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Download or read book Grab, Manipulate and Watch Single DNA Molecule Replication written by and published by . This book was released on 2023 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: DNA replication is a critical process intrinsic to the sustenance and propagation of life, involving a symphony of enzymes including helicases, polymerases, and single-stranded DNA binding proteins (SSBs). These enzymes collaboratively ensure the precise duplication of genetic information, a process that, despite significant research, has elements yet to be fully elucidated, especially intermediary steps and the dynamic engagements between replicative proteins. Single-molecule techniques have recently blossomed, offering enhanced insights into the real-time dynamics and interactions of individual molecules in their natural settings, unveiling obscured intermediate steps and enzyme conformational changes. Chapter 1 outlines the thesis's primary aim, elucidating DNA replication mechanisms. Starting with fundamental biological notions, this chapter transitions to discuss the non-equilibrium nature of living systems, emphasizing the role of single-molecule investigations. Such studies have enhanced our understanding of non-equilibrium systems, revealing cellular mechanisms and influencing factors. SSBs are crucial for maintaining genome integrity as they bind to ssDNA and coordinate with various proteins involved in DNA replication, recombination, and repair. Chapter 2 offers a comprehensive overview of recent advances in our understanding of SSBs, as elucidated by single-molecule assays such as optical tweezers, magnetic tweezers, Förster resonance energy transfer, and their combinations. These techniques have provided novel insights into the dynamics of SSB binding to ssDNA and its interactions with other proteins, emphasizing the central role of SSB in modulating the activities of other proteins. Chapter 3 presents the single-molecule observations of the T7 bacteriophage single-stranded DNA-binding protein (gp2.5) binding to ssDNA. Our experiments demonstrate the significant influence of the base sequence, ssDNA conformation, and the acidic terminal domain of T7 gp2.