变形蹄奶牛血清shotgun蛋白质组学分析  

Shotgun proteomic analysis of serum from dairy cattle with hoof deformities

作　　者：赵喜文[1] 郭东华[1] 谷城[1] 曲永利[1] 王欣宇[1] 苏明俊 魏姗[1] 王恩雨[1] 王志慧[1] 刘秋瑾[1] 辛科睿 武瑞[1] 孙东波[1] 

机构地区：[1]黑龙江八一农垦大学动物科技学院,黑龙江大庆163319

出　　处：《黑龙江畜牧兽医》2016年第5期65-69,284,共6页Heilongjiang Animal Science And veterinary Medicine

基　　金："十二五"农村领域国家科技计划项目(2012BAD12B05-2);黑龙江省高校青年学术骨干计划项目(1253G003);黑龙江省农垦总局科技计划项目(HNK125B-11-06A)

摘　　要：为了揭示变形蹄奶牛血清蛋白表达谱,探索奶牛变形蹄发生的机制,试验利用鸟枪法(shotgun)蛋白质组学技术分别对健康奶牛(UA,n=26)和变形蹄奶牛(HD,n=40)的混合血清样品进行了鉴定和生物信息学分析,进一步分析UA和HD样品的差异蛋白,并对选择的差异蛋白进行验证。结果表明:在UA样品中鉴定了393个蛋白,其中224个蛋白为高度可信蛋白;在HD样品中鉴定了409个蛋白,其中225个蛋白为高度可信蛋白。UA和HD样品中鉴定的蛋白,在分子功能、细胞进程和细胞成分分类中显示出明显的差别。在UA和HD样品中存在着24个潜在的差异蛋白,其中18个蛋白来自HD样品,6个蛋白来自UA样品;在HD样品潜在的差异蛋白中,桥粒蛋白(desmoplakin)、连接桥粒斑珠蛋白(junction plakoglobin)和钙磷脂结合蛋白A2(annexin A2)与奶牛变形蹄发生具有潜在的相关性。通过ELISA或Western-blot对UA和HD样品中的免疫球蛋白(Ig G)、牛结合珠蛋白(Bo Hp)、补体C4和补体C8进行了验证,说明变形蹄奶牛血清蛋白表达谱发生了变化。To reveal the serum protein profile ~ft dairy cattle with hoof deformities （HD） and explore the pathogenesis of HD in dairy cattle, two pooled samples from unaffected dairy cattle （ UA,n =26） and those with HD （n =40） were used for the identification and biointbrmatics analysis using shotgun proteomic techniques. Furthermore, different proteins between UA and HD samples were analyzed, and the selected proteins were verified. The results showed that a total of 393 proteins were identified in the UA serum sample, of which 224 were high -confidence proteins; a total of 409 pruteins were identified in the HD serum sample, of which 225 were high -confidence proteins. The identified proteins showed obvious differences between the UA and HD samples in molecular function, cellular processes and cellular component classification. There were twenty -four potential differential proteins in the UA and HD samples, of which 18 proteins came from the HD sample, and 6 proteins came from the UA sample. Desmoplakin, junction plakoglobin, and annexin A2 had potential relevance with HD occurrence in dairy cattle among potential differential proteins in the HD sample. The immunoglobulin （IgG） , bovine haptoglobin （BoHp） , complement C4 and C8 complement in the UA and HD samples were validated by enzyme linked immunosorbent assay （ELISA） or western blot. The results indicate that there is a change in the serum protein profiles of dairy cattle with hoof deformities.

关 键 词：奶牛 变形蹄 血清 蛋白质组学 差异蛋白 

分 类 号：S823[农业科学—畜牧学] Q816[农业科学—畜牧兽医]