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Fig. 7 | BMC Microbiology

Fig. 7

From: Activity of Vsr endonucleases encoded by Neisseria gonorrhoeae FA1090 is influenced by MutL and MutS proteins

Fig. 7

The influence of the MutS protein on DNA cleavage by V.NgoAXIII and V.NgoAXIV endonucleases. Left part of the figure – the activity of V.NgoAXIII; right part of the figure – the activity of the V.NgoAXIV endonuclease. a Examples of electrophoresis profiles of the reaction products resolved in a 10% polyacrylamide gel: (black squares) DNA cleavage by the gonococcal Vsr endonuclease in the absence of both MutS protein and 1 mM ATP; (black triangles) DNA cleavage by the gonococcal Vsr endonuclease in the presence of MutS protein and 1 mM ATP, molar ratio Vsr:MutS 1:2. These and analogous profiles were used to quantify the efficiency of cleavage of the substrate DNAs containing a T:G mismatch catalyzed by Vsr endonucleases. M - GeneRuler 50 bp DNA Ladder (Thermo Scientific). Arrows indicate substrate and reaction products obtained after DNA cleavage by a given Vsr endonuclease. P1 and P2 reaction products; S – substrate DNA. b Analysis of the data obtained and fitted to a first-order rate equation using Origin 8.5 software. The presented results were obtained using M.NgoA302P-sub (GTCGGT/ACCGGC) for V.NgoAXIII and M.NgoA1175P-sub (CTGG/CCGG) substrate DNA for V.NgoAXIV. c Histograms presenting first-order rate constants (kst) obtained for endonucleolytic reactions catalyzed by gonococcal Vsr endonucleases in the presence and absence of gonococcal MutS protein. Asterisks indicate statistically significant differences, which were calculated using the two-tailed heteroscedastic Student’s t-test (p value < 0.05). Reactions were carried out using 0.15 μM substrate DNA, 0.3 μM Vsr endonuclease and, where indicated, 0.6 μM MutS protein. The experiments were performed in triplicate, and representative images and graphs of the data fitted to a first-order rate equation are shown

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