禽痘病毒通用转移载体的构建及鉴定  

Construction of a Fowlpox Virus Transfer Vector and its Identification

作　　者：智海东[1] 王云峰[1] 王利丽[1] 王玫[1] 刘娣 童光志[1] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室,哈尔滨150001 [2]黑龙江省农业科学院,哈尔滨150001

出　　处：《中国生物工程杂志》2004年第7期89-92,共4页China Biotechnology

基　　金：国家"8 63"计划资助项目 (2 0 0 1AA2 13 0 41)

摘　　要：禽痘病毒作为活载体已经得到广泛的应用。转移载体的构建是重组禽痘病毒构建的重要环节。在分析禽痘病毒基因组的基础上 ,以FL1 1基因为插入位点 ,设计引物分别扩增 1kb的同源臂 ,将PCR扩增后的同源臂在体外连接后 ,插入到pUC1 1 9中 ,构建转移载体 ,并且以此为基础 ,构建表达绿色荧光蛋白的转移载体。含有eGFP的转移载体转染禽痘病毒感染的鸡胚成纤维细胞后 ,报告基因获得表达 ,这将为禽痘病毒载体系统的进一步开发奠定基础。Fowlpox virus have been used as safe vector to construct vector vaccine to control avian virus disease. Transfer vector are considered important step in the construction of recombinant virus. Based on the sequence analysis of the genome of fowlpox virus, the ORF F11L was chosen as the insertion site for the construction of transfer vector. The DNA of genome of Fowlpox was extracted, and the 1kb homologous flank was amplified, then the segments was ligated in vitro and subcloned into pUC119, the transfer vector was named as pSY683. Based on the transfer vector pSY683, transfer vector containing the LP2EP2 controlled EGFP was constructed, after transfection in chicken fibroblast cells infected with fowlpox virus, the reporter gene expressed 72 hours later. This results lay foundation for recombinant vaccine development. ;

关 键 词：禽疽病毒 转移栽体 绿色荧光蛋白 基因 表达 疫苗 

分 类 号：Q782[生物学—分子生物学]