These authors contributed equally to this work.
Yeast Functional Analysis Report
A suite of Gateway® cloning vectors for high-throughput genetic analysis in Saccharomyces cerevisiae
Article first published online: 21 JUN 2007
Copyright © 2007 John Wiley & Sons, Ltd.
Volume 24, Issue 10, pages 913–919, October 2007
How to Cite
Alberti, S., Gitler, A. D. and Lindquist, S. (2007), A suite of Gateway® cloning vectors for high-throughput genetic analysis in Saccharomyces cerevisiae. Yeast, 24: 913–919. doi: 10.1002/yea.1502
- Issue published online: 24 SEP 2007
- Article first published online: 21 JUN 2007
- Manuscript Accepted: 9 APR 2007
- Manuscript Received: 9 MAR 2007
- Life Sciences Research Foundation
- Deutsche Forschungsgemeinschaft (DFG)
- Udall Center. Grant Number: NS38372
In the post-genomic era, academic and biotechnological research is increasingly shifting its attention from single proteins to the analysis of complex protein networks. This change in experimental design requires the use of simple and experimentally tractable organisms, such as the unicellular eukaryote Saccharomyces cerevisiae, and a range of new high-throughput techniques. The Gateway® system has emerged as a powerful high-throughput cloning method that allows for the in vitro recombination of DNA with high speed, accuracy and reliability. Two Gateway-based libraries of overexpression plasmids containing the entire complement of yeast open reading frames (ORFs) have recently been completed. In order to make use of these powerful resources, we adapted the widely used pRS series of yeast shuttle vectors for use in Gateway-based cloning. The resulting suite of 288 yeast Gateway vectors is based upon the two commonly used GPD and GAL1 promoter expression systems that enable expression of ORFs, either constitutively or under galactose-inducible conditions. In addition, proteins of interest can be fused to a choice of frequently used N- or C-terminal tags, such as EGFP, ECFP, EYFP, Cerulean, monomeric DsRed, HA or TAP. We have made this yeast Gateway® vector kit available to the research community via the non-profit Addgene Plasmid Repository (http://www.addgene.org/yeast_gateway). Copyright © 2007 John Wiley & Sons, Ltd.