牙鲆秦皇岛弧菌HQ010712-1外膜蛋白及其抗原性分析

采用十二烷基肌氨酸钠(Sarkosyl)和苯甲基磺酰氟(PMSF)2种方法提取秦皇岛弧菌HQ010712-1(Vibrio qinhuangdaora sp.nov.)外膜蛋白,结果显示 Sarkosyl法提取效果较好,且所提取的主要外膜蛋白分子量为102kD、45 kD、39 kD、36 kD、30 kD、28 kD、24 kD、22 kD;为比较该菌株与弧菌属其他细菌外膜蛋白组分及抗原性异同,以鳗弧菌(Vibrio anguillarum)、副溶血弧菌(Vibrio parahaemolyticus)、溶藻胶弧菌(Vibrio alginolyticus)为对照,电泳图谱显示4种弧菌外膜蛋白的分子量主要集中在22～48 kD之间;利用抗秦皇岛弧菌HQ010712-1血清的免疫印迹表明菌株HQ010712-1外膜蛋白中分子量为45 kD、36 kD的蛋白条带呈现阳性反应,其他3种弧菌外膜蛋白中均有与该抗血清反应的条带,且分子量为36 kD的反应带为菌株HQ010712-1、副溶血弧菌、溶藻胶弧菌共有.本研究旨在为进一步筛选和研究致病性弧菌的共同保护性抗原提供参考.

Antigenicity of outer membrane proteins of pathogenic Vibrio qinhuangdaora isolated from flounder Paralichthys olivaceus

was reported to be a new marine pathogen that had caused vibriosis of fishes. In Qinhuangdao, Hebei Province was one of the causative agents of vibriosis that endangered the aquaculture of Japanese flounder , and no effective therapy was found other than antibiotics. Vaccines have been successfully used to control vibriosis in aquaculture, and information on the outer membrane proteins (OMPs) of several other pathogenic vibrio species have been disclosed. However, little is known about this pathogen. Outer membrane proteins (OMPs) are highly immunogenic bacterial components due to their exposed epitopes on cell surface. Interests have been attracted to understand the biological function of OMPs and their immunogencity characteristics in immune response. The present study was initiated to analysis the antigenicity of OMPs instrain HQ010712-1 and compared with three universal marine vibriosis species such as , so as to throw some insight to the protective antigens of the bacteria and facilitate vaccine development. were obtained by Sarkosyl method combining super centrifugal purification. SDS-PAGE showed that the strain HQ010712-1had 8 membrane proteins with molecular weights of 102 kD, 45 kD, 39kD, 36 kD, 30 kD, 28 kD, 24 kD and 22 kD. OMPs of the 4 strains of were analyzed and compared by using SDS-PAGE. The results showed that molecular weights focused on 22 with rabbit antiserum against strain HQ010712-1 whole cells, only 2 OMPs of strain HQ010712-1with molecular weights at 45 kD and 36 kD were detected. It showed that the 2 OMPs had some antigenicity. And the other 3 strains of all had positive reaction. The band of 36 kD had visible positive reaction in the .