Isolation of Bacteria with Membrane Proteins Homologous to Vibrio anguillarum Porin Omp35La

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Bacterial porin homologues have recently been found in seawater as a dissolved organic matter. However, the bacteria of origin have not been identified. This manuscript describes the isolation of the marine bacteria having similar antigenic proteins to porin Omp35La of Vibrio anguillarum. Colony forming bacteria from seawater on Marine Broth 2216E plate were screened to determine whether they have Omp35La-like proteins by colony western blotting with anti-Omp35La antibody. Among the 37 final positive isolates, 11 isolates were classified into the Vibrio-group, whereas 26 isolates were assigned to be non-Vibrio-group based on the biochemical properties. When outer membrane protein was purified and western blotting was performed, 8 our of 11 Vibrio-group and 10 out of 26 non-Vibrio-group isolates had antibody reactive proteins. This suggests that not only Vibrio but also other class bacteria are possibly the origin of the dissolved protein. The family Vibrionaceae specific PCR worked in 7 isolates of the Vibrio-group. However, V. anguillarum itself was not identified by PCR-RFLP assay.