Publication details
Isolation of anucleated yeast protoplasts by means of density gradient centrifugation.
| Authors | |
|---|---|
| Year of publication | 1984 |
| Type | Article in Periodical |
| Magazine / Source | J. Microbiol. Methods (Elsevier) |
| MU Faculty or unit | |
| Citation | |
| Web | http://apps.isiknowledge.com/full_record.do?product=WOS&search_mode=GeneralSearch&qid=1&SID=U1JM4IOhk@KkBjc4PDL&page=3&doc=27 |
| Field | Microbiology, virology |
| Keywords | anucleated protoplasts; density gradient centrifugation; Saccharomyces cerevisiae; yeast |
| Description | An improved method for the preparation in high yield of anucleated Saccharomyces cerevisiae has been developed. This method is based on a two-stage centrifugation of the original protoplast mixture in linear density gradients (1–10%, w/v) of Ficoll 400. The yield of anucleated protoplasts was 5–9%, its value depended on the frequency of the nucleus-free protoplasts in the original mixture. The anucleated protoplasts were characterized by RNA, DNA and protein content, and by light and electron microscopy. The protoplasts lacking nuclei had about one third the diameter of the nucleated ones, and reduced of DNA, RNA and protein in comparison to normal protoplasts. Electron microscopy showed a typical yeast ultrastructure in anucleated protoplasts except that they lacked nuclei and exhibited a higher frequency of lipid granules and exocytotic electron-dense vesicles located close to the plasmalemma. |