Project description:Transcriptomic analysis of C. difficile strain 630 subjected to a clinically relevant heat stress (41°C ) and compared to unstressed cells (37°C ). Heat stress was induced in biological triplicate broth cultures in the early exponential phase (D650nm=0.3) of growth and cells were harvested at late log phase (D650 = 1.1). Goal was to determine effects of mild heat stress on gene expression.
Project description:Transcriptomic analysis of C. difficile strain 630 subjected to a clinically relevant heat stress (41M-BM-0C ) and compared to unstressed cells (37M-BM-0C ). Heat stress was induced in biological triplicate broth cultures in the early exponential phase (D650nm=0.3) of growth and cells were harvested at late log phase (D650 = 1.1). Goal was to determine effects of mild heat stress on gene expression. Two condition experiment, heat stressed vs. control. Biological replicates: 3 control replicates, 3 heat stressed replicates, with dye flip
Project description:Clostridium difficile is a gram-positive, spore-forming enteric anaerobe which can infect humans and a wide variety of animal species. Recently, the incidence and severity of human C. difficile infection has markedly increased. In this study, we evaluated the genomic content of 73 C. difficile strains isolated from humans, horses, cattle, and pigs by comparative genomic hybridization with microarrays containing coding sequences from C. difficile strains 630 and QCD-32g58. The sequenced genome of C. difficile strain 630 was used as a reference to define a candidate core genome of C. difficile and to explore correlations between host origins and genetic diversity. Approximately 16% of the genes in strain 630 were highly conserved among all strains, representing the core complement of functional genes defining C. difficile. Absent or divergent genes in the tested strains were distributed across the entire C. difficile 630 genome and across all the predicted functional categories. Interestingly, certain genes were conserved among strains from a specific host species, but divergent in isolates with other host origins. This information provides insight into the genomic changes which might contribute to host adaptation. Due to a high degree of divergence among C. difficile strains, a core gene list from this study offers the first step toward the construction of diagnostic arrays for C. difficile.
Project description:Clostridium difficile is a gram-positive, spore-forming enteric anaerobe which can infect humans and a wide variety of animal species. Recently, the incidence and severity of human C. difficile infection has markedly increased. In this study, we evaluated the genomic content of 73 C. difficile strains isolated from humans, horses, cattle, and pigs by comparative genomic hybridization with microarrays containing coding sequences from C. difficile strains 630 and QCD-32g58. The sequenced genome of C. difficile strain 630 was used as a reference to define a candidate core genome of C. difficile and to explore correlations between host origins and genetic diversity. Approximately 16% of the genes in strain 630 were highly conserved among all strains, representing the core complement of functional genes defining C. difficile. Absent or divergent genes in the tested strains were distributed across the entire C. difficile 630 genome and across all the predicted functional categories. Interestingly, certain genes were conserved among strains from a specific host species, but divergent in isolates with other host origins. This information provides insight into the genomic changes which might contribute to host adaptation. Due to a high degree of divergence among C. difficile strains, a core gene list from this study offers the first step toward the construction of diagnostic arrays for C. difficile.investigated by determining changes in transcript profiles when aerobic steady-state cultures were depleted of air.
Project description:Grad-seq in Clostridium difficile 630. Cell lysate is analyzed in a gradient and fractionated into 21 fractions which are analysed for proteins by MS and for transcripts by RNA-sequencing.
Project description:Clostridium difficile is a gram-positive, spore-forming enteric anaerobe which can infect humans and a wide variety of animal species. Recently, the incidence and severity of human C. difficile infection has markedly increased. In this study, we evaluated the genomic content of 73 C. difficile strains isolated from humans, horses, cattle, and pigs by comparative genomic hybridization with microarrays containing coding sequences from C. difficile strains 630 and QCD-32g58. The sequenced genome of C. difficile strain 630 was used as a reference to define a candidate core genome of C. difficile and to explore correlations between host origins and genetic diversity. Approximately 16% of the genes in strain 630 were highly conserved among all strains, representing the core complement of functional genes defining C. difficile. Absent or divergent genes in the tested strains were distributed across the entire C. difficile 630 genome and across all the predicted functional categories. Interestingly, certain genes were conserved among strains from a specific host species, but divergent in isolates with other host origins. This information provides insight into the genomic changes which might contribute to host adaptation. Due to a high degree of divergence among C. difficile strains, a core gene list from this study offers the first step toward the construction of diagnostic arrays for C. difficile.investigated by determining changes in transcript profiles when aerobic steady-state cultures were depleted of air. Dye-swap experiments with genomic DNA of tested and reference strains 8383cy3.gpr- Cy3 – test, Cy5 – reference 8384cy3.gpr- Cy3 – test, Cy5 – reference 8384cy5.gpr- Cy3 – reference, Cy5 – test 8385cy3.gpr- Cy3 – test, Cy5 – reference 8385cy5.gpr- Cy3 – reference, Cy5 – test 8386cy3.gpr- Cy3 – test, Cy5 – reference 8525cy3.gpr- Cy3 – test, Cy5 – reference 8525cy5.gpr- Cy3 – reference, Cy5 – test 8527cy5.gpr- Cy3 – reference, Cy5 – test 8529cy5.gpr- Cy3 – reference, Cy5 – test 8531cy3.gpr- Cy3 – test, Cy5 – reference 8531cy5.gpr- Cy3 – reference, Cy5 – test 8533cy3.gpr- Cy3 – test, Cy5 – reference 8533cy5.gpr- Cy3 – reference, Cy5 – test 8596cy3.gpr- Cy3 – test, Cy5 – reference 8596cy5.gpr- Cy3 – reference, Cy5 – test 8694cy3.gpr- Cy3 – test, Cy5 – reference 8694cy5.gpr- Cy3 – reference, Cy5 – test 2 GPR files per Sample record except for 4 Sample records (GSM480414, GSM480417, GSM480419, and GSM480420) which have 1 GPR file each 4 GPR file for those Sample records are lost
Project description:Investigation of whole genome gene expression level changes in a Clostridium difficile fur (ferric uptake regulator) mutant, compared to the wild type strain 630 erm. The fur mutant analyzed in this study is further described in Ho and Ellermeier (2015) J. Bacteriology A microarray study using total RNA recovered from three separate wild type cultures of Clostridium difficile 630 erm strain and three separate cultures of a fur mutant strain (ltrA::ermR) were grown in Tryptone-Yeast Extract medium containing 0.25 mM ferric chloride . Each chip measures the expression level of 3,786 of the 3,787 open reading frames of the C. difficile 630 genome with 18 probes (60 oligomers each) for each gene.
Project description:The Gram-positive bacterium Clostridium difficile, a leading cause of antibiotic-associated pseudomembranous colitis, has received increasing attention due to a rising incidence of clinical C. difficile infections (CDI). Despite progress understanding bacterial factors that promote CDI-associated morbidity and mortality, many fundamental aspects of C. difficile biology remain to be explored. Compared to other Gram-positive pathogens, little is known about the bacterium’s transcriptome architecture and in particular mechanisms of post-transcriptional control. To close this knowledge gap, we have applied a suite of transcriptome-focused techniques, including transcription start site mapping (dRNA-seq), transcription termination mapping, and Hfq RIP-seq, resulting in a single-nucleotide resolution RNA map of C. difficile strain 630.