title: Systematic exploration of essential yeast gene function with temperature-sensitive mutants.
abstract: 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.
authors: Banting and Best Department of Medical Research, The Donnelly Centre, University of Toronto, Toronto, Ontario, Canada.
authors: Zhijian
authors: Z
authors: Li
authors: Franco J
authors: FJ
authors: Vizeacoumar
authors: Sondra
authors: S
authors: Bahr
authors: Jingjing
authors: J
authors: Li
authors: Jonas
authors: J
authors: Warringer
authors: Frederick S
authors: FS
authors: Vizeacoumar
authors: Renqiang
authors: R
authors: Min
authors: Benjamin
authors: B
authors: Vandersluis
authors: Jeremy
authors: J
authors: Bellay
authors: Michael
authors: M
authors: Devit
authors: James A
authors: JA
authors: Fleming
authors: Andrew
authors: A
authors: Stephens
authors: Julian
authors: J
authors: Haase
authors: Zhen-Yuan
authors: ZY
authors: Lin
authors: Anastasia
authors: A
authors: Baryshnikova
authors: Hong
authors: H
authors: Lu
authors: Zhun
authors: Z
authors: Yan
authors: Ke
authors: K
authors: Jin
authors: Sarah
authors: S
authors: Barker
authors: Alessandro
authors: A
authors: Datti
authors: Guri
authors: G
authors: Giaever
authors: Corey
authors: C
authors: Nislow
authors: Chris
authors: C
authors: Bulawa
authors: Chad L
authors: CL
authors: Myers
authors: Michael
authors: M
authors: Costanzo
authors: Anne-Claude
authors: AC
authors: Gingras
authors: Zhaolei
authors: Z
authors: Zhang
authors: Anders
authors: A
authors: Blomberg
authors: Kerry
authors: K
authors: Bloom
authors: Brenda
authors: B
authors: Andrews
authors: Charles
authors: C
authors: Boone
mesh_terms: D000483:Alleles; D030541:Databases, Genetic; D020043:Genes, Essential; D005800:Genes, Fungal; D005804:Genes, Lethal; D005818:Genetic Engineering / Q000379:methods; D056426:Genetic Loci; D016681:Genome, Fungal; D013058:Mass Spectrometry / Q000379:methods; D046228:Microarray Analysis / Q000379:methods; D018613:Microscopy, Confocal; D009154:Mutation; D010641:Phenotype; D010957:Plasmids; D012333:RNA, Messenger; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000254:growth & development; D059010:Single-Cell Analysis; D013696:Temperature; D014404:Tubulin / Q000032:analysis
chemical_list: D012333:RNA, Messenger; D014404:Tubulin