猪繁殖与呼吸综合征(PRRS)核衣壳蛋白的克隆、表达与免疫印迹  

Cloning,expression and Western blotting analysis of nucleocapsid protein gene of porcine reproductive and respiratory syndrome virus(PRRSV)

作　　者：徐丽华[1] 王一成[1] 逄春泰[1] 周永学[1] 袁秀芳[1] 张存[1] 叶伟成[1] 

机构地区：[1]浙江省农业科学院畜牧兽医研究所,浙江杭州310021

出　　处：《浙江农业学报》2005年第3期123-126,共4页Acta Agriculturae Zhejiangensis

基　　金：浙江省科技厅重点项目资助(011102102)

摘　　要：将猪繁殖与呼吸综合征病毒(PRRSV)的核衣壳蛋白(N蛋白)基因克隆到原核表达载体pET 28α(+)中,得到重组质粒pET PRRSV/N,并转化入受体菌BL21中。转化子经IPTG诱导表达4h后,表达蛋白量达到高峰。经15%SDS PAGE电泳检测和经Westernblotting分析,表达蛋白大小约为17kD,符合预期结果,并能与PRRSV阳性血清发生特异性反应,而不与阴性血清反应。这说明表达蛋白具有高度免疫反应特异性。<Abstrcat>The nucleocapsid protein(N) gene of porcine reproductive and respiratory syndrome virus(PRRSV) was cloned into the multiple cloning site of pET-28α(+) vector.The recombinant of pET-PRRSV/N was cultivated and induced with IPTG.The amount of expressed protein reached a high peak 4 hours after the induction. The examination analysis results of the protein by using SDS-PAGE and Western blotting showed that the protein was 17 kD in size and specifically reacted with PRRS-positive sera from pigs,not negative sera,indicating the recombinant protein had high specificity of immune reaction and can be used as antigen of diagnostic assay for PRRS in practice.

关 键 词：猪繁殖与呼吸综合征(PRRSV) 核衣壳蛋白 原核表 Western BLOTTING 

分 类 号：S828[农业科学—畜牧学] S852.65[农业科学—畜牧兽医]