稀有鮈鲫卵黄蛋白原基因全序列的克隆与分析  

Cloning and sequence analysis of full-length vitellogenin gene of Gobiocypris rarus

作　　者：黄婷妹[1] 蔡生力[1] 刘红[1] 

机构地区：[1]上海海洋大学农业部水产种质资源与养殖生态重点开放实验室,上海201306

出　　处：《上海海洋大学学报》2013年第5期678-683,共6页Journal of Shanghai Ocean University

基　　金：上海市教育委员会重点学科建设项目(J50701);上海市教育委员会创新项目(12YZ125)

摘　　要：稀有绚鲫（Gobiocyprisrarus）是我国特有的一种小型鲤科鱼类，目前已作为一种新型模式生物应用于实验研究。运用普通PCR方法克隆得到一段长6299bp的序列，序列分析发现编码区与稀有绚鲫卵黄蛋白原cDNA序列相似性为100％。再参考6299bp序列5’端设计两对特异性引物，进行基因组步移，获得长度为1285bp的序列。经过序列拼接后得到7573bp的序列，分析发现其中含卵黄蛋白原基因全序列长度为6595bp卵黄蛋白原基因上游调控序列长度为978bp。进一步分析开放性阅读框得知：稀有绚鲫卵黄蛋白原基因含有26个外显子和25个内含子，编码1299个氨基酸序列，CDS长3900bp。经BLAST比对发现，稀有绚鲫卵黄蛋白原上游调控序列中220～838bp区间与朝鲜鲡（Acheilognathuayamatsutae）卵黄蛋白原基因启动子序列2857～3469bp区间有80％相似性。Gobiocypris rarus is a unique species of small fish in China. At present study, a fragment of 6 299 bp was cloned by PCR method. It was found that the sequence identity of coding area of this fragment is 100% compared to the cDNA sequence of vitellogenin of Gobiocypris rarus. By using genome walking technique, a product of 1 285 bp was acquired based on the 6 299 bp sequence. Finally, a sequence of 7 573 bp was gotten by comparing the two fragments of 1 285 bp and 6 299 bp, with a vitellogenin gene of 6 595 bp and upstream sequence of 978 bp. After the ORF analysis , it was found there were 26 exons and 25 introns, the CDS was 3 900 bp which code 1 200 amino acid. Blast analysis found there existed 80% sequence identity between the upstream fragment of 220 - 838 bp of our sequence and the promoter sequence of 2 875 - 3 469 bp of vitellogenin of Acheilognathas yamatsutae.

关 键 词：卵黄蛋白原 稀有鮈鲫 基因组步移 序列分析 

分 类 号：S917[农业科学—水产科学]