安徽农业科学2009，Vol.37Issue(22)：10431-10432,10448,3.
猪轮状病毒VP4蛋白基因在大肠杆菌中的表达
Expression of Porcine Rotavirus VP4 in Escherichia coli
摘要
Abstract
[Objective] The aim was to study the expression of porcine rotavirus VP4 in Escherichia coli BL21. [Method] With the total RNA isolated from porcine lung tissues as the template, the partial genes of VP4 protein were amplified by using reverse transcription-polymerase chain reaction (RT-PCR). PCR products were cloned into the expression vector pGEX-4T-1 to construct the prokaryotic expression plasmid PGEX-4T-1-VP4 and the recombinant plasmid was made for enzyme digestion and sequencing. The recombinant vector PGEX-4T-1-VP4 was transformed into E. coli BL21 (DE3) for induction expression and SDS-PAGE analysis. The solubility analysis of recombinant E. coli was carried out. [Result] The partial genes of VP4 protein about 875 bp were obtained through RT-PCR amplification on the total RNA isolated from porcine lung tissues. After the recombinant plasmid PGEX-4T-1-VP4 was made for double enzyme digestion by BamHⅠ and XhoⅠ and sequenced, the cDNA coding sequence inserted in the porcine rotavirus VP4 had 99.5% similarity to the published nucleic acid sequence of porcine in GenBank. The recombinant plasmid pGEX-4T-1-VP4 was transformed into E. coli BL21 (DE3) and induced to express by IPTG. The result of SDS-PAGE showed that the soluble fusion protein with molecular weight of 58 kDa was observed on the SDS-PAGE. [Conclusion] The study laid the foundation for the further study on vaccine development of porcine rotavirus VP4.关键词
轮状病毒/VP4基因/克隆/原核表达Key words
Porcine rotavirus/VP4 gene/Cloning/Prokaryotic expression分类
农业科技引用本文复制引用
肖长峰,刘成倩,卢永红,陈斌,吴双林,张天庆,易建中..猪轮状病毒VP4蛋白基因在大肠杆菌中的表达[J].安徽农业科学,2009,37(22):10431-10432,10448,3.基金项目
上海市浦江人才计划(PJ[20]). （PJ[20]）
