带有小鼠CD40L基因的汉滩病毒糖蛋白表达载体的构建  

Construction of Hantaan Virus Envelope Glycoprotein Expression Vector Added with Mouse CD40L Gene

作　　者：冯伟[1] 张尧[1] 张晓晓[2] 应旗康[2] 刘梓谕[2] 吴兴安[2] 王芳[2] 

机构地区：[1]第四军医大学学员旅,陕西西安710032 [2]第四军医大学微生物学教研室,陕西西安710032

出　　处：《生物技术通讯》2017年第4期499-502,共4页Letters in Biotechnology

基　　金：国家自然科学基金(31470890;31270978);军队重点课题(BWS13G);陕西省科技统筹创新工程计划(2013KTCL03-06)

摘　　要：目的:构建汉滩病毒包膜糖蛋白基因的真核表达载体,并加入可增强免疫应答效应的细胞因子CD40L基因,检测其可否在真核细胞中表达。方法与结果:参照GenBank中汉滩病毒M基因和小鼠CD40L的全基因序列设计引物,通过聚合酶链反应(PCR)获得M和CD40L基因片段,将其与pCI-neo载体相连,测序证实该载体构建成功后,将此真核表达载体以脂质体转染法转染至哺乳动物细胞CHO-K1中,利用间接免疫荧光法(IFA)检测发现M基因和CD40L基因可以同时表达于CHO-K1细胞中。结论:构建了带有CD40L基因的汉滩病毒包膜糖蛋白重组质粒并获得表达,为深入研究汉滩病毒感染后包膜糖蛋白引起的特异性免疫应答规律奠定了实验基础。Objective： The eukaryotic expression vector of Hantaan virus glycoprotein was constructed and added with mouse CD40L gene that could strengthen mouse immune response, in order to detect whether or not it could express in the eukaryotic cell. Methods ＆ Results： Primers were designed referring to M gene sequence of Hanta- an virus and mouse CD40L gene in the GenBank. Then M gene and mouse CD40L gene were obtained through PCR, and cloned into eukaryotic expressing vector pCI-neo vector. After sequenced, the eukaryotic expressing vec- tor was transfected into eukaryotic cells CHO-K1 with liposome method. It was found that specific fluorescence appeared in the CHO-K1 after transfection by indirect immunofluoresccnce assay（IFA）. Conclusion： The results showed that the eukaryotic expressing vector of Hantaan virus glycoprotein conjugated with mouse CD40L was suc- cessfully constructed and expressed, to deeply investigate the immune response of Hantaan virus glycoprotein.

关 键 词：汉滩病毒 包膜糖蛋白 CD40L 

分 类 号：Q78[生物学—分子生物学] R392.1[医药卫生—免疫学]