牛分枝杆菌融合基因ESAT6-MPB63-HSP65的原核表达及鉴定

›› 2009, Vol. 36 ›› Issue (9): 55-59.

• 生物技术 • Previous Articles Next Articles

Prokaryotic Expression and Identification of Fused ESAT6-MPB63-HSP65 Gene of Mycobacterium bovis

HAO Hui¹， HOU Shao-hua¹， DING Jia-bo²， MAO Kai-rong²， ZHU Hong-fei¹

(1.Institute of Animal Sciences, Chinese Academy of Agricultural Sciences， Beijing 100193, China； 2.China Institute of Veterinary Drug Control， Beijing 100081， China)

Received:1900-01-01 Revised:1900-01-01 Online:2009-09-20 Published:2009-09-20
Contact: ZHU Hong-fei

Abstract

Abstract: The genomic DNA of Mycobacterium bovis Vallee Ⅲ strain was used as template and the genes of ESAT6, MPB63 and HSP65 were amplified by PCR. Then these genes were tandemly inserted into pET-32a(+) to construct recombinant pET-E6-M63-H65. After the transfromation into Escherichia coli BL21 (DE3) competent cells，the fused protein was expressed effectively with 1 mmol/L IPTG induction and purified by Ni2+ purification system for Western blotting analysis. The results showed that the fused proteins were correctly in molecular weight and present in soluble status. Meanwhile, the purified protein can interact with the positive sera to M.bovis, which support important materials for the molecular diagnosis for M.bovis in the future.

Key words: Mycobacterium bovis; fused gene; prokaryotic expression; protein purification

CLC Number:

S852.61⁺8

HAO Hui;HOU Shao-hua;DING Jia-bo;MAO Kai-rong;ZHU Hong-fei. Prokaryotic Expression and Identification of Fused ESAT6-MPB63-HSP65 Gene of Mycobacterium bovis [J]. , 2009, 36(9): 55-59.

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Prokaryotic Expression and Identification of Fused ESAT6-MPB63-HSP65 Gene of Mycobacterium bovis

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