我国D2-43病毒株PrM-E基因的重组SFV的制备及生物学鉴定  

Preparation and Biological Identification of Recombinant SFV Virus Containing the PrM-E Gene of Chinese Dengue 2 Virus

作　　者：陈水平[1] 秦鄂德[1] 于曼[1] 赵卫[1] 胡志君[1] 苑锡同[1] 范宝昌[1] 杨佩英[1] 

机构地区：[1]北京微生物流行病研究所,北京100071

出　　处：《中国生物化学与分子生物学报》2001年第6期739-742,共4页Chinese Journal of Biochemistry and Molecular Biology

基　　金：国家自然科学基金课题 (No .39770 0 36 )~~

摘　　要：为构建含PrM E基因的重组SFV病毒 ,将含我国登革 2型病毒PrM E基因的SFV表达载体和辅助载体DNA线性化后 ,进行体外转录 .然后将获得的 2种RNA转录物共转染BHK2 1细胞 ,以RT PCR法证明转染后的细胞培养上清中含有重组SFV .进一步将该重组病毒经蛋白酶激活后可使BHK2 1细胞产生细胞病变 ,并且以间接免疫荧光法在感染细胞内可检测到登革 2型病毒所特有的蛋白 .含我国登革 2型病毒PrM E基因的重组SFV的获得 ,为进一步观察该重组病毒的免疫原性奠定了基础 。To construct the recombinant SFV containing the PrM E gene, the SFV expression vector DNA, which contained the PrM E gene of Chinese dengue 2 virus, and the helper vector DNA were linearized and transcribed in vitro , respectively. Then the 2 RNA transcripts were directly coelectroporated into BHK cells. The cell supernatants were demonstrated to contain recombinant SFV virus with RT PCR. After being activated with α chymotrypsin, the recombinant virus could make BHK cells produce CPE. Furthermore, the proteins specific to dengue 2 virus could be detected in the infected cells by indirect immunofluorescence technique. The results laid a foundation for the study of the immunogenicity of the recombinant SFV virus and provided a new way to develop vaccines for dengue virus.

关 键 词：登革2型病毒 重组SFV病毒 PrM-E基因 体外转录 生物学鉴定 

分 类 号：R373.33[医药卫生—病原生物学]