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

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...

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Bibliographic Details
Published in:	Nature (London) Vol. 402; no. 6760; pp. 413 - 418
Main Authors:	Snyder, Michael, Ross-Macdonald, Petra, Coelho, Paulo S. R, Roemer, Terry, Agarwal, Seema, Kumar, Anuj, Jansen, Ronald, Cheung, Kei-Hoi, Sheehan, Amy, Symoniatis, Dawn, Umansky, Lara, Heidtman, Matthew, Nelson, F. Kenneth, Iwasaki, Hiroshi, Hager, Karl, Gerstein, Mark, Miller, Perry, Roeder, G. Shirleen
Format:	Journal Article
Language:	English
Published:	London Nature Publishing 25-11-1999 Nature Publishing Group
Subjects:	Algorithms Biological and medical sciences Classical genetics, quantitative genetics, hybrids DNA Transposable Elements Escherichia coli - genetics Fundamental and applied biological sciences. Psychology Fungal Proteins - genetics Fungal Proteins - physiology Gene Expression Profiling Genes Genetic Techniques Genetics of eukaryotes. Biological and molecular evolution Genome, Fungal Growth conditions Molecular Sequence Data Mutagenesis Oligonucleotide Array Sequence Analysis Open Reading Frames Phenotype Polymerase Chain Reaction Proteins Saccharomyces cerevisiae Saccharomyces cerevisiae - genetics Thallophyta, bryophyta Transformation, Genetic Vegetals Yeast Yeasts Yeast Cell function Targeting Transposon Gene expression Protein Fungi Phenotype Analysis method Gene Mutagenesis Insertion mutation Ascomycetes Genome Localization Saccharomyces cerevisiae Thallophyta
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Summary:	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 genome. 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.
Bibliography:	ObjectType-Article-2 SourceType-Scholarly Journals-1 ObjectType-Feature-1 content type line 23 ObjectType-Article-1 ObjectType-Feature-2
ISSN:	0028-0836 1476-4687
DOI:	10.1038/46558