title: Aptamer-regulated expression of essential genes in yeast.
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.
authors.affiliation: Department of Molecular Biosciences, Johann Wolfgang Goethe-University Frankfurt, Frankfurt, Germany. Suess@bio.uni-frankfurt.de
authors.forename: Beatrix
authors.initials: B
authors.lastname: Suess
authors.forename: Karl-Dieter
authors.initials: KD
authors.lastname: Entian
authors.forename: Peter
authors.initials: P
authors.lastname: Kötter
authors.forename: Julia E
authors.initials: JE
authors.lastname: Weigand
mesh_terms: D052157:Aptamers, Nucleotide / Q000235:genetics* / Q000378:metabolism; D015153:Blotting, Western / Q000379:methods; D015966:Gene Expression Regulation, Fungal / Q000235:genetics / Q000502:physiology*; D020043:Genes, Essential / Q000235:genetics*; D010442:Peptide Chain Initiation, Translational / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism*; D058928:Riboswitch / Q000235:genetics*; D013752:Tetracycline / Q000378:metabolism*
chemical_list: D052157:Aptamers, Nucleotide; D012333:RNA, Messenger; D058928:Riboswitch; D013752:Tetracycline