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首页> 外文期刊>African Journal of Biotechnology >Comparison of protein extraction methods suitable for proteomics analysis in seedling roots of Jerusalem artichoke under salt (NaCl) stress
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Comparison of protein extraction methods suitable for proteomics analysis in seedling roots of Jerusalem artichoke under salt (NaCl) stress

机译：盐（NaCl）胁迫下适合菊芋幼苗根部蛋白质组学分析的蛋白质提取方法的比较

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摘要

An efficient protein extraction method is a prerequisite for successful implementation of proteomics. In this study, seedling roots of Jerusalem artichoke were treated with the concentration of 250 mM NaCl for 36 h. Subsequently, six different protocols of protein extraction were applied to seedling roots of Jerusalem artichoke for comparing extraction efficiency by conducting two-dimensional electrophoresis (2-DE). The first-dimensional electrophoresis was differently performed by using tube gel and immobilized pH gradient (IPG) strips, respectively. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) in the second dimension was equally carried out. The results indicated that precipitation of ammonium acetate in methanol after phenol extraction (Method 6) provided the best effect. Less impurity was contained in extracted proteins and the 2-DE gel images showed less horizontal and vertical stripes, the most proteineous spots were extracted on the basis of the protocol, up to 903. Trichloroacetic acid (TCA) method (Method 1) exhibited relative less quality gel images. 248 protein spots were only obtained according to the protocol and were the least among six protocols. Although, Mg/Nonidet P-40 (NP-40) methods (Methods 3 and 4) could extract a large number of proteins, it exhibited the worst quality gel images among all protocols and the horizontal and vertical stripes on the gel images were the most severe. It might be caused by a high salt content in extracted proteins.
机译：有效的蛋白质提取方法是成功实施蛋白质组学的前提。在这项研究中，用250 mM NaCl的浓度处理菊芋的幼苗根部36 h。随后，将六种不同的蛋白质提取方案应用于菊芋的幼苗根部，以通过进行二维电泳（2-DE）比较提取效率。分别使用管凝胶和固定pH梯度（IPG）试纸分别进行第一维电泳。同样进行第二维十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）。结果表明，苯酚萃取（方法6）后乙酸铵在甲醇中的沉淀效果最好。提取的蛋白质中杂质较少，2-DE凝胶图像显示出较少的水平和垂直条纹，根据操作规程提取了最多的蛋白点，最高达到903。三氯乙酸（TCA）方法（方法1）相对质量较差的凝胶图像。仅根据方案获得了248个蛋白斑点，在六个方案中最少。尽管Mg / Nonidet P-40（NP-40）方法（方法3和4）可以提取大量蛋白质，但在所有实验方案中其凝胶图像质量最差，并且凝胶图像上的水平和垂直条纹都是最严重的。这可能是由于提取的蛋白质中盐含量高所致。

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