FIG. 2.
Western blot (immunoblot) analysis of HemA[WT] and HemA[KK] expressed from a plasmid-borne PBAD promoter in E. coli. The arrow indicates the HemA[WT] and HemA[KK] proteins. Lanes: b, E. coli TE2331 wild type; c, E. coli TE7202 hemA8/pTE570 (vector); d, E. coli TE7420 hemA8/pTE713 hemA[KK]; e, E. coli TE7207 hemA8/pTE694 hemA[WT]. Samples for lanes f to h were prepared as for lane e except that cultures were grown with various concentrations of the inducer arabinose: 0.001% arabinose (lane f), 0.005% arabinose (lane g), or 0.01% arabinose (lane h). Lane a contained proteins from S. typhimurium LT-2 (wild type) as a positive control. Cultures were grown to an OD600 of 0.4 in LB medium (containing ampicillin where applicable) and processed for immunoblotting with anti-HemA MAb H23 exactly as described previously (36).