FIG 1 .
Role and localization of pftAB. (A) Growth of the WT, ΔpftA PpftABpftB, ΔpftB, ΔpftAB, and ΔpftAB PhspftAB strains on M9P. (B) Cytoplasmic (C) versus membrane (M) localization of PftA-SPA (24 kDa) and PftB-SPA (32 kDa). Cells were grown in M9SE+P. Western blotting was performed using an anti-FLAG monoclonal antibody as the primary antibody and horseradish peroxidase-conjugated anti-mouse antibody as the secondary antibody. The positions of molecular mass markers (in kilodaltons) are indicated to the left of the gel. (C) Copurification of a N-terminal SPA-tagged PftA and of a C-terminal His-tagged version of PftB. The membrane fraction was first loaded onto a Ni2+ column to capture PftB-His6, and a Western blot using anti-FLAG antibodies revealed the presence of copurified SPA-PftA (F and E stand for flowthrough and eluate, respectively). The eluate was next loaded onto a CaM column to capture SPA-PftA, and a Western blot using anti-His antibodies revealed the presence of copurified PftB-His. A representative experiment is presented in each panel.