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.
Publication types
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Animals
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Baculoviridae
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Blotting, Western
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Cell Line
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Chromatography, Gel
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Chromatography, Ion Exchange
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DNA Helicases / isolation & purification
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DNA Helicases / metabolism
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DNA Primase
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DNA Replication*
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DNA, Circular / biosynthesis*
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DNA, Circular / isolation & purification
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DNA, Viral / biosynthesis*
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DNA, Viral / isolation & purification
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DNA-Binding Proteins / isolation & purification
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DNA-Binding Proteins / metabolism
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DNA-Directed DNA Polymerase / isolation & purification
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DNA-Directed DNA Polymerase / metabolism
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Electrophoresis, Agar Gel
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Exodeoxyribonucleases*
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Genome, Viral
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Herpesvirus 1, Human / enzymology*
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Herpesvirus 1, Human / genetics*
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Kinetics
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Plasmids
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RNA Nucleotidyltransferases / isolation & purification
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RNA Nucleotidyltransferases / metabolism
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Replication Origin
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Spodoptera
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Templates, Genetic
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Transfection
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Viral Proteins / biosynthesis*
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Viral Proteins / isolation & purification
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Viral Proteins / metabolism
Substances
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DNA, Circular
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DNA, Viral
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DNA-Binding Proteins
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ICP8 protein, Simplexvirus
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Viral Proteins
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DNA Primase
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RNA Nucleotidyltransferases
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DNA-Directed DNA Polymerase
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Exodeoxyribonucleases
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DNA polymerase, Simplexvirus
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DNA Helicases