Figure 4

G-DOC recombineering. The pDOC donor plasmid and the recombineering plasmid pACBSCE are co-transformed into the recipient strain. Arabinose induction promotes expression of the λ-Red gene products and I-SceI. I-SceI generates a linear DNA fragment form the donor plasmid that is a substrate for recombination with the chromosome mediated by the λ-Red system. Recombinants are selected by the ability to survive and grow on LB supplemented with kanamycin and sucrose.