中国药房2017，Vol.28Issue(13)：1751-1754,4.DOI:10.6039/j.issn.1001-0408.2017.13.07
当归种子蛋白质提取方法及SDS-PAGE技术体系研究
Study on the Protein Extraction Method in Angelicae sinensis Seed and Its SDS-PAGE Technical System
摘要
Abstract
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.关键词
当归/种子/蛋白质提取/SDS-PAGEKey words
Angelicae sinensis/Seed/Protein extraction/SDS-PAGE分类
医药卫生引用本文复制引用
张延红,高素芳,王燕,李金田,杜弢..当归种子蛋白质提取方法及SDS-PAGE技术体系研究[J].中国药房,2017,28(13):1751-1754,4.基金项目
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) （No.国中医药办规财发〔2013〕41号）
