CN1966725A - 骨形态生发蛋白(bmp)、bmp受体和bmp结合蛋白及它们在诊断和治疗青光眼中的用途 - Google Patents
骨形态生发蛋白(bmp)、bmp受体和bmp结合蛋白及它们在诊断和治疗青光眼中的用途 Download PDFInfo
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Abstract
本发明提供了诊断和治疗青光眼的方法和试剂盒。
Description
本申请是中国专利申请02820917.6的分案申请,原申请的国际申请号是PCT/US02/35251,申请日是2002年10月31日,发明名称是“骨形态发生蛋白(BMP)、BMP受体和BMP结合蛋白及它们在诊断和治疗青光眼中的用途”。
发明领域
本发明公开了用于诊断和治疗青光眼及相关疾病的方法和试剂。
相关领域的描述
“青光眼”是一组使人虚弱的眼病,它们是美国和其它发达国家不可逆失明的主要原因。原发性开角型青光眼(“POAG”)是青光眼的最常见形式,其特征为小梁网变性,从而阻塞了房水的正常性能,使眼不能关闭虹膜和角膜之间的间隙(例如“角”)(Vaughan,D.等人,(1992))。在该疾病中此阻塞的特征是眼内压(“IOP”)升高,如果不进行合适且及时的治疗,会导致进行性视力丧失和失明。该疾病估计可影响所有40岁以上成年人中的0.4%到3.3%(Leske,M.C.等人(1986);Bengtsson,B.(1989);Strong,N.P.(1992))。此外,该疾病会随年龄升高而多发,对75岁或75岁以上的人可达6%以上(Strong,N.P.,(1992)。
由于IOP升高是青光眼的一个容易测量的特征,对该疾病的诊断主要是通过测量眼内压(眼压测量法)进行筛选(Strong,N.P.(1992);Greve,M.等人(1993))。不幸的是,由于青光眼的压力范围和正常压力范围发生重叠,如果不获得多次读数,这些方法的用途是有限的(Hitchings,R.A.,(1993);Tuck,M.W.等人(1993);Vaughan,D.等人,(1992);Vernon,S.A.,(1993))。由于此原因,通常还采用其他方法如直接检查视神经乳头和测定患者视野丧失程度来提高诊断的准确性(Greve,M.等人,(1993))。
青光眼影响眼的三个单独的组织。与POAG相关的IOP升高是由于小梁网(TM)的形态学和生物化学的改变,其中小梁网组织位于虹膜和角膜间的角。大部分营养性房水通过TM存在于眼前角。TM细胞的进行性丧失和青光眼的TM中细胞外碎片的堆积导致房水流出的阻力增加(Lutjen-Drecoll和Rohen 1996;Rohen 1983;Rohen等人1993;Grierson和Calthorpe 1988),因此使IOP升高。IOP升高以及其它因素例如局部缺血引起视神经乳头(ONH)的变性性改变,导致ONH的进行性“凹陷”(Varma和Minckler 1996;Hernandez和Gong 1996;Hernandez等人1990;Hernandez和Pena 1997;Morrison等人1990)和视网膜神经节细胞丧失(Quigley等人2000;Quigley 1999;Quigley等人1995;Kerrigan等人1997)和轴突的丧失。造成TM、ONH和视网膜神经节细胞的青光眼性损伤的详细分子机制未知。
现行青光眼的治疗是降低IOP,其中所述IOP是青光眼发展和进行的主要危险因素。这些治疗降低了IOP,但它们不直接针对致病机理,且疾病继续发展。至少半数青光眼患者没被诊断出,并且当患者诊断出患有青光眼时,他们已经丧失了约40%的视网膜神经节细胞。因此,需要青光眼的早期检测和诊断方法。
鉴于青光眼的严重性和现有诊断方法中至少部分的不足,希望有改进的、更准确的方法在早期阶段诊断青光眼。并且,希望有针对青光眼致病机理的新治疗剂。
发明概述
本发明通过提供早期诊断青光眼、治疗青光眼和鉴定治疗青光眼有用的化合物的方法和试剂盒,克服了现有技术中的以上和其它不足。
在某些特定实施方案中,本发明提供了对获自细胞或体液的样本诊断青光眼的方法,该方法通过检测骨形态发生蛋白家族成员的基因表达改变而实施。该方法通常包括下列步骤:
a)从疑似患有青光眼的患者获得组织或体液样本;
b)从上达样本提取DNA;
c)获得多条PCR引物,其中每一所述引物包含的序列由来自SEQ IDNO:1、SEQ ID NO:3、SEQ ID NO:5、SEQ ID NO:7、SEQ ID NO:37、SEQ ID NO:39、SEQ ID NO:41、SEQ ID NO:43、SEQ ID NO:45、SEQ ID NO:47或SEQ ID NO:53的18至1547个连续核苷酸组成;
d)用所述引物对经提取DNA的区域进行扩增,以获得PCR产物;
e)分析PCR产物;并
f)确定PCR产物序列与正常基因序列间的差异;
其中扩增序列与正常基因序列间的差异可诊断青光眼。
一般,本发明的方法可包括从个体获得样本并从所述样本提取DNA。选择针对BMP基因家族特定成员的PCR引物,然后用于扩增所提取基因的相应区域以获得PCR产物。通过可有效确定所评估的特定BMP家族基因(所提取的DNA)的正常和突变形式间DNA序列差异的技术对PCR产物进行分析。经确定的序列间差异为青光眼的征兆。
本发明方法中应用的组织或流体样本可为血液或口细胞。
一般,引物序列长度为约10、15或18个核苷酸至约20个或约30个核苷酸。在某些实施方案中更优选较长的序列如40、50、80、90、95、100个核苷酸甚至全长序列。本领域技术人员广泛接受至少约18至20个核苷酸长度的寡核苷酸,其足以允许充分特异的杂交,因此可用作分子探针,如Lathe(1985)所述,该文献特此引用作为参考。优选地,核苷酸序列由来自SEQ ID NO:1、SEQ ID NO:3、SEQ ID NO:5、SEQ ID NO:7、SEQ ID NO:37、SEQ ID NO:39、SEQ ID NO:41、SEQ ID NO:43、SEQ ID NO:45、SEQ ID NO:47或SEQ ID NO:53的20至100个连续核苷酸组成。也认为引物序列可由来自BMP受体基因序列和BMP相关蛋白基因序列中的至少10、15或18个连续核苷酸序列组成,其中所述BMP受体基因序列和BMP相关蛋白基因序列已知。
有10、18、20、30、50、60、65或甚至高达并包括100个核苷酸左右的、与SEQ ID NO:1、SEQ ID NO:3、SEQ ID NO:5、SEQ ID NO:7、SEQ ID NO:37、SEQ ID NO:39、SEQ ID NO:41、SEQ ID NO:43、SEQID NO:45、SEQ ID NO:47或SEQ ID NO:53中的任意一个序列互补的核酸分子可用做杂交探针。本领域技术人员认为具有约18个核苷酸长度的引物或探针可提供与靶序列的高特异性杂交。片段的总计大小以及互补序列的大小最终取决于应用特定核酸片段的预期用途。较短片段一般在杂交方案中应用,其中互补区长度可不同,例如在约10、18、20或30个核苷酸至约50、60、70、80、90或100个核苷酸之间,或甚至是希望检测的互补序列的全长。
在特定优选的实施方案中,引物由来自SEQ ID NO:1、SEQ ID NO:3、SEQ ID NO:5、SEQ ID NO:7、SEQ ID NO:37、SEQ ID NO:39、SEQ ID NO:41、SEQ ID NO:43、SEQ ID NO:45、SEQ ID NO:47或SEQ ID NO:53中的连续序列组成。在其它优选的实施方案中,引物由来自BMP受体基因(公开于ten Dijke等人1993;Astrom等人1999;Nohno等人1995,所有文献在此引用作为参考)或来自BMP相关基因的连续序列组成,例如由来自chordin(NCBI NM_029130)、gremlin(Murphy等人1999;McMahon等人2000)、卵泡抑素(follistatin)(NCBINM_003892)或bambi(NCBI NM_005791)的连续序列组成。最优选地,引物由来自SEQ ID NO:3的连续序列组成。在某些方面,至少一些引物可进一步包括可检测标记。
在其它实施方案中,本发明提供了用于治疗青光眼的方法,该方法为向需要治疗的患者施用包含这样序列的组合物,所述序列由至少一个选自BMP2激动剂、BMP4激动剂、BMP5激动剂、BMP7激动剂、Smad 1/5激动剂、chordin拮抗剂、gremlin拮抗剂和卵泡抑素拮抗剂的化合物组成。
在其它方面,本发明提供了用于鉴定治疗青光眼的治疗剂的方法。例如可通过下列步骤鉴定治疗剂:
a)获得第一组合物,其包含表达BMP-2A、BMP4、BMP-5或BMP7的一群重组细胞;
b)获得候选物质;
c)孵育所述组合物和所述候选物质;
检测所述组合物启动BMP-诱导的Smad信号途径和/或受BMP-调节的基因表达的能力;以及鉴定抑制或刺激BMP的这些下游效应的候选物质。
本发明的另一方面是包含本发明序列和适当试剂例如与蛋白质、肽或抗体本身相连的可检测标记的诊断试剂盒。备选地,可检测标记可连接至与本发明序列选择性杂交的第二个序列。
相关实施方案包括治疗试剂盒,该试剂盒包括于此公开的核酸序列或肽或蛋白质序列的可药用制剂。该试剂盒可用于检测临床样本中BMP基因和蛋白质的表达改变而诊断青光眼。
附图简述
附图组成本说明书的一部分,本说明书包括附图以进一步阐述本发明的某些方面。通过参考这些附图中的一个或多个图并结合于此呈现的特定实施方案详述,可更好地理解本发明。
图1.BMP2A的核苷酸和氨基酸序列。
图2.BMP4的核苷酸和氨基酸序列。
图3.BMP5的核苷酸和氨基酸序列。
图4.BMP7的核苷酸和氨基酸序列。
图5.骨形态发生蛋白信号传导途径。骨形态发生蛋白(BMP)二聚体与包含BMP受体1和2的膜复合体结合,所述受体为丝氨酸/苏氨酸激酶。调节性Smads(Smad1/Smad5)发生磷酸化并与通用Smad(Smad4)结合。产生的Smad复合体进入核,在核内与转录因子(TF)结合并调节基因表达。BMP相关蛋白作为BMP拮抗剂通过与BMPs结合并阻止BMP与BMP受体的相互作用。
图6.人TM细胞和组织中BMP的表达。来自人TM细胞(1-5泳道)和组织(6-7泳道)中BMP表达的RT-PCR分析产生的cDNA样本BMPPCR产物的经溴化乙锭染色的琼脂糖凝胶。L=碱基对标准。C=PCR阴性对照泳道。β-肌动蛋白用作阳性RT-PCR内部对照。
图7.人TM细胞和组织中BMP受体的表达。来自人TM细胞(1-5泳道)和组织(6-7泳道)中BMP受体表达的RT-PCR分析产生的cDNA样本PCR产物的经溴化乙锭染色的琼脂糖凝胶。L=碱基对标准。C=PCR阴性对照泳道。β-肌动蛋白用作阳性RT-PCR内部对照。
图8.人ONH星形胶质细胞、ONH组织和人脑星形胶质细胞中BMP的表达。来自人ONH星形胶质细胞(1-5泳道)、ONH组织(6泳道)和人脑星形胶质细胞(7泳道)中BMP表达的RT-PCR分析产生的cDNA样本PCR产物的经溴化乙锭染色的琼脂糖凝胶。L=碱基对标准。C=PCR阴性对照泳道。β-肌动蛋白用作阳性RT-PCR内部对照。
图9.人筛板细胞系中BMP的表达。来自人筛板细胞(1-9泳道)的RT-PCR分析产生的cDNA样本PCR产物的经溴化乙锭染色的琼脂糖凝胶。L=碱基对标准。C=PCR阴性对照泳道。β-肌动蛋白用作阳性RT-PCR内部对照。
图10.人ONH星形胶质细胞、ONH组织和人脑星形胶质细胞中BMP受体的表达。来自人视神经乳头星形胶质细胞(ONA)(1-5泳道)、ONH组织(6泳道)和人脑星形胶质细胞(7泳道)中BMP受体表达的RT-PCR分析产生的cDNA样本PCR产物的经溴化乙锭染色的琼脂糖凝胶。L=碱基对标准。C=PCR阴性对照泳道。β-肌动蛋白用作阳性RT-PCR内部对照。
图11.人筛板细胞系中BMP受体的表达。来自人筛板细胞(1-9泳道)的RT-PCR分析产生的cDNA样本PCR产物的经溴化乙锭染色的琼脂糖凝胶。L=碱基对标准。C=PCR阴性对照泳道。β-肌动蛋白用作阳性RT-PCR内部对照。
图12.培养的人TM细胞、视神经乳头星形胶质细胞(ONA)和筛板细胞中BMP和BMP受体表达的Western印迹。人小梁网细胞(1-2泳道)、ONH星形胶质细胞(3-4泳道)和筛板细胞(5-6泳道)中BMP蛋白和BMP受体的化学发光检测。蛋白质大小以kDa表示。
图13.人TM细胞中BMP相关蛋白mRNA的表达。来自人TM细胞(1-5泳道)的RT-PCR分析产生的cDNA样本PCR产物的经溴化乙锭染色的琼脂糖凝胶。L=碱基对标准。C=PCR阴性对照泳道。β-肌动蛋白用作阳性RT-PCR内部对照。
图14.人筛板细胞和ONH星形胶质细胞中BMP相关蛋白mRNA的表达。来自筛板(LC)细胞(1-7泳道)和ONH星形胶质细胞(ONA)(8-11泳道)的RT-PCR分析产生的cDNA样本PCR产物的经溴化乙锭染色的琼脂糖凝胶。L=碱基对标准。C=PCR阴性对照泳道。β-肌动蛋白用作阳性RT-PCR内部对照。
图15.青光眼性TM细胞中BMP拮抗剂gremlin(CKTSF1B1)表达增加图解。用Affymetrix基因阵列(Affymetrix基因芯片U133A)评估基因表达。
优选实施方案详述
认为小梁网在房水的正常流动中起重要作用,并推断它是青光眼性眼中房水流出阻力的主要部位。人小梁网(HTM)细胞是排列于房水流出通道的特化细胞,房水通过该通道流出眼。该细胞改变的综合功能涉及到POAG、类固醇性青光眼和其它类型青光眼的发病机理。
尽管进行了多年的深入研究,参与眼的青光眼性损伤的确切分子机理还是未知的。最近的研究提出生长因子在维持青光眼相关的眼组织的正常内环境稳定中可起重要作用,生长因子/生长因子受体的改变可在青光眼的发病机理中起作用。生长因子是一个非常大的控制细胞生长和分化的多肽家族。这些分子对基因表达、细胞外基质组成和沉积、细胞骨架组织和细胞功能的调节具有很多细胞特异性效应。TM表达很多种生长因子、生长因子受体(Tripathi等人1993a;Tripathi等人1993b;Tripathi等人1994a;Tripathi等人1994b;Wordinger等人1998;Wordinger等人1999)和神经营养蛋白/神经营养蛋白因子及其受体(Liu等人2001;Wordinger等人2000)。ONH星形胶质细胞和筛板细胞为视神经乳头的两种细胞类型,它们表达生长因子、神经营养蛋白及其受体(Lambert等人2001;Pena等人1999)。房水也包含FGF2、EGF、TGFβ、HGF多种生长因子(Tripathi等人1996;Tripathi等人1991;Tripathi等人1992;Hu和Ritch 2001)和神经营养蛋白(Chundru等人2000)。有报道在PAOG患者中房水的TGFβ-2和HGF水平升高(Tripathi等人1994c;Inatani等人2001;Picht等人2001)。生长因子可通过改变TM和ONH的正常发育和/或其功能而参与到青光眼中。
本发明部分起源于对骨形态发生蛋白(BMPs)的认识,BMPs不仅仅诱导骨和软骨形成,也是对多种细胞类型具有广泛效应的多功能细胞因子(Hogan 1996;Reddi 1997),人小梁网(HTM)和视神经乳头(ONH)细胞均表达BMPs(Wordinger等人2002)。BMPs是TGFβ超家族成员,人体大约有15-20种BMPs基因、3种BMP受体和许多起到BMP拮抗剂功能的BMP相关蛋白(Yamashita等人1996)。BMPs通过由BMPR-I和BMPR-II组成的受体复合体传导信号。有报道HTM和ONH细胞均表达超家族成员TGFβ和TGFβR(Agarwal等人1997;Lambert等人1997)以及GDNF和GDNFR(Wordinger等人1999;Liu等人1999)。
眼组织表达BMPs和BMP受体(Obata等人1999;You等人1999),但以前的报道集中于眼的发育。由于小鼠中对编码BMPs的基因进行靶向性破坏导致视网膜和晶状体的严重发育缺陷,因此BMPs在眼发育中是重要的(Jena等人1997;Luo等人1995;Dudley等人1995)。BMP-2、BMP-4和BMP-7参与晶状体和视网膜的发育(Jena等人1997;Furuta和Hogan1998;Reddi 2000;Trousse等人2001)。BMP-6和BMP-7在保护神经元免受低血糖或局部缺血损伤中也显示出起作用(Nonner等人2001;Liu等人2001),并且显示BMP2增强神经节细胞神经营养蛋白的表达(Zhang等人1998)。仅Bmp4缺乏的基因敲除杂合小鼠具有包括眼前节畸形、IOP升高和视神经不正常的眼表型(Chang等人2001)。关于BMPs在人出生后眼中作用的信息公开的非常少。
Mohan和同事(1998)报道成人角膜细胞表达BMP-2和BMP-4以及BMP受体并提出BMP的功能可包括角膜的角膜细胞增生和编程性细胞死亡。You和同事(1999)证实了该项研究并且也报道在离体和培养的角膜上皮及基质细胞中BMP-3、BMP-5和BMP-7的表达。他们报道BMP转录水平在基质中较高,而在培养的角膜上皮细胞中受体水平较高。
应用RT-PCR,本发明人在HTM、筛板(LC)和ONH星形胶质细胞系和组织中发现了BMPs、BMP受体BMPR-IA、BMPR-IB和BMPR-II及BMP结合蛋白gremlin、chordin、卵泡抑素和bambi的mRNAs(Wordinger等人2002)。本发明人进一步发现HTM和ONH细胞表达BMP-2、BMP-4、BMP-5和BMP-7蛋白质。
通过以BMP信号家族成员基因的遗传变化为特征诊断青光眼。如于此应用的短语“骨形态发生蛋白家族成员基因”和“BMP信号家族”是指所有BMPs、BMP受体和相关蛋白。术语“遗传变化”是本领域技术人员众所周知的。有许多与特定基因遗传变化相关疾病的实例(例如参见Cummings 1997;Strachan等人1996;Jorde等人1999)。用本领域技术人员众所周知的多种技术,例如SSCP、DGGE、ASO、RFLP、异源双链分析、CCM、PTT和Rnase切割(参见Birren等人1998)能确定特定基因(例如BMP)的遗传变化。
青光眼可由眼中一个或多个BMP家族基因的表达改变引起,其导致IOP升高和/或青光眼性视神经病。“改变的BMP基因表达”是指该基因产物的表达与正常基因产物不同。该术语也指基因或蛋白质序列的改变。已经对正常BMP基因产物进行了描述(见上),并且已经报道在多种组织包括TM和ONH中有BMP表达。BMP家族基因编码区的遗传变化可改变这些蛋白质的功能。编码区外的遗传变化也可导致青光眼。
本领域技术人员众所周知,特定基因编码区“之外的变化”在基因表达的调节中是重要的。例如,大部分基因编码区的上游区域(5′)已知是“启动”和调控该基因表达的启动子区。启动子区包含被多种转录因子和DNA结合蛋白识别的许多核苷酸序列,其中所述转录因子和DNA结合蛋白负责基因表达的激活或抑制。基因的下游区域(3′)能决定基因产物的聚腺苷酸化,因此调节基因产物的RNA加工和翻译。
为青光眼征兆的BMP基因改变的表达或基因序列中的突变可用本领域技术人员众所周知的技术进行检测。例如,认为可使用几乎任意长度的核酸片段,其总长度优选地受限于易于制备和在预定方法中的用途。于此公开的核酸序列也可在核酸杂交实施方案中用作探针或引物。同样,认为包含以下序列区的核酸片段尤其有用,其中所述序列区由与BMP-2A(SEQ ID NO:1)、BMP-4(SEQ ID NO:3)、BMP-5(SEQ ID NO:5)、BMP-7(SEQ ID NO:7)、BMP-RIA(SEQ ID NO:37)、BMP-RIB(SEQ ID NO:39)、BMP-RII(SEQ ID NO:41)、chordin(SEQ ID NO:43)、gremlin(SEQ ID NO:45)、卵泡抑素(SEQ ID NO:47)或bambi(SEQ ID NO:53)中的14个核苷酸长的连续序列具有相同序列或与其互补的至少14个核苷酸长的连续序列组成。较长的连续相同序列或互补序列,例如约20、30、40、50、100、200、500、1000个核苷酸(包括所有中间长度),甚至约1547个核苷酸(对于BMP-2A)、1946个核苷酸(对于BMP-4)、2153个核苷酸(对于BMP-5)和1878个核苷酸(对于BMP-7)、2932个核苷酸(对于BMP-RIA)、2032个核苷酸(对于BMP-RIB)、3611个核苷酸(对于BMP-RII)、3561个核苷酸(对于chordin)、4049个核苷酸(对于gremlin)、1386个核苷酸(对于卵泡抑素)和1523个核苷酸(对于bambi)的全长序列在某些实施方案中也有应用。
容易理解“中间长度”在本文中是指在引用范围间的任意长度,例如14、15、16、17、18、19、20等;21、22、23等;30、31、32等;50、51、52、53等;100、101、102、103等;150、151、152、153等;包括200-500;500-1,000;1,000-2,000范围间的所有整数,高达并包括2,001、2002、2050、2051等长度的序列。
与BMP编码序列特异杂交的此类核酸探针和引物的能力及引物特异扩增BMP序列的能力使它们在检测特定样本中存在互补序列中有用。然而,可预见到其它用途,包括用序列信息制备突变物种的引物,或用引物制备其它遗传构件。
特别是认为具有由10、20、30、50个或甚至100-200个核苷酸左右的连续核苷酸组成的序列区的核酸分子除了用于Southern和northern印迹外,还可用作杂交探针如用于SNP评估和固相杂交试验中,其中所述连续核苷酸与BMP-2A(SEQ ID NO:1)、BMP4(SEQ ID NO:3)、BMP-5(SEQ ID NO:5)、BMP7(SEQ ID NO:7)、BMP-RIA(SEQ ID NO:37)、BMP-RIB(SEQ ID NO:39)、BMP-RII(SEQ ID NO:41)、chordin(SEQ ID NO:43)、gremlin(SEQ ID NO:45)、卵泡抑素(SEQ ID NO:47)或bambi(SEQ ID NO:53)相同或互补。这允许在组织和细胞中分析BMP结构基因或调节基因。片段的总计大小及互补片段的大小最终取决于特定核酸片段应用的预期用途。根据希望检测的互补序列的长度,较短片段一般在杂交实施方案中应用,其中连续互补区域的长度可改变,例如在约10个至约100个核苷酸之间,但可用长约1547个核苷酸(对于BMP-2A)、1946个核苷酸(对于BMP-4)、2153个核苷酸(对于BMP-5)和1878个核苷酸(对于BMP-7)、2932个核苷酸(对于BMP-RIA)、2032个核苷酸(对于BMP-RIB)、3611个核苷酸(对于BMP-RII)、3561个核苷酸(对于chordin)、4049个核苷酸(对于gremlin)、1386个核苷酸(对于卵泡抑素)和1523个核苷酸(对于bambi)的较长连续互补序列。
用长约10-14个核苷酸的杂交探针允许形成既稳定又有选择性的双链分子。一般优选具有长度大于10个碱基的连续互补序列的分子,但为了增加杂交体的稳定性和选择性并因此提高所获得的特异杂交体分子的质量和程度,一般更喜欢设计具有15至20个连续核苷酸或更长的基因互补序列的核酸分子。
杂交探针可从于此公开的任意序列的任一部分选择。所需要的是对SEQ ID NO:1、SEQ ID NO:3、SEQ ID NO:5、SEQ ID NO:7、SEQID NO:37、SEQ ID NO:39、SEQ ID NO:41、SEQ ID NO:43、SEQID NO:45、SEQ ID NO:47或SEQ ID NO:53所示序列进行检查,并选择序列的任意连续部分,该序列的连续部分来自希望用作探针或引物的长约10个核苷酸至高达并包括全长序列。探针序列和引物序列的选择可由多种因素决定,例如以举例的方式来说明,可用来自总序列末端的引物,或来自功能性结构域编码序列末端的引物,以进一步扩增DNA。
