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. 2010 Oct 21;6(10):e1001166. doi: 10.1371/journal.pgen.1001166

Figure 4. Recombination activity of P. abyssi XerA.

Figure 4

A. Temperature dependence of the reaction. XerA was incubated at different temperatures with a plasmid containing or not the predicted dif site (gray box). MW: molecular weight marker (identified in Kbp on the left). C: control without protein. The control assay was performed at 65°C. Msc: supercoiled monomer. Moc: open circular monomer. Dsc: supercoiled dimer. B. Recombination product analysis. C: control without protein. HindIII, pBend2dif full restriction by HindIII. XerA: recombination reaction, 1h at 65°C. XerA/HindIII 20°C: Recombination reaction followed by restriction with one unit of HindIII, for 1 h at 20°C. XerA/HindIII 37°C: same as before, but restriction performed at 37°C. Msc: supercoiled monomer. LM: linearized monomer (2.6 kbp). LD: linearized dimer (5.2 kbp). LT: linearized trimer (7.8 kbp). C. Resolution reaction. The substrate used is diagramed in (i) and the product in (ii). Small black arrows correspond to the hybridization position of the oligonucleotides used in the PCR assay. PCR fragments obtained on the substrate (i) or on the reaction products (ii) were analyzed by agarose gel electrophoresis (right).