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The isolation and characterisation of murine CDP-diacylglycerol synthase genes, Cds1 and Cds2.

Inglis, S.L.; (2004) The isolation and characterisation of murine CDP-diacylglycerol synthase genes, Cds1 and Cds2. Doctoral thesis , University of London. Green open access

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Abstract

Eye specific CDP-diacylglycerol synthase is a key regulator in the phototransduction pathway of the fruit fly Drosophila melanogaster. The enzyme is responsible for maintaining regeneration of phosphatidylinositol 4, 5 bisphosphate (PIP2) which is required for normal vision. The Drosophila eye-cds mutants display light dependent retinal degeneration. Mammalian homologues of Drosophila mutants such as eyