Figure 3

Complementation of the E. coli ΔtatC, pcnB strain, B1LK0-P, with tatC genes from other bacteria. Strain B1LK0-P was transformed with either: E. coli tatC encoded on pFAT417 (EcoC), P. syringae tatC on plasmid pUniprom-PC (PsyC), S. coelicolor tatC from plasmid pUniprom-SC (ScoC), A. aeolicus tatBC from plasmid pQEAQBC (AaeBC) or pBluescript (empty vector; marked as Δ in panel C). A. Growth of strains on LB medium containing 2% SDS. B. Growth of strains anaerobically on minimal glycerol TMAO medium. C. TMAO reductase activities from periplasmic fractions. *100% activity is taken as that determined from the periplasmic fraction of B1LK0-P carrying pFAT417 and corresponds to 0.79μmol benzyl viologen oxidised/min/mg protein. Error bars represent the standard error of the mean (n = 3). D. The A. aeolicus TatC protein is produced from clone pQEAQBC. Strain M15 [pREP4] harboring either pQE60 (empty vector) or pQEAQBC (AaeC) was cultured in LB medium until OD600 of 0.4 was reached, after which production of the TatChis protein was induced by addition of 1 mM isopropyl-β-D-galactopyranoside (IPTG) for a further 2 hours. Membrane fractions were prepared, proteins (50μg of total membrane protein), separated by SDS-PAGE, blotted onto nitrocellulose and developed using anti-penta-His antiserum.