利用PEG处理去除Rubisco大亚基的植物蛋白质双向电泳方法的优化
Optimized plant 2-DE protocol by eliminating Rubioco large subunit from protein sample treated by PEG fractionation
绿色植物中高丰度的Rubisco蛋白，尤其是Rubisco大亚基（rbcL）的存在，对植物蛋白质双向电泳的分析造成了很大的困扰。PEG是一种能有效除去Rubisco蛋白的试剂，本研究以拟南芥(Col)野生型为材料,通过优化结合PEG处理的植物蛋白质提取方法，筛选出一套能有效除去rbcL，从而提高低丰度蛋白检出率的植物蛋白制样方法。研究结果表明，采用工作液浓度为17.5%的PEG处理植物全蛋白溶液，利用饱和酚/醋酸铵/甲醇提取法，可有效除去Rubisco大亚基，进而获得点数较多、电泳行为较好的2-DE电泳结果。
he existence of high-abundant proteins, especially ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) in plants, is one of the main reasons for poor detection of plant proteome analysis by two-dimensional electrophoresis (2-DE). Polyethylene glycol (PEG) is an excellent differential precipitation reagent for purification of proteins from a variety of sources. In the present work, we optimized a protocol for eliminating Rubisco large subunit treated by PEG fractionations from protein samples of Arabidopsis (Col) wild type and thus visualize more proteins in 2-DE analysis of plant proteome. The results showed that the large subunit of Rubisco was effectively precipitated with 17.5% PEG. In combination with isolating plant proteins by the method of saturated Phenol / ammonium acetate / methanol, a higher 2-DE gel profile of plant proteins was achieved in terms of protein yield and protein species.
沈弘、朱学艺、许郁芬
生物科学研究方法、生物科学研究技术生物化学植物学
双向电泳拟南芥蛋白质提取方法PEG
2-DEArabidopsisprotein extraction methodPEG
沈弘,朱学艺,许郁芬.利用PEG处理去除Rubisco大亚基的植物蛋白质双向电泳方法的优化[EB/OL].(2013-05-20)[2025-08-02].http://www.paper.edu.cn/releasepaper/content/201305-310.点此复制
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