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菜籽蛋白质组双向电泳技术体系的优化研究
引用本文:王丽娟,任学敏,杜艳慧,张其彬,罗水忠,姜绍通,郑志.菜籽蛋白质组双向电泳技术体系的优化研究[J].安徽农业科学,2011(17):10178-10181.
作者姓名:王丽娟  任学敏  杜艳慧  张其彬  罗水忠  姜绍通  郑志
作者单位:合肥工业大学生物与食品工程学院安徽省农产品精深加工重点实验室
基金项目:国家863项目(2010AA101503);国家科技支撑计划项目(2006BAD05A12);合肥工业大学学生创新基金项目(XS2010100)
摘    要:目的]建立适用于菜籽蛋白质组研究的双向电泳技术。方法]以湘油17号为材料,对双向电泳技术中的样品制备方法、凝胶浓度、上样量等条件进行了优化。结果]使用TCA一丙酮法提取菜籽总蛋白最佳,所获蛋白在2-DE图谱上获得的点数最多。当采用pH3~10IPG胶条和12%的凝胶、上样量为250μg时,可得到背景清晰、重复性好、蛋白点分辨率较高的双向电泳图谱。结论]为开展油菜种子蛋白质组学研究奠定了基础。

关 键 词:菜籽  蛋白质组  双向电泳  体系优化

Optimization of Two-dimensional Electrophoresis Technology System for Rapeseed Proteome
Institution:WANG Li-juan et al (Key Laboratory for the Intensive Processing of Agricultural Products in Anhui Province, College of Biotechnology and Food Engineering, Hefei University of Technology, Hefei, Anhui 230009)
Abstract:Objective] To establish the two-dimensional electrophoresis (2D-E) technology which was suitable for the research of rapeseed proteome. Method] Xiangyou 17 as the material, the sample preparation method, gel concentration and loading amount, etc. in 2D-E technology were optimized. Result] The best extraction method of rapeseed total protein was TCA-acetone method, and the points on 2D-E map were the most. When IPG (pH 3-10) and 12% gel was used, and the loading amount was 250 μg, the two-dimensional electrophoresis map with the clear background, good repeatability and high protein point resolution was obtained. Conclusion] The research laid the foundation for carrying out the rapeseed proteomics research.
Keywords:Rapeseed  Proteome  Two-dimensional electrophoresis  System optimization
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