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Nucleic Acids Research 2005 33(9):e87; doi:10.1093/nar/gni086
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Published online 25 May 2005

© The Author 2005. Published by Oxford University Press. All rights reserved
The online version of this article has been published under an open access model. Users are entitled to use, reproduce, disseminate, or display the open access version of this article for non-commercial purposes provided that: the original authorship is properly and fully attributed; the Journal and Oxford University Press are attributed as the original place of publication with the correct citation details given; if an article is subsequently reproduced or disseminated not in its entirety but only in part or as a derivative work this must be clearly indicated. For commercial re-use, please contact journals.permissions{at}oupjournals.org

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Natural and synthetic tetracycline-inducible promoters for use in the antibiotic-producing bacteria Streptomyces

Antonio Rodríguez-García, Patricia Combes, Rosario Pérez-Redondo, Matthew C. A. Smith and Margaret C. M. Smith*

Institute of Genetics, University of Nottingham, Queens Medical Centre Nottingham NG7 2UH, UK

*To whom correspondence should be addressed at Institute of Medical Sciences, University of Aberdeen, Foresterhill, Aberdeen AB25 2ZD, UK. Tel: +44 01224 555739; Fax: +44 01224 555844; Email: Maggie.smith{at}abdn.ac.uk

Received April 15, 2005. Revised May 12, 2005. Accepted May 12, 2005.

Bacteria in the genus Streptomyces are major producers of antibiotics and other pharmacologically active compounds. Genetic and physiological manipulations of these bacteria are important for new drug discovery and production development. An essential part of any ‘genetic toolkit’ is the availability of regulatable promoters. We have adapted the tetracycline (Tc) repressor/operator (TetR/tetO) regulatable system from transposon Tn10 for use in Streptomyces. The synthetic Tc controllable promoter (tcp), tcp830, was active in a wide range of Streptomyces species, and varying levels of induction were observed after the addition of 1–100 ng/ml of anhydrotetracycline (aTc). Streptomyces coelicolor contained an innate Tc-controllable promoter regulated by a TetR homologue (SCO0253). Both natural and synthetic promoters were active and inducible throughout growth. Using the luxAB genes expressing luciferase as a reporter system, we showed that induction factors of up to 270 could be obtained for tcp830. The effect of inducers on the growth of S.coelicolor was determined; addition of aTc at concentrations where induction is optimal, i.e. 0.1–1 µg/ml, ranged from no effect on growth rate to a small increase in the lag period compared with cultures with no inducer.

Present addresses: Antonio Rodríguez-García, Instituto de Biotecnología de León (INBIOTEC), Parque Científico de León, Avda. Real no. 1, 24006 León, Spain

Patricia Combes, INSERM U384, Faculté de Médecine, 63000 Clermont-Ferrand, France

Rosario Pérez-Redondo, Área de Microbiología, Facultad de Ciencias Biológicas y Ambientales, 24071 León, Spain


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