‘四季蜜’龙眼花芽总蛋白质提取方法及双向电泳体系的优化  

Method of Total Protein Isolation from Floral Buds of Longan (Dimocarpus longan Lour. Cv Sijimi) and Optimization of Two-dimensional Electrophoresis System

作　　者：欧高政[1,2] 陈清西[1] 

机构地区：[1]福建农林大学园艺学院,福建福州350002 [2]福建农业职业技术学院,福建福州350119

出　　处：《热带作物学报》2014年第6期1095-1101,共7页Chinese Journal of Tropical Crops

摘　　要：利用TCA丙酮法、丙酮法、TCA酚抽提法和改良酚抽法等4种方法，提取‘四季蜜’龙眼花芽的总蛋白质，在蛋白产量、单mSDS—PAGE和2-DE图谱等方面进行比较，并对IPG胶条种类、蛋白质上样量及等电聚焦条件等进行优化。结果表明：采用改良酚抽法，选用24om、pH3～10的胶条，按120μg上样，在适宜的等电聚焦条件下．可获得背景清晰、重复性好、分辨率高的2-DE图谱．本研究为‘四季蜜’龙眼花芽蛋白质组学后续研究奠定了基础。The total protein in the floral buds of Dimocarpus longan Lour. Cv Sijimi was extracted with four methods, namely trichloroacetic acid （TCA） -acetone, acetone, TCA/phenol and improved phenol, and compared by protein production, one-way SDS-PAGE and 2-DE. The sample loading quantity, the gel concentration and the key process of IEF were also optimized. The result showed that the improved phenol extraction protocol was the best method for floral buds. And a high quality 2-DE map with low background was obtained using the following optimized procedure： loading 120 μg protein sample on 24 cm IPG strip with pH3-10. The study built up a good basis for the future proteomics research in D. longan （Lour. Cv Sijimi）.

关 键 词：'四季蜜’龙眼 花芽 总蛋白 提取方法 双向电泳 

分 类 号：S667.201[农业科学—果树学]