AEM Accepts, published online ahead of print on 11 April 2008
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Appl. Environ. Microbiol. doi:10.1128/AEM.00476-08
Copyright (c)2008, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

In vitro mutagenesis of Bacillus subtilis using a modified Tn7 with an outward facing inducible promoter

Christophe Bordi, Bronwyn G. Butcher, Qiaojuan Shi, Anna-Barbara Hachmann, Joseph E. Peters, and John D. Helmann*

Department of Microbiology, Cornell University, Ithaca, NY 14853-8101, USA

* To whom correspondence should be addressed. Email: jdh9{at}cornell.edu.


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Abstract

A Tn7 donor plasmid, pTn7SX, was constructed for use with the model Gram-positive bacterium Bacillus subtilis. This new mini-Tn7, mTn7SX, contains a spectinomycin resistance cassette and an outward facing xylose-inducible promoter thereby allowing for the regulated expression of genes downstream of the transposon. We demonstrate that mTn7SX inserts are obtained at a high frequency and occur randomly throughout the B. subtilis genome. The utility of this system was demonstrated by the selection of mutants with increased resistance to the antibiotics fosfomycin or duramycin.




This article has been cited by other articles:

  • Salzberg, L. I., Helmann, J. D. (2008). Phenotypic and Transcriptomic Characterization of Bacillus subtilis Mutants with Grossly Altered Membrane Composition. J. Bacteriol. 190: 7797-7807 [Abstract] [Full Text]  


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