FIG. 5.

Detection of Fpp1 in culture supernatants of F. psychrophilum. Culture supernatants (4 ml) from cultures incubated under different conditions were precipitated with trichloroacetic acid and then loaded onto a SDS–12.5% PAGE gel. Immunoblotting was carried out as described in Materials and Methods. (A) F. psychrophilum was grown until the onset of the stationary phase at 12°C (lanes 1 and 2) and 18°C (lanes 3 and 4) in NB (lanes 1 and 3) or NBF medium (lanes 2 and 4). The positions of the 55-kDa (Fpp1) and 65-kDa immunoreacting bands are indicated on the right. (B) The microorganism was grown until the stationary phase at 12°C in NB containing 0 to 20 mM CaCl₂. Lane 1, no CaCl₂; lanes 2 to 6, 0.1, 1, 5, 10, and 20 mM CaCl₂, respectively. All of the immunoblot experiments were carried out at least three times.