Figure 3
Effect of deletion of residues 206–245, 206–255 or 206–265 on FliK function. (a) Motility of TH8426 (∆fliK), SJW1103 (WT), MMK1013 (∆206–245), MMK1014 (∆206–255) and MMK1015 (∆206–265) in 0.35% soft agar. Plates were incubated at 30 °C for 8 hours. (b) Secretion assays of FliK, FlgE and FliC. Immunoblotting using polyclonal anti-FliK (1st row), anti-FlgE (2nd row) or anti-FliC (3rd row) antibody, of whole cell proteins (Cell) and culture supernatants (Sup) from the above strains. The regions of interest were cropped from original immunoblots shown in Fig. S7a in the Supplemental information using Photoshop CS6, and then the contrast and brightness were adjusted. The positions of molecular mass markers (kDa) are indicated on the left. (c) Histograms of hook length distribution of MMK1013 (∆206–245), MMK1014 (∆206–255) and MMK1015 (∆206–265). (d) Motility of TH8426 harboring pTrc99AFF4 (∆fliK), pNM201 (∆99), pMMK1028 (∆99 + ∆206–245), pMMK1029 (∆99 + ∆206–255) or pMMK1030 (∆99 + ∆206–265) in 0.35% soft agar containing 1 mM IPTG. Plates were incubated at 30 °C for 8 hours. (e) Secretion assay of FliC. Immunoblotting using polyclonal anti-FliC (1st row) or anti-FliK (2nd row) antibody, of whole cell proteins (Cell) and culture supernatants (Sup) from the above strains. The regions of interest were cropped from original immunoblots shown in Fig. S7b in the Supplemental information using Photoshop CS6, and then the contrast and brightness were adjusted. The positions of molecular mass markers (kDa) are indicated on the left.
