Open AccessA method for enucleation of Saccharomyces cerevisiae
Author(s) -
Salek Anna T.
Publication year - 1993
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1993.tb06484.x
Subject(s) - enucleation , cytoplast , saccharomyces cerevisiae , yeast , percoll , cytochalasin , nocodazole , biology , cytochalasin b , chemistry , centrifugation , biochemistry , microbiology and biotechnology , cytoplasm , in vitro , cytoskeleton , cell
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 CaCl 2 and ATP (10 μM) in the enucleation medium prevented the formation of fragile products or aggregation. Cytoplasts could be stored for at least 1 day without visible deterioration.