水稻胚乳双向电泳技术体系的优化分析

为比较同一杂交组合不同直链淀粉含量后代在灌浆期胚乳内蛋白表达差异,文章建立了一套适合灌浆期水稻胚乳蛋白的双向电泳技术,探讨了胚乳蛋白的提取、等电聚焦上样量的选择、胶条转移等双向电泳的一些关键技术。结果表明,一次溶解蛋白样品,TCA-丙酮法优于酚法蛋白提取,300μg上样量为最适上样量,能得到清晰、分离效果好、蛋白点数较多的图像。

Optimization of two-dimensional gel electrophoresis system of rice endosperm

In order to compare the difference of endosperm protein expression in grain filling period of hybrid progenies differed in amylase contents derived from the same hybrid compose,One serial of two-dimensional gel electrophoresis which was suitable to endosperm proteins of rice seed in grain filling period was set up.It was discussed on the key technologies of two-dimensional gel electrophoresis,such as the extraction,the sample loaded quantity in IEF,the transference of IPG strips and so on.The results showed that TCA-Acetone extraction was better than phenol method when the protein sample was dissolved only once,and can get the most clear and well separated two-dimensional gel electrophoresis diagram with the most protein spots,when the sample loaded quantity was 300 μg.