Department of Biotechnology, Jaypee Institute of Information Technology, A-10 Sector-62, Noida 201307, Uttar Pradesh, India.
Mol Cell Biochem. 2012 Jun;365(1-2):279-99. doi: 10.1007/s11010-012-1269-z. Epub 2012 Mar 8.
DNA-binding proteins control various cellular processes such as recombination, replication and transcription. This review is aimed to summarize some of the most commonly used techniques to determine DNA-protein interactions. In vitro techniques such as footprinting assays, electrophoretic mobility shift assay, southwestern blotting, yeast one-hybrid assay, phage display and proximity ligation assay have been discussed. The highly versatile in vivo techniques such as chromatin immunoprecipitation and its variants, DNA adenine methyl transferase identification as well as 3C and chip-loop assay have also been summarized. In addition, some in silico tools have been reviewed to provide computational basis for determining DNA-protein interactions. Biophysical techniques like fluorescence resonance energy transfer (FRET) techniques, FRET-FLIM, circular dichroism, atomic force microscopy, nuclear magnetic resonance, surface plasmon resonance, etc. have also been highlighted.
DNA 结合蛋白控制着各种细胞过程,如重组、复制和转录。本综述旨在总结一些最常用的确定 DNA-蛋白质相互作用的技术。讨论了体外技术,如足迹分析、电泳迁移率变动分析、西南印迹、酵母单杂交测定、噬菌体展示和邻近连接测定。还总结了高度多功能的体内技术,如染色质免疫沉淀及其变体、DNA 腺嘌呤甲基转移酶鉴定以及 3C 和芯片环测定。此外,还回顾了一些计算工具,为确定 DNA-蛋白质相互作用提供了计算基础。还强调了生物物理技术,如荧光共振能量转移(FRET)技术、FRET-FLIM、圆二色性、原子力显微镜、核磁共振、表面等离子共振等。