Nonselective enrichment for yeast adenine mutants by flow cytometry.
The expression of certain adenine biosynthetic mutations in the yeast Saccharomyces cerevisiae results in a red colony color. This phenomenon has historically provided an ideal genetic marker for the study of mutation, recombination, and aneuploidy in lower eukaryotes by classical genetic analysis. In this paper, it is reported that cells carrying ade1 and/or ade2 mutations exhibit primary fluorescence. Based on this observation, the nonselective enrichment of yeast cultures for viable adenine mutants by using the fluorescence-activated cell sorter has been achieved. The advantages of this approach over conventional genetic analysis of mutation, recombination, and mitotic chromosomal stability include speed and accuracy in acquiring data for large numbers of clones. By using appropriate strains, the cell sorter has been used for the isolation of both forward mutations and chromosomal loss events in S. cerevisiae. The resolving power of this system and its noninvasiveness can easily be extended to more complex organisms, including mammalian cells, in which analogous metabolic mutants are available.
D000225:Adenine / Q000378:metabolism*; D000818:Animals; D002469:Cell Separation; D002869:Chromosome Aberrations; D003116:Color; D004252:DNA Mutational Analysis; D004271:DNA, Fungal / Q000648:ultrastructure; D005434:Flow Cytometry / Q000379:methods*; D008856:Microscopy, Fluorescence; D011995:Recombination, Genetic; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*
D004271:DNA, Fungal; D000225:Adenine