Studies On The Major Outer Membrane Proteins Of Pathogenic Vibrio Anguillarum Isolated From Diseased Marine Fish In Shandong Province

Vibrio anguillarium is one of the most serious pathogens, which can infect a wide range of marine or limnetic fish species and cause mass death. Outer membrane proteins (OMP)of Vibrio anguillarum play an important role in the course of pathogenity, such as invasion, adhesion, secretion of toxins, absorption of iron and resistance to antibiotics. The present study is focused on the following aspects.Antiserum of Vibrio anguillarium CW1,CW3,CW5,CW6 and TL01 was prepared. Serological analysis showed that antiserum of Vibrio anguillarium CW1,CW3,CW5,CW6 reacted with Vibrio anguillarium strain VIB1 (serotype O1 type ), which suggested that they belonged to serotype O1. Vibrio anguillarum TL01 belonged to serotype O2 while Vibrio anguillarium CW4 belonged to serotype O3.Outer membrane proteins of 10 type strains and 6 strains of V. anguillarum were extracted by sarkosyl method and analyzed by SDS-PAGE. The OMP profiles were heterogeneous, with the molecular weight ranging from 19 kDa to 83 kDa. Major outer membrane proteins of 6 strains of V. anguillarum showed common major proteins with the sizes of 24 kDa, 38 kDa, 42 kDa and 47 kDa, which account for over 90% of total OMPs.Outer membrane protein profiles of the V. anguillarum strains were further analysed by western blotting analysis with antiserum of V. anguillarum CW1 as primary antibody. The results showed that 24 kDa, 38 kDa, 42 kDa and 47 kDa major proteins were their major common antigens. The 24kDa protein showed stronger antigenicity than others, but the OMPs whose molecular weights exceed 47 kDa did not react with the antiserum.The effect of culture temperature on expressions of OMPs were studied, results...