结核分枝杆菌Ag85B与IL-12基因真核共表达质粒的构建及表达

Construction and expression of the eukaryotic coexpression plasmid containing Ag85B gene of Mycobacterium tuberculosis and IL-12 gene

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。

AIM: To construct a eukaryotic coexpression plasmid containing Mycobacterium tuberculosis (MTB) Ag85B and IL-12 genes. METHODS: MTB Ag85B gene and IL-12 gene were cloned into pBudCE4.1 which has multiple promoters to construct recombinant plasmid pBud85B-IL12. The recombinant plasmid was transfected into COS-7 cells and the expression of target genes was assessed by RT-PCR and ELISA. (RESULTS:) The expressions of Ag85B and IL-12 could be detected in COS-7 cells. CONCLUSION: The recombinant plasmid pBud85B-IL12 was constructed and expressed successfully, which lays the foundation for further development of DNA vaccine against tuberculosis.