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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...
Autores principales: | , , , , |
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Formato: | Texto |
Lenguaje: | English |
Publicado: |
Oxford University Press
2005
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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. |
format | Text |
id | pubmed-1140374 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2005 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
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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