2D-DIGE:蛋白质组分析的强大工具

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摘要

近年来,二维凝胶电泳已成为复杂蛋白样品比较表达谱的强大分子生物学工具。它涉及到基于等电点和蛋白质分子质量的二维方法对不同蛋白质样品的分离和分辨。通过这种方式,它反映了整体蛋白质组状态的观点,包括蛋白质表达水平的分化,翻译后修饰等。此外,这允许鉴定新的生物特征,这可能为与各种类型的癌症和神经系统疾病相关的细胞或组织提供特定的病理背景。因此,利用这些工具,人们可以清楚地研究和比较特定药物对细胞或组织的影响,也可以根据广谱蛋白表达谱的变化分析疾病的影响。近年来,为了获得更准确准确的蛋白质组图谱,传统的二维凝胶电泳技术得到了改进,加入了不同类型的蛋白质标记染料,使得在一个二维凝胶中可以对不同的蛋白质样品进行更多的比较分析。在这种先进的技术(2-D-DIGE)中,蛋白质样品分别用三种不同类型的cydye (Cy2, Cy3和Cy5)标记,并结合在同一凝胶上进一步分解。这将有助于在单个凝胶平台上更准确地进行点匹配,并且还将最小化传统2d凝胶电泳中常见的实验变化。因此,在目前的蛋白质组学研究时代,2D-DIGE已被证明是一种非常强大的工具,在临床研究中对高达125 ng的蛋白质具有很高的灵敏度,特别是对神经和癌症相关疾病。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
2D-DIGE A Powerful Tool for Proteome Analysis
In the recent past, two dimensional gel electrophoresis has emerged as a powerful molecular biology tool for the comparative expression profiling of complex protein sample. It involves the separation as well as the resolution of diverse proteins sample on the basis of isoelectric points and molecular mass of protein in two dimension ways. In this way, it reflects the view of overall proteome status including differentiation in protein expression levels, post-translational modifications etc. Moreover, this allows the identification of novel biological signatures, which may give a particular identity of pathological background to cells or tissues associated with various types of cancers and neurological disorders. Therefore, by utilizing such tools, one can clearly investigate and compare the effects of particular drugs on cells of tissues and also one can analyze the effects of disease on the basis of variations in protein expression profile at broad spectrum. Recently, to get more error-less and accurate proteome profile, conventional 2-D gel electrophoresis has been enhanced with the inclusion of different types of protein labeling dyes which enables a more comparative analysis of diverse protein sample in a single 2-D gel. In this advanced technique (2-D-DIGE), protein samples are labeled with three different types of CyDyes (Cy2, Cy3, and Cy5) separately and combined and further resolved on the same gel. This will facilitate the more accurate spot matching on a single gel platform and will also minimize the experimental variations as commonly reported in the conventional 2D-gel electrophoresis. Therefore, in the present proteomic research era, 2D-DIGE has proved to be an extremely powerful tool with great sensitivity for up to 125 ng of proteins in clinical research volubility especially, neurological and cancer related disorders.
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