新疆农业科学2017，Vol.54Issue(7)：1305-1312,8.DOI:10.6048/j.issn.1001-4330.2017.07.017
独行菜属植物种子总蛋白4种提取方法的比较
Comparative Study of Four Methods of Total Protein Extraction from Seeds of Two Species of Lepidium Plants
摘要
Abstract
[Objective] To explore the extraction methods of protein samples for SDS-PAGE and two-dimensional electrophoresis (2-DE) of the stem, vegetable, and vegetable seeds, and lay the foundation for the study of seed protein proteomics of the two species of Lepidium plants.[Method]The efficiency of four protein extraction methods were compared and analyzed through protein purity detection and SDS-PAGE.[Result]Of all the four methods, TCA/acetone extraction method generated higher yield as well as highest resolution and clear estelectrophoresis bands in SDS-PAGE, lysis extraction method got the highest protein yield whereas with highest impurities, Tris-HCl method and phenol method could obtain the highest purity but lowest protein yield, meanwhile protein extracted by lysis method and phenol method had the highest level of high-abundance proteins which could cause serious interference in further SDS-PAGE and 2-DE analysis.We got 6 differential expressed bands after compared between two species of Lepidium plants by SDS-PAGE using seed total protein extracted by TCA/acetone extraction method and phenol method.[Conclusion]TCA/acetone seed total protein extraction method is an optimal sample preparation protocol for SDS-PAGE and 2-DE.Besides, the differential bands of SDS-PAGE electrophoresis of two kinds of Lepidium seed protein could provide reference for screening two seeds.关键词
独行菜/抱茎独行菜/种子总蛋白/蛋白质提取/SDS-PAGEKey words
short ultivars/growth stages/major gene plus polygene inheritance/genetic analysis/SDS-PAGE分类
农业科技引用本文复制引用
丁金鹏,张娜,李群..独行菜属植物种子总蛋白4种提取方法的比较[J].新疆农业科学,2017,54(7):1305-1312,8.基金项目
国家自然科学基因项目"新疆濒危植物高山离子芥抗冷相关功能基因组分析与关键基因克隆"(31160051) （31160051）
新疆维吾尔自治区重大科技专项"新疆早春短命植物与模式植物拟南芥的比较基因组学与功能基因组学研究"(200840102-40)National Natural Science Foundation"Functional genome analysis of cold resistant and key gene clone in Xinjiang endangered plant Chorispora bungeana"(31160051) （200840102-40）
The Key Scientific Projects for Supporting Xinjiang Uygur Autonomous Region "Comparative genomics and functional genomics of early spring ephemeral plants and model plants of Arabidopsis thaliana in Xinjiang" (200840102-40). （200840102-40）
