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电泳方法简史。

A short history of electrophoretic methods.

作者信息

Vesterberg O

机构信息

Division of Medical Chemistry, National Institute of Occupational Health, Solna, Sweden.

出版信息

Electrophoresis. 1993 Dec;14(12):1243-9. doi: 10.1002/elps.11501401188.

Abstract

High resolution separation of proteins, based on charge differences, is possible with disc electrophoresis, displacement electrophoresis (isotachophoresis) and notably isoelectric focusing (IEF). Size separation is obtained in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The combination of gel IEF, followed by SDS-PAGE in a second-dimensional slab gel, i.e. two-dimensional gel electrophoresis, affords the highest resolution with up to several thousand spots per gel. Staining of proteins gives high resolution patterns which can be scanned and stored in comprehensive databases. Over the last 10 years the electrophoretic separation in gels and subsequent visualization of nucleic acids (DNA, RNA) and even genes as well as nucleotides have been much improved, making possible efficient mapping of the genes in humans and all other organisms. This has led to the biggest concerted endeavor in the history of science, i.e. the mapping of the human genome, which will be of importance as long as mankind exists. In the last years electrophoresis in capillaries has attracted much interest because for numerous substances, such as proteins nucleic acids, pharmaceuticals, metabolites, and peptides, it offers high resolution on the analytical scale with over 1 million theoretical plates. Electrophoretic methods have unprecedented impact on life sciences, providing a basis for unique advances in biochemistry, molecular biology, genetics, gene technology and medicine.

摘要

基于电荷差异对蛋白质进行高分辨率分离,可以通过圆盘电泳、置换电泳(等速电泳),尤其是等电聚焦(IEF)来实现。在十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)中可以实现按大小分离。在凝胶IEF之后,在二维平板凝胶中进行SDS-PAGE,即二维凝胶电泳,可提供最高分辨率,每个凝胶上多达数千个斑点。蛋白质染色可得到高分辨率图谱,这些图谱可以扫描并存储在综合数据库中。在过去的10年里,凝胶中的电泳分离以及随后核酸(DNA、RNA)甚至基因和核苷酸的可视化有了很大改进,使得高效绘制人类和所有其他生物体的基因图谱成为可能。这导致了科学史上最大规模的协同努力,即人类基因组图谱绘制,只要人类存在,它都将具有重要意义。近年来,毛细管电泳引起了人们的极大兴趣,因为对于许多物质,如蛋白质、核酸、药物、代谢物和肽,它在分析规模上具有高分辨率,理论塔板数超过100万。电泳方法对生命科学产生了前所未有的影响,为生物化学、分子生物学、遗传学、基因技术和医学的独特进展提供了基础。

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