本研究运用双向电泳方法分析奶牛产前及产后1周血浆蛋白的差异表达,为理解围产期免疫抑制以及分娩前后奶牛的生理状态提供参考。血浆样品经过热SDS法处理后,使用pH4~pH7的胶条,在硝酸银染色的7.5%~17.5%的梯度聚丙烯酰胺凝胶上,利用PDQuest6.0图像分析软件检测到239个蛋白质点。比较奶牛产前及产后1周血浆蛋白差异表达情况发现,表达量变化3倍以上的点有12个。MADIL-TOF/TOF质谱鉴定出2种蛋白:白蛋白和结合珠蛋白。结合珠蛋白在产后1周的奶牛血浆中的含量显著增加,反映了产后奶牛的生理变化。但是需要结合其他标记共同作为疾病诊断的标记。
In this study two-dimensional electrophoresis was applied to analyze the difference expression of plasma protein between pre-perinatal and post-perinatal cows.The Hot-SDS method, pH 4-pH7IPG strips(17cm)and gradient gel were used for dairy cow plasma proteome analysis. Totally,239protein spots were detected using silver staining by PDQuest 6.0,which appeared repeatedly in three gels of each sample.12spots showed more than 3fold changes in abundance in plasma protein of cow.Plasma albumin and Haptoglobin were identified by MADIL-TOF/TOF. Their expressions were increased in the plasma of post-perinatal cow.Haptoglobin content ircreased in post-perinatal cow plasma in this study,which showed the physiological changes of the post-partum cows.