ROLLING CIRCLE DNA-REPLICATION IN-VITRO BY A COMPLEX OF HERPES-SIMPLEX VIRUS TYPE 1-ENCODED ENZYMES

Extracts of insect cells infected with baculoviruses recombinant for t he herpes simplex virus 1 (HSV-1)-encoded enzymes that are required fo r its replication can promote the rolling circle replication of circul ar plasmid templates. Replication is independent of a HSV-1 origin of replication (ori(s)) or the HSV-1 origin binding protein and is inhibi ted by the origin binding protein when the plasmid contains ori(s). Re plication is dependent on a complex composed of the HSV-1-encoded DNA polymerase and its processivity enhancing factor (the UL42 protein), I CP8 (the HSV-1-encoded. single-strand DNA binding protein), and the HS V-1-encoded helicase-primase. The complex can be purified by size-excl usion and anion-exchange chromatography.