Representative Results of Genome Editing following Transformation of the Single pEditing Plasmid and Verification of the Desired Construct (mexB Deletion as an Example)
(A) Plates showing the colony recovery pattern following the transformation of the control plasmid pAY5211, the pTargeting, and the pEditing. Provision of the repair donor in pEditing resumed colony recovery, indicating the success of DNA repair through the donor-mediated homologous recombination (HR) and consequently the desired mexB deletion.
(B) A representative gel image of colony PCR to verify ΔmexB in PA154197. PCR reaction using the wild-type (WT) strain as template yielded a band of 4,228 bp whereas that using recovered colonies from pEditing transformation as template yielded a band of 1,087 bp, indicating the deletion of 3,141-bp mexB gene.