【作者】 朱德康；

【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， 2003， 硕士

【摘要】 本研究采用超速离心法提取了五种血清型的鸭疫里默氏杆菌(RA)外膜蛋白，采用SDS-PAGE技术对其电泳图谱进行比较分析，并采用免疫印迹技术对其免疫原性及是否存在交叉免疫原性进行了深入研究。 对五种血清型的鸭疫里默氏杆菌外膜蛋白的SDS-PAGE图谱比较分析，结果表明其外膜蛋白的电泳图谱具有一定相似性。五种血清型的RA外膜蛋白存在7个分子量相同的条带，分子量分别为29KD、32KD、36KD、43KD、54KD、57KD和74KD。对主要外膜蛋白条带的组成进行分析后可将这五种血清型的鸭疫里默氏杆菌外膜蛋白分为三个外膜蛋白型，同时也表明不同血清型的菌株之间存在相同的外膜蛋白型。 使用抗RA外膜蛋白阳性鸭血清、抗RA阳性鸭血清和抗RA阳性鸡血清对RA外膜蛋白抗原进行免疫印迹检测，结果表明五种血清型的RA外膜蛋白均有免疫原性，且存在多种免疫原性的外膜蛋白成分；同时通过多份抗血清免疫印迹检测后的比较分析，证实7型的54KD外膜蛋白，8型的73KD外膜蛋白，21型的38KD外膜蛋白，20型的28KD外膜蛋白以及11型的28KD外膜蛋白是RA重要的免疫原性蛋白；将五种血清型的鸭疫里默氏杆菌外膜蛋白经SDS-PAGE后同时转印到硝酸纤维素膜上，再分别与7型、8型、21型、20型和11型RA外膜蛋白免疫的阳性鸭血清进行抗原交叉识别反应，结果表明不同血清型菌株的外膜蛋白之间存在交叉免疫原性，74KD和31KD外膜蛋白是重要的交叉免疫原性蛋白。更多还原

【Abstract】 The outer membrane protein (OMP) of 5 serotypes Riemerella antipastifer(RA) isolates was extracted by ultracentrifugation method. The electrophoresis profiles of OMP were gained by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The immunogenicity and antigen-cross-recognition-reaction of OMP were studied by immunoblotting test.The electrophoresis profiles showed the OMP of the 5 serotypes RA isolates had some similarities and shared 7 protein bands with molecular weight of 29KD, 32KD, 36KD, 43KD, 54KD, 57KD and 74KD. The OMP could be divided into 3 OMP patterns according to the major OMP bands. The results also indicate that there were the same OMP patterns in despite of different serotypes RA isolates.The immunogenicity of OMP was detected by using anti-OMP positive duck serum, anti-RA positive duck serum and anti-RA positive chicken serum in the immunoblotting test. The results showed that the OMP of all the 5 serotypes RA isolates had immunogenicity. The most valuable immunogenic OMP of serotype 7, 8, 21, 20 and 11 had molecular weight of 54KD, 73KD, 38KD, 28KD and 28KD respectively. The OMP antigens of all the 5 serotypes RA were transferred to the nitrocellulose membrane and reacted to the anti-OMP positive duck serum. It appeared apparently positive bands after the antigen-cross-recognition-reaction. The results indicated that there existed cross-immnuogenicity among the different serotypes RA isolates. The OMP with molecular weight of 74KD and 31KD were the most valuable components in cross-immunogenicity.更多还原