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.
A. Vannini, F. Agriesti, F. Mosca, D. Roncarati, V. Scarlato, A. Danielli (2012). A Convenient and Robust In Vivo Reporter System To Monitor Gene Expression in the Human Pathogen Helicobacter pylori. APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 78, 6524-6533 [10.1128/AEM.01252-12].
A Convenient and Robust In Vivo Reporter System To Monitor Gene Expression in the Human Pathogen Helicobacter pylori
VANNINI, ANDREA;RONCARATI, DAVIDE;SCARLATO, VINCENZO;DANIELLI, ALBERTO
2012
Abstract
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.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.