ArticleAnaerobe2016
Heat shock increases conjugation efficiency in Clostridium difficile.
Article in Anaerobe, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 56 papers.
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Who cites it
56 citing papers in PubMed.
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- Defining the order of assembly of thebioRxiv : the preprint server for biology · 2026Article
- Phase variable colony morphotypes ofbioRxiv : the preprint server for biology · 2026Article
- Nanoluciferase-Based Reporters for Clostridioides difficile.Methods in molecular biology (Clifton, N.J.) · 2026Article
- The presence and induction of regioselective dehydroxylases dictate urolithin metabolism by Enterocloster species.NPJ biofilms and microbiomes · 2025Article
- Molecular dissection of Class A PBP function uncovers novel features of the non-canonical Clostridioides difficile divisome complex.PLoS genetics · 2025Article
- Conjugation-based genetic manipulation ofmBio · 2025Article
- Deciphering the RNA-based regulation mechanism of the phage-encoded AbiF system in Clostridioides difficile.PLoS genetics · 2025Article
- Molecular dissection of Class A PBP function uncovers novel features of the non-canonicalbioRxiv : the preprint server for biology · 2025Article
- The phi027 bacteriophage influences physiology and virulence of the lysogenic strain of Clostridioides difficile.Scientific reports · 2025Article
- A thiouracil desulfurase protects Clostridioides difficile RNA from 4-thiouracil incorporation, providing a competitive advantage in the gut.Cell host & microbe · 2025Article
- Assessing horizontal gene transfer in the rhizosphere ofApplied and environmental microbiology · 2024Article
- Article
- The Impact of YabG Mutations on Clostridioides difficile Spore Germination and Processing of Spore Substrates.Molecular microbiology · 2024Article
- Identification of pathways to high-level vancomycin resistance in Clostridioides difficile that incur high fitness costs in key pathogenicity traits.PLoS biology · 2024Article
- A sequence invariable region in TcdB2 is required for toxin escape fromJournal of bacteriology · 2024Article
- Formation of the pyruvoyl-dependent proline reductase Prd fromPNAS nexus · 2024Article
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Authors and funding
2 authors.
Funding
Abstract
Clostridium difficile infection has increased in incidence and severity over the past decade, and poses a unique threat to human health. However, genetic manipulation of C. difficile remains in its infancy and the bacterium remains relatively poorly characterised. Low-efficiency conjugation is currently the only available method for transfer of plasmid DNA into C. difficile. This is practically limiting and has slowed progress in understanding this important pathogen. Conjugation efficiency varies widely between strains, with important clinically relevant strains such as R20291 being particularly refractory to plasmid transfer. Here we present an optimised conjugation method in which the recipient C. difficile is heat treated prior to conjugation. This significantly improves conjugation efficiency in all C. difficile strains tested including R20291. Conjugation efficiency was also affected by the choice of media on which conjugations were performed, with standard BHI media giving most transconjugant recovery. Using our optimised method greatly increased the ease with which the chromosome of R20291 could be precisely manipulated by homologous recombination. Our method improves on current conjugation protocols and will help speed genetic manipulation of strains otherwise difficult to work with.
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