猪附红细胞体粘附蛋白及其红细胞受体的筛选与鉴定

项目名称： 猪附红细胞体粘附蛋白及其红细胞受体的筛选与鉴定

项目编号： No.31302125

项目类型： 青年科学基金项目

立项/批准年度： 2014

项目学科： 农业科学

项目作者： 邓洪宽

作者单位： 山东理工大学

项目金额： 22万元

中文摘要： 附红细胞体病是由附红细胞体引起的一种呈全球性分布的人兽共患病，该病不仅危害人类健康，而且还能引起动物的发病和死亡。尤其是猪感染附红细胞体后，可致生长发育受阻，生产能力下降，严重时引起死亡，并且隐性感染猪和耐过猪还可长期带毒、终生散毒，因此给我国的畜牧业发展和公共卫生安全带来了巨大的损失和危害。但时至今日，尚未有效控制附红细胞体病的方法，阐明附红细胞体的致病机制对附红细胞体病的防控具有积极意义。因此本研究以猪附红细胞体为模型，首先构建猪附红细胞体信号标签突变文库，然后对其进行体内筛选和体外筛选，获得猪附红细胞体粘附蛋白基因。然后通过噬菌体展示技术、红细胞粘附抑制实验、电镜技术等对所获得的粘附蛋白基因做进一步鉴定。最后利用Far-Western blot、Pull-Down等技术筛选和鉴定附红细胞体粘附蛋白的红细胞受体。从而为阐明猪附红细胞体致病机制，进而为猪附红细胞体病的防控奠定基础。

中文关键词： 猪附红细胞体；噬菌体展示；粘附蛋白；受体；免疫学诊断

英文摘要： Eperythrozoonosis is a globally widespread zoonosis caused by Eperythrozoon, which is not only harmful to human health, but also causes animal morbidity and mortality. Especially the pigs infected with Eperythrozoon, can result in meat yield reduction, growth retardation, productivity decrease and death. And the recessive infected pigs or resistant pigs can be long-term, life-long spreading Eperythrozoon, therefore it is caused enormous loss and harm to the development of animal husbandry and public health security. To date, no method has been found to control Eperythrozoonosis, so it is important to prevent and control the disease by elucidating the pathogenic mechanism of E.suis. In this study, we use pig Eperythrozoon (E. suis) as a model to construct E.suis signature-tagged mutagenesis library, then screen the library in vivo and in vitro to obtain E.suis adhesion protein genes. After that, further identification of the adhesion protein genes were performed by phage display technology, red cell adherence inhibition assay, electron microscopy etc. In the end, the screening and identification of erythrocyte receptors were performed by Far-Western blot, Pull-Down etc. We hope this study can elucidate the pathogenic mechanism of E.suis, and lay foundation for Eperythrozoonosis prevention and control.

英文关键词： Mycoplasma suis；Phage display；Adhesion protein；Receptor；Immunodiagnosis