FASTER MT: Isolation of Pure Populations of a and α Ascospores from Saccharomyces cerevisiae
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Author(s)
Chin, Brian L.
Frizzell, Margaret A.
Timberlake, William E.
Fink, Gerald R
Date Issued
April 2012
Journal
G3: Genes-Genomes-Genetics
Publisher
Genetics Society of America
Citation
Chin, Brian L., et al. “FASTER MT : Isolation of Pure Populations of a and α Ascospores from Saccharomyces Cerevisiae.” G3: Genes|Genomes|Genetics, vol. 2, no. 4, Apr. 2012, pp. 449–52. © 2012 Chin et al.
Version
Final published version
Abstract
The budding yeast Saccharomyces cerevisiae has many traits that make it useful for studies of quantitative inheritance. Genome-wide association studies and bulk segregant analyses often serve as first steps toward the identification of quantitative trait loci. These approaches benefit from having large numbers of ascospores pooled by mating type without contamination by vegetative cells. To this end, we inserted a gene encoding red fluorescent protein into the MATa locus. Red fluorescent protein expression caused MATa and a/ a diploid vegetative cells and MATa ascospores to fluoresce; MATa cells without the gene did not fluoresce. Heterozygous diploids segregated fluorescent and nonfluorescent ascospores 2:2 in tetrads and bulk populations. The two populations of spores were separable by fluorescence-activated cell sorting with little cross contamination or contamination with diploid vegetative cells. This approach, which we call Fluorescent Ascospore Technique for Efficient Recovery of Mating Type (FASTER MT), should be applicable to laboratory, industrial, and undomesticated, strains. KEYWORDS: budding yeast, red fluorescent, protein, MATa, fluorescence-activated cell sorting, hygromycin resistance, BUD5-TAF2
MIT Department
Massachusetts Institute of Technology. Department of Biology
Whitehead Institute for Biomedical Research
Terms of Use
Creative Commons Attribution 3.0 Unported license
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DOI of Published Version
https://doi.org/10.1534/g3.111.001826
