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Integrating Optical Tweezers, DNA Tightropes, and Single-Molecule Fluorescence Imaging: Pitfalls and Traps.

Wang, Jiangzhou and Barnett, J T and Pollard, M R and Kad, Neil M (2017) Integrating Optical Tweezers, DNA Tightropes, and Single-Molecule Fluorescence Imaging: Pitfalls and Traps. In: Single-Molecule Enzymology: Nanomechanical Manipulation and Hybrid Methods. Methods in Enzymology, 582 . Elsevier, pp. 171-192. ISBN 978-0-12-809310-8. E-ISBN 978-0-12-809503-4. (doi:10.1016/bs.mie.2016.08.003) (Access to this publication is currently restricted. You may be able to access a copy if URLs are provided) (KAR id:61615)

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https://doi.org/10.1016/bs.mie.2016.08.003

Abstract

Fluorescence imaging is one of the cornerstone techniques for understanding how single molecules search for their targets on DNA. By tagging individual proteins, it is possible to track their position with high accuracy. However, to understand how proteins search for targets, it is necessary to elongate the DNA to avoid protein localization ambiguities. Such structures known as "DNA tightropes" are tremendously powerful for imaging target location; however, they lack information about how force and load affect protein behavior. The use of optically trapped microstructures offers the means to apply and measure force effects. Here we describe a system that we recently developed to enable individual proteins to be directly manipulated on DNA tightropes. Proteins bound to DNA can be conjugated with Qdot fluorophores for visualization and also directly manipulated by an optically trapped, manufactured microstructure. Together this offers a new approach to understanding the physical environment of molecules, and the combination with DNA tightropes presents opportunities to study complex biological phenomena.

Item Type: Book section
DOI/Identification number: 10.1016/bs.mie.2016.08.003
Subjects: Q Science
Divisions: Faculties > Sciences > School of Biosciences
Depositing User: N. Kad
Date Deposited: 04 May 2017 17:08 UTC
Last Modified: 06 May 2020 03:16 UTC
Resource URI: https://kar.kent.ac.uk/id/eprint/61615 (The current URI for this page, for reference purposes)
Wang, Jiangzhou: https://orcid.org/0000-0003-0881-3594
Kad, Neil M: https://orcid.org/0000-0002-3491-8595
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