Rolling circle DNA replication in vitro by a complex of herpes simplex virus type 1-encoded enzymes. |
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| Authors: | R Skaliter and I R Lehman |
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| Institution: | Department of Biochemistry, Beckman Center, Stanford University School of Medicine, CA 94305-5307. |
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| Abstract: | Extracts of insect cells infected with baculoviruses recombinant for the herpes simplex virus 1 (HSV-1)-encoded enzymes that are required for its replication can promote the rolling circle replication of circular plasmid templates. Replication is independent of a HSV-1 origin of replication (oris) or the HSV-1 origin binding protein and is inhibited by the origin binding protein when the plasmid contains oris. Replication is dependent on a complex composed of the HSV-1-encoded DNA polymerase and its processivity enhancing factor (the UL42 protein), ICP8 (the HSV-1-encoded single-strand DNA binding protein), and the HSV-1-encoded helicase-primase. The complex can be purified by size-exclusion and anion-exchange chromatography. |
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