拟南芥全细胞蛋白质样品制备及其双向电泳条件的优化

拟南芥全细胞蛋白质样品制备及其双向电泳条件的优化

席景会¹，²，殷建文¹，岳琳¹，李善玉¹，³，周欣¹，王旭¹，郝东云¹

1. 吉林大学分子酶学工程教育部重点实验室，长春 130021； 2. 吉林大学植物科学学院，长春 130062； 3. 吉林大学第一医院，长春　130021

收稿日期:2006-01-21 修回日期:1900-01-01 出版日期:2006-11-26 发布日期:2006-08-26
通讯作者: 郝东云

Optimization of Sample Preparation and Twodimensional Gel Electrophoresis for Wholecell Proteins Analysis in Arabidopsis

XI Jinghui¹,², YIN Jianwen¹, YUE Lin¹, LI Shanyu¹,³, ZHOU Xin¹, WANG Xu¹, HAO Dongyun¹

1． Key Laboratory for Molecular Enzymology & Engineering of the Ministry of Education,　Jilin University, Changchun 130021, China; 2．　College of Plant Science, Jilin University, Changchun 130062, China；　3．　The 1st Hospital of Jilin University, Changchun 130021, China

Received:2006-01-21 Revised:1900-01-01 Online:2006-11-26 Published:2006-08-26
Contact: HAO Dongyun

摘要/Abstract

摘要： 对双向电泳方法进行了改进，包括对样品的制备、双向电泳参数的选择等关键步骤进行优化. 实验发现，采用乙酸铵甲醇沉淀拟南芥全细胞蛋白质，同时结合高伏时、长时间的等电聚焦可以获得高分辨率、重复性好的蛋白质双向电泳图谱. 银染后，经Phoretix 2D v2004软件分析可分辨出1 000以上蛋白点.

关键词: 拟南芥, 样品制备, 双向电泳

Abstract: An optimized protocol was reported with special attention focused on the wholecell protein analyses based on twodimensional gel electrophoresis (2DE) and Arabidopsis as material. The optimization included sample preparation, electrophoresis conditions, gel staining, protein spots resolution and reproducibility. The results indicate that in combination with highvoltage and longrunning time in IEF, the precipitation of proteins with ammonium acetate/methanol method will give rise to satisfactory 2DE image. The maximal detectable protein spots were estimated to be more than 1 000 under silver staining. This protocol may be applicable to other plant materials for 2DE studies. 

Key words: Arabidopsis thaliana, sample preparation, twodimensional gel electrophoresis

中图分类号:

Q946

席景会，，殷建文，岳琳，李善玉，，周欣，王旭，郝东云. 拟南芥全细胞蛋白质样品制备及其双向电泳条件的优化[J]. J4, 2006, 44(06): 1011-1014.

XI Jinghui,, YIN Jianwen, YUE Lin, LI Shanyu,, ZHOU Xin, WANG Xu, HAO Dongyun. Optimization of Sample Preparation and Twodimensional Gel Electrophoresis for Wholecell Proteins Analysis in Arabidopsis[J]. J4, 2006, 44(06): 1011-1014.

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