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中国生物工程杂志

China Biotechnology
China Biotechnology  2010, Vol. 30 Issue (08): 72-75    DOI: Q786
    
Homologous Expression of M. Tuberculosis DnaA Protein in M. smegmatis
ZHOU Ai-ping,CHEN Yan-jiong,LI Wei,ZHANG Xu-yan,XU Ji-ru
Department of Medical Technology, Xi’an Jiaotong University, Xi’an 710061, China
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Abstract  

Objective To express Mycobacterium tuberculosis DnaA protein in Mycobacterium smegmatis and identify the expression product. Methods The dnaA gene of M. tuberculosis was amplified by PCR and cloned into expression vector pMF406 to generate the shuttle vector of E.coil and Mycobacterium pMF-dnaA. It was comfirmed by restriction endonuclease digestion and sequence analysis. The recombinant plasmid was transformed into M. Smegmatis mc2155 by electroporation. The recombinant M. smegmatis was induced by 0.02% acetamide and the expression product was analyzed with SDS-PAGE and Western blotting. Results The recombinant M. smegmatis was successfully constructed and the M. tuberculosis DnaA protein was expressed in the recombinant M. Smegmatis at a relatively high level identified by SDS-PAGE and Western blotting. Conclusion The successful homologous expression of the M. tuberculosis DnaA protein will be very helpful for the further study on the mechanism of M. tuberculosis DNA replication .



Key wordsM.tuberculosis      M.smegmatis      DnaA      Homologous expression     
Received: 24 March 2010      Published: 25 August 2010
Cite this article:

ZHOU Ai-Ping, CHEN Yan-Jiong, LI Wei, ZHANG Xu-Yan, XU Ji-Ru. Homologous Expression of M. Tuberculosis DnaA Protein in M. smegmatis. China Biotechnology, 2010, 30(08): 72-75.

URL:

https://manu60.magtech.com.cn/biotech/Q786     OR     https://manu60.magtech.com.cn/biotech/Y2010/V30/I08/72

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