猪流行性腹泻病毒N蛋白的原核表达及生物学活性鉴定

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.