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[Recombinant Helicobacter pylori urease B subunit and its biological properties]
Dong-shui Lu1, Xu-hu Mao, Quan-ming Zou
1Teaching and Research Section of Clinical Microbiology and Immunology, Department of Medical Laboratory, Third Military Medical University, Chongqing 400038, China. mxh95xy@mail.tmmu.com.cn
Objective:
To perform genetic recombination of the urease B subunit (UreB) of Helicobacter pylori (Hp) and examine the biological properties of the recombinant protein.
Methods:
The gene fragment encoding Hp UreB was isolated clinically from Chinese subjects by means of PCR, and cloned subsequently into an expression vector pET-11C-UreB for the non-fusion protein expression in E.coli BL21 (DE3) strain.
Results:
The expression of recombinant UreB was achieved in E.coli BL21 with a relative molecular weight of approximately 62,000 at the expression ratio of 26%, and the first 15 amino acids of recombinant UreB were MKKISREYVSMYGP. The results of peptide mapping and amino acid compositional analysis were consistent with previous theoretical prediction, and enzyme-linked immunosorbent assay together with Western blotting indicated strong immunogenicity and reactivity of the recombinant protein in BalB/c mice, which were specifically recognized by polyclonal BalB/c mice anti-Hp sera or human sera infected with Hp.
Conclusion:
The results of this study has laid an solid immunological foundation for incorporating recombinant UreB as a subunit vaccine component against Hp.