DNA replication in vitro starting with an intact phiX174 phage.
Conversion of the single-stranded DNA in the intact phiX174 phage particle to the duplex replicative form (RF) has been demonstrated in lysates form phage-sensitive cells. The conversion is resistant to rifampicin and requires participation of both a "membrane" fraction of the lysate and a multienzyme replicative system. The lipopolysaccharide phage receptor, while essential, does not replace the membrane fraction. Clear, nonsedimentable extract fractions prepared with a certain nonionic detergent can replace the membrane fraction. Purification of the activity in these extracts by adsorption to polypropylene film yields a fraction with a 5-fold increase in activity relative to lipopolysaccharide and 50-fold increase relative to protein. The low buoyant density (1.03 g/cm3) suggests a high phospholipid or detergent content in this fraction.
D002499:Centrifugation, Density Gradient; D003090:Coliphages / Q000378:metabolism*; D004261:DNA Replication; D004269:DNA, Bacterial / Q000378:metabolism; D004270:DNA, Circular / Q000096:biosynthesis; D004277:DNA, Single-Stranded / Q000378:metabolism; D004279:DNA, Viral / Q000096:biosynthesis; D004926:Escherichia coli / Q000378:metabolism; D011092:Polyethylene Glycols
D004269:DNA, Bacterial; D004270:DNA, Circular; D004277:DNA, Single-Stranded; D004279:DNA, Viral; D011092:Polyethylene Glycols