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Fig. 1 | Microbial Cell Factories

Fig. 1

From: A single-plasmid-based, easily curable CRISPR/Cas9 system for rapid, iterative genome editing in Pseudomonas putida KT2440

Fig. 1

Investigation of plasmid-curing efficiencies in P. putida KT2440 in the absence of antibiotics. (A) Schematic representation of different plasmids used for stability tests. Plasmid p1 refers to pBBR1MCS2 without additional cargo; Plasmid p2 represents pBBR-Cas9, which carries only the Cas9 gene; Plasmid p3 designates pBBR-Cas9ΔyqhD-HA500, which contains the CRISPR/Cas9 system targeting the yqhD gene. (B) Plasmid-curing tests of p1 in P. putida (Pp1) at 30 oC, 35 oC and 37 oC. (C) Plasmid-curing tests of p2 in P. putida (Pp2) at 30 oC, 35 oC and 37 oC. (D) Plasmid-curing tests of p1, p2, and p3 in P. putida (Pp1, Pp2 and Pp3) at 30 °C. (E) Verification of plasmid curing using point-to-point streaking. Pp1, P. putida carrying the plasmid p1, pBBR1MCS2; Pp2, P. putida carrying the plasmid p2, pBBR-Cas9; Pp3, P. putida carrying the plasmid p3, pBBR-Cas9ΔyqhD-HA500

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Microbial Cell Factories

ISSN: 1475-2859

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