Genetics, Vol. 162, 1091-1099, November 2002, Copyright © 2002

High-Resolution Genetic Mapping With Ordered Arrays of Saccharomyces cerevisiae Deletion Mutants

Paul Jorgensena,b, Bryce Nelsonc,d, Mark D. Robinsond, Yiqun Chend, Brenda Andrewsa, Mike Tyersa,b, and Charles Boonea,c,d
a Department of Medical Genetics and Microbiology, University of Toronto, Toronto, Ontario M5S 1A8, Canada,
b Program in Molecular Biology and Cancer, Samuel Lunenfeld Research Institute, Mt. Sinai Hospital, Toronto, Ontario M5G 1X5, Canada,
c Biology Department, Queens University, Kingston, Ontario K7L 3N6, Canada
d Banting and Best Department of Medical Research, University of Toronto, Toronto, Ontario M5G 1L6, Canada

Corresponding author: Charles Boone, Toronto, ON M5G 1L6, Canada., charlie.boone{at}utoronto.ca (E-mail)

Communicating editor: M. JOHNSTON

We present a method for high-resolution genetic mapping that takes advantage of the ordered set of viable gene deletion mutants, which form a set of colinear markers covering almost every centimorgan of the Saccharomyces cerevisiae genome, and of the synthetic genetic array (SGA) system, which automates the construction of double mutants formed by mating and meiotic recombination. The Cbk1 kinase signaling pathway, which consists minimally of CBK1, MOB2, KIC1, HYM1, and TAO3 (PAG1), controls polarized morphogenesis and activation of the Ace2 transcription factor. Deletion mutations in the Cbk1 pathway genes are tolerated differently by common laboratory strains of S. cerevisiae, being viable in the W303 background but dead in the S288C background. Genetic analysis indicated that the lethality of Cbk1 pathway deletions in the S288C background was suppressed by a single allele specific to the W303 background. SGA mapping (SGAM) was used to locate this W303-specific suppressor to the SSD1 locus, which contains a known polymorphism that appears to compromise SSD1 function. This procedure should map any mutation, dominant or recessive, whose phenotype is epistatic to wild type, that is, a phenotype that can be scored from a mixed population of cells obtained by germination of both mutant and wild-type spores. In principle, SGAM should be applicable to the analysis of multigenic traits. Large-scale construction of ordered mutations in other model organisms would broaden the application of this approach.




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