C型口蹄疫病毒VPI基因的克隆及真核表达载体的构建  

Cloning and Construction of Eukaryotic Expression Vector of VP1 Gene from Foot-and-mouth Disease Virus of Type C

作　　者：杨生海[1,2] 殷宏[2] 张杰 刘永生[2] 张继乐[2] 曾巧英[1] 陈豪泰[2] 马丽娜[2] 马艳平[2] 丁耀忠[2] 

机构地区：[1]甘肃农业大学动物医学院,甘肃兰州730070 [2]中国农业科学院兰州兽医研究所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：《安徽农业科学》2009年第10期4416-4419,共4页Journal of Anhui Agricultural Sciences

基　　金：甘肃省科技重大专项项目(0801NKDA034);兰州市科技计划项目(2008-1-167)

摘　　要：应用PT-PCR扩增C型口蹄疫病毒(FMDV)VP1基因,将其克隆到pMD18-T载体中,并对克隆载体和VP1基因进行序列分析。将VP1基因和选择标记基因——二氢叶酸还原酶(dhfr)基因插入真核表达载体pCI-neo中,对所构建的重组真核表达载体进行PCR扩增、酶切鉴定和序列分析。结果表明,VP1基因与GenBank中标准毒株(登录号EU553893)VP1的序列同源性为92.8%,VP1基因的真核表达载体pCI-VP1-D构建成功。The VP1 gene from foot-and-mouth disease virus of type C was amplified by RT-PCR,and it was cloned into pMD18-T vector.The sequences of the cloning vector and VP1 gene were analyzed.The VP1 gene and selectable marker gene dihydrofolate reductase（dhfr） were inserted into eukaryotic expression vector pCI-neo,and the constructed recombinant eukaryotic expression vector was amplified by PCR,and it was analyzed by enzyme digestion and sequence analysis.The results showed that The sequence homology of VP1 gene and standard strain（accession number EU553893） VP1 in GenBank was 92.8% and the eukaryotic expression vector pCI-VP1-D of VP1 gene was successfully constructed.

关 键 词：C型口蹄疫病毒 VPI基因 真核表达载体 

分 类 号：S188[农业科学—农业基础科学]