鸭疫里默氏杆菌血凝素基因的克隆、表达及其蛋白的定位分析  

Expression and localization of the hemagglutinin in Riemerella anatipestifer

作　　者：屠晶[1] 胡青海[1] 于圣青[1] 丁铲[1] 韩先干[1] 朱寅玉[1] 王小兰[1] 苗双[1] 卢凤英[1] 

机构地区：[1]中国农业科学院上海兽医研究所,上海200241

出　　处：《中国预防兽医学报》2012年第9期692-696,共5页Chinese Journal of Preventive Veterinary Medicine

基　　金：上海市自然科学基金(09ZR1438600);国家自然科学基金面上项目(31072156;31072161);农业部行业专项(201003012);中央科研院所公益性基础科研业务费项目(2010JB17)

摘　　要：为研究鸭疫里默氏杆菌(R.anatipestifer)血凝素蛋白的功能,本实验克隆表达了鸭疫里默氏杆菌血凝素基因,并对其蛋白定位进行了分析。根据鸭疫里默氏杆菌CH3株血凝素基因序列设计一对特异性引物,采用PCR扩增不同血清型鸭疫里默氏杆菌的血凝素基因,进行序列测定与分析。结果表明其血凝素基因全长1 059 bp,编码352个氨基酸。不同血清型鸭疫里默氏杆菌血凝素间的氨基酸同源性为94.1%～100%,与黄杆菌3519-10株血凝素的氨基酸同源性为68.0%～68.9%。采用表达的血凝素蛋白免疫鼠血清和鸭抗鸭疫里默氏杆菌阳性血清对血凝素在细菌中的定位分析表明,鸭疫里默氏杆菌血凝素可能不是一种膜蛋白,而是存在于胞浆中的一种蛋白。To study the function of hemagglutinin in Riemerella anarpestifer, a pair of specific primers were designed based on the sequence of the hemagglutinin gene of R. anatipestifer strain CH3, and the hemagglutinin gene was amplified by PCR from different serotypes of strains. The results showed that the amino acid identity of hemagglutinin among different serotypes of R. anatipestifer was 94.1% to 100%, and shared 68.0% to 68.9% identity with that of Flavobacteriaceae bacterium 3519-10 strain. The western blot and indirect immunofluorescence assay showed that R. anatipestifer was reacted positively only with the antiserum prepared from the recombinant hemagglutinin in mice, but not with duck serum against R. anatipestifer, which suggested that the hemagglutinin might be not a surface protein, but a cytoplasmic protein in R. anatipestifer.

关 键 词：鸭疫里默氏杆菌 血凝素 原核表达 免疫转印 定位 

分 类 号：S852.61[农业科学—基础兽医学]