口蹄疫病毒真核表达质粒的构建及序列分析

Construction and sequences analysis for eukaryotic expression plasmid of foot-and-mouth disease virus

根据定点突变的原理 ,获得包含有口蹄疫病毒P1、2A、3C及部分 2B编码区的目的基因片段 ,经KpnⅠ和XbaⅠ双酶切后 ,与经同样处理的真核表达质粒载体 pcDNA3.1(+ )连接。经鉴定及DNA序列分析 ,口蹄疫病毒目的基因片段已被正确克隆到真核表达质粒载体pcDNA3.1(+ )上 。

By use of site mutation strategy, the gene P12X3C of foot and mouth disease virus was obtained that includes full length P1, 2A, 3C and a part of 2B. After being digested by KpnⅠ and XbaⅡ respectively, the gene P12X3C was cloned into the pcDNA3.1 (+) expression vector that was digested by the same enzyme. The recombinant plasmid was checked by restriction enzyme analysis and nucleic acid sequencing . The result showed the gene P12X3C was cloned into eukaryotic expression plasmid correctly, and the XbaⅠ site on P12X3C genes was mutated.