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Baculovirus Replication: Purification and identification of the Trichoplusia ni Nuclear Polyhedrosis Virus-induced DNA Polymerase

NERC Institute of Virology, Mansfield Road, Oxford, OX1 3SR, U.K.

DNA polymerase activity present in Trichoplusia ni multiple nucleocapsid nuclear polyhedrosis virus-infected Spodoptera frugiperda cells has been analysed by chromatography on DNA-cellulose and phosphocellulose columns. In infected cells a new fraction of activity was found to bind to the columns more strongly that did polymerase activity in uninfected cells. The infected-cell-specific DNA polymerase was purified by a combination of DNA-cellulose chromatography of crude extracts and phospho-cellulose chromatography of the semi-purified activity. The final product contained a single polypeptide of molecular weight 126 000 which was not found in uninfected cells. The purified enzyme was inhibited by aphidicolin and [E]-5-(2-bromovinyl)-2'-deoxyuridine triphosphate, but not by bromovinyldeoxyuridine. The enzyme was shown to be an early enzyme, probably a delayed early protein, since it was present in cells inhibited by aphidicolin which were locked into the synthesis of early proteins by the drug.

Keywords: baculovirus, DNA polymerase, MNPV, BVdU, aphidicolin

Received 14 March 1983; accepted 30 June 1983.