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

Nature biotechnology | Apr 11, 2011

http://www.ncbi.nlm.nih.gov/pubmed/21441928

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 RIS Download

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

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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

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