大菱鲆致病性溶藻弧菌SR1的外膜蛋白及其抗原性分析

用十二烷基肌氨酸钠(Sarkosyl)抽提结合超速离心的方法提取了一株大菱鲆致病性溶藻弧菌(Vibrio alginolyticus)SR1和其他7株弧菌的外膜蛋白。通过SDS-PAGE图谱分析比较了这8株弧菌外膜蛋白的组成,结果表明,8株弧菌的外膜蛋白电泳一般可得到6-12条条带,其分子量多集中在65-106 kD和28-48 kD,其中36 kD的蛋白带为8株弧菌所共有。用兔抗SR1全菌血清进行Western-blot印迹显示,菌株SR1的外膜蛋白条带中有6条发生了阳性反应,其分子量分别为73 kD、48 kD4、5 kD3、9 kD、36 kD和32 kD。而其他7株弧菌的外膜蛋白与兔抗SR1血清也发生程度不等的阳性反应,这些阳性反应条带的分子量集中在65-73 kD、45-48 kD和36-41 kD之间,其中36 kD的外膜蛋白在8株弧菌中均出现明显的阳性反应,说明36 kD的外膜蛋白是这8株弧菌共有的特异性抗原。

Antigenicity of outer membrane proteins of a pathogenic Vibrio strain, Vibrio alginolyticus

A pathogenic Vibrio alginolyticus strain SR1,isolated from ascites of cultured turbot(Scophthalmus maximus) with swollen abdomen,was used to immunize New Zealand rabbit.Strain SR1 could effectively stimulate the rabbit to produce immunity response,and by enzyme-linked immunosorbent assay(ELISA) titer of the antiserum was 6 400.Strain SR1 and other seven strains of Vibrio were used for preparation of the outer membrane proteins(OMPs) with Sarkosyl method combining super centrifugal purification.SDS-PAGE showed that the pathogenic strain SR1 had 12 membrane proteins,containing 9 major protein bands with molecular weights of 106 kD,73 kD,66 kD,51 kD,48 kD,45 kD,39 kD,36 kD and 32 kD,and 3 minor protein bands with molecular weights of 92 kD,82 kD and 29 kD.OMPs of the 8 strains of Vibrio were analyzed and compared by using SDS-PAGE.The results showed that there were generally 6-12 OMP bands in each of the 8 strains.Molecular weights focused on 65-105 kD and 28-48 kD,and the protein with molecular weights of 36 kD was common for all the 8 strains.After SDS-PAGE,in the Western-blot with rabbit antiserum against SR1 whole cells,only 6 OMPs of strain SR1 with molecular weights at 73 kD,48 kD,45 kD,39 kD,36 kD and 32 kD were detected.It showed that the 6 OMPs had some antigenicity.And the other 7 strains all had positive reaction,with molecular weights of the reaction bands mainly focusing on 73-65 kD,48-45 kD and 41-36 kD.The band of 36 kD had visible positive reaction in all the 8 strains.This illuminated that the 36 kD OMP was the common special antigen of the 8 Vibrio strains.