Purification of Epstein-Barr virus DNA polymerase from P3HR-1 cells

J Virol. 1985 May;54(2):561-8. doi: 10.1128/JVI.54.2.561-568.1985.


The Epstein-Barr virus DNA polymerase was purified from extracts of P3HR-1 cells treated with n-butyrate for induction of the viral cycle. Sequential chromatography on DNA cellulose, phosphocellulose, and blue Sepharose yielded an enzyme preparation purified more than 1,300-fold. The purified enzyme was distinct from cellular enzymes but resembled the viral DNA polymerase in cells infected with herpes simplex virus type 1 or 2. The active enzyme had an apparent molecular weight of 185,000 as estimated by gel filtration on Sephacryl S-300. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed a major polypeptide corresponding to a molecular weight of ca. 110,000. This polypeptide correlated with the catalytic function of the purified enzyme, whereas the other, less abundant polypeptides did not. By immunoblotting, the 110,000-molecular-weight polypeptide could be identified as a viral polypeptide. It could not be determined whether the native enzyme was composed of more than one polypeptide.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Butyrates / pharmacology
  • Butyric Acid
  • DNA-Directed DNA Polymerase / analysis
  • DNA-Directed DNA Polymerase / biosynthesis
  • DNA-Directed DNA Polymerase / isolation & purification*
  • Herpesvirus 4, Human / enzymology*
  • Molecular Weight
  • Simplexvirus / enzymology


  • Butyrates
  • Butyric Acid
  • DNA-Directed DNA Polymerase