表达绿色荧光蛋白伪狂犬病病毒转移载体的构建及转移特性  

Construction and Characterization of Pseudorabies Virus Transfer Vector Expressing Enhanced Green Fluorescent Protein

作　　者：方六荣[1] 陈焕春[1] 肖少波[1] 熊符[1] 王革飞[1] 

机构地区：[1]华中农业大学畜牧兽医学院,湖北武汉430070

出　　处：《中国兽医学报》2001年第2期125-127,共3页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目! (39970 5 5 9)

摘　　要：对含伪狂犬病病毒 ( PRV)鄂 A株 g G全基因 ,PK、g D基因部分编码序列的质粒 p USK进行改造 ,消除其中的 Eco R 位点 ,将通过 PCR扩增的增强绿色荧光蛋白基因 ( EGFP)融合到 g G启动子下游 ,构建了由 g G启动子控制 EGFP表达的 PRV转移载体 pg G- /EGFP+。该转移载体单独转染或同 PRV基因组 DNA共转染 IBRS-2细胞 ,结果单独转染时 EGFP不能表达 ;共转染时 ,6 h就可在荧光显微镜下观察到明显的荧光。The EcoRⅠ site of plasmid pUSK containing the complete gG gene, partial PK and gD genes, was removed to result in the plasmid pUSK△E. The coding sequence of enhanced green fluorescent protein (EGFP) was amplified from pEGFP-C1 by PCR and cloned into pUSK△E under the control of the gG promoter, to obtain the transfer vector pgG -/EGFP +. When pgG -/EGFP + was transfected into IBRS-2 cells alone or cotransfected with the genome DNA of pseudorabies virus Ea strain, it is interesting that the cells which transfected pgG -/EGFP + alone could not emit green fluoresence; whereas, a green fluoresence could be observed in the cotranfected cells and as early as 6 h post-transfection.

关 键 词：绿色荧光蛋白 伪狂犬病病毒 gG基因 转移载体 转移特性 报道分子 增强绿色荧光蛋白 

分 类 号：S852.659.1[农业科学—基础兽医学]