草鱼Smad4基因的克隆、生物信息学分析及反义真核载体的构建  

Cloning of Smad4,Bioinformatics Analysis and Construction of Antisense Smad4 Gene Expression Vector from Ctenopharyngodon idellus

作　　者：吕池波[1,2] 郁二蒙[1] 谢骏[1] 王广军[1] 余德光[1] 李志斐[1] 王海英[1] 龚望宝[1] 

机构地区：[1]中国水产科学研究院珠江水产研究所,农业部热带亚热带水产资源利用与养殖重点实验室,广东广州510380 [2]上海海洋大学水产与生命学院,上海201306

出　　处：《华南农业大学学报》2013年第4期517-523,共7页Journal of South China Agricultural University

基　　金：现代农业产业体系建设专项(nycytx-49);公益性行业(农业)科研专项(201203083);广东省自然科学基金(10151038001000004);国家科技支撑计划项目(2012BAD25B04)

摘　　要：为了研究TGF-β信号传导中的关键蛋白Smad4在草鱼Ctenopharyngodon idellus发育过程中的作用,克隆出草鱼Smad4基因cDNA全序列,其碱基长度为2 974 bp,其中开放阅读框1 644 bp,编码547个氨基酸.生物信息学分析结果显示,草鱼Smad4蛋白相对分子质量为59 690.3,分子式为C2 632H4 073N753O791S24,理论等电点为6.5,含有MH1和MH2这2个高度保守的功能结构域.BLAST相似性分析显示,草鱼Smad4蛋白的氨基酸序列与鲤鱼Cyprinus carpio、斑马鱼Danio rerio的相似性较高,分别为94.23%和92.97%.还将草鱼Smad4基因的开放阅读框片段反向插入表达载体pcDNA3.1(+),成功构建了反义Smad4基因真核表达载体pcDNA3.1(+)-Anti-Smad4.Smad4 is a key protein in TGF-βsignal transduction , which can regulate the developmental processes like cell proliferation and differentiation .In order to study the role of Smad 4 in developmental process of grass carp （Ctenopharyngodon idellus）, a full length of 2 974 bp of Smad4 cDNA sequence from grass carp was obtained , a 1 644 bp CDS （ coding sequence ） encoding a polypeptide of 547 amino acids was determined in the Smad 4 of grass carp .Bioinformatics analysis showed that the relative molecu-lar mass of grass carp Smad4 protein was 59 690.3, the molecular formula was C 2 632 H4 073 N753 O791 S24 , isoelectric point was 6.5, and it contained both MH1 and MH2 highly conserved functional domains . BLAST homology analysis showed higher homology of grass carp Smad 4 protein with common carp and ze-brafish , which was 94.23%and 92.97%respectively .The open reading frame of grass carp Smad4 gene was reversely inserted into the expression vector pcDNA 3.1（＋） , which constructed the antisense Smad4 gene eukaryotic expression vector pcDNA 3.1 （＋）-Anti-Smad4 .

关 键 词：草鱼 SMAD4 反义基因 真核表达载体 胚胎发育 生物信息学分析 

分 类 号：Q78[生物学—分子生物学] S917.4[农业科学—水产科学]