Rudolph, Martin M. ; Vockenhuber, Michael-Paul ; Suess, Beatrix (2015)
Conditional Control of Gene Expression by Synthetic Riboswitches in Streptomyces coelicolor.
In: Methods in enzymology, 550
Artikel, Bibliographie
Kurzbeschreibung (Abstract)
Here we provide a step-by-step protocol for the application of synthetic theophylline-dependent riboswitches for conditional gene expression in Streptomyces coelicolor. Application of the method requires a sequence of only ~85nt to be inserted between the transcriptional start site and the start codon of a gene of interest. No auxiliary factors are needed. All tested riboswitch variants worked well in concert with the promoters galP2, ermEp1, and SF14. Moreover, they allowed theophylline-dependent expression not only of the heterologous β-glucuronidase reporter gene but also of dagA, an endogenous agarase gene. The right combination of the tested promoters with the riboswitch variants allows for the adjustment of the desired dynamic range of regulation in a highly specific and dose-dependent manner and underlines the orthogonality of riboswitch regulation. We anticipate that any additional natural or synthetic promoter can be combined with the presented riboswitches. Moreover, this system should easily be transferable to other Streptomyces species, and most likely to any other genetically manipulable bacteria.
| Typ des Eintrags: | Artikel |
|---|---|
| Erschienen: | 2015 |
| Autor(en): | Rudolph, Martin M. ; Vockenhuber, Michael-Paul ; Suess, Beatrix |
| Art des Eintrags: | Bibliographie |
| Titel: | Conditional Control of Gene Expression by Synthetic Riboswitches in Streptomyces coelicolor. |
| Sprache: | Englisch |
| Publikationsjahr: | 2015 |
| Titel der Zeitschrift, Zeitung oder Schriftenreihe: | Methods in enzymology |
| Jahrgang/Volume einer Zeitschrift: | 550 |
| Kurzbeschreibung (Abstract): | Here we provide a step-by-step protocol for the application of synthetic theophylline-dependent riboswitches for conditional gene expression in Streptomyces coelicolor. Application of the method requires a sequence of only ~85nt to be inserted between the transcriptional start site and the start codon of a gene of interest. No auxiliary factors are needed. All tested riboswitch variants worked well in concert with the promoters galP2, ermEp1, and SF14. Moreover, they allowed theophylline-dependent expression not only of the heterologous β-glucuronidase reporter gene but also of dagA, an endogenous agarase gene. The right combination of the tested promoters with the riboswitch variants allows for the adjustment of the desired dynamic range of regulation in a highly specific and dose-dependent manner and underlines the orthogonality of riboswitch regulation. We anticipate that any additional natural or synthetic promoter can be combined with the presented riboswitches. Moreover, this system should easily be transferable to other Streptomyces species, and most likely to any other genetically manipulable bacteria. |
| Fachbereich(e)/-gebiet(e): | 10 Fachbereich Biologie 10 Fachbereich Biologie > Synthetic Genetic Circuits (2020 umbenannt in "Synthetic RNA biology") |
| Hinterlegungsdatum: | 27 Jan 2015 11:34 |
| Letzte Änderung: | 27 Jan 2015 11:34 |
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