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Isolation of Sets of a, α, a/α, a/a and α/α isogenic strains inSaccharomyces cerevisiae

✍ Scribed by Charlotte Paquin; Julian Adams

Publisher
Springer-Verlag
Year
1982
Tongue
English
Weight
378 KB
Volume
6
Category
Article
ISSN
0172-8083

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✦ Synopsis

A simple, quick technique for isolating sets of a, ee, a/oe, a/a and c~/c~ isogenic strains of the yeast, Saecharomyces cerevisiae is described. Isogenic a/c~ diploids arise in haploid populations by a rare heterothallic switch of mating type followed by mating of the switched cell with one of the other cells in the population. Sucrose density gradient centrifugation was used to select for large elliptical diploid cells in a population of smaller haploid cells, since diploid cells are larger and more oval than haploid cells. From an a/ce diploid strain obtained in this manner, a/a and ~/~ cells were isolated by selecting for mating ability using a procedure similar to marker recovery. Finally isogenic a and haploids were simply obtained by sporulation and dissection of an a/c~ isogenic diploid strain.

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