水稻籽粒蛋白双向电泳条件的优化及其蛋白组学方法的比较

探讨适用于籽粒蛋白组学研究的策略，对深入研究籽粒的发育过程具有重要的意义。本文对比了3种不同的蛋白提取方法，优化了双向电泳中自制管胶一向的等电聚焦条件和IPG干胶条一向的等电聚焦时间，以MALDI-TOF/MS、Western-blot和磷酸化蛋白组学3个重要的蛋白质组学研究方法对胶内蛋白质鉴定分析。结果表明，可溶性蛋白提取法最适用于籽粒蛋白组学的研究; 电泳条件优化后，得到了较优的2-DE图谱; 胶内蛋白点适用于质谱分析、蛋白表达量验证(Western-blot)及籽粒蛋白的磷酸化组学的研究。本研究为下一步在蛋白组水平上分析籽粒发育提供了技术支持。

Optimization of Two-Dimensional Electrophoresis Condition for Rice Grain Protein and Comparison of Relevant Proteomic Methods

A suitable proteomic strategy for rice grain protein research is important for further understanding rice grain development at proteome level.In this study,three different protein extraction methods were compared and then electrophoretic conditions of hand-made gel lands and IPG lands in IEF(isoelectric focusing)were optimized.Furthermore,three important proteomic research strategies(MALDI-TOF/MS,western-blot and phosphoproteome)were used for identification and analysis of in-gel proteins.The results showed that the method of soluble protein extraction was most suitable for investigation of grain proteomics,with a well-distributed 2-DE gel profile under optimized electrophoretic conditions,and it was confirmed that MALDI-TOF/MS,western-blot and phosphoproteome were the three suitable methods for the analysis of in-gel proteins.The establishment of the suitable system for rice grain proteomic research paves the way for the next step to further analyse rice grain development at proteome level.