4种黄皮叶片蛋白质提取方法的比较.PDFVIP

第35 卷 第1 期 经 济 林 研 究 Vol. 35 No.1 2017 年3 月 Nonwood Forest Research Mar. 2017 Doi:10.14067/ki.1003-8981.2017.01.011 http: // 4种黄皮叶片蛋白质提取方法的比较 庞新华，张 宇，黄国弟，黄 强 （广西壮族自治区亚热带作物研究所，广西 南宁 530001 ） 摘 要：为了研究不同提取方法对黄皮叶片蛋白质提取质量的影响，以‘杨妃山黄皮’的叶片为材料，采用 Tris- HCl 法、三氯乙酸（TCA ）/ 丙酮法、尿素（Thi ）/ 硫脲（Urea ）法、酚（Phe ）- 甲醇/ 醋酸铵沉淀法提取叶片 组织蛋白质，对获得的蛋白质产量、单向电泳图谱和双向电泳图谱分析比较。结果显示，不同方法的提取效果各异， 就提取的蛋白质产率、单向电泳以及双向电泳结果来说，三氯乙酸（TCA ）/ 丙酮法提取效果较理想，单双向电 泳图谱辨识度强，蛋白点数多、蛋白点整洁、且纵横纹少。说明该法不但可以较好地去除黄皮叶片中存在的非 蛋白物质，而且可以获取高质量的蛋白质。 关键词：黄皮；蛋白质；Tris-HCl 法 中图分类号：S603.7 文献标志码：A 文章编号：1003—8981(2017)01—0059—05 Comparison of protein extraction methods from leaves in Clausena lansium PANG Xinhua, ZHANG Yu, HUANG Guodi, HUANG Qiang (Guangxi Subtropical Crops Research Institute, Nanning 530001, Guangxi, China) Abstract: In order to research the effects of different extraction methods on extraction quality of protein from leaves in Clausena lansium, using leaves in ‘Yangfeishan wampee’ cultivar as materials, the proteins were extracted by the methods of Tris-HCl, TCA/acetone, Thi/Urea and Phe-Methanol/ammonium acetate precipitation, and protein production, single electrophoresis and two dimensional electrophoresis map were compared. The results showed that the separation effects of the four methods were different, the production, single electrophoresis and two dimensional electrophoresis results of the extracted proteins by TCA/acetone method were better than those by the others, and the protein spots were clear, many and uniform, and the stripes were few in the single electrophoresis map. So the method not only cou