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. 2010 Apr 5;38(14):4889–4906. doi: 10.1093/nar/gkq209

Figure 1.

Figure 1.

DNA strand exchange activities of wild-type and mutant Rad51 proteins. DNA strand exchange assays were carried out using full-length φX174 DNA substrates as described under ‘Materials and Methods’ section. (A) Schematic of in vitro DNA strand exchange reaction, showing interconversion of single- and double-stranded DNA species. (B) Timecourses of DNA strand exchange reactions visualized by separating DNA substrates and products via agarose gel electrophoresis and staining with ethidium bromide. All reactions contained 33 µM each of φX174 circular ssDNA and φX174 linear dsDNA, 11 µM Rad51 enzyme, 1.65 µM RPA and 2.5 mM ATP (final concentrations). Other reaction components and assay conditions are described under ‘Materials and Methods’ section. Lanes 1–5, reaction with wild-type Rad51. Lanes 2–10, reaction with H352A Rad51 mutant. Lanes 11–15, reaction with H352Y Rad51 mutant. (C) Results of panel B quantified by fluorescence imaging. Data are averages from three separate experiments. Filled circles, wild-type Rad51. Open squares, H352A Rad51 mutant. Filled triangles, H352Y Rad51 mutant.