ArticleApplied and environmental microbiology2010
A mariner-based transposon system for in vivo random mutagenesis of Clostridium difficile.
Article in Applied and environmental microbiology, 2010. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 96 papers.
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Who cites it
96 citing papers in PubMed.
- Transposon insertion sequencing identifies novel genes involved in product synthesis in gas-fermentingSynthetic and systems biotechnology · 2026Article
- Development and Evaluation of Exosporium-Anchored Bioluminescent and Fluorescent Reporters for TrackingACS synthetic biology · 2026Article
- The CspC:CspA heterodimer transduces germinant and co-germinant signals during Clostridioides difficile spore germination.PLoS biology · 2026Article
- A set of genetic tools for use inMicrobiology (Reading, England) · 2026Article
- Nanoluciferase-Based Reporters for Clostridioides difficile.Methods in molecular biology (Clifton, N.J.) · 2026Article
- A thiouracil desulfurase protects Clostridioides difficile RNA from 4-thiouracil incorporation, providing a competitive advantage in the gut.Cell host & microbe · 2025Article
- Differential gene expression analysis shows that cephalosporin resistance is intrinsic to Clostridioides difficile strain 630.The Journal of antibiotics · 2025Article
- The adhesin RadD enhances Fusobacterium nucleatum tumour colonization and colorectal carcinogenesis.Nature microbiology · 2024Article
- Gene essentiality in the solventogenicApplied and environmental microbiology · 2024Article
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- Histidine kinase-mediated cross-regulation of the vancomycin-resistance operon in Clostridioides difficile.Molecular microbiology · 2024Article
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- Removal of mobile genetic elements from the genome ofFrontiers in microbiology · 2024Article
- RipE expression correlates with high ATP levels inFrontiers in cellular and infection microbiology · 2024Article
- Characterization of the Clostridioides difficile 630Δerm putative Pro-Pro endopeptidase CD1597.Access microbiology · 2024Article
- Engineered acetogenic bacteria as microbial cell factory for diversified biochemicals.Frontiers in bioengineering and biotechnology · 2024Review
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- Identification of novel p-cresol inhibitors that reduce Clostridioides difficile's ability to compete with species of the gut microbiome.Scientific reports · 2023Article
- Host Sorbitol and Bacterial Sorbitol Utilization Promote Clostridioides difficile Infection in Inflammatory Bowel Disease.Gastroenterology · 2023Article
- Article
36 more citing papers are in PubMed but not listed here.
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2 authors.
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Abstract
Understanding the molecular basis of Clostridium difficile infection is a prerequisite to the development of effective countermeasures. Although there are methods for constructing gene-specific mutants of C. difficile, currently there is no effective method for generating libraries of random mutants. In this study, we developed a novel mariner-based transposon system for in vivo random mutagenesis of C. difficile R20291, the BI/NAP1/027 epidemic strain at the center of the C. difficile outbreaks in Stoke Mandeville, United Kingdom, in 2003 to 2004 and 2004 to 2005. Transposition occurred at a frequency of 4.5 (+/-0.4) x 10(-4) per cell to give stable insertions at random genomic loci, which were defined only by the nucleotide sequence TA. Furthermore, mutants with just a single transposon insertion were generated in an overwhelming majority (98.3% in this study). Phenotypic screening of a C. difficile R20291 random mutant library yielded a sporulation/germination-defective clone with an insertion in the germination-specific protease gene cspBA and an auxotroph with an insertion in the pyrimidine biosynthesis gene pyrB. These results validate our mariner-based transposon system for use in forward genetic studies of C. difficile.
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