【摘要】 为评价鸭源鸡杆菌（Gallibacterium anatis,G.anatis）外膜蛋白A（Outer membrane proteins A,OmpA）的免疫原性及免疫保护力,本研究通过PCR技术扩增去除信号肽的ompA基因序列,酶切后与载体pET-32a（+）连接,构建重组表达质粒pET-32a（+）-OmpA,在IPTG诱导下成功表达出约40 ku的包涵体蛋白。Western-blot分析显示重组蛋白rOmpA具有抗原性;以纯化的rOmpA免疫小鼠,分别在一免后第2周和第4周进行加强免疫。三免后进行抗体检测并以G.anatis PDS-RZ-1-SLG株的5×LD₅₀攻毒,同时设立全菌灭活组和PBS组作为对照。结果显示,rOmpA可诱导小鼠产生较高水平的抗体,可提供50%的免疫保护力。结果表明,鸭源鸡杆菌外膜蛋白A是一种保守性共同抗原,可作为鸭源鸡杆菌疫苗的候选抗原。更多还原

【Abstract】 To evaluate immunogenicity and immune protection of Outer membrane proteins A（OmpA）of Gallibacterium anatis（G.anatis）,the ompA gene without signal peptide sequence was amplified and cloned into pET-32 a（+）.After being induced by IPTG,OmpA was successfully expressed as inclusion bodies with a size of about 40 ku.Western-blot analysis showed that the recombinant protein rOmpA had a good antigenicity.The mice were vaccined with purified OmpA,and boosted day 14 and 28 post first immunization,respectively.The antibody was examined by ELISA,and the result showed that the antibody titer was high.The rOmpA could provide 50% protection for the mice challenged with 5×LD₅₀ of G.anatis strain RZ day 14 post the third immunization.These results indicated that OmpA was a conserved common antigen of G.anatis and an apromising candidate for developing a subunit vaccine against G.anatis.更多还原