结核分枝杆菌Ag85A DNA的克隆与原核表达

Clone and Prokaryotic Expression of Mycobacterium tuberculosis Ag85A DNA

抗原85(Ag85)复合体是BCG合成分泌的可刺激机体产生细胞免疫和体液免疫应答。Ag85A是抗原85复合体组成成分之一,可显著刺激细胞免疫功能增强。研究构建了高效表达的pTrcHisB-Ag85A基因重组质粒,并将其转化至大肠埃希菌TOP10中进行重组蛋白的表达,筛选转化物,SDS-PAGE进行蛋白表达的分子量检测,并通过Western Blotting方法,应用抗Ag85A的单克隆抗体进一步确认重组Ag85A蛋白的表达,建立了Ag85A的原核表达体系,为进一步制备出有效的重组Ag85A疫苗奠定基础。

Antigen 85 （Ag85） complex is synthetic secretion BCG that can stimulate organism to produce cell immune and body fluid immune response. It can remarkably stimulate cell immune to strengthen. High-effective gene recombinant plasmid pTrcHisB-Ag85A was constructed in this study, and transferred into E. coli TOPIO to carry out recombinant protein expression, then screen for transformants, test molecularly for protein expression by SDS-PAGE, and further test for confirming the recombinant protein expression of Ag85A with its monoelonal antibody through Western Blotting, thus to construct prokaryotie expression system of Ag85A to lay a foundation for further preparation effective recombinant Ag85A vaccine.