Skip to main content
Log in

Isolation and some properties of a mammalian ribosomal DNA

  • Published:
Chromosoma Aims and scope Submit manuscript

Abstract

The DNA coding for 28 S and 18 S ribosomal RNA, including the spacer regions, has been isolated from calf (Bos taurus) thymus gland. The method used included shearing of the total DNA to a highly homogeneous size population, selective heat denaturation and S 1 nuclease treatment to remove single stranded DNA. Repeated centrifugation on density gradients yields a 140-fold purified rDNA fraction with a GC content of 61.2%. Eco RI nuclease cleaves this DNA into two fragments of 16.4 and 4.9×106 daltons. Hybridization of these fragments with 28 S and 18 S rRNA shows that the 28 S coding sequence is located mostly on the 4.9×106 dalton fragment, while both the 16.4 and 4.9×106 dalton fragments contain the 18 S sequence. The data indicate that the ribosomal RNA gene has a repeat unit of 21.3×106 daltons which includes a nontranscribed spacer of about 12.5×106 daltons.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Birnstiel, M., Speirs, J., Purdom, I., Jones, K., Loaning, U.E.: Properties and composition of the isolated ribosomal DNA satellite of Xenopus laevis. Nature (Lond.) 219, 454–463 (1968)

    Google Scholar 

  • Birnstiel, M., Telford, J., Weinberg, E., Stafford, D.: Isolation and some properties of the genes coding for histone proteins. Proc. nat. Acad. Sci. (Wash.) 71, 2900–2904 (1974)

    Google Scholar 

  • Botchan, M.R.: Bovine satellite I DNA consists of repetitive units 1,400 base pairs in length. Nature (Lond.) 251, 288–292 (1974)

    Google Scholar 

  • Bross, K., Krone, W.: On the number of ribosomal RNA genes in man. Humangenetik 14, 137–141 (1972)

    Google Scholar 

  • Brown, D.D., Wensink, P.C., Jordan, E.: Purification and some characteristics of 5S DNA from Xenopus laevis. Proc. nat. Acad. Sci. (Wash.) 68, 3175–3179 (1971)

    Google Scholar 

  • Chambon, P., Mandel, J.L., Gissinger, F., Kedinger, C., Gros-Bellard, M., Hossenloop, P.: Double-stranded viral and cellular DNA's as templates for purified mammalian DNA-dependent RNA polymerases. In: Regulation of transcription and translation in eukaryotes (E.K.F. Bautz, P. Karlson and H. Kersten, eds.), pp. 125–138. New York-Heidelberg-Berlin: Springer 1973

    Google Scholar 

  • Dawid, I.B., Brown, D.D., Reeder, R.H.: Composition and structure of chromosomal and amplified ribosomal DNA's of Xenopus laevis. J. molec. Biol. 51, 341–360 (1970)

    Google Scholar 

  • Gall, J.G.: Free ribosomal RNA genes in the macronucleus of Tetrahymena. Proc. nat. Acad. Sci. (Wash.) 71, 3078–3081 (1974)

    Google Scholar 

  • Gall, J.G., Rochaix, J.-D.: The amplified ribosomal DNA of Dytiscid beetles. Proc. nat. Acad. Sci. (Wash.) 71, 1819–1823 (1974)

    Google Scholar 

  • Greene, P.J., Betlach, M.C., Goodman, H.M., Boyer, H.W.: The EcoRI restriction endonuclease. In: Methods in molecular biology (R.B. Wickner, ed.), pp. 87–111. New York: Marcel Dekker Inc. 1974

    Google Scholar 

  • Hamkalo, B.A., Miller, O.L., Jr.: Electron microscopy of genetic activity. Ann. Rev. Biochem. 42, 379–396 (1973)

    Google Scholar 

  • Hastings, J.R.B., Kirby, K.S.: The nucleic acids of Drosophila melanogaster. Biochem. J. 100, 532–539 (1966)

    Google Scholar 

  • Hayward, G.S.: Unique double-stranded fragments of bacteriophage T5 DNA resulting from preferential shear-induced breakage at nicks. Proc. nat. Acad. Sci. (Wash.) 71, 2108–2112 (1974)

    Google Scholar 

  • Hearst, J.E., Vinograd, J.: The net hydration of T-4 bacteriophage deoxyribonucleic acid and the effect of hydration on buoyant behavior in a density gradient at equilibrium in the ultracentrifuge. Proc. nat. Acad. Sci. (Wash.) 47, 1005–1014 (1961)

    Google Scholar 

  • Joseph, D.R., Stafford, D.W.: Purification of sea urchin ribosomal RNA genes with a single-strand specific nuclease. Biochim. biophys. Acta (Amst.) 418, 167–174 (1976)

    Google Scholar 

  • Patterson, J.B., Stafford, D.W.: Sea urchin satellite deoxyribonucleic acid. Its large-scale isolation and hybridization with homologous ribosomal ribonucleic acid. Biochemistry (Wash.) 9, 1278–1283 (1970)

    Google Scholar 

  • Reeder, R.H., Roeder, R.G.: Ribosomal RNA synthesis in isolated nuclei. J. molec. Biol. 67, 433–441 (1972)

    Google Scholar 

  • Scherberg, N.H., Refetoff, S.: Hybridization of RNA labelled with 125I to high specific activity. Nature (Lond.) New Biology 242, 142–145 (1973)

    Google Scholar 

  • Scherrer, K., Darnell, J.E.: Sedimentation characteristics of rapidly labelled RNA from HeLa cells. Biochem. biophys. Res. Commun. 7, 486–490 (1962)

    Google Scholar 

  • Schildkraut, C., Lifson, S.: Dependence of the melting temperature of DNA in salt concentration. Biopolymers 3, 195–208 (1965)

    Google Scholar 

  • Southern, E.M.: Detection of specific sequences among DNA fragments separated by gel electrophoresis. J. molec. Biol. 98, 503–517 (1975)

    Google Scholar 

  • Suzuki, Y., Gage, L.P., Brown, D.D.: The genes for silk fibroin in Bombyx mori. J. molec. Biol. 70, 637–649 (1972)

    Google Scholar 

  • Vogt, V.M.: Purification and further properties of single-strand-specific nuclease from Aspergillus oryzae. Europ. J. Biochem. 33, 192–200 (1973)

    Google Scholar 

  • Wartell, R.M., Larson, J.E., Wells, R.D.: Netropsin. J. biol. Chem. 249, 6719–6731 (1974)

    Google Scholar 

  • Weinberg, E.S., Overton, G.C., Shutt, R.H., Reeder, R.H.: Histone gene arrangement in the sea urchin, Strongylocentrotus purpuratus. Proc. nat. Acad. Sci. (Wash.) 72, 4815–4819 (1975)

    Google Scholar 

  • Wellauer, P.K., Dawid, I.B., Kelley, D.W., Perry, R.: Secondary structure maps of ribosomal RNA. II. Processing of mouse L-cell ribosomal RNA and variations in the processing pathway. J. molec. Biol. 89, 397–407 (1974)

    Google Scholar 

  • Wellauer, P.K., Reeder, R.H., Carroll, D., Brown, D.D., Deutsch, A., Higashinakagawa, T., Dawid, I.B.: Amplified ribosomal DNA from Xenopus laevis has heterogeneous spacer lengths. Proc. nat. Acad. Sci. (Wash.) 71, 2823–2827 (1974)

    Google Scholar 

  • Wensink, P.C., Brown, D.D.: Denaturation map of the ribosomal DNA of Xenopus laevis. J. molec. Biol. 60, 235–247 (1971)

    Google Scholar 

  • Wikman, J., Howard, E., Busch, H.: Studies on the primary structure of ribosomal 28S ribonucleic acid and its nucleolar precursors. J. biol. Chem. 244, 5471–5480 (1969)

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Blin, N., Stephenson, E.C. & Stafford, D.W. Isolation and some properties of a mammalian ribosomal DNA. Chromosoma 58, 41–50 (1976). https://doi.org/10.1007/BF00293439

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00293439

Keywords

Navigation