Replication of Phi-X174 DNA by Escherichia coli polA- in vitro (Phi-X174 DNA-DNA replication-E. coli polA-).
Lysates of an Escherichia coli polA(-) strain convert single-stranded DNA from varphiX174 virus to the double-stranded replicative form with high efficiency on cellophane discs. The initiation of synthesis of the complementary strand appears to be rate limiting; once initiation occurs, the chain is propagated rapidly. Under these conditions, the unsealed replicative form accumulates and is slowly converted to the sealed form; this conversion requires the activity of the DPN-dependent DNA ligase.
D002499:Centrifugation, Density Gradient; D003090:Coliphages / Q000378:metabolism*; D003470:Culture Media; D004261:DNA Replication; D004279:DNA, Viral / Q000096:biosynthesis*; D004926:Escherichia coli / Q000378:metabolism*; D009693:Nucleic Acid Hybridization; D010759:Phosphorus Isotopes; D013997:Time Factors
D003470:Culture Media; D004279:DNA, Viral; D010759:Phosphorus Isotopes