Investigating Transfer of Large Chromosomal Regions Containing the Pathogenicity Locus Between Clostridium difficile Strains

Mike Brouwer, P. Mullany, E. Allan, P. Roberts

Research output: Chapter in Book/Report/Conference proceedingChapter

4 Citations (Scopus)

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
Original languageEnglish
Title of host publicationClostridium dfficile
Subtitle of host publicationMethods and protocols
PublisherSpringer
Pages215-222
ISBN (Electronic)9781493963614
ISBN (Print)9781493963591
DOIs
Publication statusPublished - 2016

Publication series

NameMethods in Molecular Biology
PublisherSpringer
Volume1476

Fingerprint

Clostridium difficile
Interspersed Repetitive Sequences
Virulence
Homologous Recombination
Cytotoxins
Genome

Cite this

Brouwer, M., Mullany, P., Allan, E., & Roberts, P. (2016). Investigating Transfer of Large Chromosomal Regions Containing the Pathogenicity Locus Between Clostridium difficile Strains. In Clostridium dfficile: Methods and protocols (pp. 215-222). (Methods in Molecular Biology; Vol. 1476). Springer. https://doi.org/10.1007/978-1-4939-6361-4_16
Brouwer, Mike ; Mullany, P. ; Allan, E. ; Roberts, P. / Investigating Transfer of Large Chromosomal Regions Containing the Pathogenicity Locus Between Clostridium difficile Strains. Clostridium dfficile: Methods and protocols. Springer, 2016. pp. 215-222 (Methods in Molecular Biology).
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Brouwer, M, Mullany, P, Allan, E & Roberts, P 2016, Investigating Transfer of Large Chromosomal Regions Containing the Pathogenicity Locus Between Clostridium difficile Strains. in Clostridium dfficile: Methods and protocols. Methods in Molecular Biology, vol. 1476, Springer, pp. 215-222. https://doi.org/10.1007/978-1-4939-6361-4_16

Investigating Transfer of Large Chromosomal Regions Containing the Pathogenicity Locus Between Clostridium difficile Strains. / Brouwer, Mike; Mullany, P.; Allan, E.; Roberts, P.

Clostridium dfficile: Methods and protocols. Springer, 2016. p. 215-222 (Methods in Molecular Biology; Vol. 1476).

Research output: Chapter in Book/Report/Conference proceedingChapter

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AB - 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

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EP - 222

BT - Clostridium dfficile

PB - Springer

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Brouwer M, Mullany P, Allan E, Roberts P. Investigating Transfer of Large Chromosomal Regions Containing the Pathogenicity Locus Between Clostridium difficile Strains. In Clostridium dfficile: Methods and protocols. Springer. 2016. p. 215-222. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-4939-6361-4_16