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. 2010 Nov 26;30(1):145–153. doi: 10.1038/emboj.2010.301

Figure 5.

Figure 5

Xer is necessary and sufficient for recombination at dif in vitro. (A) Intermolecular recombination assays. Plasmids containing a 50-bp DNA fragment encompassing DYAD3 (dIII) and DYAD2 (dII), or a 50-bp DNA fragment mutated for dIII or dII (the mutated bases are represented by bold underlined text) were incubated with Xer-6H. Recombination products (plasmid multimers) were detected by agarose gel electrophoresis (lower panel). (B) Intramolecular recombination assays. (Upper panel) Plasmids containing two test sequences in either a direct or inverted repeat orientation flanking a lacS gene cassette were incubated with Xer-6H. After digestion by SpeI, the products were analysed by agarose gel electrophoresis. For each DNA substrate tested (lower panel), the 273-bp dif3 PCR sequence is indicated by a grey box, whereas DYAD3, either in the context of the dif3 PCR fragment (within the grey box) or cloned as a 38-bp double-stranded oligonucleotide, is indicated by a white triangle. The lacS gene is shown as a white box.