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Lookup NU author(s): Dr Adam WollmanORCiD
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© Springer Science+Business Media New York 2016. Single-molecule narrow-field microscopy is a versatile tool to investigate a diverse range of protein dynamics in live cells and has been extensively used in bacteria. Here, we describe how these methods can be extended to larger eukaryotic, yeast cells, which contain subcellular compartments. We describe how to obtain single-molecule microscopy data but also how to analyze these data to track and obtain the stoichiometry of molecular complexes diffusing in the cell. We chose glucose mediated signal transduction of live yeast cells as the system to demonstrate these single-molecule techniques as transcriptional regulation is fundamentally a single-molecule problem—a single repressor protein binding a single binding site in the genome can dramatically alter behavior at the whole cell and population level.
Author(s): Wollman AJM, Leake MC
Editor(s): Mark C. Leake
Publication type: Book Chapter
Publication status: Published
Book Title: Chromosome Architecture: Methods and Protocols
Online publication date: 10/06/2016
Acceptance date: 02/04/2016
Series Title: Methods in Molecular Biology
Publisher: Humana Press Inc.
Place Published: New York
PubMed id: 27283298
Library holdings: Search Newcastle University Library for this item