Fig. 3

Schematic diagram of the essential effects among four components (Cas9 protein, gRNA cassette, homologous arms and λ-Red system) in the Pseudomonas putida KT2440 genome editing. a The strategy of plasmids with different components were transformed into KT2440. pCAS-RK2K or its derivatives was first transformed into KT2440. In the second round of electroporation, pSEVA644, pSEVA-gRic6T and its derivatives were transformed into KT2440 harboring pCAS-RK2K relevant plasmids. b Electroporation efficiency is reflected from the total number of CFU (colony-forming units); Mutation efficiency in six groups with different components. The CFU experiment is obtained from three replicates. Cells were plated on the equal concentration antibiotics plates and the amount of DNA was equivalent in each experiment