薄皮甜瓜果实总蛋白质双向电泳优化体系的建立

为建立适宜的薄皮甜瓜果实总蛋白质双向电泳的试验体系，以‘京蜜11号’薄皮甜瓜为试材，从果实总蛋白质的提取方法、等电聚焦条件、IPG胶条梯度等方面进行了探索优化。结果表明，酚-甲醇/醋酸铵沉淀法能够高效的提取薄皮甜瓜果实总蛋白质，选用24 cm pH 4~7的IPG胶条，800 μg上样量，120000 Vhs聚焦，SDS-PAGE电泳后采用胶体考马斯亮蓝染色。该优化的体系可以获得背景清晰、分辨率高、重复性好的双向电泳图谱。

The Establishment of Optimizational Two-dimensional Electrophoresis System for Melon Fruit

To establish an appropriate two- dimensional electrophoresis system of melon fruit, the variety of Cucumis melo‘Jingmi 11’was used as material, parameters including protein sample preparation, isoelectric focusing condition and the pH gradient of IPG strip were optimized. The result showed that the optimized system included the following steps: extracting the total protein from melon fruit by phenol- methanol/ ammonium acetate, separating the proteins with 24 cm pH 4-7 IPG strips, loading protein samples of 800μg, 120000 Vhs isoelectric focusing and staining the gels by colloidal coomassie brilliant blue staining after SDSPAGE electrophoresis. Clear background, reproducible profiles with high resolution were obtained by this optimized 2-DE system of melon fruit.