[Recombinant Helicobacter pylori urease B subunit and its biological properties].
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.


CONCLUSIONS
The results of this study has laid an solid immunological foundation for incorporating recombinant UreB as a subunit vaccine component against Hp.
Lu
Dong-shui
DS
Teaching and Research Section of Clinical Microbiology and Immunology, Department of Medical Laboratory, Third Military Medical University, Chongqing 400038, China. mxh95xy@mail.tmmu.com.cn
Mao
Xu-hu
XH
Zou
Quan-ming
QM
Wu
Chao
C
Yang
Jun
J
Zhang
Wei-jun
WJ
Wang
Fu-kun
FK
Xie
Qing-hua
QH
Luo
Ping
P
D000818:Animals; D001428:Bacterial Vaccines / Q000276:immunology*; D016480:Helicobacter pylori / Q000201:enzymology* / Q000276:immunology*; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D010449:Peptide Mapping; D021122:Protein Subunits; D011994:Recombinant Proteins / Q000032:analysis / Q000276:immunology; D014510:Urease / Q000032:analysis / Q000235:genetics / Q000276:immunology*; D022223:Vaccines, Subunit / Q000276:immunology; D014614:Vaccines, Synthetic / Q000276:immunology*
D001428:Bacterial Vaccines; D021122:Protein Subunits; D011994:Recombinant Proteins; D022223:Vaccines, Subunit; D014614:Vaccines, Synthetic; D014510:Urease