Skip to main content
PMC home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1971 May 1;49(2):423–429. doi: 10.1083/jcb.49.2.423

ISOLATION OF NUCLEI FROM YEAST

Madhu M Bhargava 1, Harlyn O Halvorson 1
PMCID: PMC2108327  PMID: 19866769

Abstract

A method for isolation of nuclei from Saccharomyces cervisiae in high yield is described. The DNA/protein ratio of the isolated nuclei is 10 times higher than that of whole cells. Examination of these nuclei in phase and electron microscopes has shown them to be round bodies having a double membrane, microtubules, and a dark crescent at one end. The optimum conditions for extraction and resolution of histones of these nuclei on acrylamide gels have been investigated. The nuclei have an active RNA polymerase (E.C. 2.7.7.6) and are able to synthesize RNA in vitro. They are also readily stainable with Giemsa's, Feulgen's, and acridine orange methods.

Full Text

The Full Text of this article is available as a PDF (554.2 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. ALLFREY V. G., MIRSKY A. E., STERN H. The chemistry of the cell nucleus. Adv Enzymol Relat Subj Biochem. 1955;16:411–500. doi: 10.1002/9780470122617.ch8. [DOI] [PubMed] [Google Scholar]
  2. BURTON K. A study of the conditions and mechanism of the diphenylamine reaction for the colorimetric estimation of deoxyribonucleic acid. Biochem J. 1956 Feb;62(2):315–323. doi: 10.1042/bj0620315. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. DRURY H. F. Identification and estimation of pentoses in the presence of glucose. Arch Biochem. 1948 Dec;19(3):455–466. [PubMed] [Google Scholar]
  4. Duffus J. H. The isolation of nuclei from the yeast Schizosaccharomyces pombe. Biochim Biophys Acta. 1969 Nov 19;195(1):230–233. doi: 10.1016/0005-2787(69)90619-4. [DOI] [PubMed] [Google Scholar]
  5. EATON N. R. New press for disruption of microorganisms. J Bacteriol. 1962 Jun;83:1359–1360. doi: 10.1128/jb.83.6.1359-1360.1962. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. ITIKAWA O., OGURA Y. The Feulgen reaction after hydrolysis at room temperature. Stain Technol. 1954 Jan;29(1):13–15. doi: 10.3109/10520295409115429. [DOI] [PubMed] [Google Scholar]
  7. LOVELOCK J. E. The protective action of neutral solutes against haemolysis by freezing and thawing. Biochem J. 1954 Feb;56(2):265–270. doi: 10.1042/bj0560265. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  9. Lee Y. C., Scherbaum O. H. Nucleohistone composition in stationary and division synchronized Tetrahymena cultures. Biochemistry. 1966 Jun;5(6):2067–2075. doi: 10.1021/bi00870a039. [DOI] [PubMed] [Google Scholar]
  10. MILLER G. R., MCCLARY D. O., BOWERS W. D., Jr ULTRAVIOLET AND PHASE MICROSCOPY OF SPORULATING SACCHAROMYCES. J Bacteriol. 1963 Apr;85:725–731. doi: 10.1128/jb.85.4.725-731.1963. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. MUNDKUR B. Electron microscopical studies of frozendried yeast. II. The nature of basophile particles and vesicular nuclei in Saccharomyces. Exp Cell Res. 1961 Oct;25:1–23. doi: 10.1016/0014-4827(61)90303-2. [DOI] [PubMed] [Google Scholar]
  12. Molenaar I., Sillevis Smitt W. W., Rozijn T. H., Tonino G. J. Biochemical and electron microscopic study of isolated yeast nuclei. Exp Cell Res. 1970 May;60(2):148–156. doi: 10.1016/0014-4827(70)90500-8. [DOI] [PubMed] [Google Scholar]
  13. NASH D., PLAUT W. ON THE DENATURATION OF CHROMOSOMAL DNA IN SITU. Proc Natl Acad Sci U S A. 1964 May;51:731–735. doi: 10.1073/pnas.51.5.731. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. ROBINOW C. F. Mitosis in the yeast Lipomyces lipofer. J Biophys Biochem Cytol. 1961 Apr;9:879–892. doi: 10.1083/jcb.9.4.879. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. ROZIJN T. H., TONINO G. J. STUDIES ON THE YEAST NUCLEUS. I. THE ISOLATION OF NUCLEI. Biochim Biophys Acta. 1964 Sep 11;91:105–112. doi: 10.1016/0926-6550(64)90174-4. [DOI] [PubMed] [Google Scholar]
  16. Robinow C. F., Marak J. A fiber apparatus in the nucleus of the yeast cell. J Cell Biol. 1966 Apr;29(1):129–151. doi: 10.1083/jcb.29.1.129. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. STEELE W. J., BUSCH H. STUDIES ON ACIDIC NUCLEAR PROTEINS OF THE WALKER TUMOR AND LIVER. Cancer Res. 1963 Sep;23:1153–1163. [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES