牛病毒性腹泻病毒石河子株E2基因的克隆及其表达载体的构建

Cloning of the E2 Gene of Bovine Viral Diarrhea Virus Isolated of Shihezi and Construction of Expression Vector

应用RT-PCR方法从牛病毒性腹泻病毒(BVDV)新疆石河子分离株中扩增了E2基因,命名Shihezi 148(GenBank登录号:EU159699),扩增序列分析结果表明:Shihezi 148 E2基因长度为1121 bp,编码373个氨基酸残基;同源性分析表明,Shihezi 148与澳大利亚Bega株E2基因核苷酸同源性为88.32%,与中国changchun 184株的同源性为74.42%;通过基因重组技术构建了去除C端跨膜区的截短E2基因的原核表达载体pProEX HTa-E2和包括信号肽和全长E2蛋白的真核重组质粒pVAX1-E2。经过PCR、酶切及测序结果表明pProEX HTa-E2和pVAX1-E2重组表达载体构建成功。

Bovine viral diarrhea virus （BVDV） were isolated from Shihezi Xinjiang China. The isolate strain of Shihezi 148 was obtained by reverse transcription polymerase chain reaction（RT-PCR）. Sequencing results showed that E2 gene of Shihezi148 is 1121bp nucleotides, （GenBank accession no. EU159699）. with 373 amino acid residue segment. The E2 gene of Shihezi 148 had 88.32% ,74.42~/oo homology with Bega strain and changchun184 strain; It was cloned into eukaryotic pProEX HTa and pVAX1 procaryotic expression vector. The two recombinant plasmid was identified with PCR, En zyme-cutting and sequencing. The result shows that the recombinant plasmid pproEXhta E2 and pVAX1-E2 was successfully constructed.