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Title: PrP gene expression regulation in sheep : identification of candidate transcription factors that exhibit differential binding to polymorphic variants of the ovine PrP gene promoter
Author: Burgess, Stewart Thomas George
Awarding Body: University of Edinburgh
Current Institution: University of Edinburgh
Date of Award: 2004
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Amino acid variants of PrPc particularly at codons 136, 154 & 171 have been linked to scrapie susceptibility, but do not explain all variation in disease phenotype. This study has therefore addressed the hypothesis that unexpected disease occurrence may also be linked to different levels of PrP gene expression. In order to investigate the role of the ovine PrP gene 3’UTR in the regulation of gene expression, a series of ovine PrP mini-gene constructs were produced, which differed only in their availability of previously identified polyadenylation signals. Following the transient transfection of ovine and murine cell lines with these constructs it became clear that they were being incorrectly spliced, despite the fact that sequencing confirmed the presence of all of the elements required for correct splicing to occur. Gene transcription is also regulated by the binding of sequence specific transcription factors to the promoter region and the role of the PrP gene promoter in the regulation of gene expression was investigated in the second half of the thesis. Sequence analysis using online and offline database resources revealed the presence of a number of transcription elements within the ovine PrP gene promoter. It was therefore decided to test the hypothesis that the binding of transcription factors to the ovine PrP promoter could influence the expression of the PrP gene. Using gel shift assays specific binding was observed to selected sequence elements and differential binding was demonstrated to a polymorphic variant of at least one of these motifs. In addition, binding to four motifs conserved in the mammalian PrP promoters was also analysed using a combination of gel shift assay and DNase I footprinting. Two of the four motifs showed specific binding and polymorphic variants of these motifs exhibited differential binding.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available