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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
引用本文: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.
作者姓名:刘成倩  肖长峰  易建中  关平原  吴双林  陈斌  张天庆
作者单位:内蒙古农业大学动物科学与医学学院,内蒙古呼和浩特,010018;上海水产大学生命学院,上海,200090;上海市农业科学院畜牧兽医研究所,上海,201106
基金项目:上海市科技兴农重点攻关项目 
摘    要:目的]克隆猪繁殖与呼吸综合征病毒核衣壳蛋白基因并在原核表达系统中表达。方法]根据GenBank上登录的猪繁殖与呼吸综合征病毒(PRRSV)核酸序列设计1对引物,用RT—PCR方法扩增出PRRSV的核衣壳蛋白(N)基因,并将其克隆到原核表达载体pGEX-4T.1上,得到重组质粒pGEX-4T-1-N。将重组质粒转化大肠杆菌BL31(DE3)感受态细胞,经TPTG诱导,SDS-PAGE电泳检测。结果]PRRSV肺组织所提取的RNA,经RT—PCR扩增,得到一段392bp的片段,与预期结果一致。SDS—PAGE电泳分析,所得重组蛋白的分子量约39.866Da,与预期结果相同。重组菌体超声裂解后,经10%的SDS—PAGE电泳分析,结果显示该重组蛋白为可溶性蛋白。

关 键 词:猪繁殖与呼吸综合征病毒  核衣壳蛋白  克隆  表达

Cloning of Nucleocapsid Protein Gene of Porcine Reproductive and Respiratory Syndrome Virus and Its Procaryotic Expression
LIU Cheng-qian et al.Cloning of Nucleocapsid Protein Gene of Porcine Reproductive and Respiratory Syndrome Virus and Its Procaryotic Expression[J].Journal of Anhui Agricultural Sciences,2009,37(22):10429-10430.
Authors:LIU Cheng-qian
Institution:LIU Cheng-qian et al(College of Animal Science , Medicine,Inner Mongolia Agricultural University,Hohhot,Inner Mongolia 010018)
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 compete...
Keywords:Porcine reproductive and respiratory syndrome virus  Nucleocapsid protein  Clone  Expression
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