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:Clostridioides difficile infection (CDI) represents a significant global public health concern. The Phosphatidylinositol Glycan Class L (pig-L) gene in C. difficile encodes an enzyme critical for the biosynthesis of phosphatidylinositol glycan anchors (PGAs), which play a vital role in bacterial surface protein localization and function
Project description:To reveal the effects of deleting the Δxtr gene in Clostridioides difficile, we first constructed a Δxtr knockout mutant and then examined the resulting phenotypic changes. Finally, to elucidate the underlying mechanisms, we performed RNA-sequencing-based transcriptome analysis.
Project description:Clostridioides difficile is a clinically important nosocomial pathogen whose virulence is primarily associated with toxin production and can be influenced by prophage carriage. In this study, we investigated the molecular basis of phi027-dependent phenotypic differences previously observed between the clinical RT176 strain 500/12 and its prophage-free derivative CKH08. Comparative transcriptomic analysis revealed that loss of phi027 was associated with reduced expression of the sinR/R' operon, downregulation of multiple flagellar biosynthesis genes.
Project description:The intestines house a diverse microbiota that must compete for nutrients to survive, but the specific limiting nutrients that control pathogen colonization are not clearly defined. Clostridioides difficile colonization typically requires prior disruption of the microbiota, suggesting that outcompeting commensals for resources is key in establishing C. difficile infection (CDI). The immune protein calprotectin (CP) is released into the gut lumen during CDI to chelate zinc (Zn) and other essential nutrient metals. Yet, the impact of Zn limitation on C. difficile colonization is unknown. To define C. difficile responses to Zn limitation, we performed RNA sequencing on C. difficile exposed to CP. In media with CP, C. difficile upregulated genes involved in metal homeostasis and amino acid metabolism.
Project description:Clostridioides difficile is one of the most common nosocomial pathogens and a global public health threat. Upon colonization of the gastrointestinal tract, C. difficile is exposed to a rapidly changing polymicrobial environment and a dynamic metabolic milieu. Despite the link between the gut microbiota and susceptibility to C. difficile, the impact of synergistic interactions between the microbiota and pathogens on the outcome of infection is largely unknown. Here, we show that microbial cooperation between C. difficile and Enterococcus has a profound impact on the growth, metabolism, and pathogenesis of C. difficile.. Through a process of nutrient restriction and metabolite cross-feeding, E. faecalis shapes the metabolic environment in the gut to enhance C. difficile fitness and increase toxin production. These findings demonstrate that members of the microbiota, such as Enterococcus, have a previously unappreciated impact on C. difficile behavior and virulence.
Project description:The experiment intends to reveal the difference in gene expression profiles between the wild-type strain and the ∆rpoN mutant of Clostridioides difficile. We first constructed the ∆rpoN mutant, and the phenotypic changes of the ∆rpoN mutant against the wild-type strain were studied. To further elucidate the mechanism of phenotypic changes of the ∆rpoN mutant, RNA-sequencing experiments were carried out to reveal the underlying mechanism of phenotypic changes.
Project description:The experiment intends to reveal the difference in gene expression profiles between the wild-type strain and the ∆cwp66 mutant of Clostridioides difficile. We first constructed the ∆cwp66 mutant, and the phenotypic changes of the ∆cwp66 mutant against the wild-type strain were studied. To further elucidate the mechanism of phenotypic changes of the ∆cwp66 mutant, RNA-sequencing experiments were carried out to reveal the underlying mechanism of phenotypic changes.
Project description:Fidaxomicin is considered the current gold standard antibiotic for treating Clostridioides difficile infections and kills bacterial cells by inhibition of the RNA polymerase through binding to its switch region. Although binding sites do not overlap, also Myxopyronin B inhibits the RNA polymerase by binding its switch region. The here presented data prove that there is no cross-resistance between Fidaxomicin and Myxopyronin B in a Fidaxomicin-resistant C. difficile strain. Moreover, comparative LC-MS/MS analyses of Fidaxomicin, Myxopyronin B and Rifaximin stress in C. difficile strain 630 revealed that Myxopyronin B is able to suppress early phase toxin synthesis in C. difficile to the same degree as Fidaxomicin. Conclusively, Myxopyronin B is proposed as lead structure for the design of novel antibiotics for the therapy of C. difficile infections.
Project description:To reveal the effects of deleting the ΔsigV and Δanti-sigV on clindamycin tolerance in Clostridioides difficile, we first constructed knockout mutants of the sigV and anti-sigV genes and then examined the resulting phenotypic changes, with a particular focus on alterations in clindamycin tolerance. To further elucidate the mechanism underlying the observed changes in clindamycin tolerance, we performed transcriptomic data analysis to identify how sigV and anti-sigV regulate clindamycin-related pathways.