Skip to main content
. 2017 Oct 25;7:13987. doi: 10.1038/s41598-017-14410-z

Figure 2.

Figure 2

Disruption of pafBC in M. smegmatis. (A) Schematic representation of the generation of a markerless pafBC deletion mutant in Msm. Genes encoding PafBC are shown in white, neighbouring genes in gray. The wild type locus is shown along with the targeting vector pMCS-ΔpafBC, the 3’ single crossover mutant and the Msm pafBC deletion strain. The pafA-specific probe used in the Southern blot analysis is shown in black. Grey bars and sizes in bp refer to the blot shown in panel B and assign the respective fragments. (B) Genomic DNA of Msm wild type (WT), the pafBC 3’ single-crossover strain (3sco) and the pafBC deletion mutant (ΔpafBC) was digested with KpnI. Southern blot analysis was performed using a 500-bp pafA-specific probe. (C) Anti-PafBC immunoblot analysis of Msm wild type, Msm ΔpafBC and the complemented strain Msm ΔpafBC-pafBC. The full-length blot is shown in Supplementary Fig. S6. An anti-RNA polymerase β subunit (RpoB) immunoblot served as the loading control.