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首页> 外文期刊>Separation Science Plus >Standardization of a protein extraction protocol for the proteomic analysis of the leaves of Oryza sativa L. cv. BRS AG
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Standardization of a protein extraction protocol for the proteomic analysis of the leaves of Oryza sativa L. cv. BRS AG

机译：蛋白质提取的标准化协议的蛋白质组学分析选用的叶子

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摘要

The rice cultivar BRS AG (Oryza sativa L.) is a cereal developed to serve as a raw material for ethanol production or animal feed, contributing to an increasing energy demand and the economic sustainability of the rice sector. However, only few studies have reported this new cultivar. As proteomic analysis is crucial in understanding the physiological and adaptive behavior of BRS AG rice leaves under biotic and abiotic stresses, the present study compared the following three precipitation protocols for the protein extraction of leaf tissue in two-dimensional electrophoresis analysis: phenol method, trichloroacetic acid-acetone method, and a third method developed in our laboratory (BIOPRO). The efficiency and repeatability of the protein extraction protocols were evaluated by two-dimensional electrophoresis analysis. Our results indicate that the BIOPRO protocol was the most efficient in protein extraction as compared to the efficiency of other protocols. We detected a greater amount of protein (15.44 mu g/mu L) and 414 spots, and the BIOPRO protocol provided the best results regarding reproducibility, focusing, resolved spots, the intensity of spots, and single spot detection. This novel protocol is useful for extracting high-quality proteins from leaves and presents an efficient removal of contaminants, which is directly linked to a better resolution for two-dimensional electrophoresis analysis.
机译：水稻品种BRS AG(栽培稻l .)谷物作为原材料乙醇的生产或动物饲料,做贡献日益增长的能源需求和经济大米行业的可持续发展。很少有研究报道这种新的品种。蛋白质组学分析的理解是至关重要的BRS AG)的生理适应性行为水稻叶片在生物和非生物压力,本研究比较了三种沉淀蛋白质的协议提取二维的叶组织电泳分析:苯酚的方法,三氯乙酰acid-acetone方法,三分之一我们实验室开发的方法(BIOPRO)。蛋白质的效率和可重复性提取协议进行评估二维电泳分析。结果表明,BIOPRO协议了最有效的蛋白质提取相比其他协议的效率。大量的蛋白质(15.44μg /μL)414点,BIOPRO协议提供了最好的结果重现性、聚焦,解决斑点,斑点的强度单点检测。用于提取高质量的蛋白质树叶和提供了一个有效的去除污染物,直接联系更好的解决二维电泳分析。

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