选择和制备包括来自BMP-2A(SEQ ID NO:1)、BMP4(SEQ IDNO:3)、BMP-5(SEQ ID NO:5)、BMP7(SEQ ID NO:7)、BMP-RIA(SEQ ID NO:37)、BMP-RIB(SEQ ID NO:39)、BMP-RII(SEQ IDNO:41)、chordin(SEQ ID NO:43)、gremlin(SEQ ID NO:45)、卵泡抑素(SEQ ID NO:47)或bambi(SEQ ID NO:53)中的连续序列的核酸片段的方法可另外描述为制备核酸片段。当然,也可通过其它技术获得片段,例如通过机械剪切或通过限制性酶消化。短核酸节段或短核酸片段易于制备,例如通过化学方法直接合成片段,这一般用自动寡核苷酸合成仪实行。同样,片段也可通过应用核酸增殖技术获得例如用美国专利4,683,202号和美国专利4,682,195号(此处引用作为参考)的PCRTM技术,和通过将选择的序列导入到重组载体中以进行重组生产获得,以及通过分子生物学领域技术人员一般已知的其它重组DNA技术获得。
因此,本发明的核酸序列可应用它们与BMP基因或其cDNAs的互补序列选择性形成双链分子的能力。根据预想的应用,可利用不同程度的杂交选择性获得针对靶序列的不同程度的探针选择性。对于需要高选择性的应用,一般利用相对严格条件形成杂交体,例如选择相对低盐和/或高温度条件,如NaCl为0.02M-0.15M,温度为50℃至70℃。此类选择条件几乎不容许在探针和模板或靶链间有(如果有的话)错配,因此特别适于检查BMP基因。
当然,对于某些应用,例如想用突变引物链与潜在的模板杂交制备或鉴定突变体或想从相关物种、功能等同物等中分离BMP编码序列,一般需要较不严格的杂交条件以允许异源双链的形成。在这些情况下,可采用如盐为0.15M-1.0M,温度20℃至55℃的条件。从阳性杂交信号和对照杂交信号的比较,因此能容易地确定交叉杂交物种。在任何情况下,一般认为通过降低NaCl浓度或通过加入越来越多量的甲酰胺能得到更严格的条件,甲酰胺与升高温度以同样的方式使杂种双链不稳定。因此杂交条件易于控制,并因此通常是依赖于预期结果的选择方法。
在某些实施方案中,利用本发明的核酸序列结合适当方法例如用于确定杂交的标记是有利的。本领域已知有多种合适的指示方法,包括能产生可检测信号的荧光配体、放射性配体、酶配体或其它配体,如亲和素/生物素。在优选的实施方案中,可能想利用荧光标记或酶标记物,如脲酶、碱性磷酸酶或过氧化物酶,以代替放射性或其他危害环境的试剂。当为酶标记物时,已知能用比色指示性底物提供人眼可见或分光光度计可测量的方法,以确定与包含互补核酸的样本的特异杂交。
一般,可以想象于此描述的杂交探针可用作溶液杂交中的试剂及用于使用固相的实施方案中。在涉及固相的实施方案中,待检测DNA(或RNA)吸附到或固定于选择的基质或表面。该固定的单链核酸然后与选择的探针在预定条件下进行特异杂交。选择的条件取决于具体情况,这些情况基于所需的具体指标(例如取决于G+C含量、靶核酸的类型、核酸来源、杂交探针大小,等)。洗涤杂交表面以去除非特异结合的探针分子后,检测特异杂交,或甚至通过标记进行定量。
也应理解本发明不限于BMP-2A(SEQ ID NO:1)、BMP4(SEQ IDNO:3)、BMP-5(SEQ ID NO:5)、BMP7(SEQ ID NO:7)、BMP-RIA(SEQ ID NO:37)、BMP-RIB(SEQ ID NO:39)、BMP-RII(SEQ IDNO:41)、chordin(SEQ ID NO:43)、gremlin(SEQ ID NO:45)、卵泡抑素(SEQ ID NO:47)或bambi(SEQ ID NO:53)的特定核酸序列和氨基酸序列。重组载体和分离的DNA片段因此可不同地包括BMP编码区自身、基因的上游区域或下游区域、在基本编码区中具有选择改变或修饰的编码区、或它们可编码包括BMP编码区的较大多肽或可编码具有不同氨基酸序列的生物学功能等同的蛋白质或多肽。
本发明的DNA片段包含生物学功能等同的BMP蛋白或多肽。此类序列可作为已知天然存在于核酸序列和其编码蛋白质中的密码子冗余和功能等同的结果。另外,功能等同的蛋白质或多肽可通过应用重组DNA技术来产生,其中蛋白质结构可在考虑所交换的氢基酸性质的基础上进行人工设计。人们设计的变化可通过应用定点诱变技术导入,例如提高蛋白质的抗原性或检测BMP突变以在分子水平检查结合活性。
治疗青光眼的治疗剂可为肽或蛋白质、肽模拟物、寡核苷酸或衍生的寡核苷酸、或小的药物样分子,它们均影响眼BMP途径的一个或多个方面。优选的治疗剂为:(1)BMP2、BMP4、BMP5或BMP7激动剂;(2)chordin、gremlin、卵泡抑素或bambi拮抗剂;和/或(3)Smad1、Smad5和/或Smad4激动剂。
用本领域技术人员众所周知的技术,治疗剂可直接递送至眼(例如:局部滴眼液或药膏;结膜穹窿中的缓释装置或植入邻近巩膜或眼中的缓释装置;眼周、结膜、眼球囊下、眼房内或玻璃体内注射)或肠胃外(例如:口服、静脉内注射、皮下或肌内注射;经皮递送;等)施用。以下为本发明具体化的可能制剂的实例。
(a)局部眼制剂 重量百分数
增加眼BMP-4表达的试剂 0.01-2
HPMC 0.5
氯化钠 0.8
BAC 0.01%
EDTA 0.01
NaOH/HCl 适量,调节至pH 7.4
纯水 适量,调节至100mL
(b)局部眼制剂 重量百分数
Gremlin拮抗剂 0.01-2
HPMC 0.5
氯化钠 0.8
BAC 0.01
EDTA 0.01
NaOH/HCl 适量,调节至pH 7.4
纯水 适量,调节至100mL
(c)局部眼制剂 重量百分数
Smad1/5拮抗剂 0.01-2
HPMC 0.5
氯化钠 0.8
BAC 0.01
EDTA 0.01
NaOH/HCl 适量,调节至pH 7.2
纯水 适量,调节至100mL
进一步认为本发明的化合物可制备在眼内插入装置中。
A.治疗剂的鉴定法
本发明对发现参与BMP信号途径的新抗青光眼治疗剂也有用(见图5)。选择性的BMP配体结合BMP I型和II型丝氨酸/苏氨酸激酶受体(BMP-RI和BMP-RII)并通过Smad蛋白质转导信号。BMP信号通过蛋白质-蛋白质和蛋白质-DNA相互作用由Smads传导(Attisano和TuenLee-Hoeflich 2001)。配体结合的BMP受体(通过磷酸化)激活调节性Smad1和Smad5(von Bubnoff和Cho 2001)。然后这些调节性Smads与Smad 4相互作用形成异源多聚复合体,该复合体转移到核。根据所存在的核协同因子,该复合体能激活或抑制识别该转录复合体的选择基因的转录。
BMP/Smad信号途径通过几种机制进行负调节。某些BMP-结合蛋白(例如gremlin、BAMBI或卵泡抑素)与BMPs结合并抑制它们与BMP受体的相互作用。并且,存在结合并灭活BMP受体的抑制性Smad蛋白(例如Smad6和Smad7)(Kowabata等人1998;Itoh等人2000;Miyazono2000)。本发明人发现人TM细胞、ONH星形胶质细胞和筛板细胞表达BMP受体复合体蛋白。因此,这些细胞能对内源BMP配体作出反应。
可用多种方法发现新的抗青光眼治疗剂,并且这些技术是本领域技术人员众所周知的。例如作为BMPs的激动剂或抑制剂的肽或肽模拟物能通过BMP/BMP受体结构的分子模建发现(Nickel等人2001)。BMP信号转导包括选择一系列Smad蛋白质(Kawabata等人1998;Itoh等人2000;Attiseno等人2000)。选择性BMP激动剂和Smad激动剂能用基于细胞的鉴定法发现。受检测细胞应表达适当的BMP受体并具有适当的BMP信号途径。由于BMP信号的主要效应之一是基因表达的改变,BMP激动剂和Smad激动剂能通过对BMP-诱导基因进行筛选发现。BMP调节基因的诱导也可通过用定量RT-PCR定量mRNA水平(Wang等人2001)、DNA微阵列或报道基因构建体进行分析。在BMP信号传导过程中存在天然抑制剂,为BMP结合蛋白(也称为BMP-相关蛋白)例如chordin、gremlin和卵泡抑素。所述BMP蛋白抑制剂的拮抗剂可通过配体结合试验发现。例如可将受检测试剂加入重组纯化的gremlin中,与gremlin结合的试剂用可本领域技术人员众所周知的多种技术进行鉴定。为确定这些试剂是否为gremlin拮抗剂,应用类似上述的基于细胞的试验进行确定。
认为任何已知的体外和体内模型可结合本发明用于确定针对BMP家族基因的新青光眼治疗剂。这些模型是本领域技术人员众所周知的并且它们的操作已成常规。小肽或肽模拟物可建立在BMP、BMPR和/或BMP结合蛋白基因产物的结构/功能知识的基础上进行设计。能用配体结合试验检测与BMPs、BMPRs或BMP结合蛋白结合的小分子。基于细胞的试验能检测多种试剂对BMP信号途径的效应。能产生包含BMP家族基因启动子与报道基因相连的基因敲入细胞系,用该基因敲入细胞系来寻找改变BMP家族成员基因表达的试剂。这些试验能用于鉴定激动剂和拮抗剂分子。离体试验,例如来自人眼的灌注培养的眼前节(Clark等人1995a;Pang等人2000),能用于检测试剂对TM组织中IOP和BMP信号传导的效应。能用众所周知的技术产生稳定的BMP家族成员转基因、基因敲除或敲入的小鼠和大鼠品系,从而产生青光眼的啮齿类模型。这些啮齿类模型能用于筛选改变青光眼样表型(例如眼压测量法评估对IOP的影响,组织学评估对青光眼性视神经学的影响)的试剂。
B.试剂盒
本发明为青光眼的早期检测提供了方法、组合物和试剂盒。试剂盒能包含编码BMP多肽或蛋白质的核酸片段。试剂盒进一步可包含检测样本与本发明的核酸或肽间相互作用的试剂。提供的试剂可为放射性-、荧光-或酶-标记的。试剂盒可包含能与本发明的核酸或肽或蛋白质结合或相互作用的已知放射性标记试剂。
试剂盒中的试剂可以液体溶液、结合于固相载体或以干粉提供。优选地,当试剂以液体溶液提供时,液体溶液为水溶液。优选地,当试剂以结合于固相载体提供时,固相载体可为层析介质、多孔检测板或载玻片。当试剂以干粉提供时,干粉能通过加入适当溶剂重新溶解,该溶剂是可提供的。
在更进一步的实施方案中,本发明涉及诊断青光眼的诊断方法和相关试剂盒。提出可利用本发明的BMP相关肽和核酸检测来自患者样本的BMP核酸中的多态性或突变。一般,该方法包括首先获得疑似包含此类多态性或突变的样本,在允许形成复合体的有效条件下,将样本与本发明的肽或核酸接触,然后检测复合体的存在。
一般,复合体形成的检测是本领域众所周知的并可通过应用多种方法实施。例如,本发明认为可应用ELISA、RIA、间接荧光技术等。一般,通过用标记如放射性标记或酶标记物(如碱性磷酸酶、辣根过氧化物酶或等)检测复合体形成。当然,通过应用二级结合配体可发现其它优点。
下列实施例是本发明人利用技术实现本发明各方面的代表。应当理解虽然这些技术是本发明实行中优选实施方案的范例,鉴于本公开,本领域技术人员了解在不背离本发明的精神和预期范围内可进行多种改变。
实施例1
细胞培养:如所述产生来自供体眼的人TM细胞和ONH细胞(Steely等人1992;Steely等人2000;Wilson等人1993;Clark等人1994;Clark等人1995b;Clark等人1995c;Clark等人1996;Clark等人2001a;Clark等人2001b;Dickerson等人1998;Wordinger等人1998;Wordinger等人1999;Wordinger等人2000;Wordinger等人2002;Lambert等人2001;Agarwal等人1999;Liu等人2001)。TM细胞在来自年龄为6天至90岁的供体的TM外植块上生长。人视神经乳头星形胶质细胞和筛板(LC)细胞通过小心解剖视神经乳头(供体年龄为2天至90岁)产生并根据以前报道鉴定(Lambert等人2001;Clark等人1995a)。细胞在下列培养基中生长成成片细胞:对于TM细胞,为含有10%胎牛血清(HyClone,Logan,UT)和抗生素(Gibco BRL-Life Technologies,Grand Island,NY)的Ham′s F10培养基(JRH Biosciences,Lenexa,KS);对于LC细胞,为含有10%FBS的Dulbecco′s改良Eagle′s培养基(DMEM,HyClone);对于ONH星形胶质细胞,为含有5%FBS的星形胶质细胞生长培养基(AGM,Clonetics,San Diego,CA)。
RT-PCR:也解剖来自供体眼的人TM和ONH组织(Wordinger等人1998;Wang等人2001)。用TRIzol提取液(Gibco BRL-LifeTechnologies)从TM和ONH细胞和组织中提取总RNA,并用标准方法(Wordinger等人1998;Wordinger等人1999;Wordinger等人2000;Wordinger等人2002)通过反转录制备cDNA。用Oligos 4.0软件程序设计PCR引物(见表1的引物对)。所有引物对这样设计以便潜在的杂质基因组DNA序列扩增产生的mRNA PCR产物比预计的明显地大得多,因为RNA加工期间切除的内含子序列包括在基因组DNA中。β-肌动蛋白PCR引物,AGGCCAACCGCGAGAAGATGACC(上游)和GAAGTCCAGGGCGACGTAGCAC(下游),退火温度为55℃,产生的PCR产物为350bp。
用Taq启动抗体热启动,用下列循环条件:94℃2分钟,92℃2分钟和在最佳退火温度下30秒,72℃延伸90秒和92℃变性45秒,进行40个循环,按所述(Wordinger等人1998;Wordinger等人1999;Wordinger等人2000;Lambert等人2001;Wordinger等人2002)进行PCR反应。扩增的PCR产物在1.5%琼脂糖凝胶中水平电泳进行检查。为保证RT-PCR产物的特异性,用Oligo 4.0设计的探针进行Southern印迹分析,该探针与扩增的PCR产物中的某个区域杂交。对PCR产物进行序列测定以证实PCR反应的特异性。表2列出了人TM和ONH中表达的BMP家族成员。
表1.PCR引物对、退火温度和BMPs扩增片段的大小
名称 | 登录号 | 上游PCR引物 | 下游PCR引物 | 扩增片段大小(bp) |
BMP-2A | NM_001200 | ACTGCGGTCTCCTAAAGGTCGA(SEQ ID NO:9) | GCTGACCTGAGTGCCTGCGAT(SEQ ID NO:10) | 657 |
BMP-4 | NM_001202 | GAATGCTGATGGTCGTTTTTATTATG(SEQ IDNO:11) | AGACTGAAGCCGGTAAAGAT(SEQ ID NO:12) | 348 |
BMP-5 | NM_021073 | AAGAGGACAAGAAGGACTAAAAATAT(SEQID NO:13) | GTAGAGATCCAGCATAAAGAGAGGT(SEQ IDNO:14) | 303 |
BMP-7 | NM_001719 | AGCCCGGGTAGCGCGTAGAG(SEQ ID NO:15) | GCGCCGGTGGATGAAGCTCGA(SEQ IDNO:16) | 202 |
BMPR-1A | NM_004329 | TAAAGGTGACAGTACACAGGAACA(SEQ IDNO:17) | TCTATGATGGCAAAGCAATGTCC(SEQ IDNO:18) | 298 |
BMPR-1B | NM_001203 | TACAAGCCTGCCATAAGTGAAGAAGC(SEQ IDNO:19) | ATCATCGTGAAACAATATCCGTCTG(SEQ IDNO:20) | 211 |
BMPR-II | NM_001204 | TCCTCTCATCAGCCATTTGTCCTTTC(SEQ IDNO:21) | AGTTACTACACATTCTTCATAG(SEQ ID NO:22) | 457 |
Chordin(CHRD) | AF209930 | CTCTGCTCACTCTGCACCTG(SEQ ID NO:23) | CCGGTCACCATCAAAATAGC(SEQ ID NO:24) | 198 |
Gremlin(CKTSF1B1) | NM_013372 | ATCAACCGCTTCTGTTACGG(SEQ ID NO:25) | ATGCAACGACACTGCTTCAC(SEQ ID NO:26) | 197 |
卵泡抑素(FST) | NM_006350 | TGCCACCTGAGAAAGGCTAC(SEQ ID NO:27) | ACAGACAGGCTCATCCGACT(SEQ ID NO:28) | 201 |
头蛋白(NOG) | NM_005450 | CACTACGACCCAGGCTTCAT(SEQ ID NO:29) | CTCCGCAGCTTCTTGCTTAG(SEQ ID NO:30) | 212 |
CER-1 | NM_005454 | ATAGTGAGCCCTTCCCACCT(SEQ ID NO:33) | AATGAACAGACCCGCATTTC(SEQ ID NO:34) | 294 |
NMA(BAMBI) | NM_005791 | GATCGCCACTCCAGCTACATC(SEQ ID NO:35) | GGGCACGGCAATGACC(SEQ ID NO:36) | 471 |
表2.人TM和ONH中表达的BMP家族成员
BMP家庭成员 | 小梁网 | 视神经乳头 |
BMP-2 | + | + |
BMP-4 | + | + |
BMP-5 | + | + |
BMP-7 | + | + |
BMPR-IA | + | + |
BMPR-IB | + | + |
BMPR-II | + | + |
Chordin | + | + |
Gremlin | + | + |
卵泡抑素 | + | + |
Bambi | + | + |
头蛋白 | - | - |
CER-1 | - | - |
Western印迹:用裂解缓冲液从培养细胞中提取蛋白质,并通过变性聚丙烯酰胺凝胶电泳分离蛋白质后电泳转移到硝酸纤维素膜上(Lambert等人2001)。膜用5%牛乳(对于BMPs)或3%明胶(对于BMPRs))进行封闭并与下列一级抗体孵育:BMP2、BMP4、BMP5、BMP7(它们均来自Santa Cruz,Santa Cruz,CA),或BMP-RIA、BMP-RIB、BMP-RII(来自Jackson Immuno Research,West Grove,PA)。洗涤膜,与二级抗体(对于BMPs为山羊抗-小鼠IgG-辣根过氧化物酶,Santa Cruz;对于BMP受体为驴抗-山羊-辣根过氧化物酶,Jackson Immuno Research)孵育,并用WesternBreeze化学发光免疫检测系统(Invitrogen,Carlsbad,CA)显相。
人TM细胞和组织中BMPs、BMPRs mRNA的表达:图6中显示了人TM细胞和组织中BMP-2、BMP-4、BMP-5和BMP-7引物对预期的扩增产物。用特异探针进行的Southern印迹证实其为预期的PCR产物。所有人TM细胞系和组织表达BMP-2、BMP-4和BMP-7。然而,在人TM组织样本中,BMP-5的表达低以至于检测不出来(图6,6和7泳道)。无cDNA的对照反应不出现扩增产物表明试剂和引物无DNA或RNA污染(图6,C泳道)。
图7显示了人TM细胞和组织中BMP-RIA、BMP-RIB和BMP-RII引物对预期大小的扩增产物。所有人TM细胞和组织表达BMP受体复合体。用特异探针进行的Southern印迹证实其为预期的PCR产物。在BMP-RII PCR反应中检测到另外的扩增产物(350bp)。所有人TM细胞和组织中存在该另外的扩增产物。该另外的带现在正在进行鉴定以确定其是否为受体的另一剪接形式。无cDNA的对照反应不出现扩增产物(图7,C泳道)表明试剂和引物无DNA或RNA污染。
人ONH细胞和组织中BMP和BMP受体mRNA的表达:图8中显示了人ONH星形胶质细胞和ONH组织中BMP-2、BMP-4、BMP-5和BMP-7引物对预期大小的扩增产物。所有ONH星形胶质细胞和ONH组织表达相应的BMP。人脑星形胶质细胞用作阳性对照细胞系。用特异探针进行的Southern印迹证实其为预期的PCR产物。除了BMP-2外,人脑星形胶质细胞表达所有其它BMP(图8,7泳道)。无cDNA的对照反应不出现扩增产物(图8,C泳道)表明试剂和引物无DNA或RNA污染。
图9显示了培养的人LC细胞中BMP-2、BMP-4、BMP-5和BMP-7引物对预期大小的扩增产物。所有LC细胞系表达每种BMP。用特异探针进行的Southern印迹证实其为预期的PCR产物。无cDNA的对照反应不出现扩增产物(图9,C泳道)表明试剂和引物无DNA或RNA污染。
图10显示的是人ONH星形胶质细胞和ONH组织中BMP-RIA、BMP-RIB和BMP-RII引物对预期大小的扩增产物。所有ONH星形胶质细胞系和ONH组织表达BMP-RIA和BMP-RIB。用特异探针进行的Southern印迹证实其为预期的PCR产物。除了ONH组织(图10,6泳道)外,所有ONH星形胶质细胞系表达BMP-RII。表达所有BMP受体(图10,7泳道)的人脑星形胶质细胞系用作阳性对照。显示出ONH组织和ONH细胞系中BMP-RII的表达有差异。ONH组织中降低的表达可反应低水平表达。无cDNA的对照反应不出现扩增产物(图5,C泳道)表明试剂和引物无DNA或RNA污染。
图11显示了培养的人LC细胞中BMP-RIA、BMP-RIB和BMP-RII引物对预期大小的扩增产物。所有LC细胞系均表达BMP受体。用特异探针进行的Southern印迹证实其为预期的PCR产物。无cDNA的对照反应不出现扩增产物(图11,C泳道)表明试剂和引物无DNA或RNA污染。
人TM细胞及ONH细胞和组织中BMP蛋白和BMP受体蛋白的表达:图12表示化学发光免疫印迹检测人TM及ONH细胞和组织中BMP-2、BMP-4、BMP-5、BMP-7、BMP-RIA、BMP-RIB和BMP-RII蛋白质。研究的所有细胞系表达相应的BMP蛋白。细胞系中检测的BMP蛋白具有下列分子量:BMP-2为54-56kDa,BMP-4为25-27kDa,BMP-5为55-57kDa和BMP-7为77kDa。BMP-2和BMP-4检测到多条带,如在其它研究中看到的那样它们很可能表示这些BMPs的糖基化和部分糖基化形式。然而,我们没有进行糖基化研究因为它们超出了本研究的范围。细胞系中检测的BMP受体蛋白分子量为:BMP-RIA为38kDa,BMP-RIB为64kDa,BMP-RII为57kDa。TM细胞中BMP-RIB和BMP-RII检测到多条带,如在其它研究中看到的那样它们很可能表示这些BMPs的糖基化和部分糖基化形式。显示TM细胞中BMP受体蛋白的表达水平比ONH细胞的表达水平低。例如在TM细胞中没有检测到BMP-RII并且BMP-RIB大大降低。
培养的人TM细胞和人ONH细胞中BMP相关蛋白mRNAs的表达:图13显示的是BMP相关蛋白引物对预期大小的扩增产物。人TM细胞系表达DRM(gremlin)、chordin、卵泡抑素和NMA(BAMBI)的信息。用特异探针进行的Southern印迹证实它们为预期的PCR产物。细胞系间的表达没有明显差异。受检的所有人TM细胞没有BMP相关蛋白头蛋白和Cer-1mRNA的表达。无cDNA的对照反应不出现扩增产物表明试剂和引物无DNA或RNA污染。
图14显示的是ONH星形胶质细胞和LC细胞系中BMP相关蛋白引物对预期大小的扩增产物。所有ONH星形胶质细胞和LC细胞系表达DRM(gremlin)、卵泡抑素和NMA(BAMBI)的信息。用特异探针进行的Southern印迹证实它们为预期的PCR产物。大部分LC细胞和ONH星形胶质细胞表达chordin的信息。所有受检的人ONH星形胶质细胞和LC细胞没有BMP相关蛋白头蛋白和Cer-1mRNA的表达。无cDNA的对照反应不出现扩增产物表明试剂和引物无DNA或RNA污染。
图15显示了青光眼性TM细胞中BMP拮抗剂gremlin(CKTSF1B1)增强的表达。用Affymetrix基因阵列(Affymetrix基因芯片U133A)评估了基因表达。
按照本公开于此公开和要求的所有组合物和/或方法不用过多地进行实验即可制备和实施。虽然本发明的组合物和方法以优选的实施方案进行了描述,但对本领域技术人员显然可在不背离本发明的概念、精神和范围的情况下,对此处描述的组合物和/或方法以及方法的步骤或步骤的顺序进行改变。更具体地说,显然在化学和结构上相关的某些试剂可代替于此描述的试剂以获得相同结果。如附加的权利要求中所定义,对本领域技术人员显而易见的所有此类替换和改变,认为是在本发明的精神、范围和概念内。
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序列表
<110>爱尔康公司
<120>骨形态发生蛋白(BMP)、BMP受体和BMP结合蛋白及它们在诊断和治疗青光眼中的用途
<130>2312 US
<160>54
<170>PatentIn版本3.1
<210>1
<211>1547
<212>DNA
<213>人
<400>SEQ ID NO:1
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tgccccagcg gagcctgctt cgccatctcc gagccccacc gcccctccac tcctcggcct 120
tgcccgacac tgagacgctg ttcccagcgt gaaaagagag actgcgcggc cggcacccgg 180
gagaaggagg aggcaaagaa aaggaacgga cattcggtcc ttgcgccagg tcctttgacc 240
agagtttttc catgtggacg ctctttcaat ggacgtgtcc ccgcgtgctt cttagacgga 300
ctgcggtctc ctaaaggtcg accatggtgg ccgggacccg ctgtcttcta gcgttgctgc 360
ttccccaggt cctcctgggc ggcgcggctg gcctcgttcc ggagctgggc cgcaggaagt 420
tcgcggcggc gtcgtcgggc cgcccctcat cccagccctc tgacgaggtc ctgagcgagt 480
tcgagttgcg gctgctcagc atgttcggcc tgaaacagag acccaccccc agcagggacg 540
ccgtggtgcc cccctacatg ctagacctgt atcgcaggca ctcaggtcag ccgggctcac 600
ccgccccaga ccaccggttg gagagggcag ccagccgagc caacactgtg cgcagcttcc 660
accatgaaga atctttggaa gaactaccag aaacgagtgg gaaaacaacc cggagattct 720
tctttaattt aagttctatc cccacggagg agtttatcac ctcagcagag cttcaggttt 780
tccgagaaca gatgcaagat gctttaggaa acaatagcag tttccatcac cgaattaata 840
tttatgaaat cataaaacct gcaacagcca actcgaaatt ccccgtgacc agacttttgg 900
acaccaggtt ggtgaatcag aatgcaagca ggtgggaaag ttttgatgtc acccccgctg 960
tgatgcggtg gactgcacag ggacacgcca accatggatt cgtggtggaa gtggcccact 1020
tggaggagaa acaaggtgtc tccaagagac atgttaggat aagcaggtct ttgcaccaag 1080
atgaacacag ctggtcacag ataaggccat tgctagtaac ttttggccat gatggaaaag 1140
ggcatcctct ccacaaaaga gaaaaacgtc aagccaaaca caaacagcgg aaacgcctta 1200
agtccagctg taagagacac cctttgtacg tggacttcag tgacgtgggg tggaatgact 1260
ggattgtggc tcccccgggg tatcacgcct tttactgcca cggagaatgc ccttttcctc 1320
tggctgatca tctgaactcc actaatcatg ccattgttca gacgttggtc aactctgtta 1380
actctaagat tcctaaggca tgctgtgtcc cgacagaact cagtgctatc tcgatgctgt 1440
accttgacga gaatgaaaag gttgtattaa agaactatca ggacatggtt gtggagggtt 1500
gtgggtgtcg ctagtacagc aaaattaaat acataaatat atatata 1547
<210>2
<211>396
<212>PRT
<213>人
<400>SEQID NO:2
Met Val Ala Gly Thr Arg Cys Leu Leu Ala Leu Leu Leu Pro Gln Val
1 5 10 15
Leu Leu Gly Gly Ala Ala Gly Leu Val Pro Glu Leu Gly Arg Arg Lys
20 25 30
Phe Ala Ala Ala Ser Ser Gly Arg Pro Ser Ser Gln Pro Ser Asp Glu
35 40 45
Val Leu Ser Glu Phe Glu Leu Arg Leu Leu Ser Met Phe Gly Leu Lys
50 55 60
Gln Arg Pro Thr Pro Ser Arg Asp Ala Val Val Pro Pro Tyr Met Leu
65 70 75 80
Asp Leu Tyr Arg Arg His Ser Gly Gln Pro Gly Ser Pro Ala Pro Asp
85 90 95
His Arg Leu Glu Arg Ala Ala Ser Arg Ala Asn Thr Val Arg Ser Phe
100 105 110
His His Glu Glu Ser Leu Glu Glu Leu Pro Glu Thr Ser Gly Lys Thr
115 120 125
Thr Arg Arg Phe Phe Phe Asn Leu Ser Ser Ile Pro Thr Glu Glu Phe
130 135 140
Ile Thr Ser Ala Glu Leu Gln Val Phe Arg Glu Gln Met Gln Asp Ala
145 150 155 160
Leu Gly Asn Asn Ser Ser Phe His His Arg Ile Asn Ile Tyr Glu Ile
165 170 175
Ile Lys Pro Ala Thr Ala Asn Ser Lys Phe Pro Val Thr Arg Leu Leu
180 185 190
Asp Thr Arg Leu Val Asn Gln Asn Ala Ser Arg Trp Glu Ser Phe Asp
195 200 205
Val Thr Pro Ala Val Met Arg Trp Thr Ala Gln Gly His Ala Asn His
210 215 220
Gly Phe Val Val Glu Val Ala His Leu Glu Glu Lys Gln Gly Val Ser
225 230 235 240
Lys Arg His Val Arg Ile Ser Arg Ser Leu His Gln Asp Glu His Ser
245 250 255
Trp Ser Gln Ile Arg Pro Leu Leu Val Thr Phe Gly His Asp Gly Lys
260 265 270
Gly His Pro Leu His Lys Arg Glu Lys Arg Gln Ala Lys His Lys Gln
275 280 285
Arg Lys Arg Leu Lys Ser Ser Cys Lys Arg His Pro Leu Tyr Val Asp
290 295 300
Phe Ser Asp Val Gly Trp Asn Asp Trp Ile Val Ala Pro Pro Gly Tyr
305 310 315 320
His Ala Phe Tyr Cys His Gly Glu Cys Pro Phe Pro Leu Ala Asp His
325 330 335
Leu Asn Ser Thr Asn His Ala Ile Val Gln Thr Leu Val Asn Ser Val
340 345 350
Asn Ser Lys Ile Pro Lys Ala Cys Cys Val Pro Thr Glu Leu Ser Ala
355 360 365
Ile Ser Met Leu Tyr Leu Asp Glu Asn Glu Lys Val Val Leu Lys Asn
370 375 380
Tyr Gln Asp Met Val Val Glu Gly Cys Gly Cys Arg
385 390 395
<210>3
<211>1946
<212>DNA
<213>人
<400>SEQ ID NO:3
gaaagcgagg gagggaaaga ggaggaagga agatgcgaga aggcagagga ggagggaggg 60
agggaaggag cgcggagccc ggcccggaag ctaggtgagt gtggcatccg agctgaggga 120
cgcgagcctg agacgccgct gctgctccgg ctgagtatct agcttgtctc cccgatggga 180
ttcccgtcca agctatctcg agcctgcagc gccacagtcc ccggccctcg cccaggttca 240
ctgcaaccgt tcagaggtcc ccaggagctg ctgctggcga gcccgctact gcagggacct 300
atggagccat tccgtagtgc catcccgagc aacgcactgc tgcagcttcc ctgagccttt 360
ccagcaagtt tgttcaagat tggctgtcaa gaatcatgga ctgttattat atgccttgtt 420
ttctgtcaag acaccatgat tcctggtaac cgaatgctga tggtcgtttt attatgccaa 480
gtcctgctag gaggcgcgag ccatgctagt ttgatacctg agacggggaa gaaaaaagtc 540
gccgagattc agggccacgc gggaggacgc cgctcagggc agagccatga gctcctgcgg 600
gacttcgagg cgacacttct gcagatgttt gggctgcgcc gccgcccgca gcctagcaag 660
agtgccgtca ttccggacta catgcgggat ctttaccggc ttcagtctgg ggaggaggag 720
gaagagcaga tccacagcac tggtcttgag tatcctgagc gcccggccag ccgggccaac 780
accgtgagga gcttccacca cgaagaacat ctggagaaca tcccagggac cagtgaaaac 840
tctgcttttc gtttcctctt taacctcagc agcatccctg agaacgaggc gatctcctct 900
gcagagcttc ggctcttccg ggagcaggtg gaccagggcc ctgattggga aaggggcttc 960
caccgtataa acatttatga ggttatgaag cccccagcag aagtggtgcc tgggcacctc 1020
atcacacgac tactggacac gagactggtc caccacaatg tgacacggtg ggaaactttt 1080
gatgtgagcc ctgcggtcct tcgctggacc cgggagaagc agccaaacta tgggctagcc 1140
attgaggtga ctcacctcca tcagactcgg acccaccagg gccagcatgt caggattagc 1200
cgatcgttac ctcaagggag tgggaattgg gcccagctcc ggcccctcct ggtcaccttt 1260
ggccatgatg gccggggcca tgccttgacc cgacgccgga gggccaagcg tagccctaag 1320
catcactcac agcgggccag gaagaagaat aagaactgcc ggcgccactc gctctatgtg 1380
gacttcagcg atgtgggctg gaatgactgg attgtggccc caccaggcta ccaggccttc 1440
tactgccatg gggactgccc ctttccactg gctgaccacc tcaactcaac caaccatgcc 1500
attgtgcaga ccctggtcaa ttctgtcaat tccagtatcc ccaaagcctg ttgtgtgccc 1560
actgaactga gtgccatctc catgctgtac ctggatgagt atgataaggt ggtactgaaa 1620
aattatcagg agatggtagt agagggatgt gggtgccgct gagatcaggc agtccttgag 1680
gatagacaga tatacacacc acacacacac accacataca ccacacacac acgttcccat 1740
ccactcaccc acacactaca cagactgctt ccttatagct ggacttttat ttaaaaaaaa 1800
aaaaaaaaaa atggaaaaaa tccctaaaca ttcaccttga ccttatttat gactttacgt 1860
gcaaatgttt tgaccatatt gatcatatat tttgacaaaa tatatttata actacgtatt 1920
aaaagaaaaa aataaaatga gtcatt 1946
<210>4
<211>408
<212>PRT
<213>人
<400>SEQ ID NO:4
Met Ile Pro Gly Asn Arg Met Leu Met Val Val Leu Leu Cys Gln Val
1 5 10 15
Leu Leu Gly Gly Ala Ser His Ala Ser Leu Ile Pro Glu Thr Gly Lys
20 25 30
Lys Lys Val Ala Glu Ile Gln Gly His Ala Gly Gly Arg Arg Ser Gly
35 40 45
Gln Ser His Glu Leu Leu Arg Asp Phe Glu Ala Thr Leu Leu Gln Met
50 55 60
Phe Gly Leu Arg Arg Arg Pro Gln Pro Ser Lys Ser Ala Val Ile Pro
65 70 75 80
Asp Tyr Met Arg Asp Leu Tyr Arg Leu Gln Ser Gly Glu Glu Glu Glu
85 90 95
Glu Gln Ile His Ser Thr Gly Leu Glu Tyr Pro Glu Arg Pro Ala Ser
100 105 110
Arg Ala Asn Thr Val Arg Ser Phe His His Glu Glu His Leu Glu Asn
115 120 125
Ile Pro Gly Thr Ser Glu Asn Ser Ala Phe Arg Phe Leu Phe Asn Leu
130 135 140
Ser Ser Ile Pro Glu Asn Glu Ala Ile Ser Ser Ala Glu Leu Arg Leu
145 150 155 160
Phe Arg Glu Gln Val Asp Gln Gly Pro Asp Trp Glu Arg Gly Phe His
165 170 175
Arg Ile Asn Ile Tyr Glu Val Met Lys Pro Pro Ala Glu Val Val Pro
180 185 190
Gly His Leu Ile Thr Arg Leu Leu Asp Thr Arg Leu Val His His Asn
195 200 205
Val Thr Arg Trp Glu Thr Phe Asp Val Ser Pro Ala Val Leu Arg Trp
210 215 220
Thr Arg Glu Lys Gln Pro Asn Tyr Gly Leu Ala Ile Glu Val Thr His
225 230 235 240
Leu His Gln Thr Arg Thr His Gln Gly Gln His Val Arg Ile Ser Arg
245 250 255
Ser Leu Pro Gln Gly Ser Gly Asn Trp Ala Gln Leu Arg Pro Leu Leu
260 265 270
Val Thr Phe Gly His Asp Gly Arg Gly His Ala Leu Thr Arg Arg Arg
275 280 285
Arg Ala Lys Arg Ser Pro Lys His His Ser Gln Arg Ala Arg Lys Lys
290 295 300
Asn Lys Asn Cys Arg Arg His Ser Leu Tyr Val Asp Phe Ser Asp Val
305 310 315 320
Gly Trp Asn Asp Trp Ile Val Ala Pro Pro Gly Tyr Gln Ala Phe Tyr
325 330 335
Cys His Gly Asp Cys Pro Phe Pro Leu Ala Asp His Leu Asn Ser Thr
340 345 350
Asn His Ala Ile Val Gln Thr Leu Val Asn Ser Val Asn Ser Ser Ile
355 360 365
Pro Lys Ala Cys Cys Val Pro Thr Glu Leu Ser Ala Ile Ser Met Leu
370 375 380
Tyr Leu Asp Glu Tyr Asp Lys Val Val Leu Lys Asn Tyr Gln Glu Met
385 390 395 400
Val Val Glu Gly Cys Gly Cys Arg
405
<210>5
<211>2153
<212>DNA
<213>人
<400>SEQ ID NO:5
ctggtatatt tgtgcctgct ggaggtggaa ttaacagtaa gaaggagaaa gggattgaat 60
ggacttacag gaaggatttc aagtaaattc agggaaacac atttacttga atagtacaac 120
ctagagtatt attttacact aagacgacac aaaagatgtt aaagttatca ccaagctgcc 180
ggacagatat atattccaac accaaggtgc agatcagcat agatctgtga ttcagaaatc 240
aggatttgtt ttggaaagag ctcaagggtt gagaagaact caaaagcaag tgaagattac 300
tttgggaact acagtttatc agaagatcaa cttttgctaa ttcaaatacc aaaggcctga 360
ttatcataaa ttcatatagg aatgcatagg tcatctgatc aaataatatt agccgtcttc 420
tgctacatca atgcagcaaa aactcttaac aactgtggat aattggaaat ctgagtttca 480
gctttcttag aaataactac tcttgacata ttccaaaata tttaaaatag gacaggaaaa 540
tcggtgagga tgttgtgctc agaaatgtca ctgtcatgaa aaataggtaa atttgttttt 600
tcagctactg ggaaactgta cctcctagaa ccttaggttt tttttttttt aagaggacaa 660
gaaggactaa aaatatcaac ttttgctttt ggacaaaaat gcatctgact gtatttttac 720
ttaagggtat tgtgggtttc ctctggagct gctgggttct agtgggttat gcaaaaggag 780
gtttgggaga caatcatgtt cactccagtt ttatttatag aagactacgg aaccacgaaa 840
gacgggaaat acaaagggaa attctctcta tcttgggttt gcctcacaga cccagaccat 900
tttcacctgg aaaacaagcg tcctctgcac ctctctttat gctggatctc tacaatgcca 960
tgaccaatga agaaaatcct gaagagtcgg agtactcagt aagggcatcc ttggcagaag 1020
agaccagagg ggcaagaaag ggatacccag cctctcccaa tgggtatcct cgtcgcatac 1080
agttatctcg gacgactcct ctgaccaccc agagtcctcc tctagccagc ctccatgata 1140
ccaactttct gaatgatgct gacatggtca tgagctttgt caacttagtt gaaagagaca 1200
aggatttttc tcaccagcga aggcattaca aagaatttcg atttgatctt acccaaattc 1260
ctcatggaga ggcagtgaca gcagctgaat tccggatata caaggaccgg agcaacaacc 1320
gatttgaaaa tgaaacaatt aagattagca tatatcaaat catcaaggaa tacacaaata 1380
gggatgcaga tctgttcttg ttagacacaa gaaaggccca agctttagat gtgggttggc 1440
ttgtctttga tatcactgtg accagcaatc attgggtgat taatccccag aataatttgg 1500
gcttacagct ctgtgcagaa acaggggatg gacgcagtat caacgtaaaa tctgctggtc 1560
ttgtgggaag acagggacct cagtcaaaac aaccattcat ggtggccttc ttcaaggcga 1620
gtgaggtact tcttcgatcc gtgagagcag ccaacaaacg aaaaaatcaa aaccgcaata 1680
aatccagctc tcatcaggac tcctccagaa tgtccagtgt tggagattat aacacaagtg 1740
agcaaaaaca agcctgtaag aagcacgaac tctatgtgag cttccgggat ctgggatggc 1800
aggactggat tatagcacca gaaggatacg ctgcatttta ttgtgatgga gaatgttctt 1860
ttccacttaa cgcccatatg aatgccacca accacgctat agttcagact ctggttcatc 1920
tgatgtttcc tgaccacgta ccaaagcctt gttgtgctcc aaccaaatta aatgccatct 1980
ctgttctgta ctttgatgac agctccaatg tcattttgaa aaaatataga aatatggtag 2040
tacgctcatg tggctgccac taatattaaa taatattgat aataacaaaa agatctgtat 2100
taagttttat ggctgcaata aaaagcatac tttcagacaa acagaaaaaa aaa 2153
<210>6
<211>454
<212>PRT
<213>人
<400>SEQ ID NO:6
Met His Leu Thr Val Phe Leu Leu Lys Gly Ile Val Gly Phe Leu Trp
1 5 10 15
Ser Cys Trp Val Leu Val Gly Tyr Ala Lys Gly Gly Leu Gly Asp Asn
20 25 30
His Val His Ser Ser Phe Ile Tyr Arg Arg Leu Arg Asn His Glu Arg
35 40 45
Arg Glu Ile Gln Arg Glu Ile Leu Ser Ile Leu Gly Leu Pro His Arg
50 55 60
Pro Arg Pro Phe Ser Pro Gly Lys Gln Ala Ser Ser Ala Pro Leu Phe
65 70 75 80
Met Leu Asp Leu Tyr Asn Ala Met Thr Asn Glu Glu Asn Pro Glu Glu
85 90 95
Ser Glu Tyr Ser Val Arg Ala Ser Leu Ala Glu Glu Thr Arg Gly Ala
100 105 110
Arg Lys Gly Tyr Pro Ala Ser Pro Asn Gly Tyr Pro Arg Arg Ile Gln
115 120 125
Leu Ser Arg Thr Thr Pro Leu Thr Thr Gln Ser Pro Pro Leu Ala Ser
130 135 140
Leu His Asp Thr Asn Phe Leu Asn Asp Ala Asp Met Val Met Ser Phe
145 150 155 160
Val Asn Leu Val Glu Arg Asp Lys Asp Phe Ser His Gln Arg Arg His
165 170 175
Tyr Lys Glu Phe Arg Phe Asp Leu Thr Gln Ile Pro His Gly Glu Ala
180 185 190
Val Thr Ala Ala Glu Phe Arg Ile Tyr Lys Asp Arg Ser Asn Asn Arg
195 200 205
Phe Glu Asn Glu Thr Ile Lys Ile Ser Ile Tyr Gln Ile Ile Lys Glu
210 215 220
Tyr Thr Asn Arg Asp Ala Asp Leu Phe Leu Leu Asp Thr Arg Lys Ala
225 230 235 240
Gln Ala Leu Asp Val Gly Trp Leu Val Phe Asp Ile Thr Val Thr Ser
245 250 255
Asn His Trp Val Ile Asn Pro Gln Asn Asn Leu Gly Leu Gln Leu Cys
260 265 270
Ala Glu Thr Gly Asp Gly Arg Ser Ile Asn Val Lys Ser Ala Gly Leu
275 280 285
Val Gly Arg Gln Gly Pro Gln Ser Lys Gln Pro Phe Met Val Ala Phe
290 295 300
Phe Lys Ala Ser Glu Val Leu Leu Arg Ser Val Arg Ala Ala Asn Lys
305 310 315 320
Arg Lys Asn Gln Asn Arg Asn Lys Ser Ser Ser His Gln Asp Ser Ser
325 330 335
Arg Met Ser Ser Val Gly Asp Tyr Asn Thr Ser Glu Gln Lys Gln Ala
340 345 350
Cys Lys Lys His Glu Leu Tyr Val Ser Phe Arg Asp Leu Gly Trp Gln
355 360 365
Asp Trp Ile Ile Ala Pro Glu Gly Tyr Ala Ala Phe Tyr Cys Asp Gly
370 375 380
Glu Cys Ser Phe Pro Leu Asn Ala His Met Asn Ala Thr Asn His Ala
385 390 395 400
Ile Val Gln Thr Leu Val His Leu Met Phe Pro Asp His Val Pro Lys
405 410 415
Pro Cys Cys Ala Pro Thr Lys Leu Asn Ala Ile Ser Val Leu Tyr Phe
420 425 430
Asp Asp Ser Ser Asn Val Ile Leu Lys Lys Tyr Arg Asn Met Val Val
435 440 445
Arg Ser Cys Gly Cys His
450
<210>7
<211>1878
<212>DNA
<213>人
<400>SEQ ID NO:7
gggcgcagcg gggcccgtct gcagcaagtg accgacggcc gggacggccg cctgccccct 60
ctgccacctg gggcggtgcg ggcccggagc ccggagcccg ggtagcgcgt agagccggcg 120
cgatgcacgt gcgctcactg cgagctgcgg cgccgcacag cttcgtggcg ctctgggcac 180
ccctgttcct gctgcgctcc gccctggccg acttcagcct ggacaacgag gtgcactcga 240
gcttcatcca ccggcgcctc cgcagccagg agcggcggga gatgcagcgc gagatcctct 300
ccattttggg cttgccccac cgcccgcgcc cgcacctcca gggcaagcac aactcggcac 360
ccatgttcat gctggacctg tacaacgcca tggcggtgga ggagggcggc gggcccggcg 420
gccagggctt ctcctacccc tacaaggccg tcttcagtac ccagggcccc cctctggcca 480
gcctgcaaga tagccatttc ctcaccgacg ccgacatggt catgagcttc gtcaacctcg 540
tggaacatga caaggaattc ttccacccac gctaccacca tcgagagttc cggtttgatc 600
tttccaagat cccagaaggg gaagctgtca cggcagccga attccggatc tacaaggact 660
acatccggga acgcttcgac aatgagacgt tccggatcag cgtttatcag gtgctccagg 720
agcacttggg cagggaatcg gatctcttcc tgctcgacag ccgtaccctc tgggcctcgg 780
aggagggctg gctggtgttt gacatcacag ccaccagcaa ccactgggtg gtcaatccgc 840
ggcacaacct gggcctgcag ctctcggtgg agacgctgga tgggcagagc atcaacccca 900
agttggcggg cctgattggg cggcacgggc cccagaacaa gcagcccttc atggtggctt 960
tcttcaaggc cacggaggtc cacttccgca gcatccggtc cacggggagc aaacagcgca 1020
gccagaaccg ctccaagacg cccaagaacc aggaagccct gcggatggcc aacgtggcag 1080
agaacagcag cagcgaccag aggcaggcct gtaagaagca cgagctgtat gtcagcttcc 1140
gagacctggg ctggcaggac tggatcatcg cgcctgaagg ctacgccgcc tactactgtg 1200
agggggagtg tgccttccct ctgaactcct acatgaacgc caccaaccac gccatcgtgc 1260
agacgctggt ccacttcatc aacccggaaa cggtgcccaa gccctgctgt gcgcccacgc 1320
agctcaatgc catctccgtc ctctacttcg atgacagctc caacgtcatc ctgaagaaat 1380
acagaaacat ggtggtccgg gcctgtggct gccactagct cctccgagaa ttcagaccct 1440
ttggggccaa gtttttctgg atcctccatt gctcgccttg gccaggaacc agcagaccaa 1500
ctgccttttg tgagaccttc ccctccctat ccccaacttt aaaggtgtga gagtattagg 1560
aaacatgagc agcatatggc ttttgatcag tttttcagtg gcagcatcca atgaacaaga 1620
tcctacaagc tgtgcaggca aaacctagca ggaaaaaaaa acaacgcata aagaaaaatg 1680
gccgggccag gtcattggct gggaagtctc agccatgcac ggactcgttt ccagaggtaa 1740
ttatgagcgc ctaccagcca ggccacccag ccgtgggagg aagggggcgt ggcaaggggt 1800
gggcacattg gtgtctgtgc gaaaggaaaa ttgacccgga agttcctgta ataaatgtca 1860
caataaaacg aatgaatg 1878
<210>8
<211>431
<212>PRT
<213>人
<400>SEQ ID NO:8
Met His Val Arg Ser Leu Arg Ala Ala Ala Pro His Ser Phe Val Ala
1 5 10 15
Leu Trp Ala Pro Leu Phe Leu Leu Arg Ser Ala Leu Ala Asp Phe Ser
20 25 30
Leu Asp Asn Glu Val His Ser Ser Phe Ile His Arg Arg Leu Arg Ser
35 40 45
Gln Glu Arg Arg Glu Met Gln Arg Glu Ile Leu Ser Ile Leu Gly Leu
50 55 60
Pro His Arg Pro Arg Pro His Leu Gln Gly Lys His Asn Ser Ala Pro
65 70 75 80
Met Phe Met Leu Asp Leu Tyr Asn Ala Met Ala Val Glu Glu Gly Gly
85 90 95
Gly Pro Gly Gly Gln Gly Phe Ser Tyr Pro Tyr Lys Ala Val Phe Ser
100 105 110
Thr Gln Gly Pro Pro Leu Ala Ser Leu Gln Asp Ser His Phe Leu Thr
115 120 125
Asp Ala Asp Met Val Met Ser Phe Val Asn Leu Val Glu His Asp Lys
130 135 140
Glu Phe Phe His Pro Arg Tyr His His Arg Glu Phe Arg Phe Asp Leu
145 150 155 160
Ser Lys Ile Pro Glu Gly Glu Ala Val Thr Ala Ala Glu Phe Arg Ile
165 170 175
Tyr Lys Asp Tyr Ile Arg Glu Arg Phe Asp Asn Glu Thr Phe Arg Ile
180 185 190
Ser Val Tyr Gln Val Leu Gln Glu His Leu Gly Arg Glu Ser Asp Leu
195 200 205
Phe Leu Leu Asp Ser Arg Thr Leu Trp Ala Ser Glu Glu Gly Trp Leu
210 215 220
Val Phe Asp Ile Thr Ala Thr Ser Asn His Trp Val Val Asn Pro Arg
225 230 235 240
His Asn Leu Gly Leu Gln Leu Ser Val Glu Thr Leu Asp Gly Gln Ser
245 250 255
Ile Asn Pro Lys Leu Ala Gly Leu Ile Gly Arg His Gly Pro Gln Asn
260 265 270
Lys Gln Pro Phe Met Val Ala Phe Phe Lys Ala Thr Glu Val His Phe
275 280 285
Arg Ser Ile Arg Ser Thr Gly Ser Lys Gln Arg Ser Gln Asn Arg Ser
290 295 300
Lys Thr Pro Lys Asn Gln Glu Ala Leu Arg Met Ala Asn Val Ala Glu
305 310 315 320
Asn Ser Ser Ser Asp Gln Arg Gln Ala Cys Lys Lys His Glu Leu Tyr
325 330 335
Val Ser Phe Arg Asp Leu Gly Trp Gln Asp Trp Ile Ile Ala Pro Glu
340 345 350
Gly Tyr Ala Ala Tyr Tyr Cys Glu Gly Glu Cys Ala Phe Pro Leu Asn
355 360 365
Ser Tyr Met Asn Ala Thr Asn His Ala Ile Val Gln Thr Leu Val His
370 375 380
Phe Ile Asn Pro Glu Thr Val Pro Lys Pro Cys Cys Ala Pro Thr Gln
385 390 395 400
Leu Asn Ala Ile Ser Val Leu Tyr Phe Asp Asp Ser Ser Asn Val Ile
405 410 415
Leu Lys Lys Tyr Arg Asn Met Val Val Arg Ala Cys Gly Cys His
420 425 430
<210>9
<211>22
<212>DNA
<213>人
<400>SEQ ID NO:9
actgcggtct cctaaaggtc ga 22
<210>10
<211>21
<212>DNA
<213>人
<400>SEQ ID NO:10
gctgacctga gtgcctgcga t 21
<210>11
<211>26
<212>DNA
<213>人
<400>SEQ ID NO:11
gaatgctgat ggtcgttttt attatg 26
<210>12
<211>20
<212>DNA
<213>人
<400>SEQ ID NO:12
agactgaagc cggtaaagat 20
<210>13
<211>26
<212>DNA
<213>人
<400>SEQ ID NO:13
aagaggacaa gaaggactaa aaatat 26
<210>14
<211>25
<212>DNA
<213>人
<400>SEQ ID NO:14
gtagagatcc agcataaaga gaggt 25
<210>15
<211>20
<212>DNA
<213>人
<400>SEQ ID NO:15
agcccgggta gcgcgtagag 20
<210>16
<211>21
<212>DNA
<213>人
<400>SEQ ID NO:16
gcgccggtgg atgaagctcg a 21
<210>17
<211>24
<212>DNA
<213>人
<400>SEQ ID NO:17
taaaggtgac agtacacagg aaca 24
<210>18
<211>23
<212>DNA
<213>人
<400>SEQ ID NO:18
tctatgatgg caaagcaatg tcc 23
<210>19
<211>26
<212>DNA
<213>人
<400>SEQ ID NO:19
tacaagcctg ccataagtga agaagc 26
<210>20
<211>25
<212>DNA
<213>人
<400>SEQ ID NO:20
atcatcgtga aacaatatcc gtctg 25
<210>21
<211>26
<212>DNA
<213>人
<400>SEQ ID NO:21
tcctctcatc agccatttgt cctttc 26
<210>22
<211>22
<212>DNA
<213>人
<400>SEQ ID NO:22
agttactaca cattcttcat ag 22
<210>23
<211>20
<212>DNA
<213>人
<400>SEQ ID NO:23
ctctgctcac tctgcacctg 20
<210>24
<211>20
<212>DNA
<213>人
<400>SEQ ID NO:24
ccggtcacca tcaaaatagc 20
<210>25
<211>20
<212>DNA
<213>人
<400>SEQ ID NO:25
atcaaccgct tctgttacgg 20
<210>26
<211>20
<212>DNA
<213>人
<400>SEQ ID NO:26
atgcaacgac actgcttcac 20
<210>27
<211>20
<212>DNA
<213>人
<400>SEQ ID NO:27
tgccacctga gaaaggctac 20
<210>28
<211>20
<212>DNA
<213>人
<400>SEQ ID NO:28
acagacaggc tcatccgact 20
<210>29
<211>20
<212>DNA
<213>人
<400>SEQ ID NO:29
cactacgacc caggcttcat 20
<210>30
<211>20
<212>DNA
<213>人
<400>SEQ ID NO:30
ctccgcagct tcttgcttag 20
<210>31
<211>20
<212>DNA
<213>人
<400>SEQ ID NO:31
atccttcttc atctggctgc 20
<210>32
<211>20
<212>DNA
<213>人
<400>SEQ ID NO:32
aattggtgtc ctgaggatcg 20
<210>33
<211>20
<213>人
<400>SEQ ID NO:33
atagtgagcc cttcccacct 20
<210>34
<211>20
<212>DNA
<213>人
<400>SEQ ID NO:34
aatgaacaga cccgcatttc 20
<210>35
<211>21
<212>DNA
<213>人
<400>SEQ ID NO:35
gatcgccact ccagctacat c 21
<210>36
<211>16
<212>DNA
<213>人
<400>SEQ ID NO:36
gggcacggca atgacc 16
<210>37
<211>2932
<212>DNA
<213>人
<400>SEQ ID NO:37
gctccgcgcc gagggctgga ggatgcgttc cctggggtcc ggacttatga aaatatgcat 60
cagtttaata ctgtcttgga attcatgaga tggaagcata ggtcaaagct gtttggagaa 120
aatcagaagt acagttttat ctagccacat cttggaggag tcgtaagaaa gcagtgggag 180
ttgaagtcat tgtcaagtgc ttgcgatctt ttacaagaaa atctcactga atgatagtca 240
tttaaattgg tgaagtagca agaccaatta ttaaaggtga cagtacacag gaaacattac 300
aattgaacaa tgactcagct atacatttac atcagattat tgggagccta tttgttcatc 360
atttctcgtg ttcaaggaca gaatctggat agtatgcttc atggcactgg gatgaaatca 420
gactccgacc agaaaaagtc agaaaatgga gtaaccttag caccagagga taccttgcct 480
tttttaaagt gctattgctc agggcactgt ccagatgatg ctattaataa cacatgcata 540
actaatggac attgctttgc catcatagaa gaagatgacc agggagaaac cacattagct 600
tcagggtgta tgaaatatga aggatctgat tttcagtgca aagattctcc aaaagcccag 660
ctacgccgga caatagaatg ttgtcggacc aatttatgta accagtattt gcaacccaca 720
ctgccccctg ttgtcatagg tccgtttttt gatggcagca ttcgatggct ggttttgctc 780
atttctatgg ctgtctgcat aattgctatg atcatcttct ccagctgctt ttgttacaaa 840
cattattgca agagcatctc aagcagacgt cgttacaatc gtgatttgga acaggatgaa 900
gcatttattc cagttggaga atcactaaaa gaccttattg accagtcaca aagttctggt 960
agtgggtctg gactaccttt attggttcag cgaactattg ccaaacagat tcagatggtc 1020
cggcaagttg gtaaaggccg atatggagaa gtatggatgg gcaaatggcg tggcgaaaaa 1080
gtggcggtga aagtattctt taccactgaa gaagccagct ggtttcgaga aacagaaatc 1140
taccaaactg tgctaatgcg ccatgaaaac atacttggtt tcatagcggc agacattaaa 1200
ggtacaggtt cctggactca gctctatttg attactgatt accatgaaaa tggatctctc 1260
tatgacttcc tgaaatgtgc tacactggac accagagccc tgcttaaatt ggcttattca 1320
gctgcctgtg gtctgtgcca cctgcacaca gaaatttatg gcacccaagg aaagcccgca 1380
attgctcatc gagacctaaa gagcaaaaac atcctcatca agaaaaatgg gagttgctgc 1440
attgctgacc tgggccttgc tgttaaattc aacagtgaca caaatgaagt tgatgtgccc 1500
ttgaatacca gggtgggcac caaacgctac atggctcccg aagtgctgga cgaaagcctg 1560
aacaaaaacc acttccagcc ctacatcatg gctgacatct acagcttcgg cctaatcatt 1620
tgggagatgg ctcgtcgttg tatcacagga gggatcgtgg aagaatacca attgccatat 1680
tacaacatgg taccgagtga tccgtcatac gaagatatgc gtgaggttgt gtgtgtcaaa 1740
cgtttgcggc caattgtgtc taatcggtgg aacagtgatg aatgtctacg agcagttttg 1800
aagctaatgt cagaatgctg ggcccacaat ccagcctcca gactcacagc attgagaatt 1860
aagaagacgc ttgccaagat ggttgaatcc caagatgtaa aaatctgatg gttaaaccat 1920
cggaggagaa actctagact gcaagaactg tttttaccca tggcatgggt ggaattagag 1980
tggaataagg atgttaactt ggttctcaga ctctttcttc actacgtgtt cacaggctgc 2040
taatattaaa cctttcagta ctcttattag gatacaagct gggaacttct aaacacttca 2100
ttctttatat atggacagct ttattttaaa tgtggttttt gatgcctttt tttaagtggg 2160
tttttatgaa ctgcatcaag acttcaatcc tgattagtgt ctccagtcaa gctctgggta 2220
ctgaattgcc tgttcataaa acggtgcttt ctgtgaaagc cttaagaaga taaatgagcg 2280
cagcagagat ggagaaatag actttgcctt ttacctgaga cattcagttc gtttgtattc 2340
tacctttgta aaacagccta tagatgatga tgtgtttggg atactgctta ttttatgata 2400
gtttgtcctg tgtccttagt gatgtgtgtg tgtctccatg cacatgcacg ccgggattcc 2460
tctgctgcca tttgaattag aagaaaataa tttatatgca tgcacaggaa gatattggtg 2520
gccggtggtt ttgtgcttta aaaatgcaat atctgaccaa gattcgccaa tctcatacaa 2580
gccatttact ttgcaagtga gatagcttcc ccaccagctt tattttttaa catgaaagct 2640
gatgccaagg ccaaaagaag tttaaagcat ctgtaaattt ggactgtttt ccttcaacca 2700
ccattttttt tgtggttatt atttttgtca cggaaagcat cctctccaaa gttggagctt 2760
ctattgccat gaaccatgct tacaaagaaa gcacttctta ttgaagtgaa ttcctgcatt 2820
tgatagcaat gtaagtgcct ataaccatgt tctatattct ttattctcag taacttttaa 2880
aagggaagtt atttatattt tgtgtataat gtgctttatt tgcaaatcac cc 2932
<210>38
<211>532
<212>PRT
<213>人
<400>SEQ ID NO:38
Met Thr Gln Leu Tyr Ile Tyr Ile Arg Leu Leu Gly Ala Tyr Leu Phe
1 5 10 15
Ile Ile Ser Arg Val Gln Gly Gln Asn Leu Asp Ser Met Leu His Gly
20 25 30
Thr Gly Met Lys Ser Asp Ser Asp Gln Lys Lys Ser Glu Asn Gly Val
35 40 45
Thr Leu Ala Pro Glu Asp Thr Leu Pro Phe Leu Lys Cys Tyr Cys Ser
50 55 60
Gly His Cys Pro Asp Asp Ala Ile Asn Asn Thr Cys Ile Thr Asn Gly
65 70 75 80
His Cys Phe Ala Ile Ile Glu Glu Asp Asp Gln Gly Glu Thr Thr Leu
85 90 95
Ala Ser Gly Cys Met Lys Tyr Glu Gly Ser Asp Phe Gln Cys Lys Asp
100 105 110
Ser Pro Lys Ala Gln Leu Arg Arg Thr Ile Glu Cys Cys Arg Thr Asn
115 120 125
Leu Cys Asn Gln Tyr Leu Gln Pro Thr Leu Pro Pro Val Val Ile Gly
130 135 140
Pro Phe Phe Asp Gly Ser Ile Arg Trp Leu Val Leu Leu Ile Ser Met
145 150 155 160
A1a Val Cys Ile Ile Ala Met Ile Ile Phe Ser Ser Cys Phe Cys Tyr
165 170 175
Lys His Tyr Cys Lys Ser Ile Ser Ser Arg Arg Arg Tyr Asn Arg Asp
180 185 190
Leu Glu Gln Asp Glu Ala Phe Ile Pro Val Gly Glu Ser Leu Lys Asp
195 200 205
Leu Ile Asp Gln Ser Gln Ser Ser Gly Ser Gly Ser Gly Leu Pro Leu
210 215 220
Leu Val Gln Arg Thr Ile Ala Lys Gln Ile Gln Met Val Arg Gln Val
225 230 235 240
Gly Lys Gly Arg Tyr Gly Glu Val Trp Met Gly Lys Trp Arg Gly Glu
245 250 255
Lys Val Ala Val Lys Val Phe Phe Thr Thr Glu Glu Ala Ser Trp Phe
260 265 270
Arg Glu Thr Glu Ile Tyr Gln Thr Val Leu Met Arg His Glu Asn Ile
275 280 285
Leu Gly Phe Ile Ala Ala Asp Ile Lys Gly Thr Gly Ser Trp Thr Gln
290 295 300
Leu Tyr Leu Ile Thr Asp Tyr His Glu Asn Gly Ser Leu Tyr Asp Phe
305 310 315 320
Leu Lys Cys Ala Thr Leu Asp Thr Arg Ala Leu Leu Lys Leu Ala Tyr
325 330 335
Ser Ala Ala Cys Gly Leu Cys His Leu His Thr Glu Ile Tyr Gly Thr
340 345 350
Gln Gly Lys Pro Ala Ile Ala His Arg Asp Leu Lys Ser Lys Asn Ile
355 360 365
Leu Ile Lys Lys Asn Gly Ser Cys Cys Ile Ala Asp Leu Gly Leu Ala
370 375 380
Val Lys Phe Asn Ser Asp Thr Asn Glu Val Asp Val Pro Leu Asn Thr
385 390 395 400
Arg Val Gly Thr Lys Arg Tyr Met Ala Pro Glu Val Leu Asp Glu Ser
405 410 415
Leu Asn Lys Asn His Phe Gln Pro Tyr Ile Met Ala Asp Ile Tyr Ser
420 425 430
Phe Gly Leu Ile Ile Trp Glu Met Ala Arg Arg Cys Ile Thr Gly Gly
435 440 445
Ile Val Glu Glu Tyr Gln Leu Pro Tyr Tyr Asn Met Val Pro Ser Asp
450 455 460
Pro Ser Tyr Glu Asp Met Arg Glu Val Val Cys Val Lys Arg Leu Arg
465 470 475 480
Pro Ile Val Ser Asn Arg Trp Asn Ser Asp Glu Cys Leu Arg Ala Val
485 490 495
Leu Lys Leu Met Ser Glu Cys Trp Ala His Asn Pro Ala Ser Arg Leu
500 505 510
Thr Ala Leu Arg Ile Lys Lys Thr Leu Ala Lys Met Val Glu Ser Gln
515 520 525
Asp Val Lys Ile
530
<210>39
<211>2032
<212>DNA
<213>人
<400>SEQ ID NO:39
cgcggggcgc ggagtcggcg gggcctcgcg ggacgcgggc agtgcggaga ccgcggcgct 60
gaggacgcgg gagccgggag cgcacgcgcg gggtggagtt cagcctactc tttcttagat 120
gtgaaaggaa aggaagatca tttcatgcct tgttgataaa ggttcagact tctgctgatt 180
cataaccatt tggctctgag ctatgacaag agaggaaaca aaaagttaaa cttacaagcc 240
tgccataagt gagaagcaaa cttccttgat aacatgcttt tgcgaagtgc aggaaaatta 300
aatgtgggca ccaagaaaga ggatggtgag agtacagccc ccaccccccg tccaaaggtc 360
ttgcgttgta aatgccacca ccattgtcca gaagactcag tcaacaatat ttgcagcaca 420
gacggatatt gtttcacgat gatagaagag gatgactctg ggttgcctgt ggtcacttct 480
ggttgcctag gactagaagg ctcagatttt cagtgtcggg acactcccat tcctcatcaa 540
agaagatcaa ttgaatgctg cacagaaagg aacgaatgta ataaagacct acaccctaca 600
ctgcctccat tgaaaaacag agattttgtt gatggaccta tacaccacag ggctttactt 660
atatctgtga ctgtctgtag tttgctcttg gtccttatca tattattttg ttacttccgg 720
tataaaagac aagaaaccag acctcgatac agcattgggt tagaacagga tgaaacttac 780
attcctcctg gagaatccct gagagactta attgagcagt ctcagagctc aggaagtgga 840
tcaggcctcc ctctgctggt ccaaaggact atagctaagc agattcagat ggtgaaacag 900
attggaaaag gtcgctatgg ggaagtttgg atgggaaagt ggcgtggcga aaaggtagct 960
gtgaaagtgt tcttcaccac agaggaagcc agctggttca gagagacaga aatatatcag 1020
acagtgttga tgaggcatga aaacattttg ggtttcattg ctgcagatat caaagggaca 1080
gggtcctgga cccagttgta cctaatcaca gactatcatg aaaatggttc cctttatgat 1140
tatctgaagt ccaccaccct agacgctaaa tcaatgctga agttagccta ctcttctgtc 1200
agtggcttat gtcatttaca cacagaaatc tttagtactc aaggcaaacc agcaattgcc 1260
catcgagatc tgaaaagtaa aaacattctg gtgaagaaaa atggaacttg ctgtattgct 1320
gacctgggcc tggctgttaa atttattagt gatacaaatg aagttgacat accacctaac 1380
actcgagttg gcaccaaacg ctatatgcct ccagaagtgt tggacgagag cttgaacaga 1440
aatcacttcc agtcttacat catggctgac atgtatagtt ttggcctcat cctttgggag 1500
gttgctagga gatgtgtatc aggaggtata gtggaagaat accagcttcc ttatcatgac 1560
ctagtgccca gtgacccctc ttatgaggac atgagggaga ttgtgtgcat caagaagtta 1620
cgcccctcat tcccaaaccg gtggagcagt gatgagtgtc taaggcagat gggaaaactc 1680
atgacagaat gctgggctca caatcctgca tcaaggctga cagccctgcg ggttaagaaa 1740
acacttgcca aaatgtcaga gtcccaggac attaaactct gataggagag gaaaagtaag 1800
catctctgca gaaagccaac aggtactctt ctgtttgtgg gcagagcaaa agacatcaaa 1860
taagcatcca cagtacaagc cttgaacatc gtcctgcttc ccagtgggtt cagacctcac 1920
ctttcaggga gcgacctggg caaagacaga gaagctccca gaaggagaga ttgatccgtg 1980
tctgtttgta ggcggagaaa ccgttgggta acttgttcaa gatatgatgc at 2032
<210>40
<211>502
<212>PRT
<213>人
<400>SEQ ID NO:40
Met Leu Leu Arg Ser Ala Gly Lys Leu Asn Val Gly Thr Lys Lys Glu
1 5 10 15
Asp Gly Glu Ser Thr Ala Pro Thr Pro Arg Pro Lys Val Leu Arg Cys
20 25 30
Lys Cys His His His Cys Pro Glu Asp Ser Val Asn Asn Ile Cys Ser
35 40 45
Thr Asp Gly Tyr Cys Phe Thr Met Ile Glu Glu Asp Asp Ser Gly Leu
50 55 60
Pro Val Val Thr Ser Gly Cys Leu Gly Leu Glu Gly Ser Asp Phe Gln
65 70 75 80
Cys Arg Asp Thr Pro Ile Pro His Gln Arg Arg Ser Ile Glu Cys Cys
85 90 95
Thr Glu Arg Asn Glu Cys Asn Lys Asp Leu His Pro Thr Leu Pro Pro
100 105 110
Leu Lys Asn Arg Asp Phe Val Asp Gly Pro Ile His His Arg Ala Leu
115 120 125
Leu Ile Ser Val Thr Val Cys Ser Leu Leu Leu Val Leu Ile Ile Leu
130 135 140
Phe Cys Tyr Phe Arg Tyr Lys Arg Gln Glu Thr Arg Pro Arg Tyr Ser
145 150 155 160
Ile Gly Leu Glu Gln Asp Glu Thr Tyr Ile Pro Pro Gly Glu Ser Leu
165 170 175
Arg Asp Leu Ile Glu Gln Ser Gln Ser Ser Gly Ser Gly Ser Gly Leu
180 185 190
Pro Leu Leu Val Gln Arg Thr Ile Ala Lys Gln Ile Gln Met Val Lys
195 200 205
Gln Ile Gly Lys Gly Arg Tyr Gly Glu Val Trp Met Gly Lys Trp Arg
210 215 220
Gly Glu Lys Val Ala Val Lys Val Phe Phe Thr Thr Glu Glu Ala Ser
225 230 235 240
Trp Phe Arg Glu Thr Glu Ile Tyr Gln Thr Val Leu Met Arg His Glu
245 250 255
Asn Ile Leu Gly Phe Ile Ala Ala Asp Ile Lys Gly Thr Gly Ser Trp
260 265 270
Thr Gln Leu Tyr Leu Ile Thr Asp Tyr His Glu Asn Gly Ser Leu Tyr
275 280 285
Asp Tyr Leu Lys Ser Thr Thr Leu Asp Ala Lys Ser Met Leu Lys Leu
290 295 300
Ala Tyr Ser Ser Val Ser Gly Leu Cys His Leu His Thr Glu Ile Phe
305 310 315 320
Ser Thr Gln Gly Lys Pro Ala Ile Ala His Arg Asp Leu Lys Ser Lys
325 330 335
Asn Ile Leu Val Lys Lys Asn Gly Thr Cys Cys Ile Ala Asp Leu Gly
340 345 350
Leu Ala Val Lys Phe Ile Ser Asp Thr Asn Glu Val Asp Ile Pro Pro
355 360 365
Asn Thr Arg Val Gly Thr Lys Arg Tyr Met Pro Pro Glu Val Leu Asp
370 375 380
Glu Ser Leu Asn Arg Asn His Phe Gln Ser Tyr Ile Met Ala Asp Met
385 390 395 400
Tyr Ser Phe Gly Leu Ile Leu Trp Glu Val Ala Arg Arg Cys Val Ser
405 410 415
Gly Gly Ile Val Glu Glu Tyr Gln Leu Pro Tyr His Asp Leu Val Pro
420 425 430
Ser Asp Pro Ser Tyr Glu Asp Met Arg Glu Ile Val Cys Ile Lys Lys
435 440 445
Leu Arg Pro Ser Phe Pro Asn Arg Trp Ser Ser Asp Glu Cys Leu Arg
450 455 460
Gln Met Gly Lys Leu Met Thr Glu Cys Trp Ala His Asn Pro Ala Ser
465 470 475 480
Arg Leu Thr Ala Leu Arg Val Lys Lys Thr Leu Ala Lys Met Ser Glu
485 490 495
Ser Gln Asp Ile Lys Leu
500
<210>41
<211>3611
<212>DNA
<213>人
<400>SEQ ID NO:41
cgccccccga ccccggatcg aatccccgcc ctccgcaccc tggatatgtt ttctcccaga 60
cctggatatt tttttgatat cgtgaaacta cgagggaaat aatttggggg atttcttctt 120
ggctccctgc tttccccaca gacatgcctt ccgtttggag ggccgcggca ccccgtccga 180
ggcgaaggaa cccccccagc cgcgagggag agaaatgaag ggaatttctg cagcggcatg 240
aaagctctgc agctaggtcc tctcatcagc catttgtcct ttcaaactgt attgtgatac 300
gggcaggatc agtccacggg agagaagacg agcctcccgg ctgtttctcc gccggtctac 360
ttcccatatt tcttttcttt gccctcctga ttcttggctg gcccagggat gacttcctcg 420
ctgcagcggc cctggcgggt gccctggcta ccatggacca tcctgctggt cagcactgcg 480
gctgcttcgc agaatcaaga acggctatgt gcgtttaaag atccgtatca gcaagacctt 540
gggataggtg agagtagaat ctctcatgaa aatgggacaa tattatgctc gaaaggtagc 600
acctgctatg gcctttggga gaaatcaaaa ggggacataa atcttgtaaa acaaggatgt 660
tggtctcaca ttggagatcc ccaagagtgt cactatgaag aatgtgtagt aactaccact 720
cctccctcaa ttcagaatgg aacataccgt ttctgctgtt gtagcacaga tttatgtaat 780
gtcaacttta ctgagaattt tccacctcct gacacaacac cactcagtcc acctcattca 840
tttaaccgag atgagacaat aatcattgct ttggcatcag tctctgtatt agctgttttg 900
atagttgcct tatgctttgg atacagaatg ttgacaggag accgtaaaca aggtcttcac 960
agtatgaaca tgatggaggc agcagcatcc gaaccctctc ttgatctaga taatctgaaa 1020
ctgttggagc tgattggccg aggtcgatat ggagcagtat ataaaggctc cttggatgag 1080
cgtccagttg ctgtaaaagt gttttccttt gcaaaccgtc agaattttat caacgaaaag 1140
aacatttaca gagtgccttt gatggaacat gacaacattg cccgctttat agttggagat 1200
gagagagtca ctgcagatgg acgcatggaa tatttgcttg tgatggagta ctatcccaat 1260
ggatctttat gcaagtattt aagtctccac acaagtgact gggtaagctc ttgccgtctt 1320
gctcattctg ttactagagg actggcttat cttcacacag aattaccacg aggagatcat 1380
tataaacctg caatttccca tcgagattta aacagcagaa atgtcctagt gaaaaatgat 1440
ggaacctgtg ttattagtga ctttggactg tccatgaggc tgactggaaa tagactggtg 1500
cgcccagggg aggaagataa tgcagccata agcgaggttg gcactatcag atatatggca 1560
ccagaagtgc tagaaggagc tgtgaacttg agggactgtg aatcagcttt gaaacaagta 1620
gacatgtatg ctcttggact aatctattgg gagatattta tgagatgtac agacctcttc 1680
ccaggggaat ccgtaccaga gtaccagatg gcttttcaga cagaggttgg aaaccatccc 1740
acttttgagg atatgcaggt tctcgtgtct agggaaaaac agagacccaa gttcccagaa 1800
gcctggaaag aaaatagcct ggcagtgagg tcactcaagg agacaatcga agactgttgg 1860
gaccaggatg cagaggctcg gcttactgca cagtgtgctg aggaaaggat ggctgaactt 1920
atgatgattt gggaaagaaa caaatctgtg agcccaacag tcaatccaat gtctactgct 1980
atgcagaatg aacgcaacct gtcacataat aggcgtgtgc caaaaattgg tccttatcca 2040
gattattctt cctcctcata cattgaagac tctatccatc atactgacag catcgtgaag 2100
aatatttcct ctgagcattc tatgtccagc acacctttga ctatagggga aaaaaaccga 2160
aattcaatta actatgaacg acagcaagca caagctcgaa tccccagccc tgaaacaagt 2220
gtcaccagcc tctccaccaa cacaacaacc acaaacacca caggactcac gccaagtact 2280
ggcatgacta ctatatctga gatgccatac ccagatgaaa caaatctgca taccacaaat 2340
gttgcacagt caattgggcc aacccctgtc tgcttacagc tgacagaaga agacttggaa 2400
accaacaagc tagacccaaa agaagttgat aagaacctca aggaaagctc tgatgagaat 2460
ctcatggagc actctcttaa acagttcagt ggcccagacc cactgagcag tactagttct 2520
agcttgcttt acccactcat aaaacttgca gtagaagcaa ctggacagca ggacttcaca 2580
cagactgcaa atggccaagc atgtttgatt cctgatgttc tgcctactca gatctatcct 2640
ctccccaagc agcagaacct tcccaagaga cctactagtt tgcctttgaa caccaaaaat 2700
tcaacaaaag agccccggct aaaatttggc agcaagcaca aatcaaactt gaaacaagtc 2760
gaaactggag ttgccaagat gaatacaatc aatgcagcag aacctcatgt ggtgacagtc 2820
accatgaatg gtgtggcagg tagaaaccac agtgttaact cccatgctgc cacaacccaa 2880
tatgccaatg ggacagtact atctggccaa acaaccaaca tagtgacaca tagggcccaa 2940
gaaatgttgc agaatcagtt tattggtgag gacacccggc tgaatattaa ttccagtcct 3000
gatgagcatg agcctttact gagacgagag caacaagctg gccatgatga aggtgttctg 3060
gatcgtcttg tggacaggag ggaacggcca ctagaaggtg gccgaactaa ttccaataac 3120
aacaacagca atccatgttc agaacaagat gttcttgcac agggtgttcc aagcacagca 3180
gcagatcctg ggccatcaaa gcecagaaga gcacagaggc ctaattctct ggatctttca 3240
gccacaaatg tcctggatgg cagcagtata cagataggtg agtcaacaca agatggcaaa 3300
tcaggatcag gtgaaaagat caagaaacgt gtgaaaactc cctattctct taagcggtgg 3360
cgcccctcca cctgggtcat ctccactgaa tcgctggact gtgaagtcaa caataatggc 3420
agtaacaggg cagttcattc caaatccagc actgctgttt accttgcaga aggaggcact 3480
gctacaacca tggtgtctaa agatatagga atgaactgtc tgtgaaatgt tttcaagcct 3540
atggagtgaa attatttttt gcatcattta aacatgcaga agatgtttaa aaataaaaaa 3600
aaaactgctt t 3611
<210>42
<211>1038
<212>PRT
<213>人
<400>SEQ ID NO:42
Met Thr Ser Ser Leu Gln Arg Pro Trp Arg Val Pro Trp Leu Pro Trp
1 5 10 15
Thr Ile Leu Leu Val Ser Thr Ala Ala Ala Ser Gln Asn Gln Glu Arg
20 25 30
Leu Cys Ala Phe Lys Asp Pro Tyr Gln Gln Asp Leu Gly Ile Gly Glu
35 40 45
Ser Arg Ile Ser His Glu Asn Gly Thr Ile Leu Cys Ser Lys Gly Ser
50 55 60
Thr Cys Tyr Gly Leu Trp Glu Lys Ser Lys Gly Asp Ile Asn Leu Val
65 70 75 80
Lys Gln Gly Cys Trp Ser His Ile Gly Asp Pro Gln Glu Cys His Tyr
85 90 95
Glu Glu Cys Val Val Thr Thr Thr Pro Pro Ser Ile Gln Asn Gly Thr
100 105 110
Tyr Arg Phe Cys Cys Cys Ser Thr Asp Leu Cys Asn Val Asn Phe Thr
115 120 125
Glu Asn Phe Pro Pro Pro Asp Thr Thr Pro Leu Ser Pro Pro His Ser
130 135 140
Phe Asn Arg Asp Glu Thr Ile Ile Ile Ala Leu Ala Ser Val Ser Val
145 150 155 160
Leu Ala Val Leu Ile Val Ala Leu Cys Phe Gly Tyr Arg Met Leu Thr
165 170 175
Gly Asp Arg Lys Gln Gly Leu His Ser Met Asn Met Met Glu Ala Ala
180 185 190
Ala Ser Glu Pro Ser Leu Asp Leu Asp Asn Leu Lys Leu Leu Glu Leu
195 200 205
Ile Gly Arg Gly Arg Tyr Gly Ala Val Tyr Lys Gly Ser Leu Asp Glu
210 215 220
Arg Pro Val Ala Val Lys Val Phe Ser Phe Ala Asn Arg Gln Asn Phe
225 230 235 240
Ile Asn Glu Lys Asn Ile Tyr Arg Val Pro Leu Met Glu His Asp Asn
245 250 255
Ile Ala Arg Phe Ile Val Gly Asp Glu Arg Val Thr Ala Asp Gly Arg
260 265 270
Met Glu Tyr Leu Leu Val Met Glu Tyr Tyr Pro Asn Gly Ser Leu Cys
275 280 285
Lys Tyr Leu Ser Leu His Thr Ser Asp Trp Val Ser Ser Cys Arg Leu
290 295 300
Ala His Ser Val Thr Arg Gly Leu Ala Tyr Leu His Thr Glu Leu Pro
305 310 315 320
Arg Gly Asp His Tyr Lys Pro Ala Ile Ser His Arg Asp Leu Asn Ser
325 330 335
Arg Asn Val Leu Val Lys Asn Asp Gly Thr Cys Val Ile Ser Asp Phe
340 345 350
Gly Leu Ser Met Arg Leu Thr Gly Asn Arg Leu Val Arg Pro Gly Glu
355 360 365
Glu Asp Asn Ala Ala Ile Ser Glu Val Gly Thr Ile Arg Tyr Met Ala
370 375 380
Pro Glu Val Leu Glu Gly Ala Val Asn Leu Arg Asp Cys Glu Ser Ala
385 390 395 400
Leu Lys Gln Val Asp Met Tyr Ala Leu Gly Leu Ile Tyr Trp Glu Ile
405 410 415
Phe Met Arg Cys Thr Asp Leu Phe Pro Gly Glu Ser Val Pro Glu Tyr
420 425 430
Gln Met Ala Phe Gln Thr Glu Val Gly Asn His Pro Thr Phe Glu Asp
435 440 445
Met Gln Val Leu Val Ser Arg Glu Lys Gln Arg Pro Lys Phe Pro Glu
450 455 460
Ala Trp Lys Glu Asn Ser Leu Ala Val Arg Ser Leu Lys Glu Thr Ile
465 470 475 480
Glu Asp Cys Trp Asp Gln Asp Ala Glu Ala Arg Leu Thr Ala Gln Cys
485 490 495
Ala Glu Glu Arg Met Ala Glu Leu Met Met Ile Trp Glu Arg Asn Lys
500 505 510
Ser Val Ser Pro Thr Val Asn Pro Met Ser Thr Ala Met Gln Asn Glu
515 520 525
Arg Asn Leu Ser His Asn Arg Arg Val Pro Lys Ile Gly Pro Tyr Pro
530 535 540
Asp Tyr Ser Ser Ser Ser Tyr Ile Glu Asp Ser Ile His His Thr Asp
545 550 555 560
Ser Ile Val Lys Asn Ile Ser Ser Glu His Ser Met Ser Ser Thr Pro
565 570 575
Leu Thr Ile Gly Glu Lys Asn Arg Asn Ser Ile Asn Tyr Glu Arg Gln
580 585 590
Gln Ala Gln Ala Arg Ile Pro Ser Pro Glu Thr Ser Val Thr Ser Leu
595 600 605
Ser Thr Asn Thr Thr Thr Thr Asn Thr Thr Gly Leu Thr Pro Ser Thr
610 615 620
Gly Met Thr Thr Ile Ser Glu Met Pro Tyr Pro Asp Glu Thr Asn Leu
625 630 635 640
His Thr Thr Asn Val Ala Gln Ser Ile Gly Pro Thr Pro Val Cys Leu
645 650 655
Gln Leu Thr Glu Glu Asp Leu Glu Thr Asn Lys Leu Asp Pro Lys Glu
660 665 670
Val Asp Lys Asn Leu Lys Glu Ser Ser Asp Glu Asn Leu Met Glu His
675 680 685
Ser Leu Lys Gln Phe Ser Gly Pro Asp Pro Leu Ser Ser Thr Ser Ser
690 695 700
Ser Leu Leu Tyr Pro Leu Ile Lys Leu Ala Val Glu Ala Thr Gly Gln
705 710 715 720
Gln Asp Phe Thr Gln Thr Ala Asn Gly Gln Ala Cys Leu Ile Pro Asp
725 730 735
Val Leu Pro Thr Gln Ile Tyr Pro Leu Pro Lys Gln Gln Asn Leu Pro
740 745 750
Lys Arg Pro Thr Ser Leu Pro Leu Asn Thr Lys Asn Ser Thr Lys Glu
755 760 765
Pro Arg Leu Lys Phe Gly Ser Lys His Lys Ser Asn Leu Lys Gln Val
770 775 780
Glu Thr Gly Val Ala Lys Met Asn Thr Ile Asn Ala Ala Glu Pro His
785 790 795 800
Val Val Thr Val Thr Met Asn Gly Val Ala Gly Arg Asn His Ser Val
805 810 815
Asn Ser His Ala Ala Thr Thr Gln Tyr Ala Asn Gly Thr Val Leu Ser
820 825 830
Gly Gln Thr Thr AsnIle Val Thr Hi s Arg Ala Gln Glu Met Leu Gln
835 840 845
Asn Gln Phe Ile Gly Glu Asp Thr Arg Leu Asn Ile Asn Ser Ser Pro
850 855 860
Asp Glu His Glu Pro Leu Leu Arg Arg Glu Gln Gln Ala Gly His Asp
865 870 875 880
Glu Gly Val Leu Asp Arg Leu Val Asp Arg Arg Glu Arg Pro Leu Glu
885 890 895
Gly Gly Arg Thr Asn Ser Asn Asn Asn Asn Ser Asn Pro Cys Ser Glu
900 905 910
Gln Asp Val Leu Ala Gln Gly Val Pro Ser Thr Ala Ala Asp Pro Gly
915 920 925
Pro Ser Lys Pro Arg Arg Ala Gln Arg Pro Asn Ser Leu Asp Leu Ser
930 935 940
Ala Thr Asn Val Leu Asp Gly Ser Ser Ile Gln Ile Gly Glu Ser Thr
945 950 955 960
Gln Asp Gly Lys Ser Gly Ser Gly Glu Lys Ile Lys Lys Arg Val Lys
965 970 975
Thr Pro Tyr Ser Leu Lys Arg Trp Arg Pro Ser Thr Trp Val Ile Ser
980 985 990
Thr Glu Ser Leu Asp Cys Glu Val Asn Asn Asn Gly Ser Asn Arg Ala
995 1000 1005
Val His Ser Lys Ser Ser Thr Ala Val Tyr Leu Ala Glu Gly Gly
1010 1015 1020
Thr Ala Thr Thr Met Val Ser Lys Asp Ile Gly Met Asn Cys Leu
1025 1030 1035
<210>43
<211>3561
<212>DNA
<213>人
<400>SEQ ID NO:43
cccgggtcag cgcccgcccg cccgcgctcc tcccggccgc tcctcccgcc ccgcccggcc 60
cggcgccgac tctgcggccg cccgacgagc ccctcgcggc actgccccgg ccccggcccc 120
ggccccggcc ccctcccgcc gcaccgcccc cggcccggcc ctccgccctc cgcactcccg 180
cctccctccc tccgcccgct cccgcgccct cctccctccc tcctccccag ctgtcccgtt 240
cgcgtcatgc cgagcctccc ggccccgccg gccccgctgc tgctcctcgg gctgctgctg 300
ctcggctccc ggccggcccg cggcgccggc cccgagcccc ccgtgctgcc catccgttct 360
gagaaggagc cgctgcccgt tcggggagcg gcaggctgca ccttcggcgg gaaggtctat 420
gccttggacg agacgtggca cccggaccta ggggagccat tcggggtgat gcgctgcgtg 480
ctgtgcgcct gcgaggcgac agggaccttg aggcccagag agatgaagta gcttgtctag 540
ggtcacgcag cttcctcagt ggggtcgccg taccaggggc cctggcaggg tcagctgcaa 600
gaacatcaaa ccagagtgcc caaccccggc ctgtgggcag ccgcgccagc tgccgggaca 660
ctgctgccag acctgccccc aggagcgcag cagttcggag cggcagccga gcggcctgtc 720
cttcgagtat ccgcgggacc cggagcatcg cagttatagc gaccgcgggg agccaggcgc 780
tgaggagcgg gcccgtggtg acggccacac ggacttcgtg gcgctgctga cagggccgag 840
gtcgcaggcg gtggcacgag cccgagtctc gctgctgcgc tctagcctcc gcttctctat 900
ctcctacagg cggctggacc gccctaccag gatccgcttc tcagactcca atggcagtgt 960
cctgtttgag caccctgcag cccccaccca agatggcctg gtctgtgggg tgtggcgggc 1020
agtgcctcgg ttgtctctgc ggctccttag ggcagaacag ctgcatgtgg cacttgtgac 1080
actcactcac ccttcagggg aggtctgggg gcctctcatc cggcaccggg ccctggctgc 1140
agagaccttc agtgccatcc tgactctaga aggcccccca cagcagggcg tagggggcat 1200
caccctgctc actctcagtg acacagagga ctccttgcat tttttgctgc tcttccgagg 1260
gctgctggaa cccaggagtg gggattctac accaggggca gctactgcga gaacttcagg 1320
ccaatgtctc agcccaggaa ccaggctttg ctgaggtgct gcccaacctg acagtccagg 1380
agatggactg gctggtgctg ggggagctgc agatggccct ggagtgggca ggcaggccag 1440
ggctgcgcat cagtggacac attgctgcca ggaagagctg cgacgtcctg caaagtgtcc 1500
tttgtggggc tgatgccctg atcccagtcc agacgggtgc tgccggctca gccagcctca 1560
cgctgctagg aaatggctcc ctgatctatc aggtgcaagt ggtagggaca agcagtgagg 1620
tggtggccat gacactggag accaagcctc agcggaggga tcagcgcact gtcctgtgcc 1680
acatggctgg actccagcca ggaggacaca cggccgtggg tatctgccct gggctgggtg 1740
cccgaggggc tcatatgctg ctgcagaatg agctcttcct gaacgtgggc accaaggact 1800
tcccagacgg agagcttcgg gggcacgtgg ctgccctgcc ctactgtggg catagctccc 1860
gccatgacac gctgcccgtg cccctagcag gagccctggt gctaccccct gtgaagagcc 1920
aagcagcagg gcacgcctgg ctttccttgg atacccactg tcacctgcac tatgaagtgc 1980
tgctggctgg gcttggtggc tcagaacaag gcactgtcac tgcccacctc cttgggcctc 2040
ctggaacgcc agggcctcgg cggctgctga agggattcta tggctcagag gcccagggtg 2100
tggtgaagga cctggagccg gaactgctgc ggcacctggc aaaaggcatg gcctccctga 2160
tgatcaccac caagggtagc cccagagggg agctccgagg gcaggtgcac atagccaacc 2220
aatgtgaggt tggcggactg cgcctggagg cggccggggc cgagggggtg cgggcgctgg 2280
gggctccgga tacagcctct gctgcgccgc ctgtggtgcc tggtctcccg gccctagcgc 2340
ccgccaaacc tggtggtcct gggcggcccc gagaccccaa cacatgcttc ttcgaggggc 2400
agcagcgccc ccacggggct cgctgggcgc ccaactacga cccgctctgc tcactctgca 2460
cctgccagag acgaacggtg atctgtgacc cggtggtgtg cccaccgccc agctgcccac 2520
acccggtgca ggctcccgac cagtgctgcc ctgtttgccc tgagaaacaa gatgtcagag 2580
acttgccagg gctgccaagg agccgggacc caggagaggg ctgctatttt gatggtgacc 2640
ggagctggcg ggcagcgggt acgcggtggc accccgttgt gccccccttt ggcttaatta 2700
agtgtgctgt ctgcacctgc aaggggggca ctggagaggt gcactgtgag aaggtgcagt 2760
gtccccggct ggcctgtgcc cagcctgtgc gtgtcaaccc caccgactgc tgcaaacagt 2820
gtccagtggg gtcgggggcc cacccccagc tgggggaccc catgcaggct gatgggcccc 2880
ggggctgccg ttttgctggg cagtggttcc cagagagtca gagctggcac ccctcagtgc 2940
ccccttttgg agagatgagc tgtatcacct gcagatgtgg ggcaggggtg cctcactgtg 3000
agcgggatga ctgttcactg ccactgtcct gtggctcggg gaaggagagt cgatgctgtt 3060
cccgctgcac ggcccaccgg cggccagccc cagagaccag aactgatcca gagctggaga 3120
aagaagccga aggctcttag ggagcagcca gagggccaag tgaccaagag gatggggcct 3180
gagctgggga aggggtggca tcgaggacct tcttgcattc tcctgtggga agcccagtgc 3240
ctttgctccc ctgtcctgcc tctactccca cccccactac ctctgggaac cacagctcca 3300
caagggggag aggcagctgg gccagaccga ggtcacagcc actccaagtc ctgccctgcc 3360
accctcggcc tctgtcctgg aagccccacc cctttcctcc tgtacataat gtcactggct 3420
tgttgggatt tttaatttat cttcactcag caccaagggc ccccgacact ccactcctgc 3480
tgcccctgag ctgagcagag tcattattgg agagttttgt atttattaaa acatttcttt 3540
ttcagtcaaa aaaaaaaaaa a 3561
<210>44
<211>94
<212>PRT
<213>人
<400>SEQ ID N0:44
Met Pro Ser Leu Pro Ala Pro Pro Ala Pro Leu Leu Leu Leu Gly Leu
1 5 10 15
Leu Leu Leu Gly Ser Arg Pro Ala Arg Gly Ala Gly Pro Gl u Pro Pro
20 25 30
Val Leu Pro Ile Arg Ser Glu Lys Glu Pro Leu Pro Val Arg Gly Ala
35 40 45
Ala Gly Cys Thr Phe Gly Gly Lys Val Tyr Ala Leu Asp Glu Thr Trp
50 55 60
His Pro Asp Leu Gly Glu Pro Phe Gly Val Met Arg Cys Val Leu Cys
65 70 75 80
Ala Cys Glu Ala Thr Gly Thr Leu Arg Pro Arg Glu Met Lys
85 90
<210>45
<211>4049
<212>DNA
<213>人
<400>SEQ ID NO:45
gcggccgcac tcagcgccac gcgtcgaaag cgcaggcccc gaggacccgc cgcactgaca 60
gtatgagccg cacagcctac acggtgggag ccctgcttct cctcttgggg accctgctgc 120
cggctgctga agggaaaaag aaagggtccc aaggtgccat ccccccgcca gacaaggccc 180
agcacaatga ctcagagcag actcagtcgc cccagcagcc tggctccagg aaccgggggc 240
ggggccaagg gcggggcact gccatgcccg gggaggaggt gctggagtcc agccaagagg 300
ccctgcatgt gacggagcgc aaatacctga agcgagactg gtgcaaaacc cagccgctta 360
agcagaccat ccacgaggaa ggctgcaaca gtcgcaccat catcaaccgc ttctgttacg 420
gccagtgcaa ctctttctac atccccaggc acatccggaa ggaggaaggt tcctttcagt 480
cctgctcctt ctgcaagccc aagaaattca ctaccatgat ggtcacactc aactgccctg 540
aactacagcc acctaccaag aagaagagag tcacacgtgt gaagcagtgt cgttgcatat 600
ccatcgattt ggattaagcc aaatccaggt gcacccagca tgtcctagga atgcagcccc 660
aggaagtccc agacctaaaa caaccagatt cttacttggc ttaaacctag aggccagaag 720
aacccccagc tgcctcctgg caggagcctg cttgtgcgta gttcgtgtgc atgagtgtgg 780
atgggtgcct gtgggtgttt ttagacacca gagaaaacac agtctctgct agagagcact 840
ccctattttg taaacatatc tgctttaatg gggatgtacc agaaacccac ctcaccccgg 900
ctcacatcta aaggggcggg gccgtggtct ggttctgact ttgtgttttt gtgccctcct 960
ggggaccaga atctcctttc ggaatgaatg ttcatggaag aggctcctct gagggcaaga 1020
gacctgtttt agtgctgcat tcgacatgga aaagtccttt taacctgtgc ttgcatcctc 1080
ctttcctcct cctcctcaca atccatctct tcttaagttg atagtgacta tgtcagtcta 1140
atctcttgtt tgccaaggtt cctaaattaa ttcacttaac catgatgcaa atgtttttca 1200
ttttgtgaag accctccaga ctctgggaga ggctggtgtg ggcaaggaca agcaggatag 1260
tggagtgaga aagggagggt ggagggtgag gccaaatcag gtccagcaaa agtcagtagg 1320
gacattgcag aagcttgaaa ggccaatacc agaacacagg ctgatgcttc tgagaaagtc 1380
ttttcctagt atttaacaga acccaagtga acagaggaga aatgagattg ccagaaagtg 1440
attaactttg gccgttgcaa tctgctcaaa cctaacacca aactgaaaac ataaatactg 1500
accactccta tgttcggacc caagcaagtt agctaaacca aaccaactcc tctgctttgt 1560
ccctcaggtg gaaaagagag gtagtttaga actctctgca taggggtggg aattaatcaa 1620
aaacckcaga ggctgaaatt cctaatacct ttcctttatc gtggttatag tcagctcatt 1680
tccattccac tatttcccat aatgcttctg agagccacta acttgattga taaagatcct 1740
gcctctgctg agtgtacctg acagtaagtc taaagatgar agagtttagg gactactctg 1800
ttttagcaag aratattktg ggggtctttt tgttttaact attgtcagga gattgggcta 1860
ragagaagac gacgagagta aggaaataaa gggrattgcc tctggctaga gagtaagtta 1920
ggtgttaata cctggtagaa atgtaaggga tatgacctcc ctttctttat gtgctcactg 1980
aggatctgag gggaccctgt taggagagca tagcatcatg atgtattagc tgttcatctg 2040
ctactggttg gatggacata actattgtaa ctattcagta tttactggta ggcactgtcc 2100
tctgattaaa cttggcctac tggcaatggc tacttaggat tgatctaagg gccaaagtgc 2160
agggtgggtg aactttattg tactttggat ttggttaacc tgttttcttc aagcctgagg 2220
ttttatatac aaactccctg aatactcttt ttgccttgta tcttctcagc ctcctagcca 2280
agtcctatgt aatatggaaa acaaacactg cagacttgag attcagttgc cgatcaaggc 2340
tctggcattc agagaaccct tgcaactcga gaagctgttt ttatttcgtt tttgttttga 2400
tccagtgctc tcccatctaa caactaaaca ggagccattt caaggcggga gatattttaa 2460
acacccaaaa tgttgggtct gattttcaaa cttttaaact cactactgat gattctcacg 2520
ctaggcgaat ttgtccaaac acatagtgtg tgtgttttgt atacactgta tgaccccacc 2580
ccaaatcttt gtattgtcca cattctccaa caataaagca cagagtggat ttaattaagc 2640
acacaaatgc taaggcagaa ttttgagggt gggagagaag aaaagggaaa gaagctgaaa 2700
atgtaaaacc acaccaggga ggaaaaatga cattcagaac cagcaaacac tgaatttctc 2760
ttgttgtttt aactctgcca caagaatgca atttcgttaa tggagatgac ttaagttggc 2820
agcagtaatc ttcttttagg agcttgtacc acagtcttgc acataagtgc agatttggct 2880
caagtaaaga gaatttcctc aacactaact tcactgggat aatcagcagc gtaactaccc 2940
taaaagcata tcactagcca aagagggaaa tatctgttct tcttactgtg cctatattaa 3000
gactagtaca aatgtggtgt gtcttccaac tttcattgaa aatgccatat ctataccata 3060
ttttattcga gtcactgatg atgtaatgat atattttttc attattatag tagaatattt 3120
ttatggcaag atatttgtgg tcttgatcat acctattaaa ataatgccaa acaccaaata 3180
tgaattttat gatgtacact ttgtgcttgg cattaaaaga aaaaaacaca catcctggaa 3240
gtctgtaagt tgttttttgt tactgtaggt cttcaaagtt aagagtgtaa gtgaaaaatc 3300
tggaggagag gataatttcc actgtgtgga atgtgaatag ttaaatgaaa agttatggtt 3360
atttaatgta attattactt caaatccttt ggtcactgtg atttcaagca tgttttcttt 3420
ttctccttta tatgactttc tctgagttgg gcaaagaaga agctgacaca ccgtatgttg 3480
ttagagtctt ttatctggtc aggggaaaca aaatcttgac ccagctgaac atgtcttcct 3540
gagtcagtgc ctgaatcttt attttttaaa ttgaatgttc cttaaaggtt aacatttcta 3600
aagcaatatt aagaaagact ttaaatgtta ttttggaaga cttacgatgc atgtatacaa 3660
acgaatagca gataatgatg actagttcac acataaagtc cttttaagga gaaaatctaa 3720
aatgaaaagt ggataaacag aacatttata agtgatcagt taatgcctaa gagtgaaagt 3780
agttctattg acattcctca agatatttaa tatcaactgc attatgtatt atgtctgctt 3840
aaatcattta aaaacggcaa agaattatat agactatgag gtaccttgct gtgtaggagg 3900
atgaaagggg agttgatagt ctcataaaac taatttggct tcaagtttca tgaatctgta 3960
actagaattt aattttcacc ccaataatgt tctatatagc ctttgctaaa gagcaactaa 4020
taaattaaac ctattctttc aaaaaaaaa 4049
<210>46
<211>184
<212>PRT
<213>人
<400>SEQ ID NO:46
Met Ser Arg Thr Ala Tyr Thr Val Gly Ala Leu Leu Leu Leu Leu Gly
1 5 10 15
Thr Leu Leu Pro Ala Ala Glu Gly Lys Lys Lys Gly Ser Gln Gly Ala
20 25 30
Ile Pro Pro Pro Asp Lys Ala Gln His Asn Asp Ser Glu Gln Thr Gln
35 40 45
Ser Pro Gln Gln Pro Gly Ser Arg Asn Arg Gly Arg Gly Gln Gly Arg
50 55 60
Gly Thr Ala Met Pro Gly Glu Glu Val Leu Glu Ser Ser Gln Glu Ala
65 70 75 80
Leu His Val Thr Glu Arg Lys Tyr Leu Lys Arg Asp Trp Cys Lys Thr
85 90 95
Gln Pro Leu Lys Gln Thr Ile His Glu Glu Gly Cys Asn Ser Arg Thr
100 105 110
Ile Ile Asn Arg Phe Cys Tyr Gly Gln Cys Asn Ser Phe Tyr Ile Pro
115 120 125
Arg His Ile Arg Lys Glu Glu Gly Ser Phe Gln Ser Cys Ser Phe Cys
130 135 140
Lys Pro Lys Lys Phe Thr Thr Met Met Val Thr Leu Asn Cys Pro Glu
145 150 155 160
Leu Gln Pro Pro Thr Lys Lys Lys Arg Val Thr Arg Val Lys Gln Cys
165 170 175
Arg Cys Ile Ser Ile Asp Leu Asp
180
<210>47
<211>1386
<212>DNA
<213>人
<400>SEQ ID NO:47
gctcctcgcc ccgcgcctgc ccccaggatg gtccgcgcga ggcaccagcc gggtgggctt 60
tgcctcctgc tgctgctgct ctgccagttc atggaggacc gcagtgccca ggctgggaac 120
tgctggctcc gtcaagcgaa gaacggccgc tgccaggtcc tgtacaagac cgaactgagc 180
aaggaggagt gctgcagcac cggccggctg agcacctcgt ggaccgagga ggacgtgaat 240
gacaacacac tcttcaagtg gatgattttc aacgggggcg cccccaactg catcccctgt 300
aaagaaacgt gtgagaacgt ggactgtgga cctgggaaaa aatgccgaat gaacaagaag 360
aacaaacccc gctgcgtctg cgccccggat tgttccaaca tcacctggaa gggtccagtc 420
tgcgggctgg atgggaaaac ctaccgcaat gaatgtgcac tcctaaaggc aagatgtaaa 480
gagcagccag aactggaagt ccagtaccaa ggcagatgta aaaagacttg tcgggatgtt 540
ttctgtccag gcagctccac atgtgtggtg gaccagacca ataatgccta ctgtgtgacc 600
tgtaatcgga tttgcccaga gcctgcttcc tctgagcaat atctctgtgg gaatgatgga 660
gtcacctact ccagtgcctg ccacctgaga aaggctacct gcctgctggg cagatctatt 720
ggattagcct atgagggaaa gtgtatcaaa gcaaagtcct gtgaagatat ccagtgcact 780
ggtgggaaaa aatgtttatg ggatttcaag gttgggagag gccggtgttc cctctgtgat 840
gagctgtgcc ctgacagtaa gtcggatgag cctgtctgtg ccagtgacaa tgccacttat 900
gccagcgagt gtgccatgaa ggaagctgcc tgctcctcag gtgtgctact ggaagtaaag 960
cactccggat cttgcaactg aatctgcccg taaaacctga gccattgatt cttcagaact 1020
ttctgcagtt tttgacttca tagattatgc tttaaaaaat tttttttaac ttattgcata 1080
acagcagatg ccaaaaacaa aaaaagcatc tcactgcaag tcacataaaa atgcaacgct 1140
gtaatatggc tgtatcagag ggctttgaaa acatacactg agctgcttct gcgctgttgt 1200
tgtccgtatt taaacaacag ctcccctgta ttcccccatc tagccatttc ggaagacacc 1260
gaggaagagg aggaagatga agaccaggac tacagctttc ctatatcttc tattctagag 1320
tggtaaactc tctataagtg ttcagtgttc acatagcctt tgtgcaaaaa aaaaaaaaaa 1380
aaaaaa 1386
<210>48
<211>317
<212>PRT
<213>人
<400>SEQ ID NO:48
Met Val Arg Ala Arg His Gln Pro Gly Gly Leu Cys Leu Leu Leu Leu
l 5 10 15
Leu Leu Cys Gln Phe Met Glu Asp Arg Ser Ala Gln Ala Gly Asn Cys
20 25 30
Trp Leu Arg Gln Ala Lys Asn Gly Arg Cys Gln Val Leu Tyr Lys Thr
35 40 45
Glu Leu Ser Lys Glu Glu Cys Cys Ser Thr Gly Arg Leu Ser Thr Ser
50 55 60
Trp Thr Glu Glu Asp Val Asn Asp Asn Thr Leu Phe Lys Trp Met Ile
65 70 75 80
Phe Asn Gly Gly Ala Pro Asn Cys Ile Pro Cys Lys Glu Thr Cys Glu
85 90 95
Asn Val Asp Cys Gly Pro Gly Lys Lys Cys Arg Met Asn Lys Lys Asn
100 105 110
Lys Pro Arg Cys Val Cys Ala Pro Asp Cys Ser Asn Ile Thr Trp Lys
115 120 125
Gly Pro Val Cys Gly Leu Asp Gly Lys Thr Tyr Arg Asn Glu Cys Ala
110 135 140
Leu Leu Lys Ala Arg Cys Lys Glu Gln Pro Glu Leu Glu Val Gln Tyr
145 150 155 160
Gln Gly Arg Cys Lys Lys Thr Cys Arg Asp Val Phe Cys Pro Gly Ser
165 170 175
Ser Thr Cys Val Val Asp Gln Thr Asn Asn Ala Tyr Cys Val Thr Cys
180 185 190
Asn Arg Ile Cys Pro Glu Pro Ala Ser Ser Glu Gln Tyr Leu Cys Gly
195 200 205
Asn Asp Gly Val Thr Tyr Ser Ser Ala Cys His Leu Arg Lys Ala Thr
210 215 220
Cys Leu Leu Gly Arg Ser Ile Gly Leu Ala Tyr Glu Gly Lys Cys Ile
225 230 235 240
Lys Ala Lys Ser Cys Glu Asp Ile Gln Cys Thr Gly Gly Lys Lys Cys
245 250 255
Leu Trp Asp Phe Lys Val Gly Arg Gly Arg Cys Ser Leu Cys Asp Glu
260 265 270
Leu Cys Pro Asp Ser Lys Ser Asp Glu Pro Val Cys Ala Ser Asp Asn
275 280 285
Ala Thr Tyr Ala Ser Glu Cys Ala Met Lys Glu Ala Ala Cys Ser Ser
290 295 300
Gly Val Leu Leu Glu Val Lys Hi s Ser Gly Ser Cys Asn
305 310 315
<210>49
<211>699
<212>DNA
<213>人
<400>SEQ ID NO:49
atggagcgct gccccagcct aggggtcacc ctctacgccc tggtggtggt cctggggctg 60
cgggcgacac cggccggcgg ccagcactat ctccacatcc gcccggcacc cagcgacaac 120
ctgcccctgg tggacctcat cgaacaccca gaccctatct ttgaccccaa ggaaaaggat 180
ctgaacgaga cgctgctgcg ctcgctgctc gggggccact acgacccagg cttcatggcc 240
acctcgcccc ccgaggaccg gcccggcggg ggcgggggtg cagctggggg cgcggaggac 300
ctggcggagc tggaccagct gctgcggcag cggccgtcgg gggccatgcc gagcgagatc 360
aaagggctag agttctccga gggcttggcc cagggcaaga agcagcgcct aagcaagaag 420
ctgcggagga agttacagat gtggctgtgg tcgcagacat tctgccccgt gctgtacgcg 480
tggaacgacc tgggcagccg cttttggccg cgctacgtga aggtgggcag ctgcttcagt 540
aagcgctcgt gctccgtgcc cgagggcatg gtgtgcaagc cgtccaagtc cgtgcacctc 600
acggtgctgc ggtggcgctg tcagcggcgc gggggccagc gctgcggctg gattcccatc 660
cagtacccca tcatttccga gtgcaagtgc tcgtgctag 699
<210>50
<211>232
<212>PRT
<213>人
<400>SEQ ID NO:50
Met Glu Arg Cys Pro Ser Leu Gly Val Thr Leu Tyr Ala Leu Val Val
1 5 10 15
Val Leu Gly Leu Arg Ala Thr Pro Ala Gly Gly Gln His Tyr Leu His
20 25 30
Ile Arg Pro Ala Pro Ser Asp Asn Leu Pro Leu Val Asp Leu Ile Glu
35 40 45
His Pro Asp Pro Ile Phe Asp Pro Lys Glu Lys Asp Leu Asn Glu Thr
50 55 60
Leu Leu Arg Ser Leu Leu Gly Gly His Tyr Asp Pro Gly Phe Met Ala
65 70 75 80
Thr Ser Pro Pro Glu Asp Arg Pro Gly Gly Gly Gly Gly Ala Ala Gly
85 90 95
Gly Ala Glu Asp Leu Ala Glu Leu Asp Gln Leu Leu Arg Gln Arg Pro
100 105 110
Ser Gly Ala Met Pro Ser Glu Ile Lys Gly Leu Glu Phe Ser Glu Gly
115 120 125
Leu Ala Gln Gly Lys Lys Gln Arg Leu Ser Lys Lys Leu Arg Arg Lys
130 135 140
Leu Gln Met Trp Leu Trp Ser Gln Thr Phe Cys Pro Val Leu Tyr Ala
145 150 155 160
Trp Asn Asp Leu Gly Ser Arg Phe Trp Pro Arg Tyr Val Lys Val Gly
165 170 175
Ser Cys Phe Ser Lys Arg Ser Cys Ser Val Pro Glu Gly Met Val Cys
180 185 190
Lys Pro Ser Lys Ser Val His Leu Thr Val Leu Arg Trp Arg Cys Gln
195 200 205
Arg Arg Gly Gly Gln Arg Cys Gly Trp Ile Pro Ile Gln Tyr Pro Ile
210 215 220
Ile Ser Glu Cys Lys Cys Ser Cys
225 230
<210>51
<211>804
<212>DNA
<213>人
<400>SEQ ID NO:51
atgcatctcc tcttatttca gctgctggta ctcctgcctc taggaaagac cacacggcac 60
caggatggcc gccagaatca gagttctctt tcccccgtac tcctgccaag gaatcaaaga 120
gagcttccca caggcaacca tgaggaagct gaggagaagc cagatctgtt tgtcgcagtg 180
ccacaccttg tagccaccag ccctgcaggg gaaggccaga ggcagagaga gaagatgctg 240
tccagatttg gcaggttctg gaagaagcct gagagagaaa tgcatccatc cagggactca 300
gatagtgagc ccttcccacc tgggacccag tccctcatcc agccgataga tggaatgaaa 360
atggagaaat ctcctcttcg ggaagaagcc aagaaattct ggcaccactt catgttcaga 420
aaaactccgg cttctcaggg ggtcatcttg cccatcaaaa gccatgaagt acattgggag 480
acctgcagga cagtgccctt cagccagact ataacccacg aaggctgtga aaaagtagtt 540
gttcagaaca acctttgctt tgggaaatgc gggtctgttc attttcctgg agccgcgcag 600
cactcccata cctcctgctc tcactgtttg cctgccaagt tcaccacgat gcacttgcca 660
ctgaactgca ctgaactttc ctccgtgatc aaggtggtga tgctggtgga ggagtgccag 720
tgcaaggtga agacggagca tgaagatgga cacatcctac atgctggctc ccaggattcc 780
tttatcccag gagtttcagc ttga 804
<210>52
<211>267
<212>PRT
<213>人
<400>SEQ ID NO:52
Met His Leu Leu Leu Phe Gln Leu Leu Val Leu Leu Pro Leu Gly Lys
1 5 10 15
Thr Thr Arg His Gln Asp Gly Arg Gln Asn Gln Ser Ser Leu Ser Pro
20 25 30
Val Leu Leu Pro Arg Asn Gln Arg Glu Leu Pro Thr Gly Asn His Glu
35 40 45
Glu Ala Glu Glu Lys Pro Asp Leu Phe Val Ala Val Pro His Leu Val
50 55 60
Ala Thr Ser Pro Ala Gly Glu Gly Gln Arg Gln Arg Glu Lys Met Leu
65 70 75 80
Ser Arg Phe Gly Arg Phe Trp Lys Lys Pro Glu Arg Glu Met His Pro
85 90 95
Ser Arg Asp Ser Asp Ser Glu Pro Phe Pro Pro Gly Thr Gln Ser Leu
100 105 110
Ile Gln Pro Ile Asp Gly Met Lys Met Glu Lys Ser Pro Leu Arg Glu
115 120 125
Glu Ala Lys Lys Phe Trp His His Phe Met Phe Arg Lys Thr Pro Ala
130 135 140
Ser Gln Gly Val Ile Leu Pro Ile Lys Ser His Glu Val His Trp Glu
145 150 155 160
Thr Cys Arg Thr Val Pro Phe Ser Gln Thr Ile Thr His Glu Gly Cys
165 170 175
Glu Lys Val Val Val Gln Asn Asn Leu Cys Phe Gly Lys Cys Gly Ser
180 185 190
Val His Phe Pro Gly Ala Ala Gln His Ser His Thr Ser Cys Ser His
195 200 205
Cys Leu Pro Ala Lys Phe Thr Thr Met His Leu Pro Leu Asn Cys Thr
210 215 220
Glu Leu Ser Ser Val Ile Lys Val Val Met Leu Val Glu Glu Cys Gln
225 230 235 240
Cys Lys Val Lys Thr Glu His Glu Asp Gly His Ile Leu His Ala Gly
245 250 255
Ser Gln Asp Ser Phe Ile Pro Gly Val Ser Ala
260 265
<210>53
<211>1523
<212>DNA
<213>人
<400>SEQ ID NO:53
ctggcgcggg cgggagctgc ggcggatacc cttgcgtgct gtggagaccc tactctcttc 60
gctgagaacg gccgctagcg gggactgaag gccgggagcc cactcccgac ccggggctag 120
cgtgcgtccc tagagtcgag cggggcaagg gagccagtgg ccgccgacgg gggaccggga 180
aacttttctg ggctcctggg cgcgccctgt agccgcgctc catgctccgg cagcggcccg 240
aaacccagcc ccgccgctga cggcgcccgc cgctccgggc agggcccatg ccctgcgcgc 300
tccgggggtc gtaggctgcc gccgagccgg ggctccggaa gccggcgggg gcgccgcggc 360
cgtgcggggc gtcaatggat cgccactcca gctacatctt catctggctg cagctggagc 420
tctgcgccat ggccgtgctg ctcaccaaag gtgaaattcg atgctactgt gatgctgccc 480
actgtgtagc cactggttat atgtgtaaat ctgagctcag cgcctgcttc tctagacttc 540
ttgatcctca gaactcaaat tccccactca cccatggctg cctggactct cttgcaagca 600
cgacagacat ctgccaagcc aaacaggccc gaaaccactc tggcaccacc atacccacat 660
tggaatgctg tcatgaagac atgtgcaatt acagagggct gcacgatgtt ctctctcctc 720
ccaggggtga ggcctcagga caaggaaaca ggtatcagca tgatggtagc agaaacctta 780
tcaccaaggt gcaggagctg acttcttcca aagagttgtg gttccgggca gcggtcattg 840
ccgtgcccat tgctggaggg ctgattttag tgttgcttat tatgttggcc ctgaggatgc 900
ttcgaagtga aaataagagg ctgcaggatc agcggcaaca gatgctctcc cgtttgcact 960
acagctttca cggacaccat tccaaaaagg ggcaggttgc aaagttagac ttggaatgca 1020
tggtgccggt cagtgggcac gagaactgct gtctgacctg tgataaaatg agacaagcag 1080
acctcagcaa cgataagatc ctctcgcttg ttcactgggg catgtacagt gggcacggga 1140
agctggaatt cgtatgacgg agtcttatct gaactacact tactgaacag cttgaaggcc 1200
ttttgagttc tgctggacag gagcacttta tctgaagaca aactcattta atcatctttg 1260
agagacaaaa tgacctctgc aaacagaatc ttggatattt cttctgaagg attatttgca 1320
cagacttaaa tacagttaaa tgtgttattt gcttttaaaa ttataaaaag caaagagaag 1380
actttgtaca cactgtcacc agggttattt gcatccaagg gagctggaat tgagtaccta 1440
aataaacaaa aatgtgccct atgtaagctt ctacatcttg atttattgta aagatttaaa 1500
agaaatatat atattttgtc tga 1523
<210>54
<211>260
<212>PRT
<213>人
<400>SEQ ID NO:54
Met Asp Arg His Ser Ser Tyr Ile Phe Ile Trp Leu Gln Leu Glu Leu
1 5 10 15
Cys Ala Met Ala Val Leu Leu Thr Lys Gly Glu Ile Arg Cys Tyr Cys
20 25 30
Asp Ala Ala His Cys Val Ala Thr Gly Tyr Met Cys Lys Ser Glu Leu
35 40 45
Ser Ala Cys Phe Ser Arg Leu Leu Asp Pro Gln Asn Ser Asn Ser Pro
50 55 60
Leu Thr His Gly Cys Leu Asp Ser Leu Ala Ser Thr Thr Asp Ile Cys
65 70 75 80
Gln Ala Lys Gln Ala Arg Asn His Ser Gly Thr Thr Ile Pro Thr Leu
85 90 95
Glu Cys Cys His Glu Asp Met Cys Asn Tyr Arg Gly Leu His Asp Val
100 105 110
Leu Ser Pro Pro Arg Gly Glu Ala Ser Gly Gln Gly Asn Arg Tyr Gln
115 120 125
His Asp Gly Ser Arg Asn Leu Ile Thr Lys Val Gln Glu Leu Thr Ser
130 135 140
Ser Lys Glu Leu Trp Phe Arg Ala Ala Val Ile Ala Val Pro Ile Ala
145 150 155 160
Gly Gly Leu Ile Leu Val Leu Leu Ile Met Leu Ala Leu Arg Met Leu
165 170 175
Arg Ser Glu Asn Lys Arg Leu Gln Asp Gln Arg Gln Gln Met Leu Ser
180 185 190
Arg Leu His Tyr Ser Phe His Gly His His Ser Lys Lys Gly Gln Val
195 200 205
Ala Lys Leu Asp Leu Glu Cys Met Val Pro Val Ser Gly His Glu Asn
210 215 220
Cys Cys Leu Thr Cys Asp Lys Met Arg Gln Ala Asp Leu Ser Asn Asp
225 230 235 240
Lys Ile Leu Ser Leu Val His Trp Gly Met Tyr Ser Gly His Gly Lys
245 250 255
Leu Glu Phe Val
260
Claims (1)
1.鉴定用于治疗青光眼的治疗剂的方法,所述方法包括:
a)获得第一组合物,其包含表达BMP-2A、BMP4、BMP-5或BMP7的一群重组细胞;
b)获得候选物质;
c)孵育所述组合物和所述候选物质;
d)检测所述组合物启动BMP-诱导的Smad信号途径或受BMP-调节的基因表达的能力;以及
e)鉴定抑制或刺激BMP-诱导的Smad信号途径或受BMP-调节的基因表达的候选物质。
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
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US33485201P | 2001-10-31 | 2001-10-31 | |
US60/334,852 | 2001-10-31 |
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CNA028209176A Division CN1685055A (zh) | 2001-10-31 | 2002-10-31 | 骨形态发生蛋白(bmp)、bmp受体和bmp结合蛋白及它们在诊断和治疗青光眼中的用途 |
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Publication Number | Publication Date |
---|---|
CN1966725A true CN1966725A (zh) | 2007-05-23 |
Family
ID=23309139
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CNA028209176A Pending CN1685055A (zh) | 2001-10-31 | 2002-10-31 | 骨形态发生蛋白(bmp)、bmp受体和bmp结合蛋白及它们在诊断和治疗青光眼中的用途 |
CNA2006101467420A Pending CN1966725A (zh) | 2001-10-31 | 2002-10-31 | 骨形态生发蛋白(bmp)、bmp受体和bmp结合蛋白及它们在诊断和治疗青光眼中的用途 |
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Application Number | Title | Priority Date | Filing Date |
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CNA028209176A Pending CN1685055A (zh) | 2001-10-31 | 2002-10-31 | 骨形态发生蛋白(bmp)、bmp受体和bmp结合蛋白及它们在诊断和治疗青光眼中的用途 |
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US (5) | US20050118585A1 (zh) |
EP (2) | EP1440159B1 (zh) |
JP (2) | JP4255382B2 (zh) |
KR (4) | KR101072867B1 (zh) |
CN (2) | CN1685055A (zh) |
AT (1) | ATE349554T1 (zh) |
BR (1) | BR0213738A (zh) |
CA (1) | CA2463143A1 (zh) |
DE (1) | DE60217152T2 (zh) |
ES (1) | ES2278079T3 (zh) |
HK (1) | HK1069851A1 (zh) |
MX (1) | MXPA04003697A (zh) |
PL (2) | PL373863A1 (zh) |
RU (3) | RU2336902C2 (zh) |
WO (1) | WO2003055443A2 (zh) |
ZA (1) | ZA200402662B (zh) |
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CN113811605A (zh) * | 2019-05-15 | 2021-12-17 | 诺和诺德股份有限公司 | 从人多能干细胞获得眼野祖细胞的方法 |
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