河北猪繁殖与呼吸综合征病毒HB-3(cz)株N基因变异分析及原核表达  

Sequence analysis and prokaryotic expression of ORF7 gene of porcine reproductive and respiratory syndrome virus HB-3(cz) strain

作　　者：陈赛娟[1] 刘涛[2] 曹洪战[1] 张斌[1] 付新城[3] 张丽青[1] 孙继国[1] 

机构地区：[1]河北农业大学动物科技学院,河北保定071001 [2]商务部流通产业促进中心,北京100062 [3]廊坊市畜牧水产局,河北廊坊102800

出　　处：《河北农业大学学报》2008年第5期85-88,共4页Journal of Hebei Agricultural University

基　　金：河北省科技攻关项目(06220401D-4)资助

摘　　要：为了给河北地区预防和控制猪繁殖与呼吸综合征提供理论数据。应用RT-PCR方法特异性扩增HB-3(cz)株的ORF7(N)基因片段,将扩增片段克隆入pMD-T载体后测序,应用DNAstar分析软件对序列进行分析,并与GenBank中发表的PRRSV毒株序列进行比较。结果显示:HB-3(cz)株与高热病毒株JXA1、HUB2等及传统河北分离株HB-1(sh)氨基酸同源性高达97.6%,属美洲型毒株。将目的片段克隆入原核表达载体pGEX-6P-1,重组质粒pGEX-N转化大肠杆菌BL21(DE3),在IPTG诱导下成功获得表达,经Western-blot-ting分析表明:表达的融合蛋白分子量约为39.5 kDa,能与PRRSV的阳性血清发生特异性反应,为PRRSV血清学诊断方法的建立奠定了基础。This study aims to provide theoretical evidence for prevention and control of porcine reproductive and respiratory syndrome in Hebei.The specific ORF7 gene of HB-3(cz) strain amplified by RT-PCR was cloned into pMD-T vector.The recombinant plasmid was sequenced and compared to the other published strains in GeneBank.The results showed the deduced amino acids of HB-3(cz) showed 97.6% similarity to strains JXA1,HUB2,HB-1(sh),and so on,and the HB-3(cz) strain be1ongs to North American genotype.The ORF7 gene wa...

关 键 词：猪繁殖与呼吸综合征病毒 N基因 序列分析 原核表达 

分 类 号：S852.65[农业科学—基础兽医学]