滩羊羊毛蛋白质双向电泳图谱体系的建立及优化

Establishment and Optimization of the System of Bidirectional Electrophoresis of Sheep Wool Protein

为建立宁夏滩羊羊毛蛋白质双向电泳体系,比较分析了滩羊羊毛蛋白质不同上样量、不同pH值范围的IPG胶条以及聚焦参数对羊毛蛋白质双向电泳的影响。结果表明:使用TCA/丙酮沉淀法对羊毛蛋白进行处理,上样量600μg,选择pH值为4~7的IPG胶条,聚焦时间90 000 Vhr,12%SDS-PAGE凝胶进行双向电泳,经考马斯亮蓝染色,用Images-Scanner扫描,所得到的滩羊羊毛较清晰的2-DE图谱较好,为后续滩羊羊毛的差异蛋白质组学研究建立了条件。

This experiment was designed to establish the bidirectional electrophoresis system of the sheep wool protein in Ningxia.The influence of different sample sizes and pH range of IPG tape on the bidirectional electrophoresis of sheep wool protein were compared. The experimental results showed that the optimal process were TCA/acetone precipitation at 600 mg,the IPG bar of pH4-7,focus time 90 000 Vhr or higher,12% SDS-PAGE gel for bidirectional electrophoresis and an image Scanner scan. Under the conditions above,a better pattern of 2-DE of the sheep wool could be gotten,which could be useful for the study of the different proteomics of sheep wool.