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Title: The ahp promoter of Salmonella enterica sv. typhimurium : regulation and merits for heterologous antigen expression in vivo
Author: Martin, Teresa M.
Awarding Body: University of Edinburgh
Current Institution: University of Edinburgh
Date of Award: 2002
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In this thesis, regulation of wild type and Mudlux-tagged ahp expression was examined by reverse transcription and the polymerase chain reaction (RT-PCR). Maximum induction of ahpC in MPG203 was seen in LSLB and minimal media, after treatment with both hydrogen peroxide and sodium chloride in combination, which is in agreement with the luminometer data. However, ‘alpC in MPG203 was induced to a greater degree after treatment with hydrogen peroxide compared with sodium chloride, which is in maximum induction of alpC in LSLB was recorded after treatment with salt, and after treatment with a combination of salt and hydrogen peroxide in minimal media. The basis for this aberrant osmoregulation of ahp is still not fully resolved, as to whether it is under transcriptional or post-transcriptional control. Because of its mode of activation of the ahp promoter as a tool for in vivo expression of foreign antigens (Francis et al., 1993; Taylor et al.,1998) was also examined. Two other known in vivo expressed promoters, PpagC and PspvA were included in the study for comparison. The antigens encoded on the plasmids used for immunisation, were gene fragments cloned from listeriolysin (LLO) of Listeria monocytogenes and glycoprotein 63 (Gp63) of Leishmania major which contained T cell epitopes> GFP, was used as a stable protein control was also expressed from a plasmid construct and used for immunisations. Salmonella typhimurium strain MPG424 was utilised as a vector for immunisation. It contained both a purA and an aroA mutation, leaving the strain hyper-attenuated. However, the Escherichia coli purA promoter and gene were supplied in trans on plasmids encoding antigens of interest, to ensure plasmid maintenance and to moderate attenuation by complementation (Nakayama et al. 1988; Curtiss et al. 1990).
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available