Figure 5.

Cleavage of the RecA-mediated recombination intermediate by Hjc. (A) Scheme of the formation of the Holliday junctions by RecA and the resolution of the junctions by resolvase. Asterisks show the 3′-³²P-labeled ends. (B) Recombination intermediates between gapped DNA and homologous ³²P-labeled linear duplex DNA were formed by RecA and then were treated by indicated concentration of Hjc (lanes 5–11) or Fraction V from P. furiosus (lane 12) at 55°C for 10 min or by 0.17 μM of RuvC (lane 13) at 37°C for 1 hr. The RecA-mediated products that are from incubation of the substrates with RecA for 30 min at 37°C (lane 3) and the intermediates before Hjc treatment (lane 4) were loaded as controls. ³²P-labeled linear duplex DNA (3 ng) was loaded as a marker (lane 1). Recombination products were analyzed by 1.2% agarose gel electrophoresis.