The Research on the Construction and Characteristics of Recombinant Engineering Bacteria with Multi-epitope of Helicobacter pylori

DING Na-na, YANG Jing, PAN Xing.et al

Abstract

To construct the multi-epitope prokaryotic expression plasmid and appropriate engineering bacteria expressing the multi-epitope fusion protein of urea membrane channel protein (UreI), urease B subunit (UreB) and adhesin (HpaA) ofHelicobacter pylorithen study its microbiological characteristics. Methods The target sequence contains multi-epitope gene sequence of Helicobacter pylori were designed and synthesized, subsequently; it was subcloned into the expression vector pET28a (+), confirmed by restriction enzyme digestion and DNA sequencing. The fusion protein rIBA was expressed in E.coli Rosseta (DE3) and analyzed by Western blot. Results The plasmid of pET28a(+)/IBA was constructed successfully, confirmed by endonuclease digestion and sequence analyze. The expressed rIBA protein with relative molecular mass about 40×10 3 and can be detected by Western blot. Conclusion The prokaryotic engineering bacteria expression multi-epitope of the Helicobacter pylori was constructed successfully. The recombinant protein rIBA expressed by the engineering bacteria can be identified by Sydney strain 1 of Helicobacter pylori (H. pylori SS1) specific antibody IgY, which demonstrated that the rIBA has high correlation with H.pylori SS1.

 

Keywords: Helicobacter pylori, Epitope, Urease, Adhesin, Engineering bacteria 

 

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References


Cui R, Zhou L. Helicobacter pylori infection: an overview in 2013, focus on therapy . Chin Med J (Engl), 2014; 127 (3 ) ; 568-573.

Moeller H, Correa P. Carcinogenicity of some biological agents. Cancer Epidemiol Biomarkers Prev,1994;3;627. 2012;21(8): 687-690.

Malfertheiner P. Megraud F. O, Morain CA, et al. Management of Helicobacter pylori infection-the Maastricht IvFlorence Consensus Report . Gut. 2012; 61 (5) :646-664. Backert S. Clyne M. Pathogenesis of Helicobacter pylori infection. Helicobacter, 2011; 16(Suppl 1): 19-25.

Kerstin S. Karlheinz A, Evert PB. Acid survival of Helicobacter pylori: how dose urease activity trigger cytoplasmic pH homeostasis? Trends Microbiol. 2002; 10 ( 2 ); 70-74.

Scott DR. Marcus ЕА, Weeks DL, et al. Expression of the Helicobacter pylori urel gene is required for acidic pH activation of cytoplasmic urease . Infect Irnmun, 2000; 68(2); 470-477.

Weeks DL. Scott DR. Sachs G. A H-gated urea channel; the link between Helicobacter pylori urease and gastric colonization. Science, 2000; 287(5452): 482-485.

Strugatsky D. McNulty R. Munson K. et al. Structure of the proton-gated urea channel from the gastric pathogen Helicobacter pylori. Nature.2013;493(7431):255-258.

Flach CF, Svensson N, Blomquist M, etal. A truncated form of HpaA is a promising antigen for use in a vaccine against Helicobacter pylori. Vaccine,2011;29(6) ; 1235-1241.

Huang X, Xu В, Duan G, et al. The rOmp22-HpaA fusion protein confers protective immunity against Helicobacter pylori in mice . Curr Microbiol, 2013; 67(4): 487-492.


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