文献类型：期刊文献

中文题名：当归种子蛋白质提取方法及SDS-PAGE技术体系研究

英文题名：Study on the Protein Extraction Method in Angelicae sinensis Seed and Its SDS-PAGE Technical System

作者：张延红[1];高素芳[1];王燕[1];李金田[1];杜弢[1]

第一作者：张延红

机构：[1]甘肃中医药大学药学院,兰州730000

第一机构：甘肃中医药大学药学院（西北中藏药协同创新中心办公室）

年份：2017

卷号：28

期号：13

起止页码：1751

中文期刊名：中国药房

外文期刊名：China Pharmacy

收录：CSTPCD;;北大核心:【北大核心2014】；

基金：国家中医药管理局"国家基本药物所需中药材种子种苗繁育基地建设"基金资助项目(No.国中医药办规财发[2013]41号)

语种：中文

中文关键词：当归;种子;蛋白质提取;SDS-PAGE

外文关键词：Angelicae sinensis; Seed; Protein extraction; SDS-PAGE

摘要：目的:建立适用于当归种子蛋白质提取及十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)的技术体系,为当归种子蛋白质研究和品种纯度检测提供技术支撑。方法:以蛋白质含量和电泳条带数量等为指标,采用浓缩胶法、盐溶蛋白法、电极缓冲液法、二巯基苏糖醇(DTT)法、尿素法、巯基乙醇法、三羟甲基氨基甲烷(Tris)法、丙酮沉淀法等8种方法提取当归种子蛋白质,筛选最优提取方法;再以最优提取方法为基础,考察不同料液比、样品稀释倍数(上样量)和分离胶浓度对当归种子蛋白质提取及SDSPAGE效果的影响。结果:8种提取方法中以巯基乙醇法提取蛋白质的含量较高(29.931 mg/g),且电泳条带数目多、泳道背景清晰;当以巯基乙醇法为最优提取方法时,料液比为1∶10、上样量为5倍、分离胶浓度为15%条件下电泳效果最好,共得到18条条带。结论:建立的蛋白质提取方法和SDS-PAGE技术体系适用于当归种子纯度的检测。
OBJECTIVE：To establish the technical system that is suitable for protein extracting in Angelicae sinensis seed and sodium dodecyl sulfate-polyacrylamide gel electrophoresis （SDS-PAGE）,and provide technical support for detecting the protein quality and variety purity. METHODS：Using protein content and number of electrophoretic bands as indexes,8 methods,includ-ing voncentrated gel method,salt-soluble protein method,electrode buffer method,dimercaptosyl alcohol （DTT） method,urea method,mercaptoethanol method,trimethylolaminomethane （Tris） method,and acetone precipitation method,were conducted to extract the protein in A. sinensis seed and screen the optimal extraction method. Then based on optimal extraction method,effects of different materials-lipid ratios,sample dilution times（sample volume）and separate gel concentration on SDS-PAGE were investi-gated. RESULTS：Mercaptoethanol method extracted the highest protein contents（29.931 mg/g）,with many electrophoretic bands and clear background. When mercaptoethanol method was used as optimal extraction method,electrophoresis effects were the best in the conditions of materials-lipid ratio of 1：10,sample volume of 5 times,separate gel concentration of 15%,which obtained 18 bands totally. CONCLUSIONS：Established protein extraction method and SDS-PAGE technical system are suitable for detecting the purity of A. sinensis seed.