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

Petra Ross-Macdonald, Paulo S. R. Coelho, Terry Roemer, Seema Agarwal, Anuj Kumar, Ronald Jansen, Kei-Hoi Cheung, Amy Sheehan, Dawn Symoniatis, Lara Umansky, Matthew Heidtman, F. Kenneth Nelson, Hiroshi Iwasaki, Karl Hager, Mark Gerstein, Perry Miller, G. Shirleen Roeder and Michael Snyder ()
Additional contact information
Petra Ross-Macdonald: Cellular and Developmental Biology, Yale University, PO Box 208103
Paulo S. R. Coelho: Cellular and Developmental Biology, Yale University, PO Box 208103
Terry Roemer: Cellular and Developmental Biology, Yale University, PO Box 208103
Seema Agarwal: Cellular and Developmental Biology, Yale University, PO Box 208103
Anuj Kumar: Cellular and Developmental Biology, Yale University, PO Box 208103
Ronald Jansen: Yale University, PO Box 208114
Kei-Hoi Cheung: Center for Medical Informatics, Yale University School of Medicine
Amy Sheehan: Cellular and Developmental Biology, Yale University, PO Box 208103
Dawn Symoniatis: Cellular and Developmental Biology, Yale University, PO Box 208103
Lara Umansky: Cellular and Developmental Biology, Yale University, PO Box 208103
Matthew Heidtman: Cellular and Developmental Biology, Yale University, PO Box 208103
F. Kenneth Nelson: Cellular and Developmental Biology, Yale University, PO Box 208103
Hiroshi Iwasaki: Cellular and Developmental Biology, Yale University, PO Box 208103
Karl Hager: Keck Foundation Biotechnology Resource Laboratory, 295 Congress Avenue, Yale University
Mark Gerstein: Yale University, PO Box 208114
Perry Miller: Center for Medical Informatics, Yale University School of Medicine
G. Shirleen Roeder: Cellular and Developmental Biology, Yale University, PO Box 208103
Michael Snyder: Cellular and Developmental Biology, Yale University, PO Box 208103

Nature, 1999, vol. 402, issue 6760, 413-418

Abstract: 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.

Date: 1999
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DOI: 10.1038/46558

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