牛疱疹病毒I型gB基因的原核表达  

PROKARYOTIC EXPRESSION OF BOVINE HERPESVIRUS-1 GB GENE

作　　者：李艳莉[1] 陶洁[1] 张信军[1] 王银[1] 朱国强[1] 

机构地区：[1]扬州大学兽医学院,扬州225009

出　　处：《中国动物传染病学报》2013年第5期34-39,共6页Chinese Journal of Animal Infectious Diseases

基　　金：江苏高校优势学科建设工程资助项目;教育部创新团队;江苏中美国际科技合作项目(BZ2005032);江苏省农业支撑项目(BE2008363);国家农业部948计划(2011-G24)

摘　　要：根据GenBank中BHV-1 Colorado 1毒株的全基因组序列,针对gB基因的主要B细胞免疫区域设计一对引物。提取BHV-1 Colorado 1毒株的基因组DNA,PCR扩增大小为585 bp的gB片段,将其克隆至pET-22b(+)原核表达载体并测序分析,同时用Nde I和Not I酶切鉴定。SDS-PAGE和Western blot试验结果证明,获得的可溶性gB重组蛋白能与牛传染性鼻气管炎病毒标准阳性WEI血清发生特异性反应,为ELISA等免疫诊断方法的建立奠定了物质基础。A pair of primers specific for herpesvirus-1 gB gene was designed according to whole genome sequence of BHV-1 Colorado 1 strain in GenBank. The gB gene fragment of 585 bp was amplified in PCR and inserted into prokaryotic expression vector pET-22b（＋）. The resulting recombinant plasmid pET-22b（＋）-gB was confirmed with sequencing and restriction enzyme analysis and then transformed into E. coli BL21 （DE3）. The fusion protein specifically reacted with reference serum to infectious bovine rhinotracheitis virus （IBRV） in Western blot. Therefore, the recombinant gB protein obtained in the study could be used in the development of diagnostic ELISA method, etc.

关 键 词：牛疱疹病毒 GB基因 pET-22b(+) 包涵体 蛋白免疫印迹 

分 类 号：S852.659.1[农业科学—基础兽医学]