Song Juan, Lei Xiu-juan, Yin Hong-xin, Yang Yu, Wang Ying-ping
Zhong Yao Cai. 2014 Sep;37(9):1540-5.
To establish the two-dimensional electrophoresis (2-DE)maps of callus induced from ginseng root,and to provide scientific method and data support for the study of secondary metabolites biosynthesis pathways of ginseng.
Total protein was extracted from ginseng callus by three different methods. The best protein extraction method of ginseng callus tissue was determined by the protein yield, purity and SDS-PAGE protein bands. By using 2-DE technology, the 2-DE maps of ginseng callus was established. The mass spectrometry and function on part of the protein were analyzed using MALDI-TOF-TOF and Swiss-Prot software.
Modified phenol extraction method (the third method)was the best protein extraction method of ginseng callus tissue, and it could obtain a high resolution 2-DE map. The second method was lesser. The protein quality of the first method could not be used for the analysis of 2-DE. 12 protein spots of the second and third methods were successfully identified by mass spectrometry.
This study established extraction methods suitable for ginseng callus protein and 2-DE patterns.
建立人参根愈伤组织的双向电泳（2-DE）图谱，为研究人参次生代谢产物生物合成途径提供科学方法和数据支持。
采用三种不同方法从人参愈伤组织中提取总蛋白。通过蛋白产量、纯度和SDS-PAGE蛋白条带确定人参愈伤组织最佳蛋白提取方法。利用2-DE技术建立人参愈伤组织的2-DE图谱。使用MALDI-TOF-TOF和Swiss-Prot软件对部分蛋白质进行质谱分析和功能分析。
改良酚提取法（第三种方法）是人参愈伤组织最佳蛋白提取方法，能获得高分辨率的2-DE图谱。第二种方法次之。第一种方法的蛋白质量不能用于2-DE分析。通过质谱成功鉴定出第二种和第三种方法的12个蛋白点。
本研究建立了适合人参愈伤组织蛋白的提取方法和2-DE图谱。
