4种病原弧菌外膜蛋白的提取及抗原性初步分析  

Isolation and antigenicity analysis of outer membrane proteins from 4 pathogenic Vibrio sp.

作　　者：阎斌伦[1] 张晓君[1] 秦国民[1] 毕可然[1] 徐静[1] 秦蕾[1] 

机构地区：[1]淮海工学院江苏省海洋生物技术重点实验室,江苏连云港222005

出　　处：《海洋科学》2012年第5期71-74,共4页Marine Sciences

基　　金：江苏省六大人才高峰项目(2009);连云港市科技攻关资助项目(CG0907);江苏省水产三项工程资助项目(P2009-18)

摘　　要：利用十二烷基磺酸钠抽提并结合超速离心的方法提取了鳗弧茵（Vibrio anguillarum）、河口弧菌（V．aestuarianus）、霍乱弧菌fEcholerae）和副溶血弧菌（V．parahaemolyticus）4种致病性弧茵的主要外膜蛋白，通过SDS．PAGE分析比较4种弧菌主要外膜蛋白的组分。结果表明：4株弧菌的外膜蛋白电泳图谱一般可得到5～10条蛋白带，分子量多数集中在26～40kD和48～85kD。对所提取的4种弧菌主要外膜蛋白进行SDS．PAGE后．分别与自制的兔抗鳗弧菌血清、抗河口弧菌血清、抗副溶血弧菌血清进行Western．blotting印迹分析。结果显示每种全菌抗血清可与相应菌的外膜蛋白部分组分发生免疫反应，并与其他3种弧菌外膜蛋白的部分组分产生交叉免疫反应，这些反应条带分子量主要集中于30～48kD之间。本研究为进一步研究致病性弧菌外膜蛋白免疫学特性提供参考。The outer membrane proteins of 4 pathogenic Vibrio sp. （V.anguillarum, V.aestuarianus, Echolerae and V.parahaemolyticus） were extracted using sodium laurylsulfonate and ultracentrifuging methods, and the ingredient of the outer membrane proteins were analyzed using SDS-PAGE. The SDS-PAGE results showed that five to ten proteins bands could be obtained for each vibrio with molecular weights ranging from 26 to 40 kD and from 48 to 85 kD. West- ern-blotting analysis was conducted after SDS-PAGE, the results showed that some of the outer membrane protein bands have cell antigenicity in the Western-blotting with rabbit antiserum against whole cell of V.anguillarum, V.aestuarianus, Echolerae and V.parahaemolyticus, respectively, the molecular weights were between 30 to 48 kD. This study can pro-vide reference for immunologic research of outer membrane proteins of pathogenic Vibrios.

关 键 词：鳗弧菌(Vibrio anguillarum) 河口弧菌(V.aestuarianus) 霍乱弧菌(V.cholerae) 副溶血弧菌(V.parahaemolyticus) 外膜蛋白 Western-blotting印迹 

分 类 号：S94[农业科学—水产养殖]