title: Investigating Transfer of Large Chromosomal Regions Containing the Pathogenicity Locus Between Clostridium difficile Strains.
abstract: The genomes of all sequenced Clostridium difficile isolates contain multiple mobile genetic elements. The chromosomally located pathogenicity locus (PaLoc), encoding the cytotoxins TcdA and TcdB, was previously hypothesized to be a mobile genetic element; however, mobility was not demonstrated. Here we describe the methods used to facilitate and detect the transfer of the PaLoc from a toxigenic strain into non-toxigenic strains of C. difficile. Although the precise mechanism of transfer has not yet been elucidated, a number of controls are described which indicate transfer occurs via a cell-to-cell-mediated conjugation-like transfer mechanism. Importantly, transfer of the PaLoc was shown to occur on large chromosomal fragments of variable sizes, indicating that homologous recombination is likely to be responsible for the insertion events.
authors: Department of Bacteriology and Epidemiology, Central Veterinary Institute of Wageningen UR, Lelystad, The Netherlands.
authors: Michael S M
authors: MS
authors: Brouwer
authors: Department of Microbial Diseases, UCL Eastman Dental Institute, University College London, London, UK.
authors: Peter
authors: P
authors: Mullany
authors: Department of Microbial Diseases, UCL Eastman Dental Institute, University College London, London, UK.
authors: Elaine
authors: E
authors: Allan
authors: Department of Microbial Diseases, UCL Eastman Dental Institute, University College London, London, UK. adam.roberts@ucl.ac.uk.
authors: Adam P
authors: AP
authors: Roberts
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D001427:Bacterial Toxins / Q000235:genetics / Q000378:metabolism; D001435:Bacteriophages / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D002876:Chromosomes, Bacterial / Q000737:chemistry* / Q000378:metabolism / Q000821:virology; D016360:Clostridioides difficile / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism / Q000821:virology; D003227:Conjugation, Genetic; D017931:DNA Primers / Q000737:chemistry / Q000378:metabolism; D004251:DNA Transposable Elements; D004269:DNA, Bacterial / Q000235:genetics / Q000378:metabolism; D004768:Enterotoxins / Q000235:genetics / Q000378:metabolism; D022761:Gene Transfer, Horizontal; D056426:Genetic Loci; D044404:Genomic Islands; D059014:High-Throughput Nucleotide Sequencing; D059765:Homologous Recombination; D016685:Mitomycin / Q000494:pharmacology; D016133:Polymerase Chain Reaction; D014775:Virus Activation / Q000187:drug effects
chemical_list: D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D001427:Bacterial Toxins; D017931:DNA Primers; D004251:DNA Transposable Elements; D004269:DNA, Bacterial; D004768:Enterotoxins; C041348:tcdA protein, Clostridium difficile; C031815:toxB protein, Clostridium difficile; D016685:Mitomycin
keywords: Chromosomal transfer; Conjugation; Next-generation sequencing; Pathogenicity locus; Recombination; SNP analysis; Transfer