Prior exposure strongly influences mechanisms underpinning survival of heat shock in Escherichia coli
In past, heat stress mechanisms have been identified by analysis of RNA-seq or proteomics. 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 heat stress at different temperatures. We used two approaches for heat stress, direct heat (heat shock) and stepwise heat increase (gradient heat) stress. Approximately 10^8 cells from the mutant library were inoculated into LB broth at 37C, 44C, 47C, 50C, 44C to 47C and 44C to 50C. Experiments were performed with induction using 0.2mM of Isopropyl ß-D-1-thiogalactopyranoside (IPTG). All experiments were performed in triplicate.
In past, heat stress mechanisms have been identified by analysis of RNA-seq or proteomics. 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 heat stress at different temperatures. We used two approaches for heat stress, direct heat (heat shock) and stepwise heat increase (gradient heat) stress. Approximately 10^8 cells from the mutant library were inoculated into LB broth at 37C, 44C, 47C, 50C, 44C to 47C and 44C to 50C. Experiments were performed with induction using 0.2mM of Isopropyl ß-D-1-thiogalactopyranoside (IPTG). All experiments were performed in triplicate.
European Bioinformatics Institute