Skip to main content
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1966 Apr 1;29(1):21–28. doi: 10.1083/jcb.29.1.21

INCORPORATION OF NUCLEOTIDES INTO NUCLEI OF FIXED CELLS BY DNA POLYMERASE

R C von Borstel 1, D M Prescott 1, F J Bollum 1
PMCID: PMC2106953  PMID: 4958508

Abstract

The enzyme calf thymus polymerase requires denatured or single-stranded DNA as a primer for DNA synthesis and is inactive on native DNA preparations. The enzyme and tritium-labeled deoxyribonucleoside triphosphates were incubated with alcohol-fixed and Carnoy-fixed tissue preparations to see if primer DNA could be found in several types of cells undergoing DNA synthesis. In all cases, low-pH controls were prepared for comparison. Priming activity was not found in nuclei that had been fixed in alcohol. Priming activity was found in cell nuclei that had been fixed with an acid fixative or had been treated at a low pH prior to treatment with the enzyme reaction mixture.

Full Text

The Full Text of this article is available as a PDF (574.0 KB).

Selected References

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

  1. BOLLUM F. J. Calf thymus polymerase. J Biol Chem. 1960 Aug;235:2399–2403. [PubMed] [Google Scholar]
  2. BOLLUM F. J. Thermal conversion of nonpriming deoxyribonucleic acid to primer. J Biol Chem. 1959 Oct;234:2733–2734. [PubMed] [Google Scholar]
  3. GALL J. G. Macronuclear duplication in the ciliated protozoan Euplotes. J Biophys Biochem Cytol. 1959 Mar 25;5(2):295–308. doi: 10.1083/jcb.5.2.295. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. MAYOR H. D., HILL N. O. Acridine orange staining of a single-stranded DNA bacteriophage. Virology. 1961 Jun;14:264–266. doi: 10.1016/0042-6822(61)90202-1. [DOI] [PubMed] [Google Scholar]
  5. PRESCOTT D. M., KIMBALL R. F. Relation between RNA, DNA, and protein syntheses in the replicating nucleus of Euplotes. Proc Natl Acad Sci U S A. 1961 May 15;47:686–693. doi: 10.1073/pnas.47.5.686. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Prescott D. M. Cellular sites of RNA synthesis. Prog Nucleic Acid Res Mol Biol. 1964;3:33–57. doi: 10.1016/s0079-6603(08)60738-2. [DOI] [PubMed] [Google Scholar]

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

RESOURCES