Project description:Clostridium perfringens type A is a common source of food poisoning in humans. Vegetative cells sporulate in the small intestinal tract and produce a major pathogenic factor, C. perfringens enterotoxin (CPE) during sporulation. Although sporulation plays a critical role in the pathogenesis of food poisoning, the mechanisms to induce in vivo sporulation remain unclear. Bile salts had been identified to mediate sporulation, and we have confirmed deoxycholate (DCA)-induced sporulation in C. perfringens strain NCTC8239 co-cultured with human intestinal epithelial Caco-2 cells. In this study, we performed global transcriptome analysis of strain NCTC8239 to elucidate the mechanism to induce sporulation by DCA.
Project description:Purpose: Analyze gene expression during C. perfringens colonization in the chicken Transcriptomic profile of mRNA from C. perfrinegns from in vivo and in vitro conditions were determined in biological duplicates by RNA-Seq using Illumina HiSeq 2500
Project description:Comparative transcriptome analysis by RNAseq of Necrotic Enteritis Clostridium perfringens in ligated intestinal chicken loops and in vitro conditions.
Project description:Purpose: Analyze gene expression during C. perfringens colonization in the chicken Transcriptomic profile of mRNA from C. perfrinegns from in vivo and in vitro conditions were determined in biological duplicates by RNA-Seq using Illumina HiSeq 2500 Comparison of gene expression through RNA sequencing of necrotic enteritis C. perfrinegns type A of in vivo (chicken loops) and in vitro (lab culture)
Project description:Clostridium perfringens type A is a common source of food poisoning in humans. Vegetative cells sporulate in the small intestinal tract and produce a major pathogenic factor, C. perfringens enterotoxin (CPE) during sporulation. Although sporulation plays a critical role in the pathogenesis of food poisoning, the mechanisms to induce in vivo sporulation remain unclear. Bile salts had been identified to mediate sporulation, and we have confirmed deoxycholate (DCA)-induced sporulation in C. perfringens strain NCTC8239 co-cultured with human intestinal epithelial Caco-2 cells. In this study, we performed global transcriptome analysis of strain NCTC8239 to elucidate the mechanism to induce sporulation by DCA. From the 55 contigs of C. perfringens strain NCTC8239, 2778 coding sequences were extracted. We designed a DNA probe by utilizing eArray provided by Agilent Technologies. The custom 8Ã15K oligonucleotide array, containing 60 mer oligonucleotide probes for 2,778 genes in strain NCTC8239, 2 bacterial control genes: 16S rRNA and 23S rRNA, and 3 human control genes: beta-2-microglobulin, glucuronidase beta and 18S rRNA, were ordered to Agilent Technologies. Each probe was spotted in five-fold on each microarray. Each strain was run in triplicate or quadruplicate.
Project description:To obtain an insight into host-pathogen interactions in clostridial myonecrosis, we carried out comparative RNA-seq analysis measuring host genes in a murine C. perfringens infection model. Analysis of the murine transcriptome from infected muscle tissues indicated that 270 genes were up-regulated compared to control mock-infected mice. KEGG pathway analysis of these genes revealed an enrichment of Toll-like receptor (TLR) and Nod-like receptor (NLR) signaling components.
Project description:In this study we focus on the identification of new genes tentatively involved in sporulation and those that influence properties of spores and their ability to germinate. To this end, the sporulation stages of C. perfringens enterotoxic strain SM101 were characterized based on morphological characteristics and biological indicators. Subsequently, whole genome expression profiling during key stages of the sporulation process was performed using DNA microarrays, and genes were clustered based on their time-course expression profiles during sporulation. The majority of previously characterized C. perfringens germination genes showed upregulated expression profiles in time during sporulation and mainly belonged to two clusters of genes.
Project description:We further characterize the VirSR and RevR regulatory networks by profiling the C. perfringens strain JIR325 and its isogenic virR and revR regulatory mutants using strand-specific RNA-seq. Two independent biological replicates were sequenced for each strain, generating more than 90 million sequence reads for each RNA-seq library (wild type, 97,289,148 reads; virR mutant, 116,505,992 reads and revR mutant, 131,811,486 reads). Using the edgeR analysis package, 223 genes in the virR mutant and 88 genes in the revR mutant were found to be differentially expressed. Comparative transcriptomic analysis revealed that VirR acts as a global negative regulator, whilst RevR acts as a global positive regulator. Therefore, about 88% of the differentially expressed genes were up-regulated in the virR mutant, whereas 75% of the differentially expressed genes were down-regulated in the revR mutant. Importantly, we identified 22 genes that were regulated by both VirR and RevR. Of these genes, 18 or 82%, which included the sporulation-specific spoIVA, sigG and sigF genes, were regulated positively and negatively by RevR and VirR, respectively. Furthermore, mapped RNA-seq reads visualized as a user plot showed that there were 97 previously unannotated transcripts in the intergenic regions. These transcripts may potentially encode novel genes or small RNA molecules. This study has identified genes, antisense transcripts, and transcripts within intergenic regions and on the native plasmid, which are controlled by the VirSR or RevR regulatory system. The knowledge obtained will enable a more thorough annotation of the C. perfringens genome. Comparative transcriptomic analysis on the virR and revR regulatory mutants, and the wild-type strain JIR325
Project description:RevR is a putative orphan response regulator with a high degree of similarity to YycF from Bacilus subtilis and PhoB from Clostridium kluyveri. A revR deletion mutant of C. perfringens strain 13 was generated and the transcriptome analysed using microarrays.
Project description:We further characterize the VirSR and RevR regulatory networks by profiling the C. perfringens strain JIR325 and its isogenic virR and revR regulatory mutants using strand-specific RNA-seq. Two independent biological replicates were sequenced for each strain, generating more than 90 million sequence reads for each RNA-seq library (wild type, 97,289,148 reads; virR mutant, 116,505,992 reads and revR mutant, 131,811,486 reads). Using the edgeR analysis package, 223 genes in the virR mutant and 88 genes in the revR mutant were found to be differentially expressed. Comparative transcriptomic analysis revealed that VirR acts as a global negative regulator, whilst RevR acts as a global positive regulator. Therefore, about 88% of the differentially expressed genes were up-regulated in the virR mutant, whereas 75% of the differentially expressed genes were down-regulated in the revR mutant. Importantly, we identified 22 genes that were regulated by both VirR and RevR. Of these genes, 18 or 82%, which included the sporulation-specific spoIVA, sigG and sigF genes, were regulated positively and negatively by RevR and VirR, respectively. Furthermore, mapped RNA-seq reads visualized as a user plot showed that there were 97 previously unannotated transcripts in the intergenic regions. These transcripts may potentially encode novel genes or small RNA molecules. This study has identified genes, antisense transcripts, and transcripts within intergenic regions and on the native plasmid, which are controlled by the VirSR or RevR regulatory system. The knowledge obtained will enable a more thorough annotation of the C. perfringens genome.