Oligo- and dsDNA-mediated genome editing using a tetA dual selection system in Escherichia coli
Fig 3
Genome editing using ssDNA oligos and linear dsDNA.
The lacZ segment was targeted in our genome editing method. The lacZ region was disrupted by the integration of the dual selection cassette PP3BCD2-tetA, which was rescued with either an oligo, [MG-LacZ0 (A) or MG-LacZ40 (B)] or linear dsDNA [(MG-LacZ1500 (C)]. Filled and unfilled bars represent the selection efficiency obtained with and without nickel selection, respectively. Numbers above the bar indicate the number of blue colonies (numerator) and total number of colonies (denominator) obtained by spreading the transformants on an LB plate containing X-gal/IPTG. (a), (b), and (c) refer to the editing steps followed to achieve target genome editing. Red and green in the oligos sequences represent the regions homologous to lacZ. Blue sequences represent the inserted bases. Data represent the mean value of three independent experiments and error bars represent standard deviation.