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. 2006 Jan 30;103(6):1711–1716. doi: 10.1073/pnas.0509083103

Fig. 6.

Fig. 6.

Role of the Pol activity in blunt-end NHEJ in vivo. (A) The NHEJ reporter plasmid contains a mycobacterial replication origin (OriM), a kanamycin-resistance gene (Kan), and a lacZ gene with a unique restriction site for endonuclease EcoRV, which generates the blunt DSB end structure shown. (B) NHEJ in M. smegmatis was assayed by parallel transformations with linear and circular plasmid DNA as described in ref. 2. NHEJ fidelity (percent of blue transformants) is shown for wild-type M. smegmatis, the Pol-defective ligD mutant D136A–D138A, and the ΔligD-null strain. The fidelity values are derived from pooled counts of blue and white colonies from three separate experiments entailing nine independent transformations. The number of colonies scored was 9,194 for wild-type, 4,355 for D136A–D138A, and 820 for ΔligD. (C) Relative NHEJ efficiency is the ratio of the transformation efficiencies (colonies per μg) for linear and circular DNAs normalized to the wild-type value (100%). Relative efficiency values are the mean of three independent transformations.