精粗分饲与全混合日粮饲喂方式影响牛乳蛋白组的初步研究

Changes of the Milk Protein in Dairy Cattle Feed with Total Mixed Ration and Traditional Separate Ingredients Feeding System

为了分析传统精粗分饲和全混合日粮饲喂方式影响乳蛋白的表达变化。采用二维凝胶电泳技术分离了精粗分饲和全混合日粮饲喂方式的牛乳蛋白,考马斯亮蓝G-250染色,ImageMaster 2D platinum 6.0软件分析,液相色谱串联质谱的方法鉴定凝胶中蛋白点。结果发现,以蛋白表达量变化2倍或2倍以上为判定标准时,传统精粗分饲和全混合日粮饲喂方式未能检测到乳蛋白的表达变化。乳蛋白凝胶图谱中主要蛋白点经液相色谱串联质谱鉴定为αs1-酪蛋白、αs2-酪蛋白、β-酪蛋白和κ-酪蛋白等酪蛋白及α-乳白蛋白和β-乳球蛋白等乳清蛋白,均为牛奶中的主要蛋白组分。表明饲喂方式对牛奶中主要乳蛋白成分的表达影响较小,牛奶主要乳蛋白成分存在的条件下二维凝胶电泳结合质谱的方法未能检测乳蛋白的变化。

To investigate the changes of the milk protein in dairy cattle feed with total mixed ration and traditional separate ingredients feeding system.Milk proteins were separated by two-dimensional electrophoresis,stained with Coomassie Blue G-250,detected by ImageMaster 2D platinum 6.0 software and identified by HPLC coupled with tandem ion trap spectrometry.The results showed that,changes in the stain density and intensity of twofold or more were detected to be differentially expressed spots,There were not differential spots presented in milk proteins gels of two feeding system.Spots of protein on the gels were identified as major protein in bovine milk,such as caseins and whey proteins including αs1-casein,αs2-casein,β-casein and κ-casein,as well as α-lactoalbumin and β-lactoglobin.Major milk proteins of dairy cattle feed with total mixed ration and traditional separate ingredients were not presented changes by two-dimensional electrophoresis combination mass spectrometry approach.