FIG. 3.

(A) SDS-PAGE of outer membranes of E. coli CE1248 carrying control vector pVLT35 (lane 1; C), native OprM-expressing plasmid pKW35TM (lane 2; TM), and OprM-encoding plasmids with malarial epitope insertions at sites ME1 to ME6 (lanes 3 to 8) and ME8 to ME13 (lanes 9 to 14). Each lane was loaded with 20 μg of protein, treated with 5% (vol/vol) β-mercaptoethanol, and heated at 100°C for 10 min. The molecular masses (in kilodaltons) for the prestained markers in lane M are indicated on the left. Expression of oprM and the insertion mutants was confirmed by Western immunoblotting with an anti-OprM antibody (B) and an anti-malarial epitope antibody (C). Samples were in the same order as in the gel shown in panel A, and the corresponding regions of the blots are shown, with the relevant molecular mass indicated on the left.