O型FMDV VP1基因的真核表达及其产物的生物学活性

Eukaryotic expression of the VP1 gene of foot-and-mouth disease virus type O and characterization of expressed fusion protein

根据已克隆的O型口蹄疫病毒VP1基因序列,设计了1对带有SacⅠ和HindⅢ酶切位点的引物,用其将pMD18-T-VP1质粒中的VP1基因亚克隆到真核表达载体pBlueBacHis2A中,成功地构建了重组表达质粒pBlueBacHis2A-VP1(633bp)。将pBlueBacHis2A-VP1(633 bp)质粒与Bac-N-BlueTMDNA共转染sf9昆虫细胞,蚀斑筛选重组病毒后,将纯化的重组病毒感染sf9细胞,获得了32.6 ku的目的蛋白条带。Dot-ELISA分析结果表明,该表达产物具有反应活性,可用于建立间接ELISA方法,进行口蹄疫病毒抗体检测。

According to the published VP1 gene sequence, one pair of primers with Sac Ⅰ and HindⅢ endonuclease sites was designed. A VPI gene in the plasmid pMD18-T-VPI was subcloned into an eukaryotic vector pBlueBacHis2A, and the recombinant plasmid pBlueBacHis2A-VP1 （633 bp） was constructed successfully. The plasmid pBlueBacHis2A-VPl（633 bp） and Bac-N-BlueTM DNA were co-transfected into sf9 cells. After screening the plaques, sf9 cells were infected with the recombinant virus. The target protein of 32.6 ku in size was obtained. Dot ELISA analysis showed that the protein had good antigenicity.