Systematic exploration of essential yeast gene function with temperature-sensitive mutants |
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Authors: | Li Zhijian Vizeacoumar Franco J Bahr Sondra Li Jingjing Warringer Jonas Vizeacoumar Frederick S Min Renqiang Vandersluis Benjamin Bellay Jeremy Devit Michael Fleming James A Stephens Andrew Haase Julian Lin Zhen-Yuan Baryshnikova Anastasia Lu Hong Yan Zhun Jin Ke Barker Sarah Datti Alessandro Giaever Guri Nislow Corey Bulawa Chris Myers Chad L Costanzo Michael Gingras Anne-Claude Zhang Zhaolei Blomberg Anders Bloom Kerry Andrews Brenda Boone Charles |
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Institution: | Banting and Best Department of Medical Research, The Donnelly Centre, University of Toronto, Toronto, Ontario, Canada. |
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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. |
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