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. 2007 May 3;35(10):3407–3419. doi: 10.1093/nar/gkm206

Figure 5.

Figure 5.

The rate of recombination with mutant attB sites T-15C:C+15G and G-16T:G+16A is greatly reduced. Panel (A) shows the appearance of products from recombination assays using T-15C:C+15G and G-16T:G+16A as substrates after prolonged incubation. Plasmids encoding the wild-type attB (wt) or the indicated attB mutants were incubated with pRT702 (attP) for 1, 2 or 3 h at 30°C and then the products analysed by restriction and agarose gel electrophoresis. After 2 and 3 h some product (attL) is visible in the lanes containing the −/+15 and −/+16 mutations. Panel (B) shows the time-dependent appearance of recombination intermediates when wild-type attB (wt) was used compared to C-2A:G+2T, T-15C:C+15G or G-16T:G+16A. The −/+2 mutant site rapidly forms a synapse (Int:synapse attP/B) and thereafter the reaction is blocked. The −/+15 and −/+16 attB sites slowly accumulated the cleaved intermediate (Int:cleaved attP/B) and some shifted product complexes (Int:attL/R). The remaining complexes on the gel are as described in Figure 4.