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口蹄疫病毒AF72株3D聚合酶基因的克隆及其在大肠杆菌中的表达

DOI: 10.7668/hbnxb.2008.06.007, PP. 32-35

Keywords: 口蹄疫病毒,3D聚合酶基因,原核表达

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Abstract:

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-Teasy载体,测序分析结果表明,AF723D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6ku。Westernblot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。

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