牛病毒性腹泻病毒E^(rns)基因的真核表达及抗原性检测  

Eukaryotic expression of E^(rns) gene of bovine viral diarrhea virus and antigenic detection of the expressed protein

作　　者：吴立君[1] 师东方[1] 王丹娜[1] 何海娟[1] 吴明福[1] 王君伟[1] 

机构地区：[1]东北农业大学动物医学院,黑龙江哈尔滨150030

出　　处：《中国兽医科学》2006年第6期439-443,共5页Chinese Veterinary Science

摘　　要：为研究牛病毒性腹泻病毒(BVDV)Erns基因的生物学功能,将含有牛病毒性腹泻病毒 Erns基因的质粒pMD18-T-Erns经BamH I／HindⅢ双酶切,获得了Erns片段,再与杆状病毒转移载体pBlueBacHis2A连接,构建成重组质粒。将重组质粒pBlueBacHis2A-Erns与Bac-N-BlueTM DNA 共转染至sf9昆虫细胞中,获得了重组病毒,经噬斑筛选纯化,感染sf9昆虫细胞进行表达。SDS- PAGE分析结果表明,表达的目的蛋白大小约30 ku;Western-blotting检测表明,该蛋白具有良好的抗原性。In order to study the biological functions of E^rns gene of bovine viral diarrhea virus（BVDV）, pMD18-T-E^rns was digested with BamH Ⅰ and HindⅢ, and E^rns gene fragment was obtained. The E^rns gene was subcloned into the baculovirus transfer vector pBlueBacHis2A, and the recombinant plasmid pBlueBacHis2A-E^rns was constructed, pBlueBacHis2A-E^rns and Bac-N-BlueTM DNA were co-transfected into sf9 cells. After the plaques were screened, sf9 cells were infected with the recombinant virus. SDS-PAGE analysis showed that the target protein of 30 ku in size was expressed. The protein was proved to have good antigenicity by Western-blotting analysis.

关 键 词：牛病毒性腹泻病毒 E^rns基因 重组杆状病毒 

分 类 号：S852.659.6[农业科学—基础兽医学] Q786[农业科学—兽医学]