非遷徙式單分子螢光共振能量轉移在分子生物學上的應用

Application of Immobilized Single Molecular Fluorescence Resonance Energy Transfer (FRET) in Molecular Biology

章為皓(Wei-Hau Chang)

《科儀新知》 171期 (2009/08) Pp. 14-21

https://doi.org/10.29662/IT.200908.0003

摘要

近年來單分子螢光造影和分析已經躍升為解決生物系統中的動態與非均質性的利器。其中的非遷徙式單分子螢光能共振光譜造影方法，藉著把核酸固定在玻片上，大大簡化對於蛋白質與核酸的作用的造影與分析，使得像是蛋白質的構型變化或在核酸上的遷移都可被即時觀察到。我們在此介紹單分子螢光能共振的原理簡單的全反射式螢光顯微鏡的架設，和其成功地應用於解決核醣核酸聚合上核醣核酸通道的案例。

並列摘要

In recent years, single moleucle optical imaging and analysis has emerged as a powerful way to unfold the dynamic and heterogeneous behavior of numerous biological systems. Among all single moleucle technique, single-molecule fluorescence resonance fluorescence energy transfer (smFRET) spectroscopic imaging in an immobilized fashion is particulary suitable for following the interactions between nucleic acids and proteins to 1 nm resolution with time resolution ranging from 10 ns to second. Here, we describe the principle of smFRET and a simple version of total internal reflection instrumentation and provide a successful application to the study of RNA exit channel on a transcribing RNA polymerase Ⅱ.