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. Author manuscript; available in PMC: 2010 Mar 16.
Published in final edited form as: J Biol Chem. 2002 Oct 4;277(49):47213–47224. doi: 10.1074/jbc.M206764200

Fig. 2. E. coli-produced RFC loads PCNA clamps on primed DNA.

Fig. 2

A, an SDS-PAGE analysis of S. cerevisiae RFC complex comprising five subunits in 1:1 stoichiometry. B, PCNA clamp assembly on DNA, assayed by incubating 0.03 μM 32P-PCNA with 0.02 μM RFC and 0.03 μM SSB-coated circular primed M13 ssDNA in the presence (●) or absence (○) of 0.5 mM ATP, unprimed M13 ssDNA with ATP (□), and circular duplex pBluescript DNA with ATP (△) for 5 min at 30 °C, followed by gel filtration as described under “Experimental Procedures.” PCNA loaded on DNA elutes in fractions 7–12 and free PCNA elutes in fractions 14–25.