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A fast and efficient translational control system for conditional expression of yeast genes.

Kötter, Peter and Weigand, Julia E. and Meyer, Britta and Entian, Karl-Dieter and Suess, Beatrix (2009):
A fast and efficient translational control system for conditional expression of yeast genes.
In: Nucleic acids research, 37 (18), pp. e120. ISSN 1362-4962,
[Article]

Abstract

A new artificial regulatory system for essential genes in yeast is described. It prevents translation of target mRNAs upon tetracycline (tc) binding to aptamers introduced into their 5'UTRs. Exploiting direct RNA-ligand interaction renders auxiliary protein factors unnecessary. Therefore, our approach is strain independent and not susceptible to interferences by heterologous expressed regulatory proteins. We use a simple PCR-based strategy, which allows easy tagging of any target gene and the level of gene expression can be adjusted due to various tc aptamer-regulated promoters. As proof of concept, five differently expressed genes were targeted, two of which could not be regulated previously. In all cases, adding tc completely prevented growth and, as shown for Nop14p, rapidly abolished de novo protein synthesis providing a powerful tool for conditional regulation of yeast gene expression.

Item Type: Article
Erschienen: 2009
Creators: Kötter, Peter and Weigand, Julia E. and Meyer, Britta and Entian, Karl-Dieter and Suess, Beatrix
Title: A fast and efficient translational control system for conditional expression of yeast genes.
Language: English
Abstract:

A new artificial regulatory system for essential genes in yeast is described. It prevents translation of target mRNAs upon tetracycline (tc) binding to aptamers introduced into their 5'UTRs. Exploiting direct RNA-ligand interaction renders auxiliary protein factors unnecessary. Therefore, our approach is strain independent and not susceptible to interferences by heterologous expressed regulatory proteins. We use a simple PCR-based strategy, which allows easy tagging of any target gene and the level of gene expression can be adjusted due to various tc aptamer-regulated promoters. As proof of concept, five differently expressed genes were targeted, two of which could not be regulated previously. In all cases, adding tc completely prevented growth and, as shown for Nop14p, rapidly abolished de novo protein synthesis providing a powerful tool for conditional regulation of yeast gene expression.

Journal or Publication Title: Nucleic acids research
Journal volume: 37
Number: 18
Divisions: 10 Department of Biology
10 Department of Biology > Synthetic Genetic Circuits
10 Department of Biology > RNA Biochemistry
Date Deposited: 22 Feb 2012 10:27
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