汉滩病毒76-118株核蛋白部分基因片段的克隆、表达和抗原活性测定  

Cloning and transient expression of the partial nucleocapsid gene segment of Hantaan virus 76-118 strain in fibroblast and the antigenic activities of partial nucleocapsid

作　　者：张梦寒[1] 诸葛洪祥[1] 陈明中[1] 许菊[1] 

机构地区：[1]苏州大学医学院病原生物学教研室

出　　处：《现代免疫学》2005年第4期297-299,共3页Current Immunology

基　　金：国家自然科学基金资助项目(39970654);江苏省病原生物学重点实验室开放课题资助项目

摘　　要：为了研究汉坦病毒的DNA疫苗,用PCR方法克隆汉滩病毒76-118株S片段部分基因,构建真核细胞表达质粒pEGFP-HTNV-S0.7,并在成纤维细胞L929中进行瞬时表达。成功地得到预期大小约0.7kb的重组基因,并在成纤维细胞中进行了瞬时表达。免疫印迹分析产生的相对分子质量约26000的蛋白质具有抗汉坦病毒的抗原活性。直接免疫荧光分析显示了特异性的绿色荧光。In order to prepare the DNA vaccine for Hantaan virus infection, a recombinant eukaryotic expression plasmid pEGFP-HTNV-S0.7 to clone the partial S-segment of Hantaan virus 76-118 strain was constructed by PCR, and the recombinant gene was expressed transiently in fibroblast L929 cells. It was proved that the expected recombinant gene of approximately 0.7 kb in length was obtained successfully and could be expressed in fibroblasts. The gene products of about 26 000 Mr protein with antigenicity to Hantaan virus were demonstrated by Western blotting analysis, and there was specific green fluorescence in the recombinant plasmid-transfected fibroblast L929 cells as illustrated by direct immunofluorescence analysis.

关 键 词：汉滩病毒 S片段 成纤维细胞L929 瞬时表达 

分 类 号：R373.3[医药卫生—病原生物学]