蒙古沙冬青根蛋白的提取及双向电泳体系的建立  

EXTRACTION AND TWO-DIMENSIONAL ELECTROPHORESIS OF PROTEINS FROM AMMOPIPTANTHUS MONGOLICUS ROOTS

作　　者：刘楠[1] 高飞[1] 周宜君[1] 张孜宸[1] 隋欣[1] 马冰[1] 张萌[1] 张根发[2] 

机构地区：[1]中央民族大学生命与环境科学学院,北京100081 [2]北京师范大学生命科学学院,北京100875

出　　处：《北京师范大学学报（自然科学版）》2013年第4期365-368,共4页Journal of Beijing Normal University(Natural Science)

基　　金：国家自然科学基金资助项目(31070361;31070289);中央高校基本科研业务费专项基金资助项目(1112KYQN31);中央民族大学"985工程"资助项目(MUC98504-14;MUC98507-08);国家大学生创新性实验计划资助项目(NMOE20121623)

摘　　要：以蒙古沙冬青根为实验材料,采用TCA/丙酮沉淀法和Tris-饱和酚提取法分别提取根可溶性蛋白.用Bradford法测定蛋白质浓度,计算蛋白提取率,并对所提取的蛋白样品进行双向电泳检测及电泳体系优化.结果表明,选择Tris-饱和酚法提取蛋白,采用24cm、pH5～8的IPG胶条,上样量900μg,可获得分辨率高、重现性好的双向电泳凝胶图.A procedure of root protein extraction for proteomics analysis of Ammopiptanthus mongolicus seedlings was established.Total protein extraction by TCA/acetone precipitation or by phenol extraction was compared.Protein concentration was determined by the Bradford method,proteins were separated by two-dimensional electrophoresis（2-DE）.Highly reproducible and clear 2-DE gel images were obtained with modified phenol extraction,and the following 2-DE parameters：24 cm（pH 5~8） IPG strip,900 μg protein loading volume.

关 键 词：蒙古沙冬青 根 蛋白质提取 双向电泳 

分 类 号：TS275.4[轻工技术与工程—农产品加工及贮藏工程]