Journal
APPLIED AND ENVIRONMENTAL MICROBIOLOGY
Volume 74, Issue 22, Pages 7059-7063Publisher
AMER SOC MICROBIOLOGY
DOI: 10.1128/AEM.01330-08
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Funding
- National Science Foundation (NSF) [MCB-0347317]
- National Institutes of Health (NIH) [AI50661]
- NSF Research Experience for Undergraduates site award [DBI-0453353]
- W. M. Keck Foundation
- [pRZ5412]
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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 similar to 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.
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