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首页> 外文期刊>Journal of Medicinal Plants Research >An efficient protein preparation method compatible with 2-DE analysis of Panax quinquefolius root - a tissue riches in interfering compounds
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An efficient protein preparation method compatible with 2-DE analysis of Panax quinquefolius root - a tissue riches in interfering compounds

机译：一种高效的蛋白质制备方法，可与西洋参（一种富含干扰化合物的组织）的2-DE分析相兼容

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摘要

Extraction of high-quality proteins from tissue like?Panax quinquefolius?root is difficult due to high levels of interfering compounds, mainly viscous polysaccharide, which obstructs the extraction of low abundant proteins. To establish an efficient method for the application of proteomic analysis to?P. quinquefolius?root, 4 sample preparation methods were tested: Acetone precipitation, ethanol precipitation, chloroform/isoamyl alcohol extraction and phenol/chloroform/isoamyl alcohol extraction. These methods were evaluated based on the quality of SDS-PAGE and two-dimensional electrophoresis (2-DE) patterns. In chloroform/isoamyl alcohol extraction, due to the different polarity, proteins were dispersed in water-phase and inter-phase. There were abundant water-soluble proteins in water-phase and fat-soluble proteins in inter-phase. In order to acquire an optimized 2-DE pattern containing as many protein spots as it can be, the both phase proteins were mixed together, and the results of SDS-PAGE and 2-DE showed well-resolved low-abundant proteins with low background. Some significantly changed spots on 2-DE gel of optimized method were identified and their potential relationships to root development were discussed. These results showed that optimized method gives satisfactory and reproductive 2-DE protein patterns. It is expected that this method can also be applied to other plant root tissues that are rich in interfering ingredients.
机译：由于高含量的干扰化合物（主要是粘性多糖）会阻碍低丰度蛋白质的提取，因此很难从西洋参（Panax quinquefolius）根等组织中提取高质量的蛋白质。建立有效的蛋白质组学分析方法。 quinquefolius？root，测试了4种样品制备方法：丙酮沉淀，乙醇沉淀，氯仿/异戊醇萃取和苯酚/氯仿/异戊醇萃取。这些方法是根据SDS-PAGE和二维电泳（2-DE）模式的质量进行评估的。在氯仿/异戊醇萃取中，由于极性不同，蛋白质分散在水相和中间相中。在水相中有丰富的水溶性蛋白，在相间有大量的脂溶性蛋白。为了获得包含尽可能多的蛋白质斑点的优化的2-DE模式，将两种蛋白质混合在一起，并且SDS-PAGE和2-DE的结果显示了分离度高，背景低的蛋白质。确定了优化方法的2-DE凝胶上一些明显变化的斑点，并讨论了它们与根发育的潜在关系。这些结果表明，优化方法可提供令人满意的生殖2-DE蛋白质图谱。预期该方法也可以应用于富含干扰成分的其他植物根组织。

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