FIG. 4.

Western blot analysis using polyclonal antibodies against the recombinant OmpU protein (anti-OmpU). (A) Confirmation of ompU mutant V. vulnificus by Western blot analysis. Lane 1, lysate of wild-type V. vulnificus MO6-24/O; lane 2, lysate of DK1, an ompU mutant of V. vulnificus. (B) Confirmation of the ompU complementation strain by Western blot analysis. Lane 1, lysate of DK1 harboring a broad-host-range vector, pJH0311; lane 2, lysate of DK1 harboring pSM1, a complementation plasmid of the wild-type ompU gene. Lysates of four different strains of V. vulnificus were prepared as described in Materials and Methods. Fifty micrograms of protein was subjected to SDS-PAGE per each sample and transferred onto a membrane. After being blocked with 5% nonfat dry milk, the membrane was incubated with mouse antibodies against the recombinant OmpU and was then incubated with horseradish peroxidase-conjugated anti-mouse immunoglobulin G. Immunoreactive proteins were visualized using an enhanced chemiluminescence system. MW, molecular weight (in thousands).