摘要
大肠杆菌是奶牛乳房炎的主要病原微生物之一,为了分析大肠杆菌性乳房炎奶牛血浆蛋白的表达变化,探索其防御机制。通过乳汁微生物培养的方法选择仅由大肠杆菌自然感染的隐性乳房炎奶牛。采用二维凝胶电泳技术分离大肠杆菌性乳房炎奶牛和临床健康奶牛的血浆蛋白和ProteoMiner试剂盒富集的中低丰度蛋白,凝胶用考马斯亮蓝G-250染色后,用图像分析软件检测血浆表达的差异蛋白点并用液质联用质谱鉴定。结果发现,大肠杆菌性乳房炎奶牛血浆中有19个蛋白点的表达量发生改变,其中15个蛋白点鉴定为7种蛋白质。在大肠杆菌性乳房炎奶牛血浆中结合珠蛋白、α1酸性糖蛋白和血清淀粉样蛋白A等蛋白的表达量增加。并用ELISA法测定血浆结合珠蛋白的结果也发现,大肠杆菌性乳房炎奶牛血浆结合珠蛋白水平显著高于健康牛(P<0.01)。表明大肠杆菌感染造成了奶牛血浆蛋白的表达发生了变化,对这些表达变化蛋白的解析有利于探索机体的防御机制,同时为疾病的诊断和治疗提供科学依据。
Escherichia coli are most common pathogen from a contaminated environment in lactating dairy cows.For investigating the changes of plasma protein form E.coli infected cows and exploring the mechanism of host defense,such changes were examined using an integrated proteomic approaches consisting of plasma protein and minor abundance protein enrichment by ProteoMiner kit,protein separation by two-dimensional gel electrophoresis,stained with Coomassie Blue G-250solution,and protein identification by HPLC equipped with ion trap mass spectrometer.The results showed that the nineteen protein spots were changed in cows infected with E. coli mastitis,and fifteen spots were identified as seven proteins.Haptoglobin,alpha 1acid glycoprotein,and serum amyloid protein A were up-regulated in plasma from E.coli infected cows. Expression of haptoglobin by ELISA method was significantly higher in E.coli infected cows than in healthy controls（P〈0.01）,E.coli infected cows altered the plasma protein level,and suggested that changes of proteins may be useful to clarify the host defense mechanisms and response to invasion of mammary tissues by E.coliinfection,and potential protein targets for diagnose and treatment.
出处
《畜牧兽医学报》
CAS
CSCD
北大核心
2010年第9期1191-1197,共7页
ACTA VETERINARIA ET ZOOTECHNICA SINICA
基金
安徽省自然科学基金(090411024)
安徽省博士后基金
关键词
奶牛
隐性乳房炎
大肠杆菌
血浆
蛋白质组
dairy cows
subclinical mastitis
Escherichia coli
plasma
proteome
