作　　者： ; ; ; ; ; ; ;

机构地区： 山东农业大学动物科技学院

出　　处： 《西北农业学报》 2006年第4期1-5,共5页

摘　　要： 用PCR方法从感染牛疱疹病毒Ⅰ型Bartha Nu/67株的MDBK细胞中扩增gD基因,将之克隆到pGEM-T Easy载体。根据对gD基因的结构分析,设计合成3对引物,以获得的克隆质粒为模板,PCR扩增gD胞外域基因,将之分别亚克隆到pGEX-4T-2和pET-32a表达载体中构建成表达质粒pGEX-4T-gDQ、pGEX-4T-gDQB、pGEX-4T-gDHB和pET-gDQ、pET-gDQB、pET-gDHB,利用上述质粒分别转化大肠杆菌BL21和BL21(DE3),IPTG诱导后SDS-PAGE检测,结果pGEX-4T-gDQ、pGEX-4T-gDQB、pGEX-4T-gDHB和pET-gDQ四个表达质粒出现表达且符合预期大小,Western blot表明表达产物均有抗原性。 The gene encoding gD of bovine herpesvirus 1（BHV-1） Bartha Nu/67 strain was amplified by means of PCR from virus DNA of cell cultures infected by BHV-1, and cloned into pGEX-T easy vector. Three pairs of primer were designed and synthesized according to gD gene rank of BHV-1, the genes of extracellular domain were amplified from pGEM-gD ORF plasmid and subcolned into expres- sion vector pGEX-4T-2 and pET-32a in correct reading-frames. The positive recombinant plasmids were transformed into E. coli protein were expressed in E. BL21 and BL21（DE3）and induced by IPTG. Four kinds of recombinant coli confirmed by SDS-PAGE. All of the recombinant protein could be detected by Western blot with positive serum of IBRV.

关 键 词： 牛疱疹病毒 型 糖蛋白基因 克隆 原核表达

领　　域： [农业科学] [农业科学] [农业科学]