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

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 availabilit...

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Detalles Bibliográficos
Autores principales: Rodríguez-García, Antonio, Combes, Patricia, Pérez-Redondo, Rosario, Smith, Matthew C. A., Smith, Margaret C. M.
Formato: Texto
Lenguaje:English
Publicado: Oxford University Press 2005
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1140374/
https://www.ncbi.nlm.nih.gov/pubmed/15917435
http://dx.doi.org/10.1093/nar/gni086
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author Rodríguez-García, Antonio
Combes, Patricia
Pérez-Redondo, Rosario
Smith, Matthew C. A.
Smith, Margaret C. M.
author_facet Rodríguez-García, Antonio
Combes, Patricia
Pérez-Redondo, Rosario
Smith, Matthew C. A.
Smith, Margaret C. M.
author_sort Rodríguez-García, Antonio
collection PubMed
description 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.
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spelling pubmed-11403742005-05-26 Natural and synthetic tetracycline-inducible promoters for use in the antibiotic-producing bacteria Streptomyces Rodríguez-García, Antonio Combes, Patricia Pérez-Redondo, Rosario Smith, Matthew C. A. Smith, Margaret C. M. Nucleic Acids Res Methods Online 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. Oxford University Press 2005 2005-05-25 /pmc/articles/PMC1140374/ /pubmed/15917435 http://dx.doi.org/10.1093/nar/gni086 Text en © The Author 2005. Published by Oxford University Press. All rights reserved
spellingShingle Methods Online
Rodríguez-García, Antonio
Combes, Patricia
Pérez-Redondo, Rosario
Smith, Matthew C. A.
Smith, Margaret C. M.
Natural and synthetic tetracycline-inducible promoters for use in the antibiotic-producing bacteria Streptomyces
title Natural and synthetic tetracycline-inducible promoters for use in the antibiotic-producing bacteria Streptomyces
title_full Natural and synthetic tetracycline-inducible promoters for use in the antibiotic-producing bacteria Streptomyces
title_fullStr Natural and synthetic tetracycline-inducible promoters for use in the antibiotic-producing bacteria Streptomyces
title_full_unstemmed Natural and synthetic tetracycline-inducible promoters for use in the antibiotic-producing bacteria Streptomyces
title_short Natural and synthetic tetracycline-inducible promoters for use in the antibiotic-producing bacteria Streptomyces
title_sort natural and synthetic tetracycline-inducible promoters for use in the antibiotic-producing bacteria streptomyces
topic Methods Online
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1140374/
https://www.ncbi.nlm.nih.gov/pubmed/15917435
http://dx.doi.org/10.1093/nar/gni086
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