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Title: Characterising the adsorption process of the shigatoxigenic bacteriophage, φ24B, to its bacterial host via an essential outer-membrane protein, BamA
Author: McEwen, S. A.
ISNI:       0000 0004 7428 7374
Awarding Body: University of Liverpool
Current Institution: University of Liverpool
Date of Award: 2018
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Escherichia coli is a common commensal organism that is present in all human digestive tracts. When E. coli is infected by a bacteriophage containing Shiga toxin genes (Stx phage), the E. coli cell is converted into a pathogen that can cause serious harm to humans in the form of Haemolytic Uraemic Syndrome (HUS) or Thrombotic, Thrombocytopaenic Purpura (TTP). It has been shown that ~70% of environmental Stx phages possess a tail that recognises an essential protein in the outer membrane of E. coli, BamA. This study presents the final proof required to demonstrate that BamA is the target of these short-tailed Stx phages and intends to dissect which epitopes of extracellular surface of BamA support phage adsorption by comparing E. coli to a phage resistant species, Pectobacterium atrosepticum. This resulted in the production of a library of 32 BamA mutants, which could be compared in an adsorption assay. The physiological effects of these mutants on the adsorption process were then examined in a fluorescence microscopy-based adsorption assay that allows the visualisation of temporal resolution of these processes. Given that BamA is conserved across all Enterobacteriaceae. There is potential for host range expansion, thereby converting more species that are currently harmless into potential human pathogens. The information obtained from this study may help in predicting which species is the next likely to become Stx phage susceptible.
Supervisor: Allison, Heather ; McCarthy, A. J. Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral