Geminin基因（GMNN）在抑制再复制的进程中是主要的调控者，在细胞周期中的S，G2和早中期时，它负向调节Cdt1p，阻止Mcm2–7p复合物在复制起点的装配，因此GMNN通过阻止DNA非正常地再复制来维持基因组的忠实性，确保同一个复制起点在一个细胞周期中不会活化两次。在后生动物后期生长中，GMNN在控制细胞分化的同时也控制着细胞的增殖。 GMNN分别抑制青鳉鱼视网膜中Six3和小鼠胚胎中Hox的转录因子，同时在非洲爪蟾中通过抑制Brg-1的活性来维持神经祖细胞处于未分化状态。我们从文昌鱼B.japanicum中克隆得到GMNN的cDNA克隆。运用5’RACE和3’RACE克隆得到GMN...Geminin gene (GMNN) is a central regulator of the process that inhibits re-replication. During S, G2, and early M phases of the cell cycle, it negatively regulates the Cdt1p and prevents them from loading the Mcm2–7p complex to the origins of replication. GMNN thus maintains the integrity of the genome by preventing abnormal re-replication of DNA. It ensures that, the same origin of replication is...学位：理学硕士院系专业：生命科学学院生物化学与生物技术系_化学生物学学号：2172007115411

Emmanuel MUKWAYA

Xiamen University Institutional Repository

学校编码：10384 学号：21720071154115         硕  士  学  位  论  文         文昌鱼GMNN基因的克隆、序列分析、表征及表达         Cloning, Sequencing, Characterization andExpression of GMNN Gene from Amphioxus         Emmanuel MUKWAYA         指导教师：王义权            专业名称：化学生物学              答辩日期：2009年6月   厦门大学博硕士论文摘要库厦门大学学位论文原创性声明                  本人呈交的学位论文是本人在导师指导下,独立完成的研究成果。本人在论文写作中参考其他个人或集体已经发表的研究成果，均在文中以适当方式明确标明，并符合法律规范和《厦门大学研究生学术活动规范(试行)》。                  另外，该学位论文为(                  )课题(组)的研究成果，获得(                    )课题(组)经费或实验室的资助，在(          )实验室完成。(请在以上括号内填写课题或课题组负责人或实验室名称，未有此项声明内容的，可以不作特别声明。)                           声明人(签名)：                            年   月   日厦门大学博硕士论文摘要库厦门大学学位论文著作权使用声明                  本人同意厦门大学根据《中华人民共和国学位条例暂行实施办法》等规定保留和使用此学位论文，并向主管部门或其指定机构送交学位论文(包括纸质版和电子版)，允许学位论文进入厦门大学图书馆及其数据库被查阅、借阅。本人同意厦门大学将学位论文加入全国博士、硕士学位论文共建单位数据库进行检索，将学位论文的标题和摘要汇编出版，采用影印、缩印或者其它方式合理复制学位论文。                  本学位论文属于：                  (     )1.经厦门大学保密委员会审查核定的保密学位论文，于年 月 日解密，解密后适用上述授权。                  (     )2.不保密，适用上述授权。                  (请在以上相应括号内打“√”或填上相应内容。保密学位论文应是已经厦门大学保密委员会审定过的学位论文，未经厦门大学保密委员会审定的学位论文均为公开学位论文。此声明栏不填写的，默认为公开学位论文，均适用上述授权。)                           声明人(签名)：                            年   月   日厦门大学博硕士论文摘要库摘  要                   Geminin基因（GMNN）在抑制再复制的进程中是主要的调控者，在细胞周期中的S，G2和早中期时，它负向调节Cdt1p，阻止Mcm2–7p复合物在复制起点的装配，因此GMNN通过阻止DNA非正常地再复制来维持基因组的忠实性，确保同一个复制起点在一个细胞周期中不会活化两次。在后生动物后期生长中，GMNN在控制细胞分化的同时也控制着细胞的增殖。 GMNN分别抑制青鳉鱼视网膜中Six3和小鼠胚胎中Hox的转录因子，同时在非洲爪蟾中通过抑制Brg-1的活性来维持神经祖细胞处于未分化状态。我们从文昌鱼B.japanicum中克隆得到GMNN的cDNA克隆。运用5’RACE和3’RACE克隆得到GMNN的全长cDNA 为721 bp，经进一步测序验证，然后运用Real time- PCR研究其在胚胎和组织中的表达情况。    文昌鱼GMNN序列编码一个241个氨基酸的蛋白质，它与人GMNN 基因的同源性接近48.02%，与小鼠GMNN基因的同源性达到53.73%，与大鼠GMNN基因的同源性达到62.22%，与原鸡GMNN基因的同源性达到52.4%，与倭黑猩猩GMNN基因的同源性达到62.50 %。通过对文昌鱼的GMNN基因的研究我们发现它是由8个外显子和7个内含子组成，此外，运用实时定量PCR研究GMNN基因的表达情况显示它在卵巢中高表达，而在精巢没有得到高表达。文昌鱼GMNN基因的克隆和表达结果显示文昌鱼该基因是一个编码相对短的蛋白的小基因，而与其他脊椎动物不同的是该基因并不在增生组织表达。         关键词：文昌鱼；GMNN基因；Cdt1p；Mcm2–7p复合物厦门大学博硕士论文摘要库Abstract                   Geminin gene (GMNN) is a central regulator of the process that inhibits re-replication. During S, G2, and early M phases of the cell cycle, it negativelyregulates the Cdt1p and prevents them from loading the Mcm2–7p complex to theorigins of replication. GMNN thus maintains the integrity of the genome bypreventing abnormal re-replication of DNA. It ensures that, the same origin ofreplication is not activated twice within the same cell cycle. GMNN gene controlsdifferentiation as well as cell proliferation during late metazoan development. It inhibits both Six3 and Hox transcription factors in Medaka retina and the mouseembryo, respectively, and maintains an undifferentiated state in neural progenitorcells of Xenopus laevis by antagonizing Brg-1 activity. We have cloned a cDNA clone encoding GMNN from the amphioxus, B.japanicum. The721 bp full-length GMNN gene cDNA was cloned using 5’RACEand 3’ RACE and further confirmed by nucleotide sequencing and then expressedusing Real time RT-PCR. The amphioxus GMNN gene sequence encodes a protein of 241 amino acidswhich displays approximately 48.02% homology with human GMNN gene,53.73% with the mouse GMNN gene, 62.22% with Rattus norvegicusGMNNgene, 52.4% with the Gallus gallus GMNN gene and 62.50 % with Pantroglodytes. By studying GMNN gene sequence organization we found that it is made of 8exons and 7 introns. Furthermore, the expression pattern of GMNN gene withReal Time RT-PCR reveled that GMNN gene is highly expressed in the ovary butnot in the testis. Our results on cloning and expression the GMNN gene inamphioxus revealed that the amphioxus GMNN gene is a short gene encoding arelatively short protein but unlike in other vertebrates, is not expressed   inproliferative tissues.厦门大学博硕士论文摘要库         Keywords: Amphioxus; GMNN gene; Cdt1p; Mcm2–7p complex厦门大学博硕士论文摘要库参考资料         [1] Holland, N. D. & Chen, J. Origin and early evolution of vertebrates: new insights from advances in molecularbiology, anatomy, and paleontology. 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Cloning, Sequencing, Characterization and Expression of GMNN Gene from Amphioxus

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Cloning, Sequencing, Characterization and Expression of GMNN Gene from Amphioxus

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