小麦籽粒蛋白质双向电泳体系的优化

Optimization of two-dimensional electrophresis system for grain protein in spring wheat

为建立一套适合于小麦籽粒蛋白质双向电泳的技术体系,以青海省农林科学院自育春小麦品种青春38籽粒为材料,对比了2种在植物组织中常用的蛋白质提取方法,对双向电泳过程中胶条p H值、蛋白质上样量和分离胶浓度等方面进行了优化。结果表明:与用酚提取法相比采用TCA/丙酮沉淀法提取的蛋白质所得的双向电泳图谱质量较好,蛋白质经裂解后在17 cm、p H 5~8的IPG胶条、上样量380μg和分离胶浓度12%的条件下得到了清晰、蛋白质点丰富的双向电泳图谱。

To develop a system for two-dimensional electrophresis(2-DE) of grain protein extracted from spring wheat, two protein extraction methods commonly used for plant tissues were compared. The pH range, the loading quantity of sample and the gel concentrations were optimized in the 2-DE system for the grains from Qingchun 38. The samples prepared by TCA/acetone precipitation yielded better quality 2-DE map than by phenol extraction method. The condition of 17-cm IPG with pH 5-8, 380 μg loading quantity of protein and 12% SDS-PAGE gel gave rich protein spots in the 2-DE map.