Project description:The regulatory role of the Fis protein in fis and in the transcription of several gene regions during mid-exponential and late-stationary phase, and during different growth aeration regimes, has been investigated. Studies were done during those two growth phases and in aerated and non-aerated (microaerobic) conditions, to measure Fis enrichment and binding peaks in strategic gene regions by genome-wide microarray analysis ChIP-chip. This research investigation points to central roles for SPI-1, SPI-2, DNA gyrase and topoisomerase I, the elements of the stringent response, and the regulatory function of Fis-binding patterns, in setting and re-setting the activity of the fis gene and other involved promoters as a function of the growth conditions and aeration regimes experienced by Salmonella.
Project description:Investigation of gene expression level changes in Salmonella Typhiumurium SL1344 (R27) compared to the wild-type SL1344 strain when grown at different growth temperatures and growth phases. A 24 microarray study was performed using total RNA recovered from three separate wild-type cultures of SL1344 and three separate cultures of SL1344 (R27) grown to exponential and stationary phase at 25oC and 37oC. Each microarray measured the expression level of 4,527 genes from Salmonella Typhimurium SL1344 chromosome, 103 genes from plasmid pSLT, 100 genes from plasmid pRSF, 14 genes from plasmid pCOL1B and 207 genes from plasmid R27. Seven probes were present per transcript, with two-fold technical redundancy.
Project description:Investigation of gene expression level changes in Salmonella Typhiumurium SL1344 (R27) compared to the wild-type SL1344 strain when grown at different growth temperatures and growth phases.
Project description:ChIP-chip analysis of RNA Polymerase (RNAP), RpoD, RpoE, RpoH and RpoN in exponential phase (OD 0.2) and early stationary phase (OD 2.0) Salmonella enterica serovar Typhimurium SL1344 cultures
Project description:FabR ChIP-chip on Salmonella enterica subsp. enterica serovar Typhimurium SL1344 using anti-Myc antibody against strain with chromosomally 9Myc-tagged FabR (IP samples) and wildtype strain (mock IP samples)
Project description:The regulatory role of the Fis protein in fis and in the transcription of several gene regions during mid-exponential and late-stationary phase, and during different growth aeration regimes, has been investigated. Studies were done during those two growth phases and in aerated and non-aerated (microaerobic) conditions, to measure Fis enrichment and binding peaks in strategic gene regions by genome-wide microarray analysis ChIP-chip. This research investigation points to central roles for SPI-1, SPI-2, DNA gyrase and topoisomerase I, the elements of the stringent response, and the regulatory function of Fis-binding patterns, in setting and re-setting the activity of the fis gene and other involved promoters as a function of the growth conditions and aeration regimes experienced by Salmonella. One sample with four different treatments. Three biological replicates per sample. The Aerated 2 hour sample was set as the control condition for all samples.
Project description:InvF ChIP-chip on Salmonella enterica serovar Typhimurium SL1344 using anti-Myc antibody against strain with chromosomally 9Myc-tagged InvF (IP samples) and wildtype strain (mock IP samples) Salmonella enterica serovar Typhimurium causes a range of diseases from self-limiting gastroenteritis to life-threatening systemic infections. Its complex infection process is initiated by the invasion of the intestinal epithelial monolayer by means of a type three secretion system. InvF is one of the key regulators governing the invasion of epithelial cells. By mapping the InvF regulon, i.e. locating its direct target genes, the gene network underlying invasion can be further examined, including identifying possible new effector-encoding genes. In order to map the InvF regulon, we performed chromatin immunoprecipitation combined with tiling microarray analysis (ChIP-chip) and compared expression of the identified target genes in an invF mutant and a wildtype strain. In addition, the promoter regions of these target genes were searched for the presence of an InvF recognition site. Finally, a query-driven biclustering method, combined with a microarray compendium containing publically available S. Typhimurium gene expression data, was applied as an in silico validation technique for functional relatedness between newly identified target genes and known invasion genes. As expected, under invasion inducing conditions, InvF activates the expression of invasion chaperone encoding sicA and the effector-encoding genes sopB, sopE, sopE2 and sopA by binding their promoter region. Newly identified InvF targets are steB, encoding a secreted effector, and STM1239. The presence of an InvF recognition site in the promoter regions of these target genes further supports this observation. In addition, the query-driven biclustering method revealed similarities in expression profiles between STM1239 and known InvF regulated invasion genes over a range of experimental conditions. In conclusion, we here deliver the first evidence for direct binding of InvF to the promoter regions of sopA and sopE2, and associate genes encoding a secreted effector (steB) and a putative novel effector (STM1239) with the Salmonella invasion regulator InvF.
Project description:The aim of this experiment was to determine changes in transcription profile of Salmonella SL1344 and SL1344 ∆acrB over the growth curve. Samples of both strains were taken after 1, 3 and 5 hours of growth (corresponding to early- and late-exponential phase and stationary phase) in LB media. Cells were harvested and RNA extraction, library preparation and sequencing using Illumina HiSeq were carried out by GENEWIZ Inc.