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目的 :构建结核分枝杆菌Ag85B和鼠IL 12基因的共表达载体pBud85B IL12。方法 :将结核分枝杆菌Ag85B基因和鼠IL 12基因同时克隆入含多启动子的共表达载体pBudCE4 .1中 ,构建真核共表达质粒pBud85B IL12。以pBud85B IL12转染COS 7细胞 ,通过RT PCR及ELISA方法检测目的基因的表达。结果 :在COS 7细胞中同时可检测到Ag85B和IL12的表达。结论 :pBud85B IL12共表达质粒的成功构建 ,为对其免疫原性、免疫反应性及免疫保护作用的进一步研究奠定了基础
Objective: To construct the co-expression vector pBud85B IL12 of Mycobacterium tuberculosis Ag85B and murine IL 12 gene. Methods: Mycobacterium tuberculosis Ag85B gene and murine IL-12 gene were cloned into pBudCE4. 1 containing multiple promoters simultaneously to construct eukaryotic co-expression plasmid pBud85B IL12. COS 7 cells were transfected with pBud85B IL12, and the expression of the target gene was detected by RT PCR and ELISA. Results: The expression of Ag85B and IL12 was detected simultaneously in COS 7 cells. CONCLUSION: The successful construction of the pBud85B IL12 co-expression plasmid lays the foundation for further research on its immunogenicity, immunoreactivity and immune protection