title: Gene transfer into Clostridium difficile CD630 and characterisation of its methylase genes.
abstract: Ignorance of pathogenesis in Clostridium difficile may be attributable to a lack of effective genetic tools. We have now shown that oriT-based shuttle vectors may be conjugated from Escherichia coli donors to the C. difficile strain CD630, at frequencies of around 10(-6) transconjugants per donor cell. Transfer is unaffected by either sequences present on the vector or its methylation status. Whilst the genome of this strain carries five methylase genes, there is no in silico or experimental evidence for cognate restriction enzymes. It would seem that the identified methylases do not participate in restriction-modification, and must, therefore, fulfil another role. A similar situation most likely applies to other clostridia.
authors.affiliation: Health Protection Agency, Porton Down, Salisbury, Wiltshire SP4 0JG, UK.
authors.forename: Michael
authors.initials: M
authors.lastname: Herbert
authors.forename: Triona A
authors.initials: TA
authors.lastname: O'Keeffe
authors.forename: Des
authors.initials: D
authors.lastname: Purdy
authors.forename: Michael
authors.initials: M
authors.lastname: Elmore
authors.forename: Nigel P
authors.initials: NP
authors.lastname: Minton
mesh_terms: D003001:Cloning, Molecular; D016360:Clostridioides difficile / Q000201:enzymology* / Q000235:genetics*; D003227:Conjugation, Genetic; D015254:DNA Modification Methylases / Q000235:genetics*; D018014:Gene Transfer Techniques; D010957:Plasmids
chemical_list: D015254:DNA Modification Methylases