副猪嗜血杆菌黏附基因的表达和免疫原性鉴定  

Expression and characterization of adhesin gene of Haemophilus parasuis

作　　者：耿屹[1] 曹荣峰[2] 高晓宇[3] 李守军[3] 张桂红[3] 

机构地区：[1]山西农业大学动物科技学院,山西太谷030801 [2]青岛农业大学动物科技学院,山东青岛266109 [3]华南农业大学兽医学院,广东广州510642

出　　处：《中国预防兽医学报》2010年第5期398-400,共3页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家生猪现代产业技术体系(nycytx-009)

摘　　要：为获得副猪嗜血杆菌(HPS)黏附素基因的表达产物并对其进行免疫原性鉴定,本研究采用PCR技术扩增了HPS黏附素基因序列,将扩增产物与表达载体pET-32a(+)连接,得到表达重组质粒。通过EcoRⅤ和HindⅢ双酶切鉴定和测序确认正确后,将重组质粒转化大肠杆菌BL21(DE3)中进行表达。经SDS-PAGE电泳分析,获得了59.5ku的表达产物,与预期大小的黏附素蛋白分子量一致。经western blot检测,表达产物与HPS阳性血清反应,证明表达产物具有一定的免疫活性。提示本研究所表达的蛋白将有助于HPS的血清学诊断和疫苗的研发。To express the adhesion protein of Haemophilus parasuis,the adhesin gene of H.parasuis was amplified by PCR,cloned into pET-32a（＋） and expressed in BL21（DE3）.SDS-PAGE analysis showed that the expression product had MW of 59.5 ku.Western blot assay confirmed that the protein could react with the HPS-positive serum.It is believed that the recombinant protein could be used as an effective antigen for the diagnosis of H.parasuis infection and vaccine development.

关 键 词：副猪嗜血杆菌 黏附素基因 表达 免疫原性鉴定 

分 类 号：S852.61[农业科学—基础兽医学]