安徽农业科学2009，Vol.37Issue(22)：10429-10430,10513,3.
猪繁殖与呼吸综合征病毒核衣壳蛋白基因的克隆与原核表达
Cloning of Nucleocapsid Protein Gene of Porcine Reproductive and Respiratory Syndrome Virus and Its Procaryotic Expression
摘要
Abstract
[Objective] The aim was to clone nucleocapsid protein gene of porcine reproductive and respiratory syndrome virus (PRRSV) and express it in procaryotic expression system. [Method] According to the nucleic acid sequence of PRRSV addressed in GenBank, a pair of primers was designed for amplifying the nucleoprotein (N) gene with RT-PCR. The N gene was cloned into the procaryotic expression vector pGEX-4T-1 and the recombinant plasmid pGEX-4T-1-N was obtained, then the recombinant plasmid was transformed into competent cell of Escherichia coli BL31 (DE3), after TPTG induction, it was detected by SDS-PAGE electrophoresis. [Result] A fragment of 392 bp was amplified from PRRSV RNA extracted from lung by RT-PCR, which was consistent with anticipated result. The analysis on SDS-PAGE electrophoresis showed that the molecular weight of the obtained recombinant protein was about 39.866 Da, which was consistent with anticipated result. After ultrasonic degradation, the recombinant was analyzed by 10% SDS-PAGE electrophoresis and the result showed that the recombinant protein was soluble. [Conclusion] The research laid the foundation for the purification of the recombinant protein.关键词
猪繁殖与呼吸综合征病毒/核衣壳蛋白/克隆/表达Key words
Porcine reproductive and respiratory syndrome virus/Nucleocapsid protein/Clone/Expression分类
农业科技引用本文复制引用
刘成倩,肖长峰,易建中,关平原,吴双林,陈斌,张天庆..猪繁殖与呼吸综合征病毒核衣壳蛋白基因的克隆与原核表达[J].安徽农业科学,2009,37(22):10429-10430,10513,3.基金项目
上海市科技兴农重点攻关项目[沪农科攻字(2007)第12-1号]. （2007）
