蕨菜总蛋白质双向电泳技术体系的建立

Research on Two-Dimensional Electrophoresis Conditions for Proteomic Analysis of Pteridium aquilinum

摘要:

为了建立蕨菜中总蛋白质的双向电泳技术体系,作者以新鲜蕨菜为实验材料,分别比较了不同蛋白提取方法,不同上样量以及不同pH范围IPG胶条对2-DE图谱的影响。结果表明,采用TCA-丙酮结合酚抽法提取蕨菜中的蛋白质,采用IPG胶条pH5-8、蛋白质的上样量为1 200μg、考马斯亮蓝G-250胶体考染法进行染色。在此条件下进行蕨菜全蛋白质的双向电泳所得到的图谱蛋白质点多达719个,且蛋白质点分布均匀、背景清晰、分辨率较高。

Abstract:

In order to establish two-dimensional electrophoresis technology system of total proteins of Pteridium aquilinum,2-DE maps was compared with protein extraction method,loading amount and pH ranges of IPG strips The results showed that phenol extraction protocol combined with TCA/ acetone was more favorable for total protein extraction from fresh Pteridium aquilinum.A high quality 2-DE map with more protein spots,better dispersion,clearer background and higher protein point resolution were acquired by using the following optimized parameters:loading 1200μg protein sample on 17cm IPG strip with pH 5-8,and finally detecting proteins with Coomassie brilliant blue G250 staining.Under above experimental conditions,the total number of protein spots is 719,this experimental results provides a basis for further study of post-harvested Pteridium aquilinum proteomics.