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Title: Stress and survival of Clostridium difficile
Author: Smyth, Deborah
ISNI:       0000 0004 6057 4786
Awarding Body: Ulster University
Current Institution: Ulster University
Date of Award: 2017
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CDI has been implicated in significant morbidity and mortality rates leading to complications such as pseudomembranous colitis, toxic megacolon and sepsis. Several classical symptoms include infectious and profuse diarrhoea, high grade fever, nausea and abdominal cramps. The aim of this thesis was to insert a functional dnaK heat shock gene in the ClosTron gener­ated C. difficile 630Δerm::dnaK by using a pMTL80000 series plasmid to restore the wild- type phenotype thus correcting the growth rate and temperature sensitivity. The dnaK gene, under the influence of a strong /cZv promoter, conjugated successfully into C. difficile 630Δerm::dnaK. For comparison, a novel pyrE background method established at the Uni­versity of Nottingham was used to create two further dnaK disruption mutants, C. difficile 630ΔermΔpyrE::dnaK and C. difficile R20291ΔpyrE: :dnaK. These pyrE~ dnaK disruption mutants were then complemented by the insertion of pMTL-YN series plasmids containing a functional dnaK gene. The process was approached in a logical fashion for construction and complementation of the new dnaK deletion mutants in the pyrE' background. It was verified by PCR that the dnaK gene successfully inserted into the pMTL-YN series plasmids and that these plasmids were successfully conjugated into the required C. difficile isolates. Results achieved from the growth curve experiments and motility assays showed that the dnaK dele­tion mutants in the pyrE' background and associated complements displayed no difference in their growth in comparison to the wild-type (C. difficile 630Δerm).
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available