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Applied and Environmental Microbiology, September 2008, p. 5436-5443, Vol. 74, No. 17
0099-2240/08/$08.00+0     doi:10.1128/AEM.00256-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Isolation of Predation-Deficient Mutants of Bdellovibrio bacteriovorus by Using Transposon Mutagenesis{triangledown}

John J. Tudor,* James J. Davis,{dagger} Marissa Panichella,{ddagger} and Adam Zwolak§

Department of Biology, Saint Joseph's University, Philadelphia, Pennsylvania 19131

Received 29 January 2008/ Accepted 5 July 2008

The availability of the complete genome sequence of Bdellovibrio bacteriovorus provides an opportunity for investigating genes that play a significant role in predation. Using two independently derived facultatively predatory Bdellovibrio strains, we have designed a method to cultivate and screen transposon insertion mutants in 96-well microtiter dishes. Transposon insertion mutants were produced by introducing the plasposon pRL27, which carries a mini-Tn5. Mutants have been screened for predatory activity using 96-well plates. Seventeen independent nonpredatory mutants have been isolated, and DNA flanking the insertion has been sequenced. BLAST analysis revealed that most of these interrupted DNA sequences do not code for known proteins or functions. Two of the inactivated genes were analyzed further: one was found to code for a putative serine protease and the other a probable protein involved in secretion through the outer membrane. The methods described here are the first for the generation and isolation of predation-deficient mutants using random-transposon-insertion mutagenesis. As more mutants are isolated and their gene products analyzed, more light will be shed on how this predator carries out its exclusive life processes and perhaps how these products, or the organism itself, can be used for therapeutic, agricultural, and/or other purposes.

* Corresponding author. Mailing address: Department of Biology, Saint Joseph's University, 5600 City Avenue, Philadelphia, PA 19131. Phone: (610) 660-1821. Fax: (610) 660-1832. E-mail: jtudor{at}sju.edu

{triangledown} Published ahead of print on 11 July 2008.

{dagger} Present address: Department of Microbiology, University of Illinois Urbana-Champaign, Urbana, IL.

{ddagger} Present address: Department of Microbiology and Immunology, Drexel University College of Medicine, Philadelphia, PA.

§ Present address: Laboratory of Molecular Biophysics, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD.

Applied and Environmental Microbiology, September 2008, p. 5436-5443, Vol. 74, No. 17
0099-2240/08/$08.00+0     doi:10.1128/AEM.00256-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

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