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Appl Environ Microbiol. 2008 Nov;74(22):7059-63. doi: 10.1128/AEM.01330-08. Epub 2008 Sep 19.

Effective mutagenesis of Vibrio fischeri by using hyperactive mini-Tn5 derivatives.

Author information

1
Department of Microbiology, University of Georgia, 1000 Cedar Street, Athens, GA 30602, USA.

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.

PMID:
18805998
PMCID:
PMC2583470
DOI:
10.1128/AEM.01330-08
[Indexed for MEDLINE]
Free PMC Article

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