A method for enucleation of Saccharomyces cerevisiae.
The first method for enucleation of yeast Saccharomyces cerevisiae is reported. Various strains, including some killer strain and respiratory-deficient mutants of Saccharomyces cerevisiae were enucleated after treatment with cytochalasin B. Removal of nuclei from protruding sphaeroplasts was induced by centrifugation in a Percoll density gradient. The enucleation yield (which averaged about 80%) and the quality of the cytoplasts were best when the yeast culture had been synchronized with nocodazole before the preparation. The presence of 1 mM CaCl2 and ATP (10 microM) in the enucleation medium prevented the formation of fragile products or aggregation. Cytoplasts could be stored for at least 1 day without visible deterioration.
Salek
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AT
University of Würzburg, Department of Biotechnology, FRG.
D002458:Cell Fractionation / Q000379:methods*; D002467:Cell Nucleus; D003571:Cytochalasin B / Q000494:pharmacology*; D012441:Saccharomyces cerevisiae / Q000187:drug effects*
D003571:Cytochalasin B