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.
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Acknowledgments
This work was supported by the Aventis Foundation and the Deutsche Forschungsgemeinschaft (SU402/1-2, SFB579, and the Excellence Cluster: Macromolecular Complexes).
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Suess, B., Entian, KD., Kötter, P., Weigand, J.E. (2012). Aptamer-Regulated Expression of Essential Genes in Yeast. In: Lorence, A. (eds) Recombinant Gene Expression. Methods in Molecular Biology, vol 824. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-433-9_20
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DOI: https://doi.org/10.1007/978-1-61779-433-9_20
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