牛分枝杆菌ag85b基因的瞬时表达

[目的]为牛结核病DNA疫苗的研制提供参考。[方法]利用PCR技术扩增出牛分枝杆菌ag85b基因片段,克隆到真核载体pcD-NA3.1(+)上,构建重组质粒pcAg85B,将重组质粒转染SP2/0细胞,间接免疫荧光试验检测该基因的表达情况。[结果]结果表明:在转染了重组质粒pcAg85B的SP2/0细胞中出现绿色荧光,说明ag85b基因在SP2/0细胞中成功进行了瞬时表达。[结论]为研究牛分枝杆菌ag85b DNA疫苗奠定了基础。

Transient Expression of Mycobacterium bovis ag85b Gene

[Objective]The research aimed to provide references for the DNA vaccine development of bovine tuberculosis.[Method] The ag85b gene fragment amplified by PCR from Mycobacterium bovis was cloned into the eukaryotic expression vector pcDNA3.1( ), the recombinant plasmid pcAg85B was obtained. Then the recombinant plasmid was transfected into SP2/0 cells in vitro. Indirect immunofluorescence was used to detect the expression of target protein in SP2/0 cells transfected by pcAg85B.[Result] Indirect immunofluorescence which used to detect the expression of target protein showed SP2/0 cells transfected by pcAg85B appeared fluorescence.So the target protein had been successfully expressed. [Conclusion] This study laid a foundation for ag85b gene DNA vaccine of Mycobacterium bovis.