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Title: In-vitro studies of the control of renin gene-expression in the mouse
Author: De Souza, Angus Thomas
Awarding Body: University of Leicester
Current Institution: University of Leicester
Date of Award: 1989
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Certain inbred strains of mice can be divided into two groups on the basis of their submandibular gland (SMG) renin activity. High renin producing strains have two genes Ren-1d and Ren-2d, whereas low producers have a single gene Ren-1c. To search for the presence of regulatory elements involved in renin control, approximately 1Kb of both Ren-ld and Ren-2d 5' flanking sequence containing the major transcription start-site P3 were fused to a "G-free" reporter cassette, and the resulting templates transcribed in various nuclear-protein extracts. In rat liver, mouse kidney and mouse testis extracts the Ren-2d promoter was 2-fold more active than the Ren-1d promoter whereas in mouse liver, both promoters were transcribed equally well. Deletion-mapping data are consistent with the presence of a functional negative control element(s) within Ren-1d but not Ren-2d. Polymerase catalysed chain reaction (PCR) generated deletions have shown that a fragment of 62bp, containing only the P3 promoter, was sufficient for accurate in-vitro transcription. Disruption of the P3 TATAAAA resulted in total loss of activity. Deletion of an AP-2 consensus sequence from the Ren-ld promoter had little effect on in-vitro transcription. Furthermore, sequences 3' to the Ren-1d P3 start-site when positioned either upstream or downstream to the Ren-1d promoter/G-free fusion, appeared to have a minimal effect on promoter activity suggesting that these regions are not involved in in-vitro regulation of the Ren-1d gene. Finally, patterns of specific protein-DNA interactions observed with the mobility-shift assays, are also consistent with an overall negative control hypothesis.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available
Keywords: Biochemistry Biochemistry Molecular biology Cytology Genetics