摘要
为研究猪流行性腹泻病毒(Porcine epidemic diarrhea virus,PEDV)的N蛋白特性及在诊断中的应用,本研究采用RT-PCR方法从猪流行性腹泻病毒FJ-11A株中扩增其N蛋白基因片段,并将其克隆到原核表达载体pET-32a上,构建原核表达重组质粒pET32a-PEDV-N,进行条件优化诱导表达、SDS-PAGE、Western blot和ELISA试验。结果表明,该重组目的蛋白(约60 kDa)得到表达,且具有良好的免疫学活性,该研究为开发猪流行性腹泻病毒诊断方法和研究N蛋白功能奠定基础。
In order to illustrate function of nucleoprotein of porcine epidemic diarrhea virus and its application to diagnosis,the N gene fragment was amplified in RT-PCR and cloned into the prokaryotic vector pET-32a for expression in E.coli with induction of IPTG.The expressed protein was examined in SDS-PAGE and determined to be of molecular weight of 60 kDa.The recombinant protein reacted with anti-PEDV hyperimmune serum in Western blot and indirect ELISA.Therefore,the recombinant N protein could be used for further study on PEDV diagnosis and control.
出处
《中国动物传染病学报》
CAS
2013年第2期56-60,共5页
Chinese Journal of Animal Infectious Diseases
基金
国家公益性行业(农业)科研专项(NYHYZX07-034)
福建省财政专项-福建省农业科学院科技创新团队建设项目(STIF-Y02)
关键词
猪流行性腹泻病毒
N蛋白
原核表达
Porcine epidemic diarrhea virus
N protein
prokaryotic expression
