Project description:Gene expression level of Clostridioides difficile (C. difficile) strain R20291 comparing control C. difficile carring pMTL84151 as vector plasmid with C. difficile conjugated with a pMTL84151-03890 gene. Goal was to determine the effects of 03890 gene conjugation on C. difficile strain R20291 gene expression.
Project description:Transcriptionnal profiling of C. difficile R20291 strain : a wild type strain, a fliC mutant and a fliC mutant complemented after 14h of growth in PY and a wild type strain and a fliC mutant after 14h of mouse infection in Vivo
Project description:Transcriptionnal profiling of C. difficile R20291 strain : a wild type strain, a fliC mutant and a fliC mutant complemented after 14h of growth in PY and a wild type strain and a fliC mutant after 14h of mouse infection in Vivo two-conditions experiments, R20291 strain : a wild type strain, a fliC mutant and a fliC mutant complemented in Vitro and a wild type strain and a fliC mutant in Vivo, 4 biological replicates for each condition, in an direct design.
Project description:Comparison of the expression profiles of R20291 strain : a wild type strain, a fliC mutant and a fliC mutant complemented after 14h of growth in PY and a wild type strain and a fliC mutant after 14h of mouse infection in vivo
Project description:RNA-seq was conducted to uncover the regulatory roles of RsbW in the lifestyle of Clostridioides difficile strain R20291. RNA were extracted from planktonic cultures, which were grown in BHI-S (Sigma-Aldrich, USA) to early stationary phase (10h). The cells were re-suspended in LETS buffer and lysed using Fast-prep 24 instrument (MP Bioscience) and RNA was extracted using 1 ml Trizol (Ambion, USA). Samples were cleaned and sequenced by Novogene Company Limited, PE150 with approximately 10 million reads per sample. After QC, reads were aligned to R20291 (NC_013316.1) with Bowtie2 and readcounts were generated with Samtools and Bedtools. Differential gene expression analysis was conducted with DESeq2 with a p-value of 0.05, differential gene expression was determined with ± -2 logFC.
Project description:The S-layer of C. difficile is a paracrystalline array that covers the bacterial cell but its contribution to overall disease remains unclear. A previously described spontaneous slpA-null mutant, FM2.5, with a point mutation in slpA offers the opportunity to study the role of the S-layer in vivo. Here, we confirm our previous observation that this strain is less virulent in vivo despite effectively colonising the host and producing toxin. We also show a strong in vivo selection pressure for strains that express the intact S-layer array through sequence modifications that restore slpA expression. While such modifications do not affect the overall S-layer structure, RNA-Seq analysis in vitro showed large differences in gene expression between FM2.5, the revertant strains and R20291. Despite these differences, an intact S-layer clearly provides a selective advantage of C. difficile in the host, and, for one of the revertants tested, helped restore disease severity.
