结核分支杆菌抗原ESAT-6真核表达质粒在COS-7中的表达

The Expression of Eukaryotic Expression Recombinant Plasmid of ESAT-6 from Mycobacterium Tuberculosis in COS-7 Cells

目的 探讨结核分支杆菌早期分泌性抗原靶ESAT -6基因真核表达重组质粒在真核细胞 (COS -7)中的表达。 方法 用LipofectAMINETM2 0 0 0介导真核表达重组质粒 pcDNA3 .1(+ ) -ESAT -6转染真核细胞COS -7,72h后 ,通过RT -PCR和WesternBlotting试验鉴定ESAT -6基因在COS -7中的表达。 结果 通过RT -PCR从转染了pcDNA3 .1(+ ) -ESAT -6的COS -7中检测到目的基因mRNA的转录 ;WesternBlotting试验鉴定在转染pcDNA3 .1(+ ) -ESAT -6的COS -7的细胞裂解上清液中有ESAT -6基因的表达蛋白。 结论 结核杆菌早期分泌性蛋白ESAT -6真核表达重组质粒能够在真核细胞COS -7中表达ESAT -6蛋白 ,为进一步研究其特性及为结核病的诊断。

Objective To investigate transient expression of eukaryotic expression plasmid of ESAT-6 in eukaryotic cells(COS-7). Methods Eukaryotic expression plasmid pcDNA3.1(+)-ESAT-6 were transfected into eukaryotic cells(COS-7), and the transient expression of ESAT-6 gene in eukaryotic cells was detected by RT-PCR and Western Blotting. Result Transient expression of gene ESAT-6 in eukaryotic cells(COS-7) was confirmed by RT-PCR and Western Blotting. Conclusion The recombinant eukaryotic expression plasmid pcDNA3.1(+)-ESAT-6 expressed successfully in eukaryotic cells(COS-7).It lays the foudation for diagnosis and using of new vaccine of tuberculosis and the detecting of immunity effectiveness.