Time filter

Source Type

Yang T.,Sichuan Agricultural University | Yang J.,University of Sichuan | Wang B.,University of Sichuan | Pan X.,University of Sichuan | And 5 more authors.
Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology | Year: 2015

OBJECTIVE: To prepare and purify anti-Helicobacter (H.) pylori Sydney Strain 1 (SS1) immunoglobulin Y (IgY) and anti-recombinant Ure I-UreB protein (rIB) IgY and investigate their antibacterial effects in vitro.METHODS: rIB protein and SS1 cell solution were treated by ultrasonic disruption and used as immunogens to immune Leghorn hens. SS1-IgY and IB-IgY were derived from egg yolk with method of the water dilution (WD) combined with ammonium sulfate precipitations and analyzed by growth inhibition test (GIT) and rapid urease test (RUT).RESULTS: Two specific IgY with purity of over 80% and titer over 1:12 800 were prepared successfully. Both SS1-IgY and IB-IgY inhibited the growth of H.pylori and reduced H.pylori urease activity in vitro markedly.CONCLUSION: SS1-IgY and IB-IgY can inhibit the growth of H.pylori and reduce the urease activity in vitro. Source

Wang B.-N.,University of Sichuan | Pan X.,University of Sichuan | Pan X.,Sichuan Vac Technology Co. | Huang X.-J.,University of Sichuan | And 7 more authors.
Journal of Sichuan University (Medical Science Edition) | Year: 2015

Objective: To construct the engineering bacteria with recombinant plasmid expressing the multiepitope 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 33×103 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. Conclution: 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 SSI) polyclonal antibody and anti-CTB monoclonal antibody, which demonstrated that BIB has the expected antigenicity. Source

Discover hidden collaborations