Strand-specific break near the origin of bacteriophage P2 DNA replication

Membrane-associated P2 DNA isolated early after infection under conditions that block replication (amB in phage and rep in Escherichia coli C) was analyzed by electron microscopy. Most DNA was in the form of relaxed circles (40%) and circles with short single-stranded tails (60%). When this DNA was hybridized with separate strands of linear P2 Hy dis DNA (which provides suitable reference points along the heteroduplex molecules), an interruption was located near the previously mapped origin of P2 DNA replication in one specific strand. The same strand was sometimes extended in the direction consistent with the unidirectional mode of P2 DNA replication. Similar conclusions were reached when the intracellular DNA was analyzed after partial denaturation. These results are consistent with the rolling circle mode of DNA replication.