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Fig. 1 | Microbial Cell Factories

Fig. 1

From: Genetic toolbox for Photorhabdus and Xenorhabdus: pSEVA based heterologous expression systems and CRISPR/Cpf1 based genome editing for rapid natural product profiling

Fig. 1

Single plasmid CRISPR/Cpf1 genome editing method for promoter exchange upstream of indC. A) Schematic representation of spacer sequence and homology regions selection; schematic cloning of synthetic dsDNA fragments harboring the repair templates and spacer sequence and the vanillic acid inducible promoter sequence (Pvan) into BsaI digested pAR20; schematic representation of the genome editing approach mediated by pAR20; the sequence between the homology arms is replaced by the promoter sequence; detailed genotype of pAR20 can be found in Table S1. HR-L/R = homologues region L (left) or R (right), dsDNA = synthetic double strand DNA, DR = FnCpf1 specific direct repeat, red or green diamond = spacer sequence A or B, crossing dashed lines representing crossover event after CRISPR induced double strand break. B) Phenotypic comparison of induced and non-induced mutants; DMSO extract of induced mutant; biosynthesis of 2. Figure was taken from BioRender.com and modified

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Microbial Cell Factories

ISSN: 1475-2859

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