FIGURE 4.
φ29 TP-DNA initiation and replication with φ29 TP mutants at the priming loop. The assays were carried out as described in “Experimental Procedures” using as template TP-DNA in the presence of 10 mm MgCl2. A, in vitro protein-primed initiation in the presence of TP-DNA. After 10 min of incubation at 30 °C, the reactions were stopped, processed, and analyzed by SDS-12% polyacrylamide gels and autoradiography. The position of the TP-dAMP complex is indicated. B, TP-DNA replication. After incubation for 10 min at 30 °C, the length of the synthesized DNA was analyzed by alkaline agarose gel electrophoresis. The migration position of unit length φ29 DNA is indicated.