[1]乞永艳,刘富海,Arnaud Bovy,等.二维电泳技术在大豆子叶微粒体膜结合蛋白研究中的应用[JP][J].大豆科学,2007,26(01):11-15.[doi:10.3969/j.issn.1000-9841.2007.01.004]
　QI Yong-yan,LIU Fu-hai,Arnaud Bovy,et al.TWO-DIMENSIONAL ELECTROPHORESIS OF MIC-ROSOMAL MEMBRANE BOUND PROTEINS IN SOYBEAN COTYLEDONS[J].Soybean Science,2007,26(01):11-15.[doi:10.3969/j.issn.1000-9841.2007.01.004]
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二维电泳技术在大豆子叶微粒体膜结合蛋白研究中的应用[JP]

《大豆科学》[ISSN:1000-9841/CN:23-1227/S] 卷: 第26卷 期数: 2007年01期 页码: 11-15 栏目: 出版日期: 2007-02-25
Title:
TWO-DIMENSIONAL ELECTROPHORESIS OF MIC-ROSOMAL MEMBRANE BOUND PROTEINS IN SOYBEAN COTYLEDONS
文章编号:
1000-9841(2007)01-0011-05
作者:
乞永艳1; 刘富海2; Arnaud Bovy3; Jan Cordewener3; Froukje Vander Wal3
1.中国农业科学院蜜蜂研究所，北京，100093；
2.北京天宝康高新技术开发有限公司，北京，100097；
3.Plant Research International， P.O. Box 16， 6700 AA， Wageningen， The Netherlands
Author(s):
QI Yong-yan1; LIU Fu-hai2; Arnaud Bovy3; Jan Cordewener3; Froukje Vander Wal3
1.Bee Research Institute of Chinese Academy of Agricultural Sciences， Beijing， 100093；
2. Tianbaokang High-tech Co. Ltd., Beijing, 100097;
3. Plant Research International， P.O. Box 16， 6700 AA， Wageningen， the Netherlands
关键词:
二维电泳; 大豆; 子叶; 微粒体膜结合蛋白
Keywords:
Two-dimensional electrophoresis; Glycine max (L.) Merri;  Cotyledons;  Microsomal membrane bound proteins
分类号:
S565.1
DOI:
10.3969/j.issn.1000-9841.2007.01.004
文献标志码:
A
摘要:
以硝酸银诱导的大豆子叶为材料，初步探讨了二维电泳技术在分离大豆子叶微粒体膜结合蛋白中的应用，研究了重新水化溶液的组成、加样量、等电聚焦的total voltage hours以及不同pH梯度范围IPG(固相pH梯度)胶条等因素对膜蛋白的溶解和二维电泳图像的影响。结果表明对于70×3×0.5mm IPG胶条，能够溶解较多蛋白质和获得高质量二维电泳图像的优化条件为：由尿素、硫脲、三丁基膦和C7BZO组成的重新水化溶液（TUC-7T-2）、80μg的加样量和20000vhr的total voltage hour。在此优化条件下，与宽范围pH梯度IPG胶条相比，使用窄范围pH梯度胶条可以观察到更多的和质量更好的蛋白质点，结果发现至少有13个点只存在于硝酸银诱导的样品中，而不存在于对照中，其代表的蛋白质的等电点在4.0~8.0。
Abstract:
Soybean cotyledons induced by silver nitrate were used to investigate the work of two-dimensional electrophoresis (2DE) on the separation of microsomal membrane bound proteins. The following influencing factors on the solubilization of membrane bound proteins and 2DE images were discussed， such as the composing of rehydration solution， sample amount， focusing total voltage hours and different range of immobilized pH gradients (IPG) strips. It was shown that the samples rehydrated in solution making of urea， thiourea， tributyl phosphine and (3-(4-hepty) phenyl 3-hydroxy propyl)(dimethylammonio propane sulfonate)(C-7BZO) could be seen much better 2DE images than that of the other tested rehydration solutions. At the same time， the other running conditions for 70×3×0.5mm IPG strips were optimized as that the sample amount was 80μg and the focusing total voltage hour was around 20000. Much more clear spots were received by the usage of narrow-range IPG strips than that of wide-range IPG strips. Thirteen distinguish spots were found only in the induced samples and not in the control samples， whose isoelectric points (pI) were 4.0~8.0.

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备注/Memo

基金项目：荷兰瓦赫宁根大学“交叉研究和教育基金”项目

作者简介：乞永艳（1970-），女，博士，研究方向为生物活性物质及其应用。 E-mail： piaoliang_hao@163.com

更新日期/Last Update: 2014-10-21