摘要： 采用RT－PCR方法扩增口蹄疫病毒(FMDV)AF72株的3D聚合酶基因，并将其克隆至pGEM－T easy载体，测序分析结果表明，AF72 3D聚合酶基因与GenBank中公布的其他个参考序列均具有较高的同源性。将目的基因插入原核表达载体pET－30a(+)中，构建了重组表达质粒pET－3D，鉴定后转化至大肠杆菌BL21(DE3)中，经IPTG诱导，3D聚合酶基因在大肠杆菌中获得了正确表达，目的蛋白的分子量为6 ku。Western blot检测结果显示，表达产物可以与抗A型FMDV阳性血清发生特异性反应，具有良好的反应原性。

关键词: 口蹄疫病毒, 3D聚合酶基因, 原核表达

Abstract: The 3D polymerase gene of Foot-and-mouth disease virus strain AF72 was amplified by reverse transcription polymerase chain reaction(RT-PCR),The purified PCR products were cloned into pGEM-T easy Vectors, and the nu- cleotide sequences were highly homologous to other four reference strains from GenBank. The target gene was cloned into the prokaryotic expression vector pET-30a(+), then recombinant expression plasmid pET- 3D was transformed into E. coli BL21(DE3) competent cells, the target protein was highly expressed by IPTG induct ion. The protein was about 46 ku. Western blot showed specific react ion of the expressed product with type A FMDV ant ibody- posit ive serum, and the protein had nice reaction activity.

Key words: FMDV, 3D polymerase gene, Prokaryotic expression

中图分类号: 

• Q945