title: In vitro mutagenesis of Bacillus subtilis by using a modified Tn7 transposon with an outward-facing inducible promoter.
abstract: A Tn7 donor plasmid, pTn7SX, was constructed for use with the model gram-positive bacterium Bacillus subtilis. This new mini-Tn7, mTn7SX, contains a spectinomycin resistance cassette and an outward-facing, xylose-inducible promoter, thereby allowing for the regulated expression of genes downstream of the transposon. We demonstrate that mTn7SX inserts are obtained at a high frequency and occur randomly throughout the B. subtilis genome. The utility of this system was demonstrated by the selection of mutants with increased resistance to the antibiotic fosfomycin or duramycin.
authors.affiliation: Department of Microbiology, Wing Hall, Cornell University, Ithaca, NY 14853-8101, USA.
authors.forename: Christophe
authors.initials: C
authors.lastname: Bordi
authors.forename: Bronwyn G
authors.initials: BG
authors.lastname: Butcher
authors.forename: Qiaojuan
authors.initials: Q
authors.lastname: Shi
authors.forename: Anna-Barbara
authors.initials: AB
authors.lastname: Hachmann
authors.forename: Joseph E
authors.initials: JE
authors.lastname: Peters
authors.forename: John D
authors.initials: JD
authors.lastname: Helmann
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001412:Bacillus subtilis / Q000235:genetics*; D001430:Bacteriocins / Q000494:pharmacology; D004251:DNA Transposable Elements; D024881:Drug Resistance, Bacterial / Q000235:genetics; D005578:Fosfomycin / Q000494:pharmacology; D023061:Gene Order; D016254:Mutagenesis, Insertional / Q000379:methods*; D010455:Peptides / Q000494:pharmacology; D010957:Plasmids; D011401:Promoter Regions, Genetic; D011995:Recombination, Genetic; D000198:Spectinomycin / Q000494:pharmacology; D014994:Xylose / Q000378:metabolism
chemical_list: D000900:Anti-Bacterial Agents; D001430:Bacteriocins; D004251:DNA Transposable Elements; D010455:Peptides; C040522:duramycin; D005578:Fosfomycin; D000198:Spectinomycin; D014994:Xylose