title: [Establishment of an attenuated salmonella typhimurium vaccine strain SL3261 expressing Hp-NAP gene].
abstract: OBJECTIVE
  To develop an attenuated salmonella typhimurium vaccine strain SL3261 expressing Helicobacter pylori neutrophile activation protein(Hp-NAP).
  
  
  METHODS
  The Hp-NAP gene was amplified by PCR and was cloned into prokaryotic expression plasmid pTrc99A to construct recombinant plasmid pTrc99A-NAP. After the pTrc99A-NAP was performed successively by PCR, enzyme digestion analysis and sequencing the homology was compared between the cloned Hp-NAP gene and related genes in GenBank. The vaccine strain SL3261 was transformed by pTrc99A-NAP. After cultivation, positive colonies were picked out and the plasmid was amplified by PCR and enzyme digestion analysis again. The expression of Hp-NAP protein in the SL3261 bacteria was proved by SDS-PAGE and thin layer scanning.
  
  
  RESULTS
  Sequencing and homologous analysis showed that the homology between the cloned Hp-NAP gene and related genes in GenBank reached 98% for their nucleotide and deduced amino acid sequences. The expression of Hp-NAP protein could be detected in the culture fluid of SL3261 bacteria. Relative molecular mass (M(r)) of expression product was about 15 000, and expression amount accounted for 37.5% of total bacterial protein.
  
  
  CONCLUSIONS
  An attenuated salmonella typhimurium vaccine strain SL3261 has been established successfully, which lays the foundation for further developing oral HP vaccine.
authors.affiliation: Department of Gastroenterology, The First Affiliated Hospital, Sun Yat-Sen University, Guangzhou 510080,China.
authors.forename: Zhi-Yong
authors.initials: ZY
authors.lastname: Jiao
authors.forename: Min-Hu
authors.initials: MH
authors.lastname: Chen
authors.forename: Sen-Lin
authors.initials: SL
authors.lastname: Zhu
authors.forename: Guo-Qing
authors.initials: GQ
authors.lastname: Li
authors.forename: Jie
authors.initials: J
authors.lastname: Chen
authors.forename: Pin-Jin
authors.initials: PJ
authors.lastname: Hu
mesh_terms: D001428:Bacterial Vaccines / Q000276:immunology; D004591:Electrophoresis, Polyacrylamide Gel; D016480:Helicobacter pylori / Q000276:immunology; D012486:Salmonella typhimurium / Q000235:genetics; D014613:Vaccines, Attenuated / Q000235:genetics
chemical_list: D001428:Bacterial Vaccines; D014613:Vaccines, Attenuated