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Title: Characterisation of the 5' flanking region of the murine PrP gene
Author: Cameron, John Daniel
Awarding Body: University of Edinburgh
Current Institution: University of Edinburgh
Date of Award: 1995
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Scrapie, Creutzfeldt-Jakob disease and bovine spongiform encephalopathy belong to a group of diseases known as the transmissible degenerative encephalopathies. These diseases are characterised by astrocytic gliosis, neuronal vacuolation, deposition of amyloid plaques, long incubation periods and by their transmissibility to laboratory rodents. This project sets out to examine the transcriptional regulation of the mouse PrP gene by studying the promoter region and the first intron. This was done by isolating a 3.5 kb BamHI/KpnI fragment from a murine PrP cosmid clone. This subclone contained 1.2 kb of 5' region to exon 1. This exon was shown to be separated from exon 2 by a 2.3 kb intron. Sequence analysis of the 5' region showed that it was highly G+C rich, contained on TATA box and had potential binding sites for many transcription factors including Sp1, AP1, AP2 and EGR1. In a transient transfection assay this 3.5 kb BamHI / KpnI fragment was shown to act as a promoter by linking it to the bacterial reporter gene - chloramphenicol acetyl transferase and introducing it into neuro2a cells. Subsequent deletions both in the 5' flanking region and in the intron revealed positive regulatory elements in both. The transcriptional start sites of the PrP gene were mapped using the primer extension technique with an oligonucleotide whose target sequence resides in exon 2. These reporter gene constructs will be used to generate transgenic mice and to assess their role, more fully, in the regulation of the PrP gene.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available