Figure 2
The plug in the periplasmic region of stator is crucial for FliL polar localization. (A) A scheme of the plasmids. eGFP, FliL and PomA constructs are labeled in dashed lines, and they do not reflect the real length. PomB is shown in the gray line and an in-frame deletion of the periplasmic plug region (a.a. 44–58) is shown in dash lines. (B) Motility assay of ZSW2 strains harboring different plasmids. Vector, pZSW7 encoding both pomAB and fliL and pZSW81 series of plasmids were introduced into ZSW2. Overnight cultures of each strain were spotted on the soft agar plate containing 0.02% arabinose and 2.5 μg/mL chloramphenicol and incubated for 6 hours at 30 °C. (C) Representative fluorescent microscopic images of ZSW2 cell expressed eGFP-pFliL in presence of stator (pZSW81) or of plug deleted stator (pZSW81-B1). Scale bar = 2 μm. (D) Protein detection of eGFP-FliL, stator and stator with a plug deletion. Protein detection from whole cell extracts by immunoblot using anti-PomBc antibody is indicated by a hollow triangle. Protein detection of eGFP-FliL by immunoblot using anti-FliLc antibody is indicated by a solid triangle. Full-length blot is shown in Fig. S2.
