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- Title
Systematic exploration of essential yeast gene function with temperature-sensitive mutants.
- Authors
Zhijian Li; Vizeacoumar, Franco J.; Bahr, Sondra; Jingjing Li; Warringer, Jonas; Vizeacoumar, Frederick S.; Renqiang Min; VanderSluis, Benjamin; Bellay, Jeremy; DeVit, Michael; Fleming, James A.; Stephens, Andrew; Haase, Julian; Zhen-Yuan Lin; Baryshnikova, Anastasia; Hong Lu; Zhun Yan; Ke Jin; Barker, Sarah; Datti, Alessandro
- 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.
- Publication
Nature Biotechnology, 2011, Vol 29, Issue 4, p361
- ISSN
1087-0156
- Publication type
Academic Journal
- DOI
10.1038/nbt.1832