title: Effective mutagenesis of Vibrio fischeri by using hyperactive mini-Tn5 derivatives.
abstract: We have developed a transposon mutagenesis system for Vibrio fischeri ES114 that utilizes a hyperactive mutant Tn5 transposase (E54K and M56A) and optimized transposon ends. Using a conjugation-based procedure, we obtained independent single-insertion mini-Tn5 mutants at a rate of approximately 10(-6). This simple and inexpensive technique represents a significant improvement over previous methods for transposon mutagenesis of V. fischeri and should be applicable to many other bacteria.
authors.affiliation: Department of Microbiology, University of Georgia, 1000 Cedar Street, Athens, GA 30602, USA.
authors.forename: Noreen L
authors.initials: NL
authors.lastname: Lyell
authors.forename: Anne K
authors.initials: AK
authors.lastname: Dunn
authors.forename: Jeffrey L
authors.initials: JL
authors.lastname: Bose
authors.forename: Susan L
authors.initials: SL
authors.lastname: Vescovi
authors.forename: Eric V
authors.initials: EV
authors.lastname: Stabb
mesh_terms: D048908:Aliivibrio fischeri / Q000235:genetics*; D015139:Blotting, Southern; D004251:DNA Transposable Elements; D004269:DNA, Bacterial / Q000235:genetics; D016254:Mutagenesis, Insertional / Q000379:methods*
chemical_list: D004251:DNA Transposable Elements; D004269:DNA, Bacterial