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Comparison of protein extraction methods for 2DE-based proteomic analysis of duckweed Spirodela polyrhiza, a small aquatic model plant
Aquatic Botany ( IF 1.9 ) Pub Date : 2020-02-18 , DOI: 10.1016/j.aquabot.2020.103216
Yibo Wang , Xiuyun Wang , Ruixue Yang , Liangjie Niu , Wei Wang

Duckweed, a small floating aquatic plant in the family Lemnaceae, has considerable potential for agricultural and environmental applications. Proteomic analysis will facilitate the study of plant biology and plant-environment interactions. A reliable and reproducible protein extraction method is critical for successful proteomic analysis. To date, there have been no evaluation studies of extraction methods for duckweed proteomics. In this study, we developed a trichloroacetic acid (TCA)/acetone/TCA precipitation method (TAT) for Spirodela polyrhiza and compared it with three extraction methods, primarily by two-dimensional gel electrophoresis (2DE). These methods were based on TCA/acetone precipitation, phenol extraction or their combination to concentrate proteins and remove interfering substances. As a result, these methods produced significant differences in protein yield, and each revealed a unique set of proteins. In general, TAT produced better 2DE protein patterns with high reproducibility regarding the number, coverage and abundance of protein spots. Our results provide useful information for selecting suitable extraction methods for proteomic analysis of duckweed plants.



中文翻译:

小型水生模型植物浮萍Spirodela polyrhiza基于2DE蛋白质组分析的蛋白质提取方法的比较

浮萍是Lemnaceae家族中的一种小型漂浮水生植物,在农业和环境应用方面具有相当大的潜力。蛋白质组学分析将有助于植物生物学和植物-环境相互作用的研究。可靠且可重现的蛋白质提取方法对于成功进行蛋白质组学分析至关重要。迄今为止,还没有关于浮萍蛋白质组学提取方法的评价研究。在这项研究中,我们开发了用于螺旋藻的三氯乙酸(TCA)/丙酮/ TCA沉淀法(TAT)并将其与三种提取方法(主要是通过二维凝胶电泳(2DE))进行比较。这些方法基于三氯乙酸/丙酮沉淀,苯酚萃取或它们的组合以浓缩蛋白质并去除干扰物质。结果,这些方法在蛋白质产量上产生了显着差异,并且每种方法都揭示了一套独特的蛋白质。总的来说,TAT产生了更好的2DE蛋白质模式,在蛋白质斑点的数量,覆盖范围和丰度方面具有很高的可重复性。我们的结果为选择适合浮萍植物蛋白质组学分析的提取方法提供了有用的信息。

更新日期:2020-02-18
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