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Figure 3. Effect of RPA on end unwinding. (A) Unwinding assay. Thin line, normal nucleotides; thick line, thio nucleotides; *, 32P label. (B) The thio 5′ oligonucleotide duplex precoated onto Streptavidin magnetic beads was incubated in RPA- or mock-depleted NPE. Samples were separated into bead and supernatant fractions and analyzed on a 10% native TAE-PAGE. B, beads; S, supernatant. (C) Rescue of the unwinding defect by the purified RPA protein. The thio 5′ oligonucleotide duplex precoated onto Streptavidin magnetic beads was incubated in RPA-depleted NPE supplemented with either 0.25 µM of the purified RPA protein or buffer. After 30-min incubation, the reactions were terminated and analyzed in the same way as in A. The arrowhead indicates the released product. (D) Differential activities of RPA and gp32 in supporting end unwinding. The final concentrations for RPA and gp32 are 0.25 and 1 µM, respectively.

Image published in: Yan H et al. (2011)

© 2011 Yan et al. Creative Commons Attribution-NonCommercial-ShareAlike license

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