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【目的】探索适用于阳春砂种子蛋白质的提取方法,为研究阳春砂种子休眠与萌发过程中的差异蛋白质组学奠定基础。【方法】分别采用4种较常用的蛋白质提取方法 [Tris-HCl提取法、三氯乙酸(TCA)-丙酮沉淀法、直接裂解法以及改良Tris-饱和酚法]提取,对得到的蛋白质提取液进行定量及十二烷基磺酸钠—聚丙烯酰胺凝胶(SDS-PAGE)电泳分析。【结果】直接裂解法蛋白质提取率最大,小分子蛋白较多;Tris-HCl法提取率最小,且条带扩散;TCA-丙酮沉淀法和改良Tris-饱和酚法提取率居中,但是TCA-丙酮沉淀法条带少,饱和酚法条带多,且可以得到大分子蛋白。【结论】可以综合直接裂解法和改良Tris-饱和酚法,为阳春砂种子双向电泳的蛋白质提取方法提供参考。
【Objective】 The objective of this study was to explore the extraction methods of seeds suitable for the seeds of Populus euphratica and lay the foundation for the study of differential proteomics in the process of dormancy and germination of Poplar sand. 【Method】 Four kinds of protein extraction methods (Tris-HCl extraction, TCA-acetone precipitation, direct pyrolysis and modified Tris-saturated phenol method) were used to extract the protein extracts Quantitative and sodium dodecyl sulfate - polyacrylamide gel (SDS-PAGE) electrophoresis analysis. 【Result】 The results showed that protein extracted by direct lysis method had the highest protein extraction rate and the small protein was the most. The extraction rate of Tris-HCl was the smallest and the band was diffused. The TCA-acetone precipitation and modified Tris- Precipitation method with less band, saturated phenol method and more, and can get macromolecular protein. 【Conclusion】 This study provides a reference for the extraction of protein from the two-dimensional gel electrophoresis of Populus euphratica by combining the direct lysis method and the modified Tris-saturated phenol method.