Project description:Young adult fer-15;fem-1 Caenorhabditis elegans were infected with Staphylococcus aureus for 8 h to determine the transcriptional host response to Staphylococcus aureus. Analysis of differential gene expression in C. elegans young adults exposed to two different bacteria: E. coli strain OP50 (control), wild-type Staphylococcus aureus RN6390. Samples were analyzed at 8 hours after exposure to the different bacteria. These studies identified C. elegans genes induced by pathogen infection. Keywords: response to pathogen infection, innate immunity, host-pathogen interactions Overall design: There are 6 samples total: 3 biological replicates of Staphylococcus aureus-infected animals, and 3 biological replicates of E. coli-fed controls. Three independent biological replicates were isolated for each treatment. All treatments were performed in parallel.
Project description:Investigation of mRNA expression level changes in a Staphylococcus aureus Mu50 delta-SAV1322 mutant, compared to the wild-type strain. A comparison of the wild-type and the mutant transcription profiles Overall design: Total RNA recovered from four separate wild-type culture of Staphylococcus aureus Mu50 and four separate culture of a mutant strain, Mu50 delta-SAV1322, in which 689bp deleted of SAV1322 locus. Each chip measures the expression level of 967 genes from Staphylococcus aureus Mu50 with 11 probe pairs per genes (total probes 10,637).
Project description:To study the roles of NWMN_0641, we used microarray to compare the transcriptome of the NWMN_0641 deletion strain with that of the wild-type Staphylococcus aureus Newman strain. Transcriptome of the NWMN_0641 deletion mutant strain and the wild-type Newman strain We sought to obtain the transcriptomes of the NWMN_0641 deletion mutant strain and the wild-type Newman strain when bacteria were grown TSB medium (without glucose)
Project description:Methicillin-resistant Staphylococcus aureus (MRSA) infections result in more than 200,000 hospitalizations and 10,000 deaths in the United States each year and remain an important medical challenge. To better understand the transcriptome of Staphylococcus aureus USA300 NRS384, a community-acquired MRSA strain, we have conducted an RNA-Seq experiment on WT samples. Overall design: RNA-Seq analysis was performed on log phase WT Staphylococcus aureus USA300 NRS384 grown at 30 degrees C and grown at 37 degrees C. Each condition was performed in triplicate thereby yielding a total of 6 samples for RNA-Seq analysis.
Project description:Vancomycin-intermediate Staphylococcus aureus (VISA) evolve in a strain-specific manner and acquire mutations that lead to alterations in cell wall metabolism that reduce susceptibility to vancomycin. We had earlier isolated several VISA mutant strains of the clinical hVISA strain MM66. This study is aimed at analyzing the metabolome of these mutants in comparison to the parent strain.
Project description:Staphylococcus aureus is a gram-positive cocci and an important human commensal bacteria and pathogen. S. aureus infections are increasingly difficult to treat because of the emergence of highly resistant MRSA (Methicillin-resistant S. aureus) strains. Here we present a method to study differential gene expression in S. aureus using high-throughput RNA-sequencing (RNA-seq). We use RNA-seq to examine the differential gene expression in S. aureus RN4220 cells containing an exogenously expressed transcription factor and between two S. aureus strains (RN4220 and NCTC8325-4). The information provided by RNA-seq was a significant advance over previously described microarray based techniques. We investigated the sequence and gene expression differences between RN4220 and NCTC8325-4 and used the RNA-seq data to identify S. aureus promoters suitable for in vitro analysis. We used RNA-seq to describe, on a genome wide scale, genes positively and negatively regulated by a phage encoded transcription factor, gp67. RNA-seq offers the ability to study differential gene expression with single-nucleotide resolution, and is a considerable improvement over the predominant genome-wide transcriptome technologies used in S. aureus. RNA-seq analysis of Staphylococcus aureus RN4220 (electrocompetent strain) carrying either empty pRMC2 (inducible expression vector) or pRMC2 carrying the ORF67 gene (encodes gp67). Both strains were grown to OD 0.2 and transgene expression was induced with 100ng/ml anhydrotetracycline. As a control, Staphylococcus aureus strain NCTC8325-4 (non-electrocompetent strain) was grown under identical conditions except without the addition of anhydrotetracycline.
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 Overall design: 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:Staphylococcus aureus is one of the most important pathogens in humans and animals, multiply resistant strains are increasingly widespread, new agents are needed for the treatment of S. aureus. Rhein, a natural plant product, has potential antimicrobial activity against Staphylococcus aureus. We employed Affymetrix Staphylococcus aureus GeneChipsTM arrays to investigate the global transcriptional profiling of Staphylococcus aureus ATCC25923 treated with rhein. Results provided insight into mechanisms involved in rhein - Staphylococcus aureus interactions. Keywords: rhein response Staphylococcus aureus cells were exposed for 45 minutes to rhein at concentration of 8 µg/ml (1/2× MIC), 6 samples including 3 control samples are analyzed.
Project description:Accurate annotation of regulatory RNAs is a complex task but nevertheless essential as sRNA molecular and functional studies ensue from it. Several formerly considered small RNAs (sRNA) are now known to be parts of UTR transcripts. In light of experimental data, we review hundreds of Staphylococcus aureus putative regulatory RNAs. We pinpoint those that are likely acting in trans and are not expressed from the opposite strand of a coding gene. We conclude that HG003, a NCTC8325 derivative strain, has about 50 bona fide sRNAs, indicating that these RNAs are less numerous than commonly stated. Overall design: Total RNA samples were extracted from HG003 grown in various biological conditions. All 16 RNA samples were pooled together for RNA sequencing. Pooled RNAs were processed using the MICROBExpress kit (Ambion, AM1905) as recommended by the suppliers, for rRNAs removal and before high-throughput sequencing using Illumina Genome Analyzer IIx generating single-end 40-nt reads.