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

1985; American Society for Microbiology; Volume: 54; Issue: 2 Linguagem: Inglês

10.1128/jvi.54.2.561-568.1985

1098-5514

Bengt Kallin, Lars Sternås, A K Saemundssen, János Luka, Hans Jörnvall, B. Eriksson, Pei‐Zhen Tao, Mikael Nilsson, George Klein,

Herpesvirus Infections and Treatments

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.