histocompatibility complex, class II antigen-associated)（BG333618）；유전자 adducin 3 (gamma)（AL135243）；유전자 solute carrier family 16 (monocarboxylic acid transporters), member 4（NM_004696）；유전자 protein S (alpha)（NM_000313）；유전자 phosphatidylinositol (4,5) bisphosphate 5-phosphatase homolog; phosphatidylinositol polyphosphate 5-phosphatase type IV（AF187891）；유전자 progesterone membrane binding protein（BE858855）；유전자 brain-derived neurotrophic factor（X60201）；또는 유전자 apolipoprotein E（BF967316）인 것을 특징으로 하는 간엽계 간세포 검출용 유전자 마커.
- 유전자 HOMO SAPIENS WNT INHIBITORY FACTOR-1 (WIF-1), MRNA（NM_007191）；유전자 HOMO SAPIENS HYPOTHETICAL PROTEIN FLJ11175 (FLJ11175), MRNA（NM_018349）；유전자 HOMO SAPIENS RHO GDP DISSOCIATION INHIBITOR (GDI) BETA (ARHGDIB), MRNA（NM_001175）；유전자 HOMO SAPIENS POTASSIUM INTERMEDIATE/SMALL CONDUCTANCE CALCIUM-ACTIVATE（NM_021614）；유전자 HOMO SAPIENS NUCLEAR FACTOR (ERYTHROID-DERIVED 2)-LIKE 3 (NFE2L3), MRNA（NM_004289）；유전자 HOMO SAPIENS GS3955 PROTEIN (GS3955), MRNA（NM_021643）；유전자 HOMO SAPIENS CDNA 3' END SIMILAR TO SIMILAR TO GLYCOPROTEIN MUC18（AA302605）；유전자 HOMO SAPIENS HYPOTHETICAL PROTEIN FLJ21841 (FLJ21841), MRNA（NM_024609）；유전자 HOMO SAPIENS ZINC FINGER PROTEIN 185 (LIM DOMAIN) (ZNF185), MRNA（NM_007150）；유전자 SC1=PUTATIVE TRANS-ACTING FACTOR INVOLVED IN CELL CYCLE CONTROL [HUMAN, MRNA, 24]（S53374）；유전자 HOMO SAPIENS NADH DEHYDROGENASE (UBIQUINONE) 1 ALPHA SUBCOMPLEX, 8 (19K)（NM_014222）；유전자 HOMO SAPIENS HOMOLOG OF YEAST MCM10; HYPOTHETICAL PROTEIN PRO2249 (PRO2)（NM_018518）；유전자 HOMO SAPIENS COLLAGEN, TYPE VII, ALPHA 1 (EPIDERMOLYSIS BULLOSA, DYSTRO（NM_000094）；유전자 HOMO SAPIENS AUTOCRINE MOTILITY FACTOR RECEPTOR (AMFR), MRNA（NM_001144）；유전자 HOMO SAPIENS CLONE 24421 MRNA SEQUENCE /CDS=UNKNOWN /GB=AF070641 /GI=3283914 /UG=（AF070641）；유전자 HOMO SAPIENS MRNA;유전자 CDNA DKFZP586E1621 (FROM CLONE DKFZP586E1621) /CDS=UNKNOWN /GB（AL080235）인 것을 특징으로 하는 간엽계 간세포 검출용 유전자 마커.
- 유전자 HOMO SAPIENS RESERVED (KCNK12), MRNA（NM_022055）；유전자 HOMO SAPIENS UBIQUITIN-CONJUGATING ENZYME E2C (UBE2C), MRNA（NM_007019）；유전자 HOMO SAPIENS CDNA FLJ10517 FIS, CLONE NT2RP2000812 /CDS=(61,918) /GB=AK001379 /G（AK001379）；유전자 HOMO SAPIENS CARTILAGE LINKING PROTEIN 1 (CRTL1), MRNA（NM_001884）；유전자 HOMO SAPIENS MRNA; CDNA DKFZP434F2322 (FROM CLONE DKFZP434F2322) /CDS=UNKNOWN /GB（AL133105）；유전자 HOMO SAPIENS, CLONE MGC:9549 IMAGE:3857382, MRNA, COMPLETE CDS /CDS=(92,1285) /G（BC012453）；유전자 HOMO SAPIENS HYPOTHETICAL PROTEIN DKFZP586J1119 (DKFZP586J1119), MRNA（NM_032270）；유전자 HOMO SAPIENS GLIA MATURATION FACTOR, GAMMA (GMFG), MRNA（NM_004877）；유전자 HOMO SAPIENS CDNA: FLJ23095 FIS, CLONE LNG07413 /CDS=UNKNOWN /GB=AK026748 /GI=10（AK026748）；유전자 HOMO SAPIENS HYPOTHETICAL PROTEIN DKFZP762E1312 (DKFZP762E1312), MRNA（NM_018410）；유전자 HOMO SAPIENS IROQUOIS-CLASS HOMEODOMAIN PROTEIN (IRX-2A), MRNA（NM_005853）；또는 유전자 HOMO SAPIENS HYPOTHETICAL PROTEIN FLJ10540 (FLJ10540), MRNA（NM_018131）인 것을 특징으로 하는 간엽계 간세포 검출용 유전자 마커.
- 유전자 HOMO SAPIENS MRNA; CDNA DKFZP434C1915 (FROM CLONE DKFZP434C1915); PARTIAL CDS /C（AL137698）；유전자 HOMO SAPIENS MINICHROMOSOME MAINTENANCE DEFICIENT (S. CEREVISIAE) 7 (M)（NM_005916）；유전자 HOMO SAPIENS DYNEIN, CYTOPLASMIC, INTERMEDIATE POLYPEPTIDE 1 (DNCI1), M（NM_004411）；유전자 HUMAN MRNA FOR PROTEIN GENE PRODUCT (PGP) 9.5.（X04741）；유전자 HOMO SAPIENS RAD51-INTERACTING PROTEIN (PIR51), MRNA（NM_006479）；유전자 HOMO SAPIENS BACULOVIRAL IAP REPEAT-CONTAINING 5 (SURVIVIN) (BIRC5), MR（NM_001168）；유전자 HOMO SAPIENS UDP-N-ACETYL-ALPHA-D-GALACTOSAMINE:POLYPEPTIDE N-ACETYLGAL（NM_004482）；유전자 HOMO SAPIENS CYCLIN B1 (CCNB1), MRNA（NM_031966）；유전자 HOMO SAPIENS FLAP STRUCTURE-SPECIFIC ENDONUCLEASE 1 (FEN1), MRNA（NM_004111）；유전자 HOMO SAPIENS SERINE/THREONINE KINASE 15 (STK15), MRNA（NM_003600）；유전자 HOMO SAPIENS MAD2 (MITOTIC ARREST DEFICIENT, YEAST, HOMOLOG)-LIKE 1 (MA)（NM_002358）；유전자 HOMO SAPIENS NUCLEOLAR PROTEIN ANKT (ANKT), MRNA（NM_018454）；유전자 HOMO SAPIENS HSPC150 PROTEIN SIMILAR TO UBIQUITIN-CONJUGATING ENZYME (H)（NM_014176）；유전자 HOMO SAPIENS CYTOCHROME P450 RETINOID METABOLIZING PROTEIN (P450RAI-2),（NM_019885）；유전자 HOMO SAPIENS NIMA (NEVER IN MITOSIS GENE A)-RELATED KINASE 2 (NEK2), MR（NM_002497）；유전자 HOMO SAPIENS BONE MORPHOGENETIC PROTEIN 4 (BMP4), MRNA（NM_001202）；유전자 HOMO SAPIENS CDNA FLJ10674 FIS, CLONE NT2RP2006436 /CDS=UNKNOWN /GB=AK001536 /GI（AK001536）；또는 유전자 HOMO SAPIENS CENTROMERE PROTEIN A (17KD) (CENPA), MRNA（NM_001809）인 것을 특징으로 하는 간엽계 간세포 검출용 유전자 마커.
- 유전자 HOMO SAPIENS MRNA; CDNA DKFZP564P116 (FROM CLONE DKFZP564P116) /CDS=UNKNOWN /GB=A（AL049338）；유전자 HOMO SAPIENS TTK PROTEIN KINASE (TTK), MRNA（NM_003318）；유전자 HOMO SAPIENS KARYOPHERIN ALPHA 2 (RAG COHORT 1, IMPORTIN ALPHA 1) (KPNA（NM_002266）；유전자 HOMO SAPIENS POLYMERASE (DNA DIRECTED), THETA (POLQ), MRNA（NM_006596）；유전자 HOMO SAPIENS ETS VARIANT GENE 5 (ETS-RELATED MOLECULE) (ETV5), MRNA（NM_004454）；유전자 HOMO SAPIENS TRANSFORMING, ACIDIC COILED-COIL CONTAINING PROTEIN 3 (TAC)（NM_006342）；유전자 HOMO SAPIENS KINESIN-LIKE 1 (KNSL1), MRNA（NM_004523）；유전자 HOMO SAPIENS CENTROMERE PROTEIN E (312KD) (CENPE), MRNA（NM_001813）；유전자 HOMO SAPIENS CDNA, 3' END /CLONE=IMAGE:1651289 /CLONE_END=3' /GB=AI12（AI123815）；유전자 HOMO SAPIENS DISTAL-LESS HOMEO BOX 5 (DLX5), MRNA（NM_005221）；유전자 HOMO SAPIENS V-MYB AVIAN MYELOBLASTOSIS VIRAL ONCOGENE HOMOLOG-LIKE 2 （NM_002466）；또는 유전자 HOMO SAPIENS SERUM DEPRIVATION RESPONSE (PHOSPHATIDYLSERINE-BINDING PRO（NM_004657）인 것을 특징으로 하는 간엽계 간세포 검출용 유전자 마커.
- 유전자 HOMO SAPIENS HYPOTHETICAL PROTEIN FLJ10604 (FLJ10604), MRNA（NM_018154）；유전자 HOMO SAPIENS RIBONUCLEASE HI, LARGE SUBUNIT (RNASEHI), MRNA（NM_006397）；유전자 HOMO SAPIENS ANILLIN (DROSOPHILA SCRAPS HOMOLOG), ACTIN BINDING PROTEIN（NM_018685）；유전자 HOMO SAPIENS H4 HISTONE FAMILY, MEMBER G (H4FG), MRNA（NM_003542）；유전자 HOMO SAPIENS G PROTEIN-COUPLED RECEPTOR 37 (ENDOTHELIN RECEPTOR TYPE B-（NM_005302）HOMO SAPIENS UBIQUITIN CARRIER PROTEIN (E2-EPF), MRNA（NM_014501）；유전자 HOMO SAPIENS, SIMILAR TO TUMOR DIFFERENTIALLY EXPRESSED 1, CLONE IMAGE:3639252, （BC007375）；유전자 HOMO SAPIENS HYPOTHETICAL PROTEIN MGC2577 (MGC2577), MRNA（NM_031299）；유전자 HOMO SAPIENS HYPOTHETICAL PROTEIN FLJ13912 (FLJ13912), MRNA（NM_022770）；유전자 HOMO SAPIENS ANNEXIN A3 (ANXA3), MRNA（NM_005139）；유전자 HOMO SAPIENS STATHMIN 1/ONCOPROTEIN 18 (STMN1), MRNA（NM_005563）HOMO SAPIENS, SIMILAR TO RIBOSOMAL PROTEIN L39, CLONE MGC:20168 IMAGE:4555759, M（BC012328）；유전자 HOMO SAPIENS POLO (DROSOPHIA)-LIKE KINASE (PLK), MRNA（NM_005030）인 것을 특징으로 하는 간엽계 간세포 검출용 유전자 마커.
- 유전자 HOMO SAPIENS PROTEASOME (PROSOME, MACROPAIN) 26S SUBUNIT, ATPASE, 5 (PS)（NM_002805）；유전자 HOMO SAPIENS EUKARYOTIC TRANSLATION INITIATION FACTOR 4 GAMMA, 2 (EIF4G)（NM_001418）；유전자 HOMO SAPIENS DIHYDROPYRIMIDINASE-LIKE 3 (DPYSL3), MRNA）（NM_001387）；유전자 HOMO SAPIENS ADAPTOR-RELATED PROTEIN COMPLEX 1, SIGMA 2 SUBUNIT (AP1S2)（NM_003916）；유전자 HOMO SAPIENS PLECTIN 1, INTERMEDIATE FILAMENT BINDING PROTEIN, 500KD (P)（NM_000445）；유전자 HOMO SAPIENS HETEROGENEOUS NUCLEAR RIBONUCLEOPROTEIN U (SCAFFOLD ATTAC（NM_031844）；유전자 HOMO SAPIENS HYPOTHETICAL PROTEIN MGC5306 (MGC5306), MRNA（NM_024116）；유전자 HOMO SAPIENS MHC CLASS I POLYPEPTIDE-RELATED SEQUENCE A (MICA), MRNA（NM_000247）；유전자 HOMO SAPIENS PROTEIN ASSOCIATED WITH PRK1 (AWP1), MRNA（NM_019006）；유전자 HOMO SAPIENS PROGESTERONE RECEPTOR MEMBRANE COMPONENT 2 (PGRMC2), MRNA（NM_006320）；유전자 HOMO SAPIENS POLYGLUTAMINE BINDING PROTEIN 1 (PQBP1), MRNA（NM_005710）；유전자 HOMO SAPIENS S-ADENOSYLMETHIONINE DECARBOXYLASE 1 (AMD1), MRNA（NM_001634）； HOMO SAPIENS INTERFERON-INDUCED, HEPATITIS C-ASSOCIATED MICROTUBULAR AG（NM_006417）；유전자 HOMO SAPIENS PROTEIN KINASE, AMP-ACTIVATED, GAMMA 1 NON-CATALYTIC SUBUN（NM_002733）；HOMO SAPIENS PROCOLLAGEN-PROLINE, 2-OXOGLUTARATE 4-DIOXYGENASE (PROLINE（NM_004199）；유전자 HOMO SAPIENS ENDOTHELIAL DIFFERENTIATION, LYSOPHOSPHATIDIC ACID G-PROTE（NM_012152）；또는 유전자 HOMO SAPIENS KIAA0127 GENE PRODUCT (KIAA0127), MRNA（NM_014755）인 것을 특징으로 하는 간엽계 간세포 검출용 유전자 마커.
- 제 1항 내지 제 9항 중의 어느 한 항에 기재된 간엽계 간세포 검출용 마커 유전자와 스트린젠트한 조건하에서 하이브리다이즈하는 DNA배열을 갖는 간엽계 간세포 마커 유전자 검출용 프로브.
- 제 10항에 있어서, 제 1항 내지 제 9항 중의 어느 한 항에 기재된 염기배열의 안티센스사슬의 전부 또는 일부로 이루어지는 간엽계 간세포 마커 유전자 검출용 프로브.
- 제 10항 또는 제 11항 기재의 DNA의 적어도 하나 이상을 고정화시킨 것을 특징으로 하는 간엽계 간세포 마커 유전자 검출용 마이크로어레이 또는 DNA칩.
- 제 12항에 있어서,배열표의 배열번호 1, 2, 4, 6, 7, 9, 10, 11, 12, 14, 15, 17, 19 또는 46에 나타내는 염기배열을 갖는 유전자의 그룹, 및 제 2항 내지 제 9항 중의 어느 한 항에 기재된 유전자 그룹의 2 이상의 유전자를 검출하는 프로브를 고정화시킨 것을 특징으로 하는 간엽계 간세포 마커 유전자 검출용 마이크로어레이 또는 DNA칩.
- 배열표의 배열번호 3, 5, 8, 13, 16 또는 18에 나타내는 아미노산 배열을 갖는 폴리펩티드인 것을 특징으로 하는 간엽계 간세포 검출용 폴리펩티드 마커.
- 제 14항에 기재된 폴리펩티드를 사용하여 유도되며, 상기 폴리펩티드에 특이적으로 결합하는 것을 특징으로 하는 항체.
- 제 15항에 있어서,상기 항체가 모노클로널항체인 것을 특징으로 하는 항체.
- 제 15항에 있어서,상기 항체가 폴리클로널항체인 것을 특징으로 하는 항체.
- 피검세포에 있어서의 제 1항 내지 제 9항 중의 어느 한 항에 기재된 간엽계 간세포 마커 유전자의 발현을 검출하는 것을 특징으로 하는 간엽계 간세포의 식별방법.
- 제 18항에 있어서,상기 간엽계 간세포 마커 유전자 발현의 검출을, 노던 블로팅법을 사용하여 실시하는 것을 특징으로 하는 간엽계 간세포의 식별방법.
- 제 18항에 있어서,상기 간엽계 간세포 마커 유전자 발현의 검출을, 제 10항 또는 제 11항에 기재된 간엽계 간세포 마커 유전자 검출용 프로브를 사용하여 실시하는 것을 특징으로 하는 간엽계 간세포의 식별방법.
- 제 18항에 있어서,상기 간엽계 간세포 마커 유전자 발현의 검출을, 제 12항 또는 제 13항에 기재된 간엽계 간세포 마커 유전자 검출용 마이크로어레이 또는 DNA칩을 사용하여 실시하는 것을 특징으로 하는 간엽계 간세포의 식별방법.
- 제 18항 내지 제 21항 중의 어느 한 항에 기재된 간엽계 간세포 마커 유전자 발현의 검출이, 정량적 또는 반정량적 PCR의 사용을 포함하고 있는 것을 특징으로 하는 간엽계 간세포의 식별방법.
- 제 22항에 있어서,상기 정량적 또는 반정량적 PCR의 사용이, RT-PCR법인 것을 특징으로 하는 간엽계 간세포의 식별방법.
- 배열표의 배열번호20에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호21에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호22에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호23에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호24에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호25에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호26에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호27에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호28에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호29에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호30에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호31에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호32에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호33에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호34에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호35에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호36에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호37에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호38에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호39에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호40에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호41에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호42에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호43에 나타내는 염기배열을 갖는 안티센스프라이머, 또는 배열표의 배열번호44에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호45에 나타내는 염기배열을 갖는 안티센스프라이머로 이루어지는 간엽계 간세포 마커 유전자 증폭을 위한 RT-PCR용 프라이머.
- 배열표의 배열번호47에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호48에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호49에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호50에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호51에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호52에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호53에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호54에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호55에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호56에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호57에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호58에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호59에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호60에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호61에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호62에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호63에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호64에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호65에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호66에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호67에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호68에 나타내는 염기배열을 갖는 안티센스프라이머, 또는 배열표의 배열번호69에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호70에 나타내는 염기배열을 갖는 안티센스프라이머로 이루어지는 간엽계 간세포 마커 유전자 증폭을 위한 실시간 PCR용 프라이머.
- 배열표의 배열번호71에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호72에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호73에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호74에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호75에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호76에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호77에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호78에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호79에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호80에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호81에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호82에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호83에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호84에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호85에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호86에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호87에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호88에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호89에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호90에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호91에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호92에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호93에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호94에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호95에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호96에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호97에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호98에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호99에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호100에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호101에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호102에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호103에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호104에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호105에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호106에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호107에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호108에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호109에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호110에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호111에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호112에 나타내는 염기배열을 갖는 안티센스프라이머, 배열표의 배열번호113에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호114에 나타내는 염기배열을 갖는 안티센스프라이머, 또는 배열표의 배열번호115에 나타내는 염기배열을 갖는 센스프라이머와 배열표의 배열번호116에 나타내는 염기배열을 갖는 안티센스프라이머로 이루어지는 간엽계 간세포 마커 유전자 증폭을 위한 RT-PCR용 프라이머.
- 제 18항에 있어서,피검세포에 있어서의 유전자를, 제 24항 내지 제 26항 중의 어느 한 항에 기재된 센스프라이머 및 안티센스프라이머의 적어도 1쌍 이상의 프라이머를 사용하여 증폭시키고, 증폭한 유전자를 제 10항 또는 제 11항에 기재된 간엽계 간세포 마커 유전자 검출용 프로브를 사용하여 검출하는 것을 특징으로 하는 간엽계 간세포의 식별방법.
- 제 15항 내지 제 17항 중의 어느 한 항에 기재된 항체를 사용하여, 피검세포에 있어서의 간엽계 간세포 마커 폴리펩티드의 발현을 검출하는 것을 특징으로 하는 간엽계 간세포의 식별방법.
- 피검세포에 있어서의 배열표의 배열번호 1, 11 및 19로 이루어지는 간엽계 간세포 마커 유전자 그룹의 1유전자 이상의 발현과, 배열표의 배열번호 2, 4, 6, 7, 9, 10, 12, 14, 15 및 17로 이루어지는 간엽계 간세포 마커 유전자 그룹의 1유전자 이상의 발현을 조합하여 평가하는 것을 특징으로 하는 간엽계 간세포의 식별방법.
- 제 10항 또는 제 11항에 기재된 간엽계 간세포 마커 유전자 검출용 프로브 및/또는 제 15항 내지 제 17항 중의 어느 한 항에 기재된 항체를 사용하여, 피검세포에 있어서의 간엽계 간세포 검출용 마커 유전자 및/또는 간엽계 간세포 검출용 마커 폴리펩티드의 발현을 검출하여, 식별한 간엽계 간세포를 분리하는 것을 특징으로 하는 간엽계 간세포의 식별분리방법.
- 제 30항에 있어서,제 15항 내지 제 17항 중의 어느 한 항에 기재된 항체를 사용한 형광항체법에 의하여 피검세포 중에 있어서의 간엽계 간세포를 표지화하고, 상기 표지화한 간엽계 간세포를 분리하는 것을 특징으로 하는 간엽계 간세포의 식별분리방법.
- 제 10항 또는 제 11항 기재의 간엽계 간세포 마커 유전자 검출용 프로브, 상기 프로브를 고정한 제 12항 또는 제 13항에 기재된 간엽계 간세포 마커 유전자 검출용 마이크로어레이 또는 DNA칩, 및 제 15항 내지 제 17항 중의 어느 한 항에 기재된 항체의 적어도 하나 이상을 장비하여 이루어지는 간엽계 간세포의 식별용 키트.
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| WO (1) | WO2004081174A2 (ko) |
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| WO2006112390A1 (ja) * | 2005-04-14 | 2006-10-26 | Japan As Represented By General Director Of Agency Of National Cardiovascular Center | 脂肪由来前駆細胞およびその利用 |
| US20060246498A1 (en) * | 2005-04-28 | 2006-11-02 | Ramot At Tel Aviv University Ltd. | HNRPLL polypeptides, polynucleotides encoding same and compositions and methods of using same |
| JP4958094B2 (ja) * | 2005-09-22 | 2012-06-20 | 国立大学法人広島大学 | 間葉系幹細胞の均質性識別方法、その方法を利用して得られる均質間葉系幹細胞 |
| CN101360761B (zh) | 2005-12-08 | 2012-09-12 | 米德列斯公司 | 抗蛋白质酪氨酸激酶7（ptk7）的人单克隆抗体及其用途 |
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| WO2007085266A1 (en) * | 2006-01-30 | 2007-08-02 | Dako Denmark A/S | High-speed quantification of antigen specific t-cells in whole blood by flow cytometry |
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| EP2167536A1 (en) * | 2007-07-03 | 2010-03-31 | Dako Denmark A/S | Mhc multimers, methods for their generation, labeling and use |
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| CA2898601C (en) | 2013-02-01 | 2021-05-18 | Tohoku University | Method for separating cell from biological tissue |
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| CN104655857B (zh) * | 2015-02-10 | 2017-05-31 | 南京大学 | 一种微生物细胞内多聚磷酸盐的定量检测方法 |
| US10539553B2 (en) | 2015-03-17 | 2020-01-21 | National Institute Of Advanced Industrial Science And Technology | Method and kit for detecting stem cell |
| WO2017004592A1 (en) | 2015-07-02 | 2017-01-05 | Terumo Bct, Inc. | Cell growth with mechanical stimuli |
| CN105018524B (zh) * | 2015-07-13 | 2020-03-27 | 广州赛琅生物技术有限公司 | 一种细胞寿命延长及成血管能力增强的人干细胞的制备方法及其试剂盒 |
| US11965175B2 (en) | 2016-05-25 | 2024-04-23 | Terumo Bct, Inc. | Cell expansion |
| US11685883B2 (en) | 2016-06-07 | 2023-06-27 | Terumo Bct, Inc. | Methods and systems for coating a cell growth surface |
| US11104874B2 (en) | 2016-06-07 | 2021-08-31 | Terumo Bct, Inc. | Coating a bioreactor |
| CN106771256B (zh) * | 2017-01-23 | 2019-02-01 | 福建医科大学 | 骨骼肌组织冰冻切片肌球蛋白三磷酸腺苷酶染色方法 |
| WO2018159431A1 (ja) | 2017-03-03 | 2018-09-07 | ロート製薬株式会社 | 間葉系幹細胞及び肝疾患治療剤 |
| US11624046B2 (en) | 2017-03-31 | 2023-04-11 | Terumo Bct, Inc. | Cell expansion |
| EP3656841A1 (en) | 2017-03-31 | 2020-05-27 | Terumo BCT, Inc. | Cell expansion |
| SG11201909153UA (en) * | 2017-04-10 | 2019-10-30 | Immatics Biotechnologies Gmbh | Peptides and combination thereof for use in the immunotherapy against cancers |
| PL241520B1 (pl) * | 2020-03-27 | 2022-10-17 | Polski Bank Komorek Macierzystych Spolka Akcyjna | Zastosowanie panelu genów do określania potencjału teratogennego komórek mezenchymalnych i pochodzenia perinatalnego |
| US12043823B2 (en) | 2021-03-23 | 2024-07-23 | Terumo Bct, Inc. | Cell capture and expansion |
| EP4408979A1 (en) * | 2021-09-30 | 2024-08-07 | Polski Bank Komórek Macierzystych S.A. | Expression analysis of a specific gene pool to determine whether the population of adipose tissue-derived mesenchymal cells (at-msc) selected for clinical application can undergo a transformation into neoplastic cells |
| CN114622019A (zh) * | 2022-03-23 | 2022-06-14 | 新乡医学院 | Birc5基因作为人骨髓间充质干细胞成骨分化抑制剂的应用 |
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| US5340739A (en) * | 1988-07-13 | 1994-08-23 | Brigham & Women's Hospital | Hematopoietic cell specific transcriptional regulatory elements of serglycin and uses thereof |
| US5965436A (en) * | 1996-11-15 | 1999-10-12 | Osiris Therapeutics, Inc. | Method of isolating mesenchymal stem cells associated with isolated megakaryocytes by isolating megakaryocytes |
| AU2180799A (en) * | 1998-02-13 | 1999-08-30 | Peter Sonderegger | Neuroserpin |
| ATE328067T1 (de) * | 2000-04-28 | 2006-06-15 | Childrens Medical Center | Isolierung von mesenchymalen stammzellen und deren verwendung |
| WO2001094629A2 (en) * | 2000-06-05 | 2001-12-13 | Avalon Pharmaceuticals | Cancer gene determination and therapeutic screening using signature gene sets |
| WO2002028999A2 (en) * | 2000-10-03 | 2002-04-11 | Gene Logic, Inc. | Gene expression profiles in granulocytic cells |
| JP2003052365A (ja) * | 2001-08-20 | 2003-02-25 | Japan Science & Technology Corp | 哺乳動物からの間葉系幹細胞の分離及びその利用方法 |
| EP1288293A1 (en) * | 2001-08-30 | 2003-03-05 | Norio Sakuragawa | Human neural stem cells originated from human amniotic mesenchymal cell layer |
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2004
- 2004-02-27 AU AU2004219820A patent/AU2004219820B2/en not_active Ceased
- 2004-02-27 US US10/548,681 patent/US20060166214A1/en not_active Abandoned
- 2004-02-27 EP EP04715524A patent/EP1605044A4/en not_active Withdrawn
- 2004-02-27 KR KR1020057016952A patent/KR100818689B1/ko not_active IP Right Cessation
- 2004-02-27 AT AT07017847T patent/ATE488588T1/de not_active IP Right Cessation
- 2004-02-27 DE DE602004030172T patent/DE602004030172D1/de not_active Expired - Lifetime
- 2004-02-27 CN CNA200810081304XA patent/CN101265503A/zh active Pending
- 2004-02-27 EP EP07017847A patent/EP1860186B1/en not_active Expired - Lifetime
- 2004-02-27 WO PCT/JP2004/002457 patent/WO2004081174A2/ja not_active Application Discontinuation
- 2004-02-27 CN CNA2004800075500A patent/CN1761749A/zh active Pending
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| CN1761749A (zh) | 2006-04-19 |
| WO2004081174A2 (ja) | 2004-09-23 |
| EP1860186B1 (en) | 2010-11-17 |
| AU2004219820A1 (en) | 2004-09-23 |
| ATE488588T1 (de) | 2010-12-15 |
| CN101265503A (zh) | 2008-09-17 |
| AU2004219820B2 (en) | 2008-06-12 |
| EP1860186A3 (en) | 2008-03-12 |
| WO2004081174A3 (ja) | 2004-12-02 |
| KR100818689B1 (ko) | 2008-04-02 |
| JP2004290189A (ja) | 2004-10-21 |
| EP1605044A4 (en) | 2006-12-06 |
| US20060166214A1 (en) | 2006-07-27 |
| EP1860186A2 (en) | 2007-11-28 |
| DE602004030172D1 (de) | 2010-12-30 |
| EP1605044A2 (en) | 2005-12-14 |
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