title: Large-scale analysis of the yeast genome by transposon tagging and gene disruption.
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 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.
authors.affiliation: Department of Molecular, Cellular and Developmental Biology, Yale University, New Haven, Connecticut 06520-8103, USA.
authors.forename: P
authors.initials: P
authors.lastname: Ross-Macdonald
authors.forename: P S
authors.initials: PS
authors.lastname: Coelho
authors.forename: T
authors.initials: T
authors.lastname: Roemer
authors.forename: S
authors.initials: S
authors.lastname: Agarwal
authors.forename: A
authors.initials: A
authors.lastname: Kumar
authors.forename: R
authors.initials: R
authors.lastname: Jansen
authors.forename: K H
authors.initials: KH
authors.lastname: Cheung
authors.forename: A
authors.initials: A
authors.lastname: Sheehan
authors.forename: D
authors.initials: D
authors.lastname: Symoniatis
authors.forename: L
authors.initials: L
authors.lastname: Umansky
authors.forename: M
authors.initials: M
authors.lastname: Heidtman
authors.forename: F K
authors.initials: FK
authors.lastname: Nelson
authors.forename: H
authors.initials: H
authors.lastname: Iwasaki
authors.forename: K
authors.initials: K
authors.lastname: Hager
authors.forename: M
authors.initials: M
authors.lastname: Gerstein
authors.forename: P
authors.initials: P
authors.lastname: Miller
authors.forename: G S
authors.initials: GS
authors.lastname: Roeder
authors.forename: M
authors.initials: M
authors.lastname: Snyder
mesh_terms: D000465:Algorithms; D004251:DNA Transposable Elements; D004926:Escherichia coli / Q000235:genetics; D005656:Fungal Proteins / Q000235:genetics / Q000502:physiology; D020869:Gene Expression Profiling; D005821:Genetic Techniques; D016681:Genome, Fungal; D008969:Molecular Sequence Data; D016296:Mutagenesis; D020411:Oligonucleotide Array Sequence Analysis; D016366:Open Reading Frames; D010641:Phenotype; D016133:Polymerase Chain Reaction; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D014170:Transformation, Genetic
chemical_list: D004251:DNA Transposable Elements; D005656:Fungal Proteins