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

被引：295

作者：

Li, Zhijian ^{[2
]}

Vizeacoumar, Franco J. ^{[2
]}

Bahr, Sondra ^{[2
]}

Li, Jingjing ^{[2
]}

Warringer, Jonas ^{[3
]}

Vizeacoumar, Frederick S. ^{[4
]}

Min, Renqiang ^{[2
]}

VanderSluis, Benjamin ^{[5
]}

Bellay, Jeremy ^{[5
]}

DeVit, Michael ^{[6
]}

Fleming, James A. ^{[6
]}

Stephens, Andrew ^{[7
]}

Haase, Julian ^{[7
]}

Lin, Zhen-Yuan ^{[2
]}

Baryshnikova, Anastasia ^{[2
]}

Lu, Hong ^{[2
]}

Yan, Zhun ^{[2
]}

Jin, Ke ^{[2
]}

Barker, Sarah ^{[2
]}

Datti, Alessandro ^{[4
,8
]}

Giaever, Guri ^{[2
]}

Nislow, Corey ^{[2
]}

Bulawa, Chris ^{[6
]}

Myers, Chad L. ^{[5
]}

Costanzo, Michael ^{[2
]}

Gingras, Anne-Claude ^{[2
]}

Zhang, Zhaolei ^{[2
]}

Blomberg, Anders ^{[3
]}

Bloom, Kerry ^{[7
]}

Andrews, Brenda ^{[1
,2
]}

Boone, Charles ^{[2
]}

机构：

[1] Univ Toronto, Banting & Best Dept Med Res, Toronto, ON, Canada

[2] Univ Toronto, Dept Mol Genet, Donnelly Ctr, Toronto, ON, Canada

[3] Univ Gothenburg, Dept Cell & Mol Biol, Gothenburg, Sweden

[4] Mt Sinai Hosp, Samuel Lunenfeld Res Inst, Toronto, ON M5G 1X5, Canada

[5] Univ Minnesota, Dept Comp Sci & Engn, Minneapolis, MN USA

[6] FoldRx Pharmaceut Inc, Cambridge, MA USA

[7] Univ N Carolina, Dept Biol, Chapel Hill, NC USA

[8] Univ Perugia, Dept Expt Med & Biochem Sci, I-06100 Perugia, Italy

来源：

NATURE BIOTECHNOLOGY | 2011年 / 29卷 / 04期

基金：

加拿大健康研究院;

关键词：

CHROMOSOME CONDENSATION; SPINDLE CHECKPOINT; DELETION MUTANTS; PROTEIN-KINASE; ANAPHASE; COMPLEX; PATHWAY; IDENTIFICATION; LOCALIZATION; ORGANIZATION;

D O I：

10.1038/nbt.1832

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

Conditional temperature-sensitive (ts) mutations are valuable reagents for studying essential genes in the yeast Saccharomyces cerevisiae. We constructed 787 ts strains, covering 497 (similar to 45%) of the 1,101 essential yeast genes, with similar to 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.

引用

页码：361 / U105

页数：9

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