副结核分枝杆菌SOD基因在BHK-21细胞中的表达

Expression of Mycobacterium paratuberculosis SOD Gene in BHK-21 Cells

将重组质粒pGEM-T-SOD中的副结核分枝杆菌SOD基因亚克隆至真核表达载体pVAX1中,构建真核表达重组质粒pVAX1-SOD,以脂质体介导法转染至BHK-21细胞中,并采用RT-PCR和间接免疫荧光技术检测SOD基因在BHK-21细胞中的表达。结果显示,成功地构建了副结核分枝杆菌SOD基因的真核表达载体,且SOD基因在哺乳动物细胞中获得了表达。为研究副结核分枝杆菌SOD基因的免疫效果及作为牛副结核病DNA疫苗奠定基础。

Mycobacterium paratuberculosis SOD gene of the recombinant plasmid pGEM-T-SOD was subcloned into eukaryotic expression vector pVAX1 and construct the recombinant plasmid pVAX1-SOD.Transfect BHK-21 cells with the constructed recombinant plasmid in mediation of liposome,and observe the expression of SOD gene in BHK-21 cells by RT-PCR and indirect immunofluorescence technique.The results show that the eukaryotic expression vector for Mycobacterium paratuberculosis SOD gene was successfully constructed and expressed in mammalian cells,which laid a foundation of developing immune effect of Mycobacterium paratuberculosis SOD gene and DNA vaccine for bovine paratuberculosis.