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两相法分离地被菊叶片质膜的研究
引用本文:沈漫.两相法分离地被菊叶片质膜的研究[J].植物学报,2004,21(1):66-73.
作者姓名:沈漫
作者单位:(北京农学院植物科学技术系北京102206)
基金项目:北京市科技新星计划项目
摘    要:以地被菊(Dendranthema×grandiflorum Kitamura)叶片为材料,通过水溶性聚合物Dextran T-500和PEG 3350所构成的两相分配体系制备质膜。在一定盐浓度(5 mmol.L-1 NaCl)下选用5种不同的聚合物浓度(5.8%、6.0%、6.2%、6.4%、6.6%,W/W),研究了地被菊叶片质膜在两相体系中的分配情况,在此基础上进一步研究了不同盐浓度(2、4、5、10、20 mmol.L-1 NaCl)对地被菊叶片质膜的纯度及蛋白产率的影响。标志酶鉴定及磷钨酸染色电镜检测的结果表明,地被菊叶片选用6.2%(W/W)聚合物浓度和7 mmol.L-1 NaCl组成的两相分配体系可获得较高纯度的密实正向型质膜囊泡。

关 键 词:两相分配法  质膜  地被菊  标志酶  磷钨酸染色
收稿时间:2002-12-11
修稿时间:2003-05-19

Preparation of Plasma Membrane from Leaves of Grand-cover Chrysanthemum by Aqueous Polymer Two-phase Partition
SHEN Man.Preparation of Plasma Membrane from Leaves of Grand-cover Chrysanthemum by Aqueous Polymer Two-phase Partition[J].Bulletin of Botany,2004,21(1):66-73.
Authors:SHEN Man
Institution:( Department of Plant Science & Technology, Beijing Agriculture College, Beijing 102206 )
Abstract:Plasma membranes were prepared from grand-cover chrysanthmum (Dendranthema×grandiflorum Kitamura) leaves by aqueous polymer two-phase partitioning system which was composed of Dextran T-500 and PEG 3350. The effect of the concentration of polymer and salt on partition of plasma membrane in aqueous polymer system was studied. The results from the electron micrograph staining with phosphotungstic acid and the mark enzyme activity analyses showed that the preparation of the right-side-out plasma membrane vesicles from grand-cover chrysanthmum leaves had obtained high purity and yield by the system which consisted of 6.2%(W/W) Dextran T-500 and PEG 3350, 7 mmol.L-1 NaCl.
Keywords:,
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