FIGURE 5.

Anchoring of PknB to the cell membrane is essential for cell survival. A, schematic representation of PknB-TM deletion and swap mutations. B, mc²ΔpknB was transformed with pNit vector, pNit-PknB, pNit-PknB-(A-TM), pNit-PknB-(H-TM), or pNit-PknB-ΔTM constructs. Cultures were seeded at an initial A₆₀₀ of 0.2 and grown in the presence or absence of ATc for 6 h. WCLs prepared were subjected to Western blotting with α-PknB and α-GroEL1 antibodies. C, in vitro growth analysis of mc²ΔpknB transformants performed in the absence of inducer ATc. Cultures were seeded at an initial A₆₀₀ of 0.02. D, growth analysis of Rv-pptr-B transformants. Rv-pptr-B transformants were streaked on 7H10 agar plates in the presence or absence of 2 μg/ml pristinamycin 1A. Error bars, S.E.