Division of Biological Sciences, University of Michigan, 48109, Ann Arbor, Michigan, USA.
Curr Genet. 1982 Oct;6(1):21-4. doi: 10.1007/BF00397636.
A simple, quick technique for isolating sets of a, α, a/α, a/a and α/α isogenic strains of the yeast, Saccharomyces cerevisiae is described. Isogenic a/α 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/α diploid strain obtained in this manner, ala and α/α cells were isolated by selecting for mating ability using a procedure similar to marker recovery. Finally isogenic a and α haploids were simply obtained by sporulation and dissection of an a/α isogenic diploid strain.
介绍了一种简单、快速的方法，用于分离酵母 Saccharomyces cerevisiae 的 a、α、a/α、a/a 和 α/α 同系菌株。同系的 a/α 二倍体是通过罕见的异配性交配型转换在单倍体群体中产生的，然后将转换的细胞与群体中的另一个细胞交配。通过蔗糖密度梯度离心从较小的单倍体细胞群体中选择大的椭圆形二倍体细胞，因为二倍体细胞比单倍体细胞更大、更椭圆形。从以这种方式获得的 a/α 二倍体菌株中，通过使用类似于标记恢复的程序选择交配能力来分离 ala 和 α/α 细胞。最后，通过 a/α 同系二倍体菌株的孢子形成和解剖，简单地获得了同系的 a 和 α 单倍体。
