奶牛初乳SIgA的提纯与电泳分析  

The Purification and Electrophoretic Analysis of Siga in Bovine Colostrum

作　　者：梁靖坤[1] 王继芳[1] 曹荣峰[1] 

机构地区：[1]青岛农业大学动物科技学院,山东青岛266109

出　　处：《中国农学通报》2009年第15期6-8,共3页Chinese Agricultural Science Bulletin

摘　　要：为了建立提取奶牛初乳中SIgA的方法，取分娩当日的奶牛乳汁，经10000r／min离心10min，O．2mol／L（pH=6．0）PBS透析，上清液加0．06mol／L硫酸锌处理后，调pH为6．8-6．9，加入饱和硫酸铵，取沉淀溶于水，并在0．02mol／L（pH=8．0）PBS中透析至无NH4^＋，样品浓缩后上DEAEScpharosc（Fast Flow）柱，用O．02mol/L（pH=8．0）至O．3mol／L（pH=8．0）PBS梯度洗脱；收集峰蛋白上葡聚糖凝胶-200层析柱，用0．01mol／L（pH=7．4）PBS洗脱；收集出现蛋白峰的洗脱液，经SDS-PAGE电泳鉴定，为SIgA单体。经初步测定奶牛初乳中SIgA含量为3．47mg／ml。此试验已成功建立提取奶牛初乳中SIgA的方法。The experiment had established the extraction method of SIgA in bovine colostrum. The milk of accouchement within one day was centrifuged 10 min by 10 000 r/min. After dialyzed by 0.2 mol/L（pH=6.0） PBS, accommodated the supernatant fluid pH to 6.8-6.9 after disposal of 0.06 mol/L Zn SO4. Then added saturated ammonium sulfate to produce sedimentum, sedimentum dissolved in water. The supernatant fluid were dialyzed by 0.02 mol/L （pH=8.0）PBS to no NH4^＋. Specimen was concentrated, then it was purified by DEAE Sepharose FF chromato bar, gradient elutioned by 02 mol/L（pH=8.0）to reach 0.3 mol/L（pH=8.0）PBS. Hump proteinum was collected, and then it was purified by Sephadex G-200. The elutriant was 0.01 mol/L （pH=7.4） PBS. The elutriant appearance hump proteinum was collected, and then it was appraised through electrophoresis of SDS-PAGE. The results of the electrophoresis showed that SIgA was metamer. The results of rough determination showed that the content of SIgA was 3.47 mg/ml in bovine Colostrum. This experiment had established the method for extract SIgA in bovine colostrum.

关 键 词：奶牛 初乳 SIGA 提纯 电泳分析 

分 类 号：S852.43[农业科学—基础兽医学]