2种弧菌鞭毛蛋白的分离纯化与鉴定

Separation,Purification and Identification of Two Vibrio Flagellin

尝试分离提取2种弧菌的鞭毛蛋白，采用垂直板聚丙烯酰胺凝胶电泳对所提蛋白进行分离纯化，Western免疫印迹检测，电镜鉴定。结果显示：实验已经成功分离提纯了哈维氏弧菌的鞭毛蛋白，获得了溶藻弧菌鞭毛蛋白的初提液。提纯的哈维氏弧菌鞭毛蛋白经SDS-PAGE显示3条清晰蛋白带（相对分子量分别为39.7，41.9和42.7KDa），透射电镜观察表明该蛋白呈经典的丝状，可以认为哈维氏弧菌的鞭毛丝是由相对分子量分别为39.7，41.9和42.7KDa的3种鞭毛蛋白亚基构成。溶藻弧菌鞭毛蛋白的初提液经SDS-PAGE后显示5-9条蛋白带，其中36.8KDa的蛋白带有强的免疫原性。

The vibro flagellin of Vibro alginolyticus and Vibro harveyi was pxtracted and then detected by high-speed centrifugal, cycles of differential centrifugation, vertical slab gel electrophoresis, western blotting and electronic microscopy identification. The result shows that the experiment has already separated the purificated flagellin of Vibro harveyi and the original flagellin extracts of Vibro alginolyticus succesfully. The purification of Vibro harveyi has three clear protein lines by SDS-PAGE （relative molecular weight are 39.7,41.9 and 42.7 KDa）. Transmission electronic microscopy discovers that the protein assumes classical filamentous. This suggests that the flagellar filament of Vibro harveyi is made up of three subunit of flagellin whose molecular weight are 39.7,41.9 and 42.7 KDa respectively. The original flagellin extracts of Vibro alginolyticus have five to nine protein lines by SDS-PAGE, and the protein whose molecular weight are 36.8 KDa have remarkable immuno-genicity.