title: Behavior and target site selection of conjugative transposon Tn916 in two different strains of toxigenic Clostridium difficile.
abstract: The insertion sites of the conjugative transposon Tn916 in the anaerobic pathogen Clostridium difficile were determined using Illumina Solexa high-throughput DNA sequencing of Tn916 insertion libraries in two different clinical isolates: 630ΔE, an erythromycin-sensitive derivative of 630 (ribotype 012), and the ribotype 027 isolate R20291, which was responsible for a severe outbreak of C. difficile disease. A consensus 15-bp Tn916 insertion sequence was identified which was similar in both strains, although an extended consensus sequence was observed in R20291. A search of the C. difficile 630 genome showed that the Tn916 insertion motif was present 100,987 times, with approximately 63,000 of these motifs located in genes and 35,000 in intergenic regions. To test the usefulness of Tn916 as a mutagen, a functional screen allowed the isolation of a mutant. This mutant contained Tn916 inserted into a gene involved in flagellar biosynthesis.
authors.affiliation: Department of Microbial Diseases, UCL Eastman Dental Institute, University College London, London, UK. p.mullany@ucl.ac.uk
authors.forename: Peter
authors.initials: P
authors.lastname: Mullany
authors.forename: Rachel
authors.initials: R
authors.lastname: Williams
authors.forename: Gemma C
authors.initials: GC
authors.lastname: Langridge
authors.forename: Daniel J
authors.initials: DJ
authors.lastname: Turner
authors.forename: Rachael
authors.initials: R
authors.lastname: Whalan
authors.forename: Chris
authors.initials: C
authors.lastname: Clayton
authors.forename: Trevor
authors.initials: T
authors.lastname: Lawley
authors.forename: Haitham
authors.initials: H
authors.lastname: Hussain
authors.forename: Katherine
authors.initials: K
authors.lastname: McCurrie
authors.forename: Nicky
authors.initials: N
authors.lastname: Morden
authors.forename: Elaine
authors.initials: E
authors.lastname: Allan
authors.forename: Adam P
authors.initials: AP
authors.lastname: Roberts
mesh_terms: D001412:Bacillus subtilis / Q000235:genetics; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D002874:Chromosome Mapping; D016360:Clostridioides difficile / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification / Q000648:ultrastructure; D003227:Conjugation, Genetic; D004251:DNA Transposable Elements / Q000235:genetics*; D004269:DNA, Bacterial / Q000235:genetics; D005407:Flagella / Q000378:metabolism; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D016254:Mutagenesis, Insertional; D009154:Mutation; D021521:Ribotyping; D013045:Species Specificity
chemical_list: D001426:Bacterial Proteins; D004251:DNA Transposable Elements; D004269:DNA, Bacterial