Suess, Beatrix ; Entian, Karl-Dieter ; Kötter, Peter ; Weigand, Julia E. (2012)
Aptamer-regulated expression of essential genes in yeast.
In: Methods in molecular biology (Clifton, N.J.), 824
Artikel, Bibliographie
Kurzbeschreibung (Abstract)
Conditional gene expression systems are important tools for the functional analysis of essential genes. Tetracycline (tc)-binding aptamers can be exploited as artificial riboswitches for the efficient control of gene expression by inserting them into the 5' untranslated region of an mRNA. The ligand-bound form of those mRNAs inhibits gene expression by interfering with translation initiation. In contrast to previous tc-dependent regulatory systems, where tc inhibits or activates transcription upon binding to the repressor protein TetR, the tc-binding aptamer system inhibits translation of the respective mRNA. We describe here a simple and powerful PCR-based strategy which allows easy tagging of any target gene in yeast using a tc aptamer-containing insertion cassette. The expression window can be adjusted with different promoters and protein synthesis is rapidly switched off.
| Typ des Eintrags: | Artikel |
|---|---|
| Erschienen: | 2012 |
| Autor(en): | Suess, Beatrix ; Entian, Karl-Dieter ; Kötter, Peter ; Weigand, Julia E. |
| Art des Eintrags: | Bibliographie |
| Titel: | Aptamer-regulated expression of essential genes in yeast. |
| Sprache: | Englisch |
| Publikationsjahr: | 2012 |
| Titel der Zeitschrift, Zeitung oder Schriftenreihe: | Methods in molecular biology (Clifton, N.J.) |
| Jahrgang/Volume einer Zeitschrift: | 824 |
| Kurzbeschreibung (Abstract): | Conditional gene expression systems are important tools for the functional analysis of essential genes. Tetracycline (tc)-binding aptamers can be exploited as artificial riboswitches for the efficient control of gene expression by inserting them into the 5' untranslated region of an mRNA. The ligand-bound form of those mRNAs inhibits gene expression by interfering with translation initiation. In contrast to previous tc-dependent regulatory systems, where tc inhibits or activates transcription upon binding to the repressor protein TetR, the tc-binding aptamer system inhibits translation of the respective mRNA. We describe here a simple and powerful PCR-based strategy which allows easy tagging of any target gene in yeast using a tc aptamer-containing insertion cassette. The expression window can be adjusted with different promoters and protein synthesis is rapidly switched off. |
| Fachbereich(e)/-gebiet(e): | 10 Fachbereich Biologie 10 Fachbereich Biologie > Synthetic Genetic Circuits (2020 umbenannt in "Synthetic RNA biology") 10 Fachbereich Biologie > RNA Biochemie |
| Hinterlegungsdatum: | 22 Feb 2012 09:57 |
| Letzte Änderung: | 06 Nov 2020 07:16 |
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