Preparative-scale isolation and purification of procaryotic and eucaryotic ribosomal 5 S RNA: Bacillus subtilis, Neurospora crassa, and wheat germ

https://doi.org/10.1016/0003-2697(84)90841-8Get rights and content

Abstract

Ribosomal 5 S RNA from three different organisms has been isolated in high yield and purity. Without prior isolation of ribosomes, a presoak in buffer followed by phenol extraction, DE-32 ion-exchange chromatography, and Sephadex G-75 gel-permeation chromatography yields at least 5–10 mg of electrophoretically homogeneous 5 S RNA from 100 g of cells. Ribonuclease activity is eliminated by various combinations of low temperature, sodium dodecyl sulfate, phenol, and bentonite. High-molecular-weight contaminants are suppressed by either 65°C heat treatment or lowered sodium dodecyl sulfate concentration. For the eucaryotes, 5.8 S RNA contamination is reduced either by low temperature in the initial solubilization or by postponing 65°C heat treatment until after the phenol extraction step.

References (32)

  • G. Rubin

    Methods Cell. Biol

    (1975)
  • G.A. Luoma et al.

    J. Mol. Biol

    (1978)
  • P.D. Burns et al.

    Biochem. Biophys. Res. Commun

    (1980)
  • K. Morikawa et al.

    FEBS Lett

    (1982)
  • A. Maxam et al.
  • E. Martegani et al.

    Biochim. Biophys. Acta

    (1980)
  • C.A. Marotta et al.

    J. Biol. Chem

    (1976)
  • V.A. Erdmann

    Nucleic Acids Res

    (1982)
  • R.P. Singhal et al.

    Prog. Nucleic Acid Res. Mol. Biol

    (1983)
  • M.E. Geroch et al.

    Eur. J. Biochem

    (1968)
  • K.O. Burkey et al.

    Biochemistry

    (1983)
  • S.-J. Li et al.

    Biochemistry

    (1984)
  • L.-H. Chang et al.

    Biochemistry

    (1984)
  • G.A. Luoma et al.
  • G.A. Luoma et al.

    Biochemistry

    (1980)
  • P.J.M. Salemink et al.

    Biochemistry

    (1981)
  • Cited by (12)

    • Nuclease properties of two putative zinc finger peptides

      1993, International Journal of Biological Macromolecules
    View all citing articles on Scopus
    View full text