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. 2018 Mar 26;200(8):e00743-17. doi: 10.1128/JB.00743-17

FIG 3.

FIG 3

Regulation of the cas genes by CnpB. (A) Results of RT-PCR of csm3 in M. tuberculosis WT, ΔRv1357c, ΔcnpB expressing orn (+orn), and ΔcnpB harboring the control vector (+vector) strains. The PCR bands were quantitatively analyzed using ImageJ. The relative expression of csm3 was normalized by that of sigA. (B) β-Galactosidase assays of cas6 promoter-reporter fusion in orn-expressing (+orn) BCG WT and ΔcnpB strains. The BCG ΔcnpB strain bearing an expression vector (+vector) was used as a control. The cas6 promoter fusion was constructed based on our mapping results. Note that the expression of the promoter fusion in BCG WT harboring the vector control was indistinguishable from that in the WT expressing orn, and therefore, it is not shown. (C) Determination of intrabacterial c-di-AMP levels in M. tuberculosis WT, ΔRv1357c, ΔcnpB expressing orn (+orn), and ΔcnpB harboring the control vector (+vector) strains. Samples were prepared from 7-day cultures and analyzed using ELISA. The data shown are the means of three repeat experiments. The error bars indicate SEM. *, P < 0.05; ***, P < 0.001.