Fig. 5
The presence of MutLNgo protein reduces the amount of Vsr endonuclease required for DNA digestion. The presented results were obtained (a) using M.NgoA302P-sub (GTCGGT/ACCGGGC) (lane 2, 3, 4), M.NgoAI-sub (AGTGCT/AGCGCT) (lane 5, 6, 7) and M.NgoAIV-sub (GTCGGC/GCCGGC) (lane 8, 9, 10) for V.NgoAXIII and (b) M.NgoA1175P-sub (CTGG/CCGG) (lane 2, 3, 4), M.NgoAIII “second”-sub (CTGCGG/CCGCGG) (lane 5, 6, 7) and M.NgoA302P-sub (GTCGGT/ACCGGC) (lane 8, 9, 10) for V.NgoAXIV. Reactions were carried out using 0.15 μM substrate DNA, 0.3 μM or 1.5 μM Vsr endonuclease and, where indicated, 0.6 μM MutLNgo protein. M – marker 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. The experiments were performed in triplicate, and representative images are shown