期刊
WORLD JOURNAL OF GASTROENTEROLOGY
卷 11, 期 3, 页码 454-456出版社
BAISHIDENG PUBL GRP CO LTD
DOI: 10.3748/wjg.v11.i3.454
关键词
Helicobacter pylori; Neutrophil-activating protein; Maltose-binding protein; E. coli
资金
- Medical Science Foundation for Distinguished Scholars of Henan Province [200084]
AIM: To produce a recombinant protein rMBP-NAP, which was fusionally expressed by Helicobacter pylori (H pylori) neutrophil-activating protein (NAP) and E. coli maltose-binding protein (MBP) and to evaluate its immunoreactivity and immunogenicity. METHODS: Neutrophil-activating protein gene of H pylori (HP-napA) was subcloned from the recombinant plasmid pNEB-napA, and fused to MalE gene of expressing vector pMAL-c2x. The recombinant plasmid pMAL-c2x-napA was confirmed by restriction enzyme digestion, and then transformed into E. coli TB1. Fusion protein rMBP-NAP was induced by IPTG and identified by SDS-PAGE analysis. Soluble rMBP-NAP was purified by amylose affinity chromatography. Immunoreactivity and immunogenicity of the fusion protein were evaluated by animal experiment, Western blotting with human H pylori anti-sera. RESULTS: E. coli TB1 carrying recombinant plasmid pMAL-c2x-napA was constructed and led to a high efficiency cytosol expression of fusion protein rBMP-NAP when induced by IPTG. The molecular weight of rBMP-NAP was about 57 kD, accounting for 37.55% of the total protein in the sonicated supernatant of E. coli TB1 (pMAL-c2x-napA). The purity of the fusion protein after one-step affinity chromatography was 94% and the yield was 100 mg per liter of bacterial culture. The purified fusion protein could be specifically recognized by both human anti-sera from clinical patients with H pylori infection and rabbit sera immunized by rMBP-NAP itself. CONCLUSION: Recombinant protein rMBP-NAP might be a novel antigen for vaccine development against H pylori. (C) 2005 The WJG Press and Elsevier Inc. All rights reserved.
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