Purification of avian myeloblastosis virus DNA polymerase by affinity chromatography on polycytidylate-agarose

J Virol. 1974 Oct;14(4):853-9. doi: 10.1128/JVI.14.4.853-859.1974.

Abstract

Polycytidylic acid [poly(rC)] covalently linked to cyanogen bromide-activated agarose is an effective affinity matrix for the RNA-dependent DNA polymerase from avian myeloblastosis virus. Poly(rC)-agarose is capable of binding large quantities of avian myeloblastosis DNA polymerase, which is then eluted by using a linear KCl gradient of increasing concentration. The DNA polymerase isolated from crude, detergent-disrupted virions by a single pass through columns of poly(rC)-agarose appears nearly homogeneous (approximately 90% pure) as determined by sodium dodecyl sulfate-polyacrylamide disc gel electrophoresis. Complete recovery of input enzymatic activity was obtained. Results suggest that polyribonucleotide columns may provide a high-yield, rapid method for the purification of oncornaviral DNA polymerase.

MeSH terms

  • Avian Leukosis Virus / enzymology*
  • Cell-Free System
  • Centrifugation, Density Gradient
  • Chromatography, Affinity*
  • Cytosine Nucleotides
  • Electrophoresis, Polyacrylamide Gel
  • Guanosine Triphosphate / metabolism
  • Molecular Weight
  • Polynucleotides
  • Polysaccharides
  • Potassium Chloride
  • RNA-Directed DNA Polymerase / analysis
  • RNA-Directed DNA Polymerase / isolation & purification*
  • RNA-Directed DNA Polymerase / metabolism
  • Templates, Genetic
  • Thymine Nucleotides / metabolism
  • Tritium

Substances

  • Cytosine Nucleotides
  • Polynucleotides
  • Polysaccharides
  • Thymine Nucleotides
  • Tritium
  • Potassium Chloride
  • Guanosine Triphosphate
  • RNA-Directed DNA Polymerase