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Applied and Environmental Microbiology, February 2009, p. 1211-1214, Vol. 75, No. 4
0099-2240/09/$08.00+0     doi:10.1128/AEM.02139-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Cytosine Deaminase as a Negative Selection Marker for Gene Disruption and Replacement in the Genus Streptomyces and Other Actinobacteria {triangledown} ,{dagger}

Marie-Pierre Dubeau,1,{ddagger} Mariana Gabriela Ghinet,1,{ddagger} Pierre-Étienne Jacques,2 Nancy Clermont,1 Carole Beaulieu,1 and Ryszard Brzezinski1*

Centre d'Étude et de Valorisation de la Diversité Microbienne, Département de Biologie, Faculté des Sciences, Université de Sherbrooke, Sherbrooke, Québec, Canada,1 Institut de Recherches Cliniques de Montréal, Montréal, Québec, Canada2

Received 15 September 2008/ Accepted 12 December 2008

We developed a novel negative selection system for actinobacteria based on cytosine deaminase (CodA). We constructed vectors that include a synthetic gene encoding the CodA protein from Escherichia coli optimized for expression in Streptomyces species. Gene disruption and the introduction of an unmarked in-frame deletion were successfully achieved with these vectors.

* Corresponding author. Mailing address: Département de Biologie, Faculté des Sciences, Université de Sherbrooke, 2500 Boul. de l'Université, Sherbrooke, QC J1K 2R1, Canada. Phone: (819) 821-8000, ext. 61077. Fax: (819) 821-8049. E-mail: Ryszard.Brzezinski{at}USherbrooke.ca

{triangledown} Published ahead of print on 19 December 2008.

{dagger} Supplemental material for this article may be found at http://aem.asm.org/.

{ddagger} These authors contributed equally to this work.

Applied and Environmental Microbiology, February 2009, p. 1211-1214, Vol. 75, No. 4
0099-2240/09/$08.00+0     doi:10.1128/AEM.02139-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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