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. 2010 May 10;285(28):21698–21707. doi: 10.1074/jbc.M110.125724

FIGURE 4.

FIGURE 4.

Complementation of M. bovis BCG ΔpcaA M. smegmatis ΔMSMEG_1351 mutants. Complementation of M. bovis BCG ΔpcaA by MSMEG_1351. One-dimensional (A) and two-dimensional (B) argentation TLC of 14C-radiolabeled mycolic acids from M. bovis BCG Pasteur and BCG mc22801 (BCG Pasteur pcaA::Tn5370) transformed with various vectors and grown at 37 °C. The arrowhead indicates the position of the hybrid mycolate accumulating in the ΔpcaA mutant, with a cis double bond at the proximal position in place of a cis-cyclopropane present in the wild-type α-mycolate as determined previously (5). α, α-mycolates; K, keto-mycolates; WT, wild type. C, single BCG colonies grown on cord reading agar and photographed after 15 days of incubation at 37 °C. Magnification, ×100. D, complementation of M. smegmatis ΔMSMEG_1351 by pcaA. Two-dimensional TLC analysis of 14C-radiolabeled mycolic acids from M. smegmatis ΔMSMEG_1351 transformed either with pMV261, pMV306_1351, or pMV261_pcaA grown at 25 °C. Arrows indicate the position of the cyclopropanated mycolate missing in the ΔMSMEG_1351 mutant.