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A systematic library for comprehensive overexpression screens in Saccharomyces cerevisiae

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Abstract

Modern genetic analysis requires the development of new resources to systematically explore gene function in vivo. Overexpression screens are a powerful method to investigate genetic pathways, but the goal of routine and comprehensive overexpression screens has been hampered by the lack of systematic libraries. Here we describe the construction of a systematic collection of the Saccharomyces cerevisiae genome in a high-copy vector and its validation in two overexpression screens.

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Figure 1: Construction of a systematic yeast overexpression library.
Figure 2: Validation of the systematic library in overexpression screens.

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Acknowledgements

We thank T. Moore at Open Biosystems for production of library plasmid DNA. This work was supported by grant GM52486 to G.P. from the US National Institutes of Health, a National Institutes of Health Research Supplement to Promote Diversity in Health-Related Research to G.M.J., a Dean's Pilot Project grant from the Albert Einstein College of Medicine, and the Wellcome Trust.

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Authors and Affiliations

  1. Department of Molecular Genetics, Albert Einstein College of Medicine, 1300 Morris Park Avenue, Bronx, 10461, New York, USA

    Grace Marie Jones & Gregory Prelich

  2. The Welcome Trust Sanger Institute, Welcome Trust Genome Campus, Hinxton, CB10 1SA, Cambridge, UK

    Jim Stalker, Sean Humphray, Anthony West, Tony Cox, Jane Rogers & Ian Dunham

Authors
  1. Grace Marie Jones
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  2. Jim Stalker
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  3. Sean Humphray
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  4. Anthony West
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  5. Tony Cox
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  6. Jane Rogers
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  7. Ian Dunham
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  8. Gregory Prelich
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Contributions

G.M.J. performed experiments. T.C. and J.S., bioinformatics; S.H., clone management; A.W., sequencing. J.R. and I.D. directed and managed sequencing and bioinformatics; G.P. oversaw the project; G.M.J. and G.P. wrote the manuscript.

Corresponding author

Correspondence to Gregory Prelich.

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Competing interests

G.P. and his lab receive compensation from Open Biosystems for the distribution of their library.

Supplementary information

Supplementary Text and Figures

Supplementary Figure 1, Supplementary Methods (PDF 698 kb)

Supplementary Table 1

Yeast Genomic Tiling Collection. (XLS 2214 kb)

Supplementary Table 2

YGPM plasmids that confer a sick or lethal phenotype. (XLS 15 kb)

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Jones, G., Stalker, J., Humphray, S. et al. A systematic library for comprehensive overexpression screens in Saccharomyces cerevisiae. Nat Methods 5, 239–241 (2008). https://doi.org/10.1038/nmeth.1181

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  • DOI: https://doi.org/10.1038/nmeth.1181

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