论文部分内容阅读
AIM:In vivo induced genes are thought to play an importantrole during infection of host.AlkA was identified as an invivo-induced gene by in vivo expression technology (IVET),but its virulence in Shigella flexneri was not reported.Thepurpose of this study was to identify the role of alkA gene inthe pathogenesis of S.flexneri.METHODS:PCR was used to amplify alkA gene of S.flexneri2a and fragment 028pKm.The fragment was thentransformed into 2457T05 strain,a S flexneri 2a straincontaining Red recombination system,which was constructedwith a recombinant suicide plasmid pXLkd46.By in vivohomologous recombination,alkA mutants were obtained andverified by PCR and sequencing.Intracellular survival assayand virulence assay were used to test the intracellular survivalability in HeLa cell model and the virulence in mice lunginfection model respectively.RESULTS:Deletion mutant of S.flexneri 2a alkA wassuccessfully constructed by λ Red recombination system.The mutant exhibited significant survival defects and muchsignificant virulence defects in mice infection assay.CONCLUSION: AlkA gene plays an important role in theof epithelial cells and is a virulent gene of Shigella spp.
AIM: In vivo induced genes are thought to play an importantrole during infection of host. AlkA was identified as an invivo-induced gene by in vivo expression technology (IVET), but its virulence in Shigella flexneri was not reported .purpose of this study was to identify the role of alkA gene inthe pathogenesis of S. flexneri. METHODS: PCR was used to amplify alkA gene of S.flexneri 2a and fragment 028pKm.The fragment was thentransformed into 2457T05 strain, aS flexneri 2a straincontaining Red recombination system, which was constructed with a recombinant suicide plasmid pXLkd46.By in vivohomologous recombination, alkA mutants were obtained and verified by PCR and sequencing. Intracellular survival assay and virulence assay were used to test the intracellular survivability in HeLa cell model and the virulence in mice lung infection model respectively .RESULTS: Deletion mutant of S.flexneri 2a alkA wassuccessfully constructed by λ Red recombination system. mutantImmatory highlights survival defects and much more than virulence defects in mice infection assay. CONCLUSION: AlkA gene plays an important role in the of epithelial cells and is a virulent gene of Shigella spp.