Quantitative phenotypic analysis of yeast deletion mutants using a highly parallel molecular bar-coding strategy.
A quantitative and highly parallel method for analysing deletion mutants has been developed to aid in determining the biological function of thousands of newly identified open reading frames (ORFs) in Saccharomyces cerevisiae. This approach uses a PCR targeting strategy to generate large numbers of deletion strains. Each deletion strain is labelled with a unique 20-base tag sequence that can be detected by hybridization to a high-density oligonucleotide array. The tags serve as unique identifiers (molecular bar codes) that allow analysis of large numbers of deletion strains simultaneously through selective growth conditions. Hybridization experiments show that the arrays are specific, sensitive and quantitative. A pilot study with 11 known yeast genes suggests that the method can be extended to include all of the ORFs in the yeast genome, allowing whole genome analysis with a single selective growth condition and a single hybridization.
Shoemaker
D D
DD
Department of Biochemistry, Beckman Center, Stanford University Medical Center, CA 94305, USA.
Lashkari
D A
DA
Morris
D
D
Mittmann
M
M
Davis
R W
RW
D004247:DNA; D004252:DNA Mutational Analysis / Q000379:methods; D005453:Fluorescence; D005821:Genetic Techniques; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D016366:Open Reading Frames; D010641:Phenotype; D010865:Pilot Projects; D016133:Polymerase Chain Reaction / Q000379:methods; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D017384:Sequence Deletion
D004247:DNA