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G3 (Bethesda). 2012 Jul;2(7):753-60. doi: 10.1534/g3.112.002550. Epub 2012 Jul 1.

A noncomplementation screen for quantitative trait alleles in saccharomyces cerevisiae.

Author information

  • 1Department of Genetics, Washington University, St. Louis, Missouri 63108, USA.

Abstract

Both linkage and linkage disequilibrium mapping provide well-defined approaches to mapping quantitative trait alleles. However, alleles of small effect are particularly difficult to refine to individual genes and causative mutations. Quantitative noncomplementation provides a means of directly testing individual genes for quantitative trait alleles in a fixed genetic background. Here, we implement a genome-wide noncomplementation screen for quantitative trait alleles that affect colony color or size by using the yeast deletion collection. As proof of principle, we find a previously known allele of CYS4 that affects colony color and a novel allele of CTT1 that affects resistance to hydrogen peroxide. To screen nearly 4700 genes in nine diverse yeast strains, we developed a high-throughput robotic plating assay to quantify colony color and size. Although we found hundreds of candidate alleles, reciprocal hemizygosity analysis of a select subset revealed that many of the candidates were false positives, in part the result of background-dependent haploinsufficiency or second-site mutations within the yeast deletion collection. Our results highlight the difficulty of identifying small-effect alleles but support the use of noncomplementation as a rapid means of identifying quantitative trait alleles of large effect.

KEYWORDS:

complex trait; copper sulfate; hydrogen sulfide

PMID:
22870398
PMCID:
PMC3385981
DOI:
10.1534/g3.112.002550
[PubMed - indexed for MEDLINE]
Free PMC Article
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