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Evaluation of the Tat export pathway for the production of recombinant proteins in Escherichia coli

Malherbe, Gilles (2019) Evaluation of the Tat export pathway for the production of recombinant proteins in Escherichia coli. Doctor of Philosophy (PhD) thesis, University of Kent,. (KAR id:81263)

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Abstract

Recently, the twin-arginine translocation (Tat) pathway has raised interest due to reports of its unique proofreading ability to export correctly folded proteins. However, only a few recombinant proteins fused to an N-terminal Tat signal peptide have been reported to be exported via Tat. The hypothesis for this thesis was to evaluate whether the range of proteins of interest that can be exported by the Tat pathway can be extended by C-terminally fusing them to a natural Tat substrate (NTS). Thus, the NTS would act as a soluble carrier to improve cytoplasmic solubility and facilitate Tat export. Moreover, the CyDisCo technology enabling cytoplasmic formation of disulphide bonds was investigated to reach this goal.

However, this project revealed unexpected export of sfGFP, hGH and FABP4 including the recombinant proteins fused to hGH without a signal peptide. This unidentified translocation mechanism(s) needs further characterisation which can lead to the discovery of a new export pathway(s) and may highlight a promising alternative way to target proteins of interest to the periplasm.

Item Type: Thesis (Doctor of Philosophy (PhD))
Thesis advisor: Robinson, Colin
Uncontrolled keywords: Tat export, CyDisCo
Subjects: Q Science > QH Natural history > QH301 Biology
Divisions: Divisions > Division of Natural Sciences > School of Biosciences
SWORD Depositor: System Moodle
Depositing User: System Moodle
Date Deposited: 15 May 2020 14:10 UTC
Last Modified: 20 May 2021 13:24 UTC
Resource URI: https://kar.kent.ac.uk/id/eprint/81263 (The current URI for this page, for reference purposes)
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