【作者】 刘兆宇；

【导师】 程安春； 汪铭书；

【作者基本信息】 四川农业大学 ， 预防兽医学， 2002， 硕士

【摘要】 本文对4种血清型的(1型RA、2型RA、4型RA、5型RA)的外膜蛋白及其免疫原性进行了探讨。 结果表明采用超速离心法提取细菌的外膜蛋白，在电镜下观察都呈典型的双层泡状结构，但是形态有一定差异，有的呈“O”形，有的呈牙齿状。 所提取的4种血清型RA外膜蛋白经SDS-PAGE凝胶电泳分析结果表明，其外膜蛋白电泳图谱基本相同，但是分子量范围有一定差异，并且他们之间的主要条带也有差异。1型RA的分子量范围在25-143KD之间，其中主要蛋白带分子量分别是31KD，44KD，69KD，77KD，82KD，91KD和107KD，另外还有一些轻染的次要蛋白带；2型RA的分子量范围是26KD-135KD之间，其中主要蛋白带分子量分别是31KD，33KD，75KD，101KD和116KD；4型RA的分子量范围是26KD-124KD之间，其中主要蛋白带分子量是32KD，38KD，80KD，114KD；5型RA的分子量范围是28-121KD之间，其中主要蛋白带分子量是31KD，43KD，70KD，89KD，92KD，111KD。 经过免疫印迹证实，1型RA的44KD外膜蛋白与免疫鸭血清出现较强的阳性反应，是1型RA的主要免疫源；2型RA的40KD外膜蛋白与免疫鸭血清出现较强的阳性反应，是2型RA的主要免疫源；4型RA的75KD外膜蛋白与免疫鸭血清出现较强的阳性反应，是4型RA的主要免疫源；5型RA的39KD外膜蛋白与免疫鸭血清出现较强的阳性反应，是5型RA的主要免疫源。 4种血清型的外膜蛋白免疫小鸭后都能诱导机体产生较强的抗体反应，对同源细菌攻击的保护率为100％。更多还原

【Abstract】 Four major serotypes of Riemerella anatipestifer-serotype 1,2,4,5 are chosed for the test, we extract their outer membrane protein and study their immunogenicity.Ultracentrifugationg-Sarkosyl extraction was chosen for the isolation of the outer membrane proteins (OMP) of Riemerella antipestifer (RA). Under the electron microscopy, the ultra structure of RA was observed. The OMP of RA showed the typical double round vesicules with negative stain under electronic microscopy; however, there are some differences among the morphologic. Some are O form, some are tooth form.The electrophoretogram of the four serotypes is some similarity after SDS-PAGE, but there are many differences among them. The molecular weight of OMP of serotype 1 RA ranges from 25kd to 143kd, among them, 31KD,44KD,69KD,77KD,83KD, 91KD and 107KD proteins are more heavily stained by Commassie blue; serotype 2 RA ranges from 26kd to 135kd, among them, 31kd,33kd,75kd, 101kd and 116kd are more heavily stained by Commassie blue; serotype 4 RA ranges from 26kd to 124kd, among them, 32kd,28kd,80kd,114kd are more heavily stained by Commassie blue; serotype 5 ranges from 28kd to 121kd,among them, 31kd, 43kd,70kd,89kd,92kd,lllkd are more heavily stained by Commassie blue.The 44KD of OMP of serotype 1 RA have strong masculine with duck serum reaction by WESTERN-BLOTTING; the 40KD of OMP of serotype 2 RA appears obvious reaction with duck serum by WESTERN-BLOTTING; the 75KD of OMP of serotype 4 RA appears obvious reaction with duck serum by WESTERN-BLOTTING; the 39KD of OMP of serotype 5 RA appears obvious reaction with duck serum by WESTERN-BLOTTING;From the result of duckling immured by outer membrane protein, we know that OMP of all the four serotypes can induce obvious antibody reaction, and the protective ratio of the attack from the RA of the same bacteria was 100%.更多还原