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The journal of microbiology v.47 no.4, 2009년, pp.498 - 505  

Fusion Expression and Immunogenicity of EHEC EspA-Stx2A1 Protein: Implications for the Vaccine Development

Cheng, Yan    (Department of Clinical Microbiology and Immunology, College of Medical Laboratory, The Third Military Medical University   ); Feng, Youjun    (Department of Clinical Microbiology and Immunology, College of Medical Laboratory, The Third Military Medical University   ); Luo, Ping    (Department of Clinical Microbiology and Immunology, College of Medical Laboratory, The Third Military Medical University   ); Gu, Jiang    (Department of Clinical Microbiology and Immunology, College of Medical Laboratory, The Third Military Medical University   ); Yu, Shu    (Department of Clinical Microbiology and Immunology, College of Medical Laboratory, The Third Military Medical University   ); Zhang, Wei-Jun    (Department of Clinical Microbiology and Immunology, College of Medical Laboratory, The Third Military Medical University   ); Liu, Yan-Qing    (Department of Clinical Microbiology and Immunology, College of Medical Laboratory, The Third Military Medical University   ); Xang, Qing-Xu    (Department of Clinical Microbiology and Immunology, College of Medical Laboratory, The Third Military Medical University   ); Zou, Quan-Ming    (Department of Cl  ); Mao, Xu-Hu  
  • 초록

    Shiga toxin 2 (Stx2) is a major virulence factor for enterohemorrhagic Escherichia coli (EHEC), which is encoded by ${\lambda}$ lysogenic phage integrated into EHEC chromosome. Stx2A1, A1 subunit of Stx2 toxin has gathered extensive concerns due to its potential of being developed into a vaccine candidate. However, the substantial progress is hampered in part for the lack of a suitable in vitro expression system. Here we report use of the prokaryotic system pET-28a::espA-Stx2A1/BL21 to carry out the fusion expression of Stx2A1 which is linked to E. coli secreted protein A (EspA) at its N-terminus. Under the IPTG induction, EspAStx2A1 fusion protein in the form of inclusion body was obtained successfully, whose expression level can reach about 40% of total bacterial protein at $25^{\circ}C$ , much higher than that at $37^{\circ}C$ . Western blot test suggested the refolded fusion protein is of excellent immuno-reactivity with both monoclonal antibodies, which are specific to EspA and Stx2A1, respectively. Anti-sera from Ralb/c mice immunized with the EspA-Stx2A1 fusion protein were found to exhibit strong neutralization activity and protection capability in vitro and in vivo. These data have provided a novel feasible method to produce Stx2A1 in large scale in vitro, which is implicated for the development of multivalent subunit vaccines candidate against EHEC O157:H7 infections.


  • 주제어

    EHEC O157:H7 .   EspA .   Stx2A1 .   fusion protein .   vaccine.  

  • 참고문헌 (31)

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