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Updated: May 26, 2026

Rapid Synthesis and Screening of Chemically Activated Transcription Factors with GFP-based Reporters
Published on: November 26, 2013
Aptamer-regulated expression of essential genes in yeast
Beatrix Suess1, Karl-Dieter Entian, Peter Kötter
1Department of Molecular Biosciences, Johann Wolfgang Goethe-University Frankfurt, Frankfurt, Germany. Suess@bio.uni-frankfurt.de
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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