Figure 5.

Archaeal RER reconstituted in vitro. A, reaction schematic. The substrate depicted in Fig. 3 is incubated with purified Thermococcus sp. 9°N proteins including PCNA, RFC, and different combinations of PolB, PolD, Fen1, and DNA ligase. Reactions were incubated over a time course from 0 to 30 min at 60 °C and repair was monitored by appearance of the 134-nt FAM/MAX-labeled DNA product by CE. B, representative CE traces for each reaction condition at the 30-min time point are shown. C, quantification of the conversion of 90-nt FAM substrate to 134-nt FAM/MAX product at the 30-min time point. Data are the average of three independent experiments with S.D.