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Single-molecule fluorescence imaging has recently developed into a powerful method for studying biophysical and biochemical phenomena (Moerner, 2007), including DNA metabolism (Ha, 2004; Hilario & Kowalczykowski, 2010; Zlatanova & van Holde, 2006). While classical biochemical methods yield parameters that are ensemble averaged, singlemolecule fluorescence imaging can be used to observe real-time behavior of individual biomolecules, allowing us to study their dynamic characteristics in great detail. Among the various biomolecules, protein molecules, which play central roles in many biological functions, are the prime targets for single-molecule imaging. Direct observations of singlemolecule DNA-binding proteins acting on their DNA targets in real time have provided new insights into DNA metabolism. In this chapter, I focus primarily on recent advances in direct visualization of single-molecule DNA-binding proteins in vitro, especially on the key techniques employed for this visualization.

Direct Visualization of Single-Molecule DNA-Binding Proteins Along DNA to Understand DNA-Protein Interactions