鲮Myf5基因克隆及其SNPs分析  

Cloning and SNPs analysis of Myf5 gene from Cirrhinus molitorella

作　　者：钟茂春[1,2] 郑光明[1] 赵建[1] 朱新平[1] 马丽莎[1] 潘德博[1] 陈昆慈[1] 谢文平[1] 史燕[1] 

机构地区：[1]中国水产科学研究院珠江水产研究所,广东广州510380 [2]上海海洋大学生命科学与技术学院,上海200090

出　　处：《中国水产科学》2010年第4期681-688,共8页Journal of Fishery Sciences of China

基　　金：国家自然科学基金资助项目(30271022);国家公益性行业(农业)科研专项(200903045);国家重点基础研究发展计划项目(2004CB117401);广东省科技计划(2005B20301007);广东省科技厅农业攻关子项目(2005A20105001);国家科技基础条件平台建设子项目(2006DKA30470-008)

摘　　要：提取新鲜鲮(Cirrhinus molitorella)肌肉总RNA,采用RT-PCR技术分段克隆鲮Myf5基因核心序列,获得1104bp的目的片段,其中开放阅读框为723bp,编码240个氨基酸。分析表明,鲮Myf5蛋白具有MRF家族基因的典型性碱性bHLH螺旋-环-螺旋(Basic helix-loop-helix,bHLH)结构。设计特异性引物扩增获得2个内含子,大小分别为1311bp和90bp。用单链构向多态性SSCP的方法分析了Myf5基因在珠江水系3个不同江段鲮中的遗传变异,分布及群体杂合性等群体遗传信息。发现其编码区非常保守,仅在外显子412位点处发生突变C→T。对该位点进行基因型分析,结果表明:珠江北江段鲮中该位点符合Hardy-Weinberg平衡,而东江和西江段种群不符合Hardy-Weinberg平衡,3个江段种群总体上符合Hardy-Weinberg平衡,并存在较大基因流。以Myf5基因该位点的多态性分析表明,3个种群未出现明显分化;通过该位点的遗传结构分析表明:3个江段种群杂合度都较高,中度信息含量(PIC)及多态信息指数,即该位点有较好的变异性及遗传多态性。3个江段种群总体上该位点在不符合Hardy-Weinberg的情况下AA>BB,并由该位点群体杂合性及中性分析表明该位点突变群体具体良好的多态性且不遵循中性选择模式,说明BB型个体受到自然选择。本研究结果表明,鲮Myf5基因的C412T与个体生长有相关性,可作为候选基因标记,用于鲮生长相关分子标记辅助育种研究。Several molecular markers were used in mud carp genetic diversity research. There is no choiceness varietal mud carp（ Cirrhinus molitorella） in present culture. Three part of Pearl-river drainage wild mud carp （Cirrhinus molitorella） were sampled as experimentation fish,and PCR-SSCP（single strand conformation polymorphism,SSCP）was used to screen Myf5 ORF（open reading frame） DNA sequence,expecting to find SNPs relative to growth and trait. Mud carp（ Cirrhinus molitorella） belongs to Cypriniformes,Cyprinidae,distributing in Pearl River,Ming River,Han River,Hainan. Taiwan and southeast of Asian. It is an important fishery production in Pearl-river drainage area. Myf5 gene（ myogenic factor 5） is a member of MRFs gene family,which include Myf5,MyoG,MyoD and MRF4. The primary function of the gene family is advance myoblast multiplication and differentiation. The function of MyoD and Myf5 genes is determined to initiate the differentiation process of that muscular satellite cells to be muscle derived stem cell. And that of MyoG and MRF4 genes was thought to be involved in regulating the differentiation process of muscle derived stem cell to be myotube and muscle fibers. All of those functions were implementing by the typical trait b-HLH structure of MRF family,which form dimeric by combining with E-protein. Then Key gene is activated and pressed by the dimeric recognize upstream E-box. Many species of Myf5 gene or cDNA have been cloned. Such as Mus musculus,Bos Taurus,Sus scrofa,Ovis ammon,Gallus gallus, Danio rerio,Cyprinus carpio,Micropterus salmoide and Oncorhynchus mykiss. Myf5 gene of mud carp（ Cirrhinus molitorella） has been cloned,sequenced and analyzed by PCR-SSCP. Mud carp Genome DNA and total RNA was extracted from flesh muscle of mud carp. Myf5 gene was cloned including three exons and two introns of which the length is 453 bp,76 bp,194 bp and 1 311 bp and 90 bp（GeneBank accession： GU289508）. The cDNA sequence of Myf5 gene was obtained by RT-PCR. The length of the cDNA is 1 1

关 键 词：鲮鱼 Myf5基因 SSCP 遗传结构 

分 类 号：Q959[生物学—动物学] S96[农业科学—水产养殖]