猪繁殖与呼吸障碍综合征病毒ORF5基因的克隆与表达  

Cloning and Expression of the ORF5 Gene of Porcine Reproductive and Respiratory Syndrome Virus(PRRSV)

作　　者：田小辉[1,2] 王爱萍[1] 乔松林[2] 张改平[2] 肖治军[2] 冯丽丽[2] 刘运超[1] 李乔木[1] 李卓阳[2] 

机构地区：[1]郑州大学生物工程系,河南郑州450001 [2]河南省农业科学院,河南省动物免疫学重点实验室,河南郑州450002

出　　处：《河南农业科学》2009年第2期103-106,共4页Journal of Henan Agricultural Sciences

基　　金：国家自然科学基金项目(30730068)

摘　　要：用RT-PCR方法从河南分离的猪繁殖与呼吸障碍综合征病毒(PRRSV)变异株(HN07-1)中扩增ORF5全长基因,将其克隆至pMD19-T载体。测序结果表明,该序列与标准美洲株VR-2332同源性为89%,与我国早期分离的BJ-4株同源性为88%,与HB-1、JXAI变异株同源性为96%和99%。将该基因亚克隆到原核表达载体pGEX-6p-1后,转化大肠杆菌BL21,IPTG诱导表达。SDS-PAGE和Western-Blot结果表明,成功表达了41.5 kD的融合蛋白。The full-length of ORF5 gene was amplified from the genomic RNA of Henan variant PRRSV strain HN07 - 01 by RT - PCR. The amplified fragment was cloned into pMD19 - T vector and then sequenced. Compared with the ORF5 gene sequences of BJ - 4, classical American genotype VR - 2332, HB - 1 and JXA - 1, the homology of HN07 - 01 ORF5 gene was 88 %, 89 %, 96 % and 99 %, respectively. The ORF5 gene was subcloned into the prokaryotic expression vector pGEX - 6p - 1 ,transformed into E. coli BL21 and induced with IPTG. The results of SDS - PAGE and Western-Blot confirmed that the recombinant in E. coli BL21. protein of 41.5 kD was successfully expressed

关 键 词：猪繁殖与呼吸障碍综合征病毒 变异株 ORF5 原核表达 

分 类 号：S852.65[农业科学—基础兽医学]