猪瘟病毒E2蛋白BC抗原域的酿酒酵母表面展示  

Display of Classical Swine Fever Virus E2 Protein BC Antigenic Domain on the Cell Surface of Saccharomyces cerevisiae

作　　者：刘小凤 汪倩 罗明阳 孙金福 Xiaofeng Liu;Qian Wang;Mingyang Luo;Jinfu Sun(Institute of Biotechnology, College of Life and Health Sciences, Northeastern University, Shenyang Liaoning)

机构地区：[1]东北大学生命科学与健康学院生物技术研究所,辽宁沈阳

出　　处：《微生物前沿》2017年第3期72-78,共7页Advances in Microbiology

基　　金：国家自然基金重点项目(31130052).

摘　　要：为构建猪瘟病毒(CSFV)E2蛋白的B、C抗原域(E2BC)酿酒酵母表面展示体系,以pVEXE2为模板,用PCR克隆了E2BC基因,并插入酵母展示表达载体p1v5AG的BamH I和EcoRI位点,构建重组酵母展示表达载体,用LiAC方法转化酿酒酵母细胞W303,构建重组酵母菌。重组菌经培养后,对酵母菌细胞进行针对His标签的间接免疫荧光染色,检测His-E2BC融合表达蛋白。结果显示,重组酵母菌表面有绿色荧光,表明E2BC蛋白成功表达于酵母表面,为开发酵母载体口服疫苗奠定了基础。To construct the yeast display system of classical swine fever virus E2 glycoprotein BC antigenic domain, gene fragment was cloned by PCR using pVEXE2 as a template. E2BC was inserted into the site of BamH I and EcoR I of p1v5AG to produce recombinant surface displaying vector of p1E2BCAG. p1E2BCAG was transformed into Saccharomyces cerevisiae by LiAC method. After recombinant cells were cultivated, indirect immunofluorescence was used to detect E2BC expression. The results indicate that E2BC protein was displayed on the surface of Saccharomyces cerevisiae cell successfully. This work provides a foundation for the development of Saccharomyces cerevisiae vector oral vaccine of CSFV E2BC protein.

关 键 词：酵母表面展示 猪瘟病毒 E2BC抗原域 口服疫苗 

分 类 号：Q7[生物学—分子生物学]