山羊痘弱毒疫苗株TK基因的克隆及序列分析  

Cloning and sequence analysis of TK gene of the attenuated capripoxvirus vaccine strain

作　　者：鞠厚斌[1] 张强[2] 袁明[3] 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,甘肃兰州730046 [3]甘肃省医学科学研究院,甘肃兰州730050

出　　处：《中国兽医科学》2007年第3期190-194,共5页Chinese Veterinary Science

基　　金：国家重点基础研究发展计划(973)项目(2005CB523201);国家基础平台项目(2005DKA21205-3)

摘　　要：以提取的山羊痘弱毒疫苗株HLJ—V9 DNA为模板，设计特异性引物并进行PCR扩增，获得了2078bp的DNA片段，并将PCR产物克隆至pGEM—T Easy载体。酶切鉴定、PCR鉴定和测序结果表明，成功获得了TK基因，获得的TK基因内存在一个Kpn Ⅰ酶切位点和编码区的早期转录终止信号TTTTTN（T）T。核苷酸和氨基酸同源性分析表明，弱毒疫苗毒株HLJ—V9 TK基因与参考毒株的同源性在95．5％～100％之间，说明羊痘病毒TK基因具有高度的保守性。The genomic DNA was extracted from the capripoxvirus live attenuated vaccine strain HLJ- V9,and a pair of specific primers were designed in order to amplify a TK gene. The PCR product approximately 2 078 bp in size was cloned into pGEM-T Easy vector. Restriction enzyme assay,PCR and sequencing confirmed that the TK gene was obtained successfully. Only one Kpn I site and a transcription termination signal TTTTTN（T）T were found in the gene. Analysis showed that HLJ-V9 strain shared 95.5% --100% identity rates with the reference strains in levels of nucleotide and amino acid,indicating that TK gene is very conservative.

关 键 词：山羊痘病毒 TK基因 序列分析 

分 类 号：S852.654[农业科学—基础兽医学]