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A new family of yeast vectors and S288C‐derived strains for the systematic analysis of gene function
Author(s) -
Tomlin Gregory C.,
Wixon Joanne L.,
BolotinFukuhara Monique,
Oliver Stephen G.
Publication year - 2001
Publication title -
yeast
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.923
H-Index - 102
eISSN - 1097-0061
pISSN - 0749-503X
DOI - 10.1002/yea.703
Subject(s) - ura3 , biology , plasmid , genetics , gene , shuttle vector , auxotrophy , genome , saccharomyces cerevisiae , yeast , escherichia coli , gene family , computational biology , vector (molecular biology) , recombinant dna
The yeast genome has been shown to contain a significant number of gene families with more than three members. In order to study these families it is often necessary to generate strains carrying deletions of all members of the family, which can require a wide range of auxotrophic markers. To facilitate such studies, we have generated yeast strains containing deletions of a selection of nutritional marker genes ( ade2 , ade4 , ade8 , met3 and met14 ). We have also cloned the corresponding cognate genes, allowing their use in PCR‐based gene disruptions. Two new pRS family Saccharomyces cerevisiae–Escherichia coli shuttle vectors containing ADE8 (one low‐copy, pRS4110, and one high‐copy, pRS4210) have been produced for use in conjunction with the new strains. A system for easier synthetic lethal screening using one of these new markers is also presented. The ADE8 and HIS3 genes have been cloned together on a high‐copy vector (pRS4213), providing a plasmid for red–white colour screening in the ade2 Δ 0 ade8 Δ 0 strains we have generated. In contrast to some conventional systems, this plasmid allows for screening using gene libraries constructed in URA3 plasmids. Copyright © 2001 John Wiley & Sons, Ltd.