Figure 7
Rationally designed Cre mutants show increased recombination activity in E.coli. (a) Schematic drawing of the plasmid assay. Important regions in the plasmids are indicated. Note the reduced size of the plasmid after recombination. The restrictions sites (BsrGI and XbaI) used for cloning indicated Cre-recombinase variants are depicted. The rox target sites are shown as red triangles. CmR, chloramphenicol resistance gene; ori, origin of replication; AraC, arabinose operon regulatory gene. (b) Agarose gels of three independently picked clones showing BsrGI and XbaI digested plasmids carrying indicated recombinases. The amount of arabinose added to the growth medium is presented below each band in μg/ml l+-arabinose. The line with two triangles indicates no recombination, whereas the line with one triangle marks the recombined band. Quantifications of the ratios of band intensities (in percent of recombination) are shown to the right for each mutant. The amount of arabinose added to the growth medium is shown on the X-axis. Error bars depict standard deviation from the three independent experiments.
