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 .
出处
《华南农业大学学报》
CAS
CSCD
北大核心
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)
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
Ctenopharyngodon idella
Smad4
antisense
eukaryotic expression vector
embryonic development
bioinformatics analysis
