UCL Discovery
UCL home » Library Services » Electronic resources » UCL Discovery

Control of B2 bradykinin receptor gene expression

Willer, Elizabeth; (1997) Control of B2 bradykinin receptor gene expression. Doctoral thesis (Ph.D), UCL (University College London). Green open access

[thumbnail of Control_of_B2_bradykinin_recep.pdf] Text
Control_of_B2_bradykinin_recep.pdf

Download (11MB)

Abstract

In order to determine those factors which direct expression of the B2 bradykinin receptor, we have analysed the structure of the gene and its promoter. To characterise the regulatory elements which control B2 bradykinin receptor gene expression, a mouse cosmid clone was isolated containing exon 1 (an untranslated exon), approximately 35kb of sequence upstream as well as approximately 5kb of sequence downstream to transcription initiation site 1. This clone was mapped and a 2.6kb region of upstream sequence, together with transcription initiation site 1 and 31bp of exon 1 was subcloned into a luciferase reporter vector (pGL2_Basic). Transient transfection studies demonstrated that this construct was capable of driving reporter gene expression in neuroblastoma x glioma NG108-15, (a cell line which expresses an endogenous B2 bradykinin receptor). Deletional analysis of this construct demonstrated that a region encompassing 162bp of 5' sequence, transcription initiation site 1, and 31bp of exon 1 contained an active promoter. The presence of two additional downstream transcription initiation sites was shown to have no effect upon reporter gene expression in NG108-15 cells. Electromobility shift assay demonstrated two protein/DNA interactions in the region from-203bp to +31bp using nuclear protein extracted from NG108-15 cells, whereas only a single protein/DNA interaction was shown using nuclear protein extracted from CHO cells (which do not express an endogenous B2 bradykinin receptor). The NG108-15 specific protein/DNA interaction was localised to a 44bp region, 47bp upstream from transcription initiation site 1. The protein/DNA complex running at the same position for both NG108-15 and CHO nuclear protein was localised to a region within 20bp of transcription initiation site 1. Southwestern analysis of the -203bp to +31bp region of the promoter demonstrated two protein/DNA interactions of identical molecular weights (115kDa and 106kDa) using nuclear protein extracted from NG108-15 and CHO cell lines.

Type: Thesis (Doctoral)
Qualification: Ph.D
Title: Control of B2 bradykinin receptor gene expression
Open access status: An open access version is available from UCL Discovery
Language: English
Additional information: Thesis digitised by ProQuest.
Keywords: Health and environmental sciences
URI: https://discovery.ucl.ac.uk/id/eprint/10104971
Downloads since deposit
48Downloads
Download activity - last month
Download activity - last 12 months
Downloads by country - last 12 months

Archive Staff Only

View Item View Item