Applied and Environmental Microbiology · 2012 · 13 citations · 29 references
Life SciencesBiotechnologyGeneticsGene ExpressionGene RegulationMicrobial GeneticsBacterial Gene RegulationGene Expression ImagingGene Expression ProfilingPathogen DetectionPromoter EngineeringPromoter RegulationBioluminescent Gene ExpressionBioluminescent ReportersFluorescent Reporter SystemsGene Expression ReportersGenetically Encoded Fluorescent ReportersLuminescent Reporter SystemsPromotersReporter Gene Assays
Thirty years of intensive research have significantly contributed to our understanding of Helicobacter pylori biology and pathogenesis. However, the lack of convenient genetic tools, in particular the limited effectiveness of available reporter systems, has notably limited the toolbox for fundamental and applied studies. Here, we report the construction of a bioluminescent H. pylori reporter system based on the Photorhabdus luminescens luxCDABE cassette. The system is constituted of a promoterless lux acceptor strain in which promoters and sequences of interest can be conveniently introduced by double homologous recombination of a suicide transformation vector. We validate the robustness of this new lux reporter system in noninvasive in vivo monitoring of dynamic transcriptional responses of inducible as well as repressible promoters and demonstrate its suitability for the implementation of genetic screens in H. pylori.
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Molecular cloning: A laboratory manual
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FACS-optimized mutants of the green fluorescent protein (GFP)
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The primary transcriptome of the major human pathogen Helicobacter pylori
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