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. 2011 Apr;193(8):1981–1990. doi: 10.1128/JB.01402-10

Fig. 2.

Fig. 2.

One-dimensional SDS-PAGE analysis. (A) Overexpression of ∼15-kDa protein in M. smegmatis. Std, molecular mass standards (kDa); lane 1, wild-type mc2155; lane 2, mshA::Tn5 strain; lane 3, strain 49 [mshA(G32D) chemical mutant]; lane 4, mshA(G32D) strain with site-directed mutation; lane 5, mshD::Tn5 strain (MshD-deficient transposon mutant). (B) Regulation of ohr expression by mshA. Std, molecular mass standards (kDa); lane 1, wild-type mc2155; lane 2, mshA::Tn5 strain; lane 3, mshA::Tn5c (mshA::Tn5 mutant complemented with pcv125 containing M. tuberculosis mshA); lane 4, mshA::Tn5(pcv125) (empty vector) strain. (C) Purification of Ohr: Std, molecular mass standards (kDa); lane 1, crude extract run with 2-mercaptoethanol; lane 2, purified Ohr run with 2-mercaptoethanol; lane 3, purified Ohr run without 2-mercaptoethanol; lane 4, Ohr treated with diamide; lane 5, Ohr treated with dithiothreitol and then with N-ethylmaleimide. For the assays whose results are shown in panels A and B, cells were grown on Middlebrook 7H9 agar with 1% glucose and 0.05% Tween 80 for 1 day at 37°C followed by 10 days at 23°C.