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. 2023 May 1;51(11):5714–5742. doi: 10.1093/nar/gkad310

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© The Author(s) 2023. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 6. — RecF over-expression stimulates Tet recombination events through its ATPase activity and recF deletion decreases their occurence. The effect of RecF ATPase over-expression on Tet repeat recombination events (Tet^R) was examined using a plasmid-based assay. Strains carrying the indicated plasmid were grown 16 h in the presence of the indicated concentration of arabinose (or 10% if not specified) and plated on Amp and Tet/Amp plates to determine the recombination frequency. The percentage of Tet^R recombination events relative to the total c.f.u. of at least six biological replicates is represented as a dot plot, for each strain. The mean and the median are respectively represented by a square or a line, respectively. (A). Tet deletions events was tested upon increased concentration of arabinose. Wild type (P_wt-recF), EAW1130 (P_BAD-recF) and EAW1148 (P_BAD-recF_K36R) strains were transformed with the pSTL78 (upper panel), pSTL74 (medium panel) or pMB302 (lower panel). (B). The requirement for RecF ATPase for the Tet repeats recombination events was tested for deletion and point mutation strains using a plasmid-based assay. The wt, EAW629 (ΔrecF), EAW1190 (recF_K36R), EAW114 (ΔrecO), ZJR04 (ΔradD Δuup), EAW1063 (ΔradD Δuup ΔrecF), CJH0115 (ΔradD Δuup recF_K36R), and EAW1064 (ΔradD Δuup ΔrecO), strains were transformed with the pSTL78 (upper panel) or pMB302 (lower panel). Significant difference compared to the parental strain (wt) was tested by Mann-Whitney and are indicated in black (* for P = 0.05, ** for P = 0.005 or *** for P = 0.0005), an additional Kruskal–Wallis test was realized to compare the effect of increased concentration of arabinose for each strains and significance is indicated in grey.