金钗石斛花芽蛋白质提取及其双向电泳体系优化

Optimization of Protein Extraction and Two-Dimensional Electrophoresis of the Dendrobium nobile. Flower Bud

摘要: 通过对蛋白质提取方法、水化方式、等点聚焦程序、IPG胶条选择等方面的优化，建立金钗石斛花芽蛋白质双向电泳体系.结果表明，采用酚抽提法提取蛋白，样品与水化液1∶〖KG-*2/3〗40复溶；被动水化，等点聚焦程序B2；选用24 cm pH 3～10NL 胶条和12% 的凝胶；采用考马斯亮蓝G-250染色，获得了可识别1 422个蛋白质斑点的双向电泳图谱，且重复性好、分辨率高，为进一步开展金钗石斛花芽在蛋白质组学水平上的分析提供了技术支持.

Abstract: To establish the two-dimensional gel electrophoresis system of Dendrobium nobile flower bud proteins and gain a higher reproducibility and resolution, the proteins of D. nobile flower bud were extracted by optimized phenol method. The sample was passively rehydrated by dissolving in 40-fold (w/v) rehydration solution. Isoelectric focusing was performed in Program B2, using 24 cm pH 3-10NL gradient IPG strip and 12% SDS-PAGE. The gel was stained with Coomassie brilliant blue G-250. By this modified method, the 2-D electrophoretic maps of the D. nobile flower bud proteins showed 1 422 protein spots, with high repeatability and high resolution. This work provides a good method for proteomic analysis of D. nobile flower buds.