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Helicobacter pylori  is a Gram-negative, microaerophilic bacterium associated with human gastric diseases. Further investigations on virulence genes are still required to clarify the pathogenic mechanism of  H. pylori  and the heterogeneous problem of infection. In order to develop an efficient and accurate method to study gene functions in  H. pylori  pathogenesis, an unmarked deletion method for both a single gene and a large fragment was established based on the FLP-FRT recombination system. Using this method, the gene  hp0788 , encoding an outer membrane protein (HofF), was deleted. Deletion of  hp0788  did not affect growth or motility of  H. pylori , but reduced the adherence of the bacteria to gastric epithelial cells. The apoptosis of GES-1 cells caused by  H. pylori  infection was also reduced by the defection of  hp0788 . These suggest that  hp0788  takes part in the bacterium-host interaction and plays an important role in  H. pylori  infection. Furthermore, a large genomic fragment deletion from  hp0541  to  hp0547  in  cag  pathogenicity island was also successfully achieved using FLP-FRT method. The innovative application of the FLP-FRT recombination system in  H. pylori  to construct unmarked deletion would provide a helpful tool for further function research of putative pathogenic genes and contribute to the understanding of  H. pylori  pathogenesis.

Application of FLP-FRT System to Construct Unmarked Deletion in Helicobacter pylori and Functional Study of Gene hp0788 in Pathogenesis