猪繁殖与呼吸综合征病毒河南分离株GP5基因的克隆及在293T细胞中表达  

Cloning and Expression in 293T Cell of GP5 Gene of Porcine Reproductive and Respiratory Syndrome Virus Isolated in Henan

作　　者：党占国[1] 夏平安[1] 高进勇[2] 尹彦涛[1] 王建举[1] 崔保安[1] 陈溥言[3] 

机构地区：[1]河南农业大学牧医工程学院,河南郑州450002 [2]福州大学,福建福州350002 [3]南京农业大学,江苏南京210095

出　　处：《河南农业大学学报》2008年第2期196-199,共4页Journal of Henan Agricultural University

基　　金：中国博士后基金项目(20060390944)

摘　　要：根据GenBank公布的猪繁殖与呼吸综合征病毒(PRRSV)ATCC VR-2332株GP5基因序列设计1对特异性引物,应用RT-PCR方法扩增PRRSV Hn/06-1分离株GP5基因片段.将扩增片段克隆到真核表达载体pcDNA3.0中,测序后用DNAstar序列分析.构建的pcDNA-GP5重组质粒瞬时转染293T细胞,48 h后用Western-blot检测,证明GP5基因在293T细胞中获得了表达,可与PRRSV阳性血清发生特异性反应.According to the gene sequence of American standard strain of ATCC VR-2332, a pair of specific primers against the GP5 gene of PRRSV was designed. GP5 gene of porcine reproductive and respiratory syndrome virus isolated in Henan Province from suspected pigs was amplified with RT-PCR method. The amplified GP5 gene was then cloned into the eukaryotic expression vector pcDNA3.0, and the constructed pc DNA-GP5 recombinant plasmids was instantly transfected in 293T cell. Western blotting analysis indicated that the expressed recombinant protein could vigorously react against the positive serum of PRRSV.

关 键 词：猪繁殖与呼吸综合征病毒 GP5基因 克隆 真核表达 

分 类 号：S852.65[农业科学—基础兽医学]