摘要： 为了获得甜菜叶片双向电泳中蛋白质最佳提取方法，建立甜菜叶片蛋白质双向电泳最佳的双向电泳体系。以甜菜叶片为材料，通过比较分析，研究甜菜叶片总蛋白提取方法、IPG胶条pH值及其对应上样量等因素对甜菜叶片蛋白质双向电泳结果的影响。结果表明：酚提取法、TCA/丙酮提取法和可溶性蛋白提取法3种蛋白提取方法中TCA/丙酮法提取效果较好，pH值4~7与pH值5~8的双向蛋白电泳比较发现：pH值4~7胶条蛋白点清晰且在图谱上的蛋白点分布均匀，无蛋白点集中现象，更适合甜菜叶片的蛋白质组分析。采用7 cm线性固相IPG胶条进行双向电泳，150 μg上样量蛋白点明显增多，双向电泳效果更好，pH值4~7的17 cm胶条300 μg上样量胶点更多更清晰，试验结果为甜菜蛋白质组学的研究提供了最佳的试验方法。

关键词: 甜菜, 蛋白质提取方法, 双向电泳, 体系优化

Abstract: In order to obtain the best extraction method of protein in two-dimensional electrophoresis of beet leaves and establish the best two-dimensional electrophoresis system. It took beet leaves as material. To study the effects of the extraction methods of total protein,the pH value of IPG strip,and the corresponding amount of sample on the results of two-dimensional electrophoresis of sugar beet leaves through comparative analysis.The results showed that TCA/acetone extraction method was better than the phenol extraction method and soluble protein extraction method. Comparing IPG strip of pH 4-7 with pH 5-8 in two-dimensional electrophoresis:Protein spots with pH 4-7 were clear and distributed uniform in the spectra,no protein concentrated.It was more suitable for proteomics analysis of sugar beet leaves.Using the 7 cm linear phase IPG strip,protein spots with 150 μg sample volume increased obviously and was better. Using the 17 cm strip of pH 4-7 with 300 μg sample volume protein spots were more clear.The study provided the theoretical basis and reference for the study of sugar beet proteomics.

Key words: Sugar beet, Protein extraction method, Two-dimensional electrophoresis, System optimization

中图分类号: 

• S566.3