title: [The New Bacteria Expressing Recombinant Multi-epitope Vaccine against Helicobacter pylori and Its Microbiological Characteristics].
abstract: OBJECTIVE
  To construct the engineering bacteria with recombinant plasmid expressing the multi-epitope vaccine which composed of Helicobacter pylori urea membrane channel protein (UreI), Helicobacter pylori urease B subunit (UreB) and cholera toxin B subunit (CTB), and then to study it's microbiological characteristics.
  
  
  METHODS
  The sequence contains some dominant epitopes of Helicobacter pylori UreI and UreB was designed, and ctB was added at the N-terminal, all the sequence were linked by flexible linkers. Codon optimization was done according to Escherichia coli (E. coli) BL21 (DE3) bias, the optimized sequence was designated BIB. BIB sequence was synthesized and cloned into plasmid pET28a(+). The recombinant plasmid was confirmed by restriction enzyme digestion and DNA sequencing. The recombinant protein BIB was expressed in E. coli BL21 (DE3) and analyzed by Western blot.
  
  
  RESULTS
  The plasmid of pET28a(+)/BIB was constructed successfully, confirmed by restriction enzyme digestion and DNA sequencing. The recombinant protein BIB with relative molecular mass about 33 x 10(3) could be produced by E. coli BL21 (DE3) and was detected by Western blot. The relative molecular mass and N-terminal amino acid sequence of BIB were 100% identity with the design.
  
  
  CONCLUSIONS
  The engineering bacteria with recombinant plasmid expressing the multi-epitope vaccine against Helicobacter pylori was constructed successfully. The recombinant protein BIB can be identified by anti-Sydney strain 1 of Helicobacter pylori (H. pylori SS1) polyclonal antibody and anti-CTB monoclonal antibody, which demonstrated that BIB has the expected antigenicity.
authors.forename: Bao-ning
authors.initials: BN
authors.lastname: Wang
authors.forename: Xing
authors.initials: X
authors.lastname: Pan
authors.forename: Xiao-jun
authors.initials: XJ
authors.lastname: Huang
authors.forename: Yong-jun
authors.initials: YJ
authors.lastname: Zhou
authors.forename: Jie
authors.initials: J
authors.lastname: Zhu
authors.forename: Li-zhen
authors.initials: LZ
authors.lastname: Gao
authors.forename: Xiao-juan
authors.initials: XJ
authors.lastname: Niu
authors.forename: Wan-yi
authors.initials: WY
authors.lastname: Li
authors.forename: Ming-yuan
authors.initials: MY
authors.lastname: Li
authors.forename: Hong-ren
authors.initials: HR
authors.lastname: Wang
mesh_terms: D001426:Bacterial Proteins / Q000276:immunology; D001428:Bacterial Vaccines / Q000096:biosynthesis / Q000276:immunology*; D002772:Cholera Toxin / Q000276:immunology; D000939:Epitopes / Q000276:immunology*; D004926:Escherichia coli; D016480:Helicobacter pylori; D026901:Membrane Transport Proteins / Q000276:immunology; D010957:Plasmids / Q000096:biosynthesis; D014614:Vaccines, Synthetic / Q000096:biosynthesis / Q000276:immunology
chemical_list: D001426:Bacterial Proteins; D001428:Bacterial Vaccines; D000939:Epitopes; D026901:Membrane Transport Proteins; C114527:UreI protein, Helicobacter pylori; D014614:Vaccines, Synthetic; D002772:Cholera Toxin