创伤弧菌外膜蛋白的分离及其抗原性分析

分别采用Sarkosyl和PMSF法分离、提取鳗源创伤弧菌FJ03-X2的外膜蛋白,并应用SDS-PAGE和Western-blotting分析比较两种方法提取的外膜蛋白的组分和抗原性。SDS-PAGE分析结果表明,Sarkosyl法分离的主要外膜蛋白分子量集中在14～66 kDa之间;而PMSF法分离的主要外膜蛋白分子量分布在14～92 kDa之间。在两种方法中,分子量38 kDa、36 kDa的外膜蛋白均为高丰度蛋白。两种方法提取的创伤弧菌外膜蛋白经SDS-PAGE后,用兔抗创伤弧菌FJ03-X2血清进行Western-blotting,结果显示,兔抗FJ03-X2高免血清能识别绝大多数外膜蛋白组分,说明FJ03-X2菌株的外膜蛋白具有良好的抗原性。其中,分子量分别为66kDa、47 kDa、44 kDa、38 kDa、36 kDa、34 kDa的外膜蛋白呈强阳性反应,是主要的免疫原组分。同时,经比较分析认为PMSF法提取创伤弧菌外膜蛋白的效果更好。

Antigenicity and identification of Vibrio vulnificus outer membrane proteins

The outer membrane proteins(OMPs) are the major components in outer membrane,which is unique to the gram-negative bacteria and plays an important role in bacterial life functions.This study focuses on the antigenicity and identification of OMPs of Vibrio vulnificus FJ03-X2,a pathogenic stain isolated from European eel(Anguilla anguilla).By applying Sarkosyl and PMSF methods,the major OMPs(MOMPs) of V.vulnificus were obtained.The constituents and antigenicities of MOMPs were analyzed by means of SDS-PAGE and...