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| Recombinant Expression of Mycobacterium tuberculosis Protein ESAT-6 and the Study about Its Binding to Cell Membrane |
| LI Hao1, YIN Ying1, MAO Ya-li2, DONG Da-yong1, ZHANG Jun1, FU Ling1, GUO Ji-hong2, XU Jun-jie1, CHEN Wei1 |
1. State Key Laboratory of Pathogen and Biosecurity, Institute of Microbiology and Epidemiology, Beijing 100071,China;
2. Department of Pharmacy, Air Force General Hospital, Beijing 100142,China |
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Abstract ESAT-6 gene was amplified by PCR from the genome of Mycobacterium tuberculosis H37Rv, and then cloned into pET21a(+) plasmid. The recombinant plasmid that was successfully constructed was transformed into E.coli BL21(DE3). After induced with IPTG, the expressed recombinant protein was confirmed by SDS-PAGE and Western blot. The vector yielded satisfactory levels of recombinant ESAT-6 protein expressed as a soluble protein in E. coli. After ultrasonication, the recombinant ESAT-6 protein was firstly purified by a column packed with Ni-NTA Resin and then a column packed with DEAE-SepharoseTM Fast Flow matrix. The purity of the purified protein was about 95%. The purified ESAT-6 protein was incubated with RAW264.7 cells, and the result got by Immunofluorescence showed that ESAT-6 could directly bind to the macrophage membrane.
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Received: 10 November 2010
Published: 27 May 2011
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Cite this article:
LI Hao, YIN Ying, MAO Ya-li, DONG Da-yong, ZHANG Jun, FU Ling, GUO Ji-hong, XU Jun-jie, CHEN Wei. Recombinant Expression of Mycobacterium tuberculosis Protein ESAT-6 and the Study about Its Binding to Cell Membrane. China Biotechnology, 2011, 31(5): 55-59.
URL:
https://manu60.magtech.com.cn/biotech/ OR https://manu60.magtech.com.cn/biotech/Y2011/V31/I5/55
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