Project description:MepR is a substrate-responsive repressor of mepR and mepA, which encode itself and a MATE family multidrug efflux pump. Microarray analyses of Staphylococcus aureus SH1000 and its mepR-disrupted derivative revealed changes in expression of many genes in addition to mepR and mepA, notably several involved in virulence Keywords: Staphylococcus aureus, MATE efflux pump, MepR Staphylococcus aureus strains SH1000 wildtype and mepR were grown in duplicate to exponential and post-exponential phase (corresponding to an A550 nm of 0/4 and 2.0 respectively). RNA was harvested, converted to cDNA, labelled with Biotin and used to probe custom-designed Affymetrix antisense S.aureus GeneChips. Eight samples in total were prepared and analyzed.
Project description:MepR is a substrate-responsive repressor of mepR and mepA, which encode itself and a MATE family multidrug efflux pump. Microarray analyses of Staphylococcus aureus SH1000 and its mepR-disrupted derivative revealed changes in expression of many genes in addition to mepR and mepA, notably several involved in virulence Keywords: Staphylococcus aureus, MATE efflux pump, MepR
Project description:We have demonstrated previously that high-level resistance to nisin can occur in Staphylococcus aureus as a consequence of a single non-synonymous mutation in nsaS, which encodes a putative sensor kinase. To explore the mechanism by which this mutation confers high-level resistance we compared global transcriptomes of SH1000 and SH1000 (NsaS A208E) using RNAseq. This process identified several genes to be upregulated in SH1000 (NsaS A208E), including members of the NsaRS regulon which encode VraDE and BraDE, two putative ABC-transporters and are known to provide intrinsic nisin resistance. Gene deletion and complementation experiments revealed that both BraDE and VraDE are essential to high-level nisin resistance, with BraDE required for signal transduction through NsaRS, and VraDE directly responsible for nisin detoxification.
Project description:A library of transposon mutants were generated in S.aureus strain SH1000 using a custom Mariner transposon with an outward-facing T7 promoter. Genomic DNA was extracted from the library and digested with an appropriate restriction enzyme (AluI or RsaI). Labelled RNA run-offs were produced from the T7 promoter and hybridised to a tiling microarray to determine the position of the mutant. This was done using a library of a million S. aureus mutants, and genes not disrupted by a transposon were inferred to be essential for replication and growth in vitro.
Project description:The transcription level of a rex-deficient S. aureus mutant in comparison to its parental strain S. aureus SH1000 was analyzed using DNA microarrays.