Figure 2.

Products of leading and lagging strand synthesis. Products formed by rolling circle DNA replication in vitro were analyzed by alkaline agarose gel electrophoresis and restriction enzyme cleavage as described in Materials and Methods. The reaction mixtures contained 50 fmol of DNA polymerase-UL42, 350 fmol of the UL5/UL52/UL8 heterotrimer, 500 fmol of ICP8, and 35 fmol of the minicircle template. (A) Autoradiograph of an alkaline agarose gel. First lane, DNA size markers; second lane, products labeled by [α-³²P]dGTP; third lane, products labeled by [α-³²P]dCTP. (B) Autoradiograph of Southern blot analyses of replication products. First lane, the oligonucleotide probe is complementary to the lagging strand; second lane, the oligonucleotide probe is complementary to the leading strand; third lane, DNA size markers. (C) Products labeled with [α-³²P]dCTP and [α-³²P]dGTP were cleaved with MboI and were subjected to polyacrylamide gel electrophoresis as described in Materials and Methods.