猪流行性腹泻病毒安徽分离株N基因的克隆与表达  

Cloning and Expression of N Gene of Porcine Epidemic Diarrhea Virus from Anhui Province

作　　者：潘孝成[1,2] 沈学怀 赵瑞宏[1,2] 戴银 胡晓苗[1,2] 侯宏艳 周学利[1,2] 张丹俊 PAN Xiao-cheng;SHEN Xue-huai;ZHAO Rui-hong(Institute of Animal Husbandry and Veterinary Science,Anhui Academy of Agricultural Sciences,Hefei,Anhui 230031;Livestock and Poultry Epidemic Diseases Research Center of Anhui Province,Hefei,Anhui 230031)

机构地区：[1]安徽省农业科学院畜牧兽医研究所,安徽合肥230031 [2]安徽省畜禽疫病研究中心,安徽合肥230031

出　　处：《安徽农业科学》2019年第17期91-93,共3页Journal of Anhui Agricultural Sciences

基　　金：安徽省科技重点研发项目(1704a07020066);安徽省农业科学院平台项目(2019YL065);安徽省农业科学院创新团队项目(13C0405);安徽省生猪产业技术体系项目(AHCYTX-05-09);安徽省科技重大专项(17030701008)

摘　　要：[目的]克隆和表达猪流行性腹泻病毒(PEDV)的N基因。[方法]采用RT-PCR方法对PEDVFD株的N基因进行扩增,将扩增产物连接pET-28B载体,阳性质粒转入大肠杆菌BL21感受态细胞,诱导表达目的蛋白。[结果]PEDVFD株N基因序列全长为1326个核苷酸,编码441个氨基酸;重组N蛋白为可溶性表达,大小约58ku;Westernblot检测结果表明,重组N蛋白与PEDV抗体阳性血清发生特异性反应。[结论]该试验制备的PEDV重组N蛋白具有较好的免疫原性,可用于PEDV诊断方法的开发及相关研究。[Objective]To clone and express the N gene of porcine epidemic diarrhea virus(PEDV).[Method]N gene of PEDV FD strain was amplified by RT-PCR and clone into pET-28B vector,the positive plasmid was transformed into BL21 competent cell of Escherichia coli to express N protein under IPTG induction.[Result]N gene of PEDV FD strain contained 1 326 nucleotides in whole length,encoding 441 amino acids.The expressed recombinant N protein was soluble and it was about 58 ku.Western-blot detection results showed that the N protein and PEDV positive sera reacted specifically.[Conclusion]The recombinant N protein of PEDV had a good immunogenicity,which could be used for the diagnosis method development and research of PEDV.

关 键 词：猪流行性腹泻病毒 N蛋白 原核表达 

分 类 号：S852.657[农业科学—基础兽医学]