Isolation and characteristics of major outer membrane proteins of aquatic pathogens Vibrio anguillarum and Vibrio alginolyticus

Three different extraction methods (SDS, Sarkosyl and PMSF) were used for preparing major outer membrane proteins (MOMP) from the cell membrane fraction of Vibrio anguillarum and Vibrio alginolyticus. The MOMP of the two vibrios were analyzed by using SDSPAGE. The membrane proteins of molecular weight at 27-138 kD can be shown in OMP extracted from Vibrio anguillarum and 27-104 kD in OMP isolated from Vibrio alginolyticus. Both of the two strains contain the protein bands of molecular weights at 45 kD, 30 kD and 27 kD. By comparing the results of the three methods, Sarkosyl method was the best in evidence in extracting the OMPs of vibrios. Three MOMPs with molecular weights at 97 kD, 51 kD and 30 kD were distinctly detected in the Westernblotting with rabbit antiserum against V.anguillarum whole cell, indicating the three MOMPs were involved in the determination of the cell antigenicity.Two MOMPs with molecular weights at 51kD and 27 kD in the OMPs of Vibrio alginolyticus also presented strong antigens. The MOMP at 51 kD was a weak antigen common to both Vibrio species