血清2型鸭疫里默氏杆菌外膜蛋白多肽的组成及免疫原性  

Outer Membrane Protein Polypeptide Component and Immunogenicity of Serotype 2 Riemerella Anatipestifer

作　　者：程安春[1] 汪铭书[1] 刘兆宇[1] 周毅[1] 陈孝跃[1] 方鹏飞[1] 郭宇飞[1] 

机构地区：[1]四川农业大学动物科技学院禽病防治研究中心,四川雅安625014

出　　处：《中国兽医学报》2004年第4期329-332,共4页Chinese Journal of Veterinary Science

基　　金：教育部重点项目 ( 0 110 2 );四川省"十五"攻关项目( 0 10 0 2 40 0 13 )

摘　　要：采用超声波裂解和超速离心法提取血清 2型鸭疫里默氏杆菌 (RA)的外膜蛋白 ,在透射电镜下观察 ,外膜蛋白呈典型的双层泡状结构 ,形态主要呈 O型或牙齿状或不规则的小碎片。SDS- PAGE电泳分析 ,血清 2型 RA的外膜蛋白由 11条多肽组成 ,相对分子质量 (Mr)在 2 6 0 0 0～ 135 0 0之间 ,主要蛋白为 310 0 0、330 0 0、75 0 0 0、10 10 0 0和 116 0 0 0 ,其相对百分含量分别为 16 .97%、15 .14 %、13.97%、19.83%和 12 .6 9%。经免疫印迹证实 ,血清 2型 RA的 4 0 0 0 0外膜蛋白与免疫鸭血清呈较强的阳性反应 ,是主要免疫原蛋白。用分离的外膜蛋白加上弗氏佐剂制备的亚单位疫苗免疫雏鸭后 ,可诱导产生较强的抗体 ,免疫鸭对同源 RA菌株的攻击可产生 10 0Ultrasonic cracked and ultracentrifugation-sarkosyl extraction was chosen for the isolation of the(OMP) of serotype 2 Riemerella antipestifer(RA).The ultra structure of RA was observed with the electron microscopy.The OMP of RA showed the typical double round vesicules with “O” shape or “teeth” shape and unequal fractionlet when be negative stained under electronic microscopy.By SDS-PAGE,11 polypeptides are make up of OMPs from serotype 2 RA,their molecular weight ranges from (26 000) to (135 000),molecular weight of mainly polypeptides are (31 000),(33 000),(75 000),(101 000) and (116 000),and their percent contents are 16.97,15.14,13.97,19.83 and 12.69 percent separately.(40 000) polypeptide from OMPs of serotype 2 RA appears obvious reaction with vaccined duck serum by Western-blotting,they are important immunogenic proteins.Ducklings immuned with the vaccines made of OMPs from serotype 2 RA with Fruend′s complete adjuvant can produce high ELISA antibody titers and against the same serotype RA infected.

关 键 词：2型鸭疫里默氏杆菌 外膜蛋白 多肽 免疫原性 菌株 

分 类 号：S852.61[农业科学—基础兽医学]