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目的:为了获得清晰的蛋白凝胶图谱,建立适用于大头典竹蛋白组学双向电泳的优化体系。方法:对大头典竹蛋白提取方法、胶条的水化运行方式、等电聚焦(IEF)程序的设置、蛋白上样量等步骤进行优化。结果:大头典竹的蛋白斑点主要分布在pH5~8的范围;三氯乙酸-丙酮法与酚抽结合法更适合大头典竹蛋白的提取;IPG胶条胶面向上、被动水化方式、蛋白上样量为50μL,效果更好。结论:通过建立双向电泳(2-DE)的优化体系,获得的蛋白点均匀、清晰、横纵条纹少的凝胶图谱,提高了分辨率,为大头典竹差异蛋白组学研究奠定了基础。
OBJECTIVE: To establish a clear gel map and establish an optimized system for two-dimensional electrophoresis of Proteus tuberosum. Methods: The extraction methods of Radix et Rhizoma Coptidis, the hydration mode of tape, the setting of IEF program and the sample loading of protein were optimized. Results: The protein spots of Radix Codonopsis were mainly distributed in the range of pH5 ~ 8. The combination of trichloroacetic acid-acetone and phenol was more suitable for the extraction of Radix Cynanchi paniculatum. IPG gel surface was upward and passive hydration, The amount of sample is 50μL, the effect is better. Conclusion: By establishing a two-dimensional electrophoresis (2-DE) optimization system, the obtained gel spots with uniform, clear and horizontal stripes can improve the resolution, which lays the foundation for the differential proteomics research of Radix Cynanchi.