FIG. 4.

Protein immunoblots of recombinant MBP-AnkA (A), total lysate of E. coli transformed by ee-o (B), and whole HGE agent (C). (A) Recombinant MBP-AnkA probed with the following primary antibodies: mouse polyclonal anti-MBP-AnkA (lane 1), mouse polyclonal anti-HGE agent (BDS strain) (lane 2), nonimmune mouse serum (lane 3), rabbit anti-MBP (lane 4), nonimmune rabbit serum (lane 5), mouse monoclonal IE3 [IgG1(κ)] to MBP-AnkA (lane 6), and mouse monoclonal IgG1(κ) control antibody (lane 7). (B) Lysates of E. coli transformed by ee-o (lanes 1 and 3) or not transformed (lanes 2 and 4) were processed as described for panel A and probed with the following primary antibodies: mouse monoclonal IE3 [IgG1(κ)] to MBP-AnkA (lanes 1 and 2) and mouse polyclonal anti-HGE agent (lanes 3 and 4). (C) Whole HGE agent was processed as for panels A and B and probed with the following primary antibodies: rabbit anti-HGE agent (Webster strain) (lane 1), rabbit anti-MBP (lane 2), nonimmune rabbit serum (lane 3), mouse polyclonal anti-HGE agent (BDS strain) (lane 4), mouse polyclonal anti-MBP-AnkA (lane 5), and nonimmune mouse serum (lane 6). The numbers on the right represent the molecular masses (MWs) of HGE agent proteins that contain AnkA antigens (150, 90, 75, and 51 kDa) and the 42-kDa immunodominant HGE agent antigen that reacts only with polyclonal HGE agent antibodies.