1887

Abstract

Summary: A method has been developed for isolating nuclei from the filamentous fungus . In this procedure, the mycelia from 14 to 16 h spore-derived cultures of NR1, a stable diploid strain, were frozen with liquid nitrogen and homogenized in a Waring blender. After homogenization, the mycelia were warmed to 4 °C and the nuclei were purified from the homogenate by differential centrifugation followed by sedimentation through 2·1 -sucrose. The final nuclear yield was 15 to 20%, based on DNA estimations. The purified nuclear pellet was free of whole cells. The morphology of the isolated nuclei resembled that of the nuclei; they contained a nucleolus, chromatin, and had a surrounding double membrane. The purified nuclei were characterized by a DNA:RNA: protein ratio of 1:2.8:8·7.

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/content/journal/micro/10.1099/00221287-94-1-204
1976-05-01
2021-05-14
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References

  1. Bhargava M. M., Halvorson H. O. 1971; Isolation of nuclei from yeast. Journal of Cell Biology 49:423–429
    [Google Scholar]
  2. Bracker C. E. 1967; Ultrastructure of fungi. Annual Review of Phytopathology 5:343–374
    [Google Scholar]
  3. Clutterbuck A. J., Roper J. A. 1966; A direct determination of nuclear distribution in heterokaryotes of Aspergillus nidulans. Genetical Research, Cambridge 7:185–194
    [Google Scholar]
  4. Doi K., Doi A. 1974; Isolation of nuclei from a tetraploid strain of Saccharomyces cerevisiae. Journal of Biochemistry 75:1017–1026
    [Google Scholar]
  5. Felden R. A., Sanders M. M., Morris N. R. 1976; The presence of histones in Aspergillus nidulans. Journal of Cell Biology 68:430–439
    [Google Scholar]
  6. Giles K. W., Myers A. 1965; An improved diphenylamine method for the estimation of deoxyribonucleic acid. Nature; London: 20693
    [Google Scholar]
  7. Hsiang M. W., Cole R. D. 1973; The isolation of histone sfrom Neurospora crassa. Journal of Biological Chemistry 248:2007–2013
    [Google Scholar]
  8. Jaworski A. J., Horgen P. A. 1973; The ribosomal cistrons of the water mold Achylabisexualis. Archives of Biochemistry and Biophysics 157:260–267
    [Google Scholar]
  9. Lowry O. H., Rosebrough N. J., Farr A.L, Randall R. J. 1951; Protein measurement with the Folin phenol reagent. Journal of Biological Chemistry 193:265–275
    [Google Scholar]
  10. Minssen M., Munkres K. D. 1973; Preparation of mitochondrial membrane proteins from Neurosporacrassa. Prevention of lipid autoxidation by an anti-oxidant. Biochimica et biophysica acta 291:398–410
    [Google Scholar]
  11. Morris N. R. 1976; A temperature sensitive mutant of Aspergillus nidulans reversibly blocked in nuclear division. Experimental Cell Research in the Press
    [Google Scholar]
  12. Peberdy J. F., Gibson R. K. 1971; Regeneration of Aspergillus nidulans protoplasts. Journal of General Microbiology 69:325–330
    [Google Scholar]
  13. Reich E., Tsuda S. 1961; Isolation of nuclei from Neurosporacrassa. Biochimica et biophysica acta 53:574–575
    [Google Scholar]
  14. Robinow C. F., Caten C. E. 1969; Mitosis in Aspergillus nidulans. Journal of Cell Science 5:403–431
    [Google Scholar]
  15. Rozijn R. H., Tonino G. J. M. 1964; Studies on the yeast nucleus. I. The isolation of nuclei. Biochimica et biophysica acta 91:105–112
    [Google Scholar]
  16. Sargent M. L. 1973; Use of liquid nitrogen and high ionic strength for the isolation of functional polyribosomes from Neurosporacrassa. Biochimicaet biophysica acta 324:267–274
    [Google Scholar]
  17. Schneider W. C. 1957; Determination of nucleic acids by pentose analysis. In Methods in Enzymology 3 pp. 680–684 Colowick S. P., Kaplan N. O. Edited by New York: Academic Press;
    [Google Scholar]
  18. Tata J. R. 1974; Isolation of nuclei from liver and other tissues. In Methods in Enzymology 31 pp. 253–262 Fleisher S., Packer L. Edited by New York: Academic Press;
    [Google Scholar]
  19. Wintersberger V., Smith P., Letnansky K. 1973; Yeast chromatin. Preparation from isolated nuclei, histone composition and transcription capacity. European Journal of Biochemistry 33:123–130
    [Google Scholar]
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