山羊痘病毒通用转移载体的构建及鉴定  

Construction of a Goatpox Virus Transfer Vector and Its Identification

作　　者：鞠厚斌[1] 张强[2] 郑海学[2] 施程洪[2] 韩若婵[2] 谢芳[2] 吴国华[2] 颜新敏[2] 李健[2] 朱海霞[2] 朱彩珠[2] 卫广森[1] 

机构地区：[1]沈阳农业大学畜牧兽医学院,辽宁沈阳110161 [2]中国农业科学院兰州兽医研究所家畜疫病病原生物学国家重点实验室农业部畜禽病毒学重点开放实验室,甘肃兰州730046

出　　处：《中国兽药杂志》2007年第2期1-4,9,共5页Chinese Journal of Veterinary Drug

摘　　要：转移载体的构建是重组羊痘病毒构建的重要环节,在分析羊痘病毒基因组的基础上,以TK基因为插入靶序列,经PCR、酶切鉴定,获得了带有EcoRⅠ、BamHⅠ酶切位点的TK基因片段。将此片段与经相同酶切的pUC119载体连接,构建转移载体pUC119-TK,并且以此为基础构建表达绿色荧光蛋白山羊痘病毒转移载体pTK-P7.5-GFP。将pTK-P7.5-GFP转染羊痘病毒感染的BHK21细胞,48 h后报告基因得到表达,这为羊痘病毒载体系统的进一步开发奠定了基础。Transfer vector is considered important step in the construction of recombinant virus. Based on the sequence analysis of the genome of goatpox virus,TK gene was chosen as the insertion site for the construction of transfer vector. TK gene with EcoR ⅠBamH Ⅰ restriction sites was got by PCR, cloning and restricting. Then the segments was ligated into pUCll9. The transfer vector was named as pUCll9-TK. Based on the pUCll9-TK, transfer vector with the P7.5 controlled EGFP was constructed. After transfection in BHK21 cells infected with goatpox virus, the reporter gene was expressed 48 hours later. This results lay foundation for recombinant vaccine development.

关 键 词：羊痘病毒 TK基因 转移载体 构建 鉴定 

分 类 号：S852.654[农业科学—基础兽医学]