Skip to main content
NCBI home page
Biochemical Journal logoLink to Biochemical Journal
. 1978 Jul 1;173(1):145–153. doi: 10.1042/bj1730145

Preparation of chromatin containing ribosomal deoxyribonucleic acid from the macronucleus of Tetrahymena pyriformis.

R W Jones
PMCID: PMC1185757  PMID: 99140

Abstract

A method is described that enables a chromatin fraction containing ribosomal DNA (DNA containing sequences coding for rRNA) to be prepared from the macronuclei of growing or stationary cultures of Tetrahymena pyriformis. This material is obtained in yields of between 25 and 75% of the theoretical maximum. The DNA in this fraction was identified as ribosomal DNA on the basis of its density and molecular weight, and it appears not to be appreciably contaminated by other DNA. The method relies on the approximate assumption that ribosomal DNA is the smallest species of DNA in chromatin in the nucleus, and avoids the use of mechanical force, or enzyme action, to fractionate chromatin.

Full text

PDF
145

Selected References

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

  1. Cameron I. L., Jeter J. R., Jr Synchronization of the cell cycle of Tetrahymena by stravation and refeeding. J Protozool. 1970 Aug;17(3):429–431. doi: 10.1111/j.1550-7408.1970.tb04708.x. [DOI] [PubMed] [Google Scholar]
  2. Engberg J., Andersson P., Leick V., Collins J. Free ribosomal DNA molecules from Tetrahymena pyriformis GL are giant palindromes. J Mol Biol. 1976 Jun 25;104(2):455–470. doi: 10.1016/0022-2836(76)90281-3. [DOI] [PubMed] [Google Scholar]
  3. Engberg J., Christiansen G., Leick V. Autonomous rDNA molecules containing single copies of the ribosomal RNA genes in the macronucleus of Tetrahymena pyriformis. Biochem Biophys Res Commun. 1974 Aug 19;59(4):1356–1365. doi: 10.1016/0006-291x(74)90463-x. [DOI] [PubMed] [Google Scholar]
  4. Gall J. G. Free ribosomal RNA genes in the macronucleus of Tetrahymena. Proc Natl Acad Sci U S A. 1974 Aug;71(8):3078–3081. doi: 10.1073/pnas.71.8.3078. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Gorovsky M. A. Studies on nuclear structure and function in Tetrahymena pyriformis. II. Isolation of macro- and micronuclei. J Cell Biol. 1970 Dec;47(3):619–630. doi: 10.1083/jcb.47.3.619. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Jones R. W. Histone composition of a chromatin fraction containing ribosomal deoxyribonucleic acid isolated from the macronucleus of Tetrahymena pyriformis. Biochem J. 1978 Jul 1;173(1):155–164. doi: 10.1042/bj1730155. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Karrer K. M., Gall J. G. The macronuclear ribosomal DNA of Tetrahymena pyriformis is a palindrome. J Mol Biol. 1976 Jun 25;104(2):421–453. doi: 10.1016/0022-2836(76)90280-1. [DOI] [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. Leer J. C., Nielsen O. F., Piper P. W., Westergaard O. Isolation of the ribosomal RNA gene from Tetrahymena in the state of transcriptionally active chromatin. Biochem Biophys Res Commun. 1976 Sep 20;72(2):720–731. doi: 10.1016/s0006-291x(76)80099-x. [DOI] [PubMed] [Google Scholar]
  10. Mathis D. J., Gorovsky M. A. Subunit structure of rDNA-containing chromatin. Biochemistry. 1976 Feb 24;15(4):750–755. doi: 10.1021/bi00649a005. [DOI] [PubMed] [Google Scholar]
  11. Nilsson J. R., Leick V. Nucleolar organization and ribosome formation in Tetrahymena pyriformis GL. Exp Cell Res. 1970 Jun;60(3):361–372. doi: 10.1016/0014-4827(70)90529-x. [DOI] [PubMed] [Google Scholar]
  12. Noll M., Thomas J. O., Kornberg R. D. Preparation of native chromatin and damage caused by shearing. Science. 1975 Mar 28;187(4182):1203–1206. doi: 10.1126/science.187.4182.1203. [DOI] [PubMed] [Google Scholar]
  13. Thomas M., Davis R. W. Studies on the cleavage of bacteriophage lambda DNA with EcoRI Restriction endonuclease. J Mol Biol. 1975 Jan 25;91(3):315–328. doi: 10.1016/0022-2836(75)90383-6. [DOI] [PubMed] [Google Scholar]
  14. Wood P., English J., Chakraborty J., Hinton R. The use of non-ionic industrial detergents in liquid scintillation counting. Lab Pract. 1975 Nov;24(11):739–740. [PubMed] [Google Scholar]
  15. Yao M. C., Gorovsky M. A. Comparison of the sequences of macro- and micronuclear DNA of Tetrahymena pyriformis. Chromosoma. 1974;48(1):1–18. doi: 10.1007/BF00284863. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES