Purification of biologically active globin messenger RNA by chromatography on oligothymidylic acid-cellulose

Proc Natl Acad Sci U S A. 1972 Jun;69(6):1408-12. doi: 10.1073/pnas.69.6.1408.

Abstract

A convenient technique for the partial purification of large quantities of functional, poly(adenylic acid)-rich mRNA is described. The method depends upon annealing poly(adenylic acid)-rich mRNA to oligothymidylic acid-cellulose columns and its elution with buffers of low ionic strength. Biologically active rabbit globin mRNA has been purified by this procedure and assayed for its ability to direct the synthesis of rabbit globin in a cell-free extract of ascites tumor. Inasmuch as various mammalian mRNAs appear to be rich in poly(adenylic acid) and can likely be translated in the ascites cell-free extract, this approach should prove generally useful as an initial step in the isolation of specific mRNAs.

MeSH terms

  • Acrylamides
  • Animals
  • Carcinoma, Krebs 2 / metabolism
  • Cell-Free System
  • Centrifugation, Density Gradient
  • Chromatography*
  • Chromatography, Ion Exchange
  • Electrophoresis
  • Globins / analysis*
  • Leucine / metabolism
  • Methods
  • Neoplasm Proteins / biosynthesis
  • Polynucleotides
  • RNA, Messenger / isolation & purification*
  • RNA, Messenger / metabolism
  • RNA, Ribosomal / isolation & purification
  • Rabbits
  • Reticulocytes / analysis
  • Sodium Dodecyl Sulfate
  • Thymine Nucleotides
  • Tritium

Substances

  • Acrylamides
  • Neoplasm Proteins
  • Polynucleotides
  • RNA, Messenger
  • RNA, Ribosomal
  • Thymine Nucleotides
  • Tritium
  • Sodium Dodecyl Sulfate
  • Globins
  • Leucine