Project description:TraDIS study on Salmonella Typhi subjected to serum bactericidal assays.These data are part of a pre-publication release. For information on the proper use of pre-publication data shared by the Wellcome Trust Sanger Institute (including details of any publication moratoria), please see http://www.sanger.ac.uk/datasharing/
Project description:We used TraDIS-Xpress to determine the mechanism of action of a novel antimicrobial compound. We found that it inhibits lipid IVA biosynthesis in both Escherichia coli and Salmonella enterica serovar Typhimurium. We also were able to determine mechanisms of synergy with colistin, through ATP biosynthesis and the BasSR signalling system.
Project description:Salmonella Typhimurium is a major human pathogen, and additionally serves as a model system for the study of both intracellular pathogens and bacterial small non-coding RNAs (sRNAs). Over the last decade, it has become increasingly clear that sRNAs play central roles in the regulation of a number of cellular processes, including quorum sensing, metabolism, and various stress responses. However, the contribution of sRNAs to virulence remains largely unexplored. Recent and preliminary RNA-seq and TraDIS data from our lab and others suggests that a number ofsRNAs are induced during infection of both epithelial and immune cells, and furthermore that their deletion leads to measurable replication phenotypes in whole animal model systems. However, these experiments leave the mechanisms underlying these phenotypes largely unexplained.We have used the TraDIS method to perform genetic interaction screens for S. Typhimurium sRNAs in cell models of infection, which will provide direct insight in to the processes affected by sRNA regulation during infection. This data is part of a pre-publication release. For information on the proper use of pre-publication data shared by the Wellcome Trust Sanger Institute (including details of any publication moratoria), please see http://www.sanger.ac.uk/datasharing/
Project description:TraDIS study to identify novel immunity proteins and their effector proteins associated with the Type VI secretion system (T6SS) in Pseudomonas aeruginosaThese data are part of a pre-publication release. For information on the proper use of pre-publication data shared by the Wellcome Trust Sanger Institute (including details of any publication moratoria), please see http://www.sanger.ac.uk/datasharing/
Project description:In past, resistance mechanisms have been identified by analysis of resistant isolates or defined mutants. Recently, high-throughput transposon mutagenesis coupled with sequencing (TraDIS-Xpress) is another approach proving useful for elucidating the roles of genes involved in the overall cellular response to a particular stress. In this study, we used TraDIS-Xpress to determine the role played by genes following exposure to colistin stress. Approximately 10^7 cells from the mutant library were inoculated into LB broth at a range of doubling concentrations of colistin ( 0.25 x MIC, 0.5 x MIC, 1 x MIC, 2 X MIC). Experiments were performed with no induction, or with induction using 0.2 or 1 mM of Isopropyl β-D-1-thiogalactopyranoside (IPTG). All experiments were performed in duplicate.
Project description:This study investigates the mechanisms employed by Salmonella to colonise and establish itself on fresh produce at critical timepoints following infection. We established an alfalfa infection model and compared the findings to those obtained from glass surfaces. Our research revealed dynamic changes in the pathways associated with biofilm formation over time, with distinct plant-specific and glass-specific mechanisms for biofilm formation, alongside the identification of shared genes playing pivotal roles in both contexts.