慢病毒载体介导的西尼罗河病毒prME蛋白的表达  

Lentiviral vector-mediated expression of West Nile virus prME protein

作　　者：徐敏[1] 杨晓红[1] 陈龙[1] 李么明[1] 叶静[1] 陈焕春[1] 毕丁仁[1] 曹胜波[1] 

机构地区：[1]华中农业大学动物医学院农业微生物学国家重点实验室,湖北武汉430070

出　　处：《中国兽医科学》2010年第8期778-782,共5页Chinese Veterinary Science

基　　金：农业公益性行业科研专项(200903037)

摘　　要：将西尼罗河病毒(WNV)的主要免疫原性基因prME插入HIV-1慢病毒载体pHR中,构建了转移质粒pHR-WNV/prME。在脂质体介导下,将该质粒与p△8.2、pVSV/G共转染293T细胞,48h后收获细胞上清。将上清转导正常的293T细胞,通过报告基因荧光观察、间接免疫荧光检测。结果表明,利用HIV-1慢病毒载体系统包装得到了假型病毒,可以高效转导293T细胞,并实现了WNVprME基因在被转导细胞中的表达,从而为基于慢病毒载体的WNV新型疫苗的研制奠定基础。prME,the major antigenic gene of West Nile virus（WNV）,was inserted into HIV-1 lentiviral vector pHR,and the constructed transfer plasmid pHR-WNV/prME was co-transfected with p△8.2 and pVSV/G into 293T cells by Lipofectin.At hour 48 post-cotransfection,the supernatant of 293T cells was harvested and transducted into normal 293T cells.Subsequently,the expression of target protein was detected by indirect immunofluorescence assay.The results showed that the pseudotype virus pHR-WNV prME was efficiently transducted into 293T cells,and WNV prME protein gene was successfully expressed,providing the experimental basis for the development of new WNV-vaccines in the future.

关 键 词：西尼罗河病毒 prME蛋白 慢病毒载体 表达 

分 类 号：S852.659.6[农业科学—基础兽医学]