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Large-scale analysis of the yeast genome by transposon tagging and gene disruption

Nature volume 402, pages 413–418 (1999)Cite this article

Abstract

Economical methods by which gene function may be analysed on a genomic scale are relatively scarce. To fill this need, we have developed a transposon-tagging strategy for the genome-wide analysis of disruption phenotypes, gene expression and protein localization, and have applied this method to the large-scale analysis of gene function in the budding yeast Saccharomyces cerevisiae. Here we present the largest collection of defined yeast mutants ever generated within a single genetic background—a collection of over 11,000 strains, each carrying a transposon inserted within a region of the genome expressed during vegetative growth and/or sporulation. These insertions affect nearly 2,000 annotated genes, representing about one-third of the 6,200 predicted genes in the yeast genome1,2. We have used this collection to determine disruption phenotypes for nearly 8,000 strains using 20 different growth conditions; the resulting data sets were clustered to identify groups of functionally related genes. We have also identified over 300 previously non-annotated open reading frames and analysed by indirect immunofluorescence over 1,300 transposon-tagged proteins. In total, our study encompasses over 260,000 data points, constituting the largest functional analysis of the yeast genome ever undertaken.

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Figure 1: The mTn insertion project.
Figure 2: Distribution and phenotypic analysis of 1,917 mTn-mutagenized ORFs within the S. cerevisiae genome.
Figure 3: Phenotypic macroarray analysis.
Figure 4: Graphical representation of clustered phenotypic data.
Figure 5: Immunolocalization of epitope-tagged proteins.

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Acknowledgements

P.S.R.C. is supported by the Fundacao de Amparo a Pesquisa do Estado de Sao Paulo, Brazil. This work was supported by an NIH grant (to G.S.R. and M.S.).

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Authors and Affiliations

  1. Department of Molecular, Cellular and Developmental Biology, Yale University, PO Box 208103, New Haven, 06520-8103, Connecticut, USA

    Petra Ross-Macdonald, Paulo S. R. Coelho, Terry Roemer, Seema Agarwal, Anuj Kumar, Amy Sheehan, Dawn Symoniatis, Lara Umansky, Matthew Heidtman, F. Kenneth Nelson, Hiroshi Iwasaki, G. Shirleen Roeder & Michael Snyder

  2. Department of Molecular Biophysics and Biochemistry, Yale University, PO Box 208114, New Haven, 06520-8114, Connecticut, USA

    Ronald Jansen, Mark Gerstein & Michael Snyder

  3. Department of Anesthesiology, Center for Medical Informatics, Yale University School of Medicine, New Haven, 06510, Connecticut, USA

    Kei-Hoi Cheung & Perry Miller

  4. Keck Foundation Biotechnology Resource Laboratory, 295 Congress Avenue, Yale University, New Haven, 06520, Connecticut, USA

    Karl Hager

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  1. Petra Ross-Macdonald
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Correspondence to Michael Snyder.

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Ross-Macdonald, P., Coelho, P., Roemer, T. et al. Large-scale analysis of the yeast genome by transposon tagging and gene disruption. Nature 402, 413–418 (1999). https://doi.org/10.1038/46558

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