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Engineering bacteriophage K1F to develop a novel point-of-care Escherichia coli K1F detection system
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Wheatley, Joseph (2022) Engineering bacteriophage K1F to develop a novel point-of-care Escherichia coli K1F detection system. PhD thesis, University of Warwick.
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WRAP_THESIS_Wheatley_2022.pdf - Submitted Version - Requires a PDF viewer. Download (215Mb) | Preview |
Official URL: http://webcat.warwick.ac.uk/record=b3878427~S15
Abstract
The internal capsid proteins that reside within the phage K1F capsid structure hold high potential for being used as sensitive and reliable diagnostic tools. The concealed nature of the capsid interior ensures that any encapsulated signal or signal-generating enzyme, e.g. fused to an internal capsid protein, is suppressed whilst the phage is unaccompanied by its host. Furthermore, the only naturally-occurring mechanism for releasing the internal capsid proteins and therefore exposing their amalgamated signal/enzyme, is for them to be passed through the tail and subsequently ejected out of the phage - a post-adsorption phenomenon which exclusively occurs when the host is present – thus presenting a precise model for signal/enzyme release only upon pathogen presence. Here, a small N-terminal subunit of the NanoLuc luciferase is fused and incorporated into the K1F internal capsid structure using a simple, non-genomic method.
This internalised subunit is exposed to the test solution containing its C-terminal counterpart (spontaneous complementation immediately forms the full NanoLuc enzyme) and substrate (furimazine) only when the K1F host, E. coli K1 or lab strain EV36, is present – thereby presenting a novel method for rapidly detecting this disease-causing pathogen. Finally, it is expected that by building upon this internal capsid protein engineering approach, which completely bypasses the time-consuming processes of intracellular nucleic acid transcription and translation, an unprecedentedly rapid detection device can be developed for an array of bacterial pathogens.
Item Type: | Thesis (PhD) | ||||
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Subjects: | Q Science > QR Microbiology R Medicine > RC Internal medicine |
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Library of Congress Subject Headings (LCSH): | Bacteriophages, Escherichia coli -- Detection, Point-of-care testing, Capsid proteins | ||||
Official Date: | June 2022 | ||||
Dates: |
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Institution: | University of Warwick | ||||
Theses Department: | School of Engineering | ||||
Thesis Type: | PhD | ||||
Publication Status: | Unpublished | ||||
Supervisor(s)/Advisor: | Kulkarni, Vishwesh ; Sagona, Antonia | ||||
Extent: | xv, 196 pages : illustrations, charts | ||||
Language: | eng |
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