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 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.
Ross-Macdonald
P
P
Department of Molecular, Cellular and Developmental Biology, Yale University, New Haven, Connecticut 06520-8103, USA.
Coelho
P S
PS
Roemer
T
T
Agarwal
S
S
Kumar
A
A
Jansen
R
R
Cheung
K H
KH
Sheehan
A
A
Symoniatis
D
D
Umansky
L
L
Heidtman
M
M
Nelson
F K
FK
Iwasaki
H
H
Hager
K
K
Gerstein
M
M
Miller
P
P
Roeder
G S
GS
Snyder
M
M
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
D004251:DNA Transposable Elements; D005656:Fungal Proteins