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Characterisation and functional analysis of actin cytoskeleton components and development of a novel hydrostatic pressure live cell imaging system

Brooker, Holly (2017) Characterisation and functional analysis of actin cytoskeleton components and development of a novel hydrostatic pressure live cell imaging system. Doctor of Philosophy (PhD) thesis, University of Kent,. (KAR id:62466)

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Abstract

The main aims of this project were to use a variety of techniques to study various components of the actin cytoskeleton using Caenorhabditis elegans and Schizosaccharomyces pombe, in addition to developing a novel hydrostatic pressure imaging system.

In vitro and in vivo studies were next undertaken to explore the impact of fluorescent protein fusions and temperature sensitive mutations upon the the stability and function of S. pombe tropomyosin. While both Ts mutants, Cdc8-27 and Cdc8-110, had a two-step thermal unfolding transition lower than that of wild-type Cdc8, fluorescent fusions did not impact stability. Amino- terminal fusions however, mimic acetylation whereas carboxyl terminal fusions abolished polymerisation, localisation and functionality. The final part of this project used a methodical approach to develop a hydrostatic imaging system that could be used to follow fluorescently labelled protein dynamics in a live cell context. Preliminary data revealed the application of 100 bar of pressure induced a cell cycle delay in both S. pombe and Candida albicans, however the current limitations of the system make it incompatible with fluorescence microscopy.

Item Type: Thesis (Doctor of Philosophy (PhD))
Thesis advisor: Mulvihill, Daniel
Divisions: Divisions > Division of Natural Sciences > School of Biosciences
SWORD Depositor: System Moodle
Depositing User: System Moodle
Date Deposited: 28 Jul 2017 12:47 UTC
Last Modified: 20 May 2021 13:30 UTC
Resource URI: https://kar.kent.ac.uk/id/eprint/62466 (The current URI for this page, for reference purposes)
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