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.
出处
《安徽农业科学》
CAS
北大核心
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)
关键词
C型口蹄疫病毒
VPI基因
真核表达载体
Foot-and-mouth disease virus of type C
VP1 gene
Eukaryotic expression vector
