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Systematic exploration of essential yeast gene function with temperature-sensitive mutants.

Conditional temperature-sensitive (ts) mutations are valuable reagents for studying essential genes in the yeast Saccharomyces cerevisiae. We constructed 787 ts strains, covering 497 (∼45%) of the 1,101 essential yeast genes, with ∼30% of the genes represented by multiple alleles. All of the alleles are integrated into their native genomic locus in the S288C common reference strain and are linked to a kanMX selectable marker, allowing further genetic manipulation by synthetic genetic array (SGA)-based, high-throughput methods. We show two such manipulations: barcoding of 440 strains, which enables chemical-genetic suppression analysis, and the construction of arrays of strains carrying different fluorescent markers of subcellular structure, which enables quantitative analysis of phenotypes using high-content screening. Quantitative analysis of a GFP-tubulin marker identified roles for cohesin and condensin genes in spindle disassembly. This mutant collection should facilitate a wide range of systematic studies aimed at understanding the functions of essential genes.

Pubmed ID: 21441928


  • Li Z
  • Vizeacoumar FJ
  • Bahr S
  • Li J
  • Warringer J
  • Vizeacoumar FS
  • Min R
  • Vandersluis B
  • Bellay J
  • Devit M
  • Fleming JA
  • Stephens A
  • Haase J
  • Lin ZY
  • Baryshnikova A
  • Lu H
  • Yan Z
  • Jin K
  • Barker S
  • Datti A
  • Giaever G
  • Nislow C
  • Bulawa C
  • Myers CL
  • Costanzo M
  • Gingras AC
  • Zhang Z
  • Blomberg A
  • Bloom K
  • Andrews B
  • Boone C


Nature biotechnology

Publication Data

April 11, 2011

Associated Grants

  • Agency: Canadian Institutes of Health Research, Id: MOP-97939
  • Agency: NIGMS NIH HHS, Id: R01 GM032238
  • Agency: NIGMS NIH HHS, Id: R01 GM032238-24
  • Agency: NHGRI NIH HHS, Id: R01 HG005084
  • Agency: NIGMS NIH HHS, Id: R37 GM032238

Mesh Terms

  • Alleles
  • Databases, Genetic
  • Genes, Essential
  • Genes, Fungal
  • Genes, Lethal
  • Genetic Engineering
  • Genetic Loci
  • Genome, Fungal
  • Mass Spectrometry
  • Microarray Analysis
  • Microscopy, Confocal
  • Mutation
  • Phenotype
  • Plasmids
  • RNA, Messenger
  • Saccharomyces cerevisiae
  • Single-Cell Analysis
  • Temperature
  • Tubulin