Xie X Sunney, Choi Paul J, Li Gene-Wei, Lee Nam Ki, Lia Giuseppe
Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts 02138, USA.
Annu Rev Biophys. 2008;37:417-44. doi: 10.1146/annurev.biophys.37.092607.174640.
Recent developments on fluorescent proteins and microscopy techniques have allowed the probing of single molecules in a living bacterial cell with high specificity, millisecond time resolution, and nanometer spatial precision. Recording movies and analyzing dynamics of individual macromolecules have brought new insights into the mechanisms of many processes in molecular biology, such as DNA-protein interactions, gene regulation, transcription, translation, and replication, among others. Here we review the key methods of single-molecule detection and highlight numerous examples to illustrate how these experiments are contributing to the quantitative understanding of the fundamental processes in a living cell.
荧光蛋白和显微镜技术的最新进展使得在活细菌细胞中以高特异性、毫秒级时间分辨率和纳米级空间精度探测单个分子成为可能。记录单个大分子的动态过程并进行分析,为深入了解分子生物学中许多过程的机制带来了新的见解,比如DNA-蛋白质相互作用、基因调控、转录、翻译和复制等。在这里,我们回顾单分子检测的关键方法,并列举大量实例来说明这些实验如何有助于定量理解活细胞中的基本过程。