May 3, 2014

Mapping small effect mutations in Saccharomyces cerevisiae: impacts of experimental design and mutational properties

G3 : Genes - Genomes - Genetics
Fabien DuveauPatricia J Wittkopp

Abstract

Genetic variants identified by mapping are biased toward large phenotypic effects because of methodologic challenges for detecting genetic variants with small phenotypic effects. Recently, bulk segregant analysis combined with next-generation sequencing (BSA-seq) was shown to be a powerful and cost-effective way to map small effect variants in natural populations. Here, we examine the power of BSA-seq for efficiently mapping small effect mutations isolated from a mutagenesis screen. Specifically, we determined the impact of segregant population size, intensity of phenotypic selection to collect segregants, number of mitotic generations between meiosis and sequencing, and average sequencing depth on power for mapping mutations with a range of effects on the phenotypic mean and standard deviation as well as relative fitness. We then used BSA-seq to map the mutations responsible for three ethyl methanesulfonate-induced mutant phenotypes in Saccharomyces cerevisiae. These mutants display small quantitative variation in the mean expression of a fluorescent reporter gene (-3%, +7%, and +10%). Using a genetic background with increased meiosis rate, a reliable mating type marker, and fluorescence-activated cell sorting to efficiently s...Continue Reading

Mentioned in this Paper

Microorganism
Bacterial Proteins
Saccharomyces cerevisiae allergenic extract
Mutagenesis, Site-Directed
Fluorescence-Activated Cell Sorting
Genome Mapping
Fluorescent stain
Isolate compound
Sequencing
Increased Meiosis [PE]

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