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Nucleic Acids Research Advance Access originally published online on July 10, 2009
Nucleic Acids Research 2009 37(18):e120; doi:10.1093/nar/gkp578
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Nucleic Acids Research, 2009, Vol. 37, No. 18 e120
© 2009 The Author(s)
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Methods Online

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

Peter Kötter1,2, Julia E. Weigand1,3, Britta Meyer1,2, Karl-Dieter Entian1,2,* and Beatrix Suess1,3,*

1Institut für Molekulare Biowissenschaften, 2Cluster of Excellence: Macromolecular Complexes, Johann Wolfgang Goethe-Universität Frankfurt, Max-von-Laue-Str. 9, D-60438 Frankfurt/M., Germany and 3Aventis Foundation Endowed Professorship

*To whom correspondence should be addressed. Tel: +49 69 798 29785; Fax: +49 69 798 29323; Email: suess{at}bio.uni-frankfurt.de

Correspondence may also be addressed to Karl-Dieter Entian. Tel: +49 69 798 29525; Fax: +49 69 798 29323; Email: entian{at}bio.uni-frankfurt.de

Received January 28, 2009. Revised June 23, 2009. Accepted June 23, 2009.

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.


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