Fig. 2.

CRISPR–Cas9 mediated nicC gene deletion in the Pseudomonas putida KT2440. a The phenotypes of pSEVA-gRNAT derivatives transformed into KT2440 harboring pCAS-RK2K. All plasmids were electrotransformed into pCAS-RK2K cells with an equal amount of DNA. b The schematic represents the design of identification primers for nicC gene deletion. Yellow arrow means the location of N20 sequence in nicC gene. Blue arrow represents the location of identification primers NT-JF and NT-JR. c Agarose gel electrophoresis shows the result of colony PCR to confirm nicC gene editing efficiency. d DNA sequencing proves that the 1149-nt nicC gene have been successfully deleted