【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。更多还原

【Abstract】 In order to study the biological functions of Erns gene of bovine viral diarrhea virus(BVDV), pMD18-T-Erns was digested with Bam H I and HindⅢ,and Erns gene fragment was obtained. The Erns gene was subcloned into the baculovirus transfer vector pBlueBacHis2A,and the recombinant plasmid pBlueBacHis2A-Erns was constructed. pBlueBacHis2A-Erns and Bac-N-Blue?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.更多还原