Engineering Vibrio fischeri for Inducible Gene Expression



Jakob M Ondrey , Karen L Visick*
Department of Microbiology and Immunology, Health Sciences Division, Loyola University Chicago, 2160 S. First Avenue Building 105 Room 3936, Maywood, IL 60153, USA


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© Ondrey and Visick; Licensee Bentham Open.

open-access license: This is an open access article licensed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted, non-commercial use, distribution and reproduction in any medium, provided the work is properly cited.

* Address correspondence to this author at the Department of Microbiology and Immunology, Health Sciences Division, Loyola University Chicago, 2160 S. First Avenue Building 105 Room 3936, Maywood, IL 60153, USA; Tel: (708)216-0869; Fax: (708)216-9574; E-mail: kvisick@luc.edu


Abstract

The marine bacterium Vibrio fischeri serves as a model organism for a variety of natural phenomena, including symbiotic host colonization. The ease with which the V. fischeri genome can be manipulated contributes greatly to our ability to identify the factors involved in these phenomena. Here, we have adapted genetic tools for use in V. fischeri to promote our ability to conditionally control the expression of genes of interest. Specifically, we modified the commonly used mini-Tn5 transposon to contain an outward-facing, LacI-repressible/IPTG-inducible promoter, and inserted the lacI gene into the V. fischeri chromosome. Used together, these tools permit the identification and induction of genes that control specific phenotypes. To validate this approach, we identified IPTG-controllable motility mutants. We anticipate that the ability to randomly insert an inducible promoter into the genome of V. fischeri will advance our understanding of various aspects of the physiology of this microbe.

Keywords: Genetics, inducible gene expression, motility, mutagenesis, Vibrio fischeri.