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目的:建立人参根愈伤组织双向电泳图谱,为人参蛋白质组学分析及次生代谢产物合成途径研究提供方法参考和数据支撑。方法:采用3种不同方法提取人参根愈伤组织总蛋白;利用蛋白产率、纯度、SDS-PAGE蛋白条带确定其最佳提取方法;利用双向电泳技术建立人参根愈伤组织双向电泳图谱;利用MALDI-TOF-TOF和SwissProt软件对部分蛋白点进行质谱鉴定和功能分析。结果:改良酚抽提法(方法三)是最佳人参根愈伤组织蛋白提取方法,可获得高分辨率双向电泳图谱,方法二次之,方法一获得的蛋白不能用于双向电泳分析;经质谱成功鉴定了方法二和方法三12个蛋白点。结论:本实验建立了适合人参根愈伤组织蛋白质提取的方法和双向电泳图谱。
OBJECTIVE: To establish a two-dimensional gel electrophoresis map of root callus of ginseng and provide reference and data support for the ginseng proteomics analysis and secondary metabolite synthesis pathway. Methods: Three kinds of different methods were used to extract the total callus of callus of Ginseng root. The best extraction method was determined by protein yield, purity and SDS-PAGE protein band. Two-dimensional gel electrophoresis of ginseng root callus was established by two- Some protein spots were identified by MALDI-TOF-TOF and SwissProt software for mass spectrometry and functional analysis. Results: The modified phenol extraction method (Method 3) was the best method for extracting callus from Ginseng root, which could obtain high-resolution two-dimensional gel electrophoresis. The method was twice, and the protein obtained by method one could not be used for two-dimensional electrophoresis analysis. Mass spectrometry successfully identified method two and method three twelve protein spots. Conclusion: This experiment established a method suitable for the extraction of protein from root callus of Ginseng and two-dimensional electrophoresis.