›› 2012, Vol. 32 ›› Issue (11): 1444-.doi: 10.3969/j.issn.1674-8115.2012.11.011

• Monographic report (Pathogenic microbiology) • Previous Articles     Next Articles

Cloning, expression and immune response in mice of specific antigen Rv3117 from M. tuberculosis

TANG Jun-ming1, CHEN Cui-cui2, WANG Xue-cai1, ZHAO Jun-wei3, ZHANG Shu-lin3   

  1. 1.Yixing People's Hospital, Yixing 214200, China；2.The First Affiliated Hospital of Zhengzhou University, Zhengzhou 450001, China；3.Department of Microbiology and Parasitology, Shanghai Jiaotong University, Shanghai 200025, China
  • Online:2012-11-28 Published:2012-11-30
  • Supported by:

    National Natural Science Foundation of China, 81271794；Shanghai Science and Technology Committee Foundation, 12441903300

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Abstract:

Objective To clone recombinant protein Rv3117 from M. tuberculosis, and investigate its immune response in mice. Methods The gene encoding Rv3117 protein was amplified by PCR from genome DNA of M. tuberculosis, and was then cloned into corresponding site of the expression vector pET-32a. The recombinant plasmid pET32a-Rv3117 was transformed into E.coli BL21 plysS (DE3), induced with isopropyl-β-D-thiogalactopyranoside (IPTG), and purified by Ni-NTA purification system. The expression of recombinant protein was identified by SDS-PAGE analysis. Western blotting was performed on sera from Rv3117-immunized C57BL/6 mice. Results The prokaryotic expression vector pET32a-Rv3117 was successfully constructed. The target protein was expressed in soluble form in E.coli BL21 plysS (DE3), with the relative molecular weight of 48 100, which was in line with the expectations. Western blotting revealed a positive stripe at the destination. Conclusion The recombinant protein Rv3117 has been successfully cloned from M. tuberculosis, which could induce immune response in mice.

Key words: M. tuberculosis, Rv3117, clone, Western blotting