Effective mutagenesis of Vibrio fischeri by using hyperactive mini-Tn5 derivatives.
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.
Lyell
Noreen L
NL
Department of Microbiology, University of Georgia, 1000 Cedar Street, Athens, GA 30602, USA.
Dunn
Anne K
AK
Bose
Jeffrey L
JL
Vescovi
Susan L
SL
Stabb
Eric V
EV
D048908:Aliivibrio fischeri / Q000235:genetics*; D015139:Blotting, Southern; D004251:DNA Transposable Elements; D004269:DNA, Bacterial / Q000235:genetics; D016254:Mutagenesis, Insertional / Q000379:methods*
D004251:DNA Transposable Elements; D004269:DNA, Bacterial