A. Production of the cellulase BcsZ in the ΔbcsG mutants of the cellulose synthase-positive strain MAE97. MAE97 bcsZ deletion strains with pBAD30 vector control (ΔbcsZ-VC) and with overexpressed BcsZ (ΔbcsZ-pBcsZ) were used as negative and positive controls, respectively. Detection of BcsZ production was by Western blot using a rabbit anti-BcsZ antibody.
B. Production of the putative outer membrane pore BcsC in S. typhimurium wild type and bcsG derivatives. Wild-type S. typhimurium UMR1 with BcsC-3xFLAG and its ΔbcsG mutant were complemented by the same plasmids as in panel A with the addition of the second truncated variant, pBcsG1–165. Detection of the 132.7 kDa BcsC-3xFLAG was by Western blot using a mouse anti-FLAG-tag antibody. UMR1 BcsC-3xFLAG with pBAD30 (vector control, left lane) and UMR1 without the FLAG tag (right lane) were used as positive and negative controls, respectively.
Cells were grown on salt-free LB agar plates for 24 h at either 28 °C (A, top lane) or 37 °C (A, bottom lane), or for 16 h at 28 °C (panel B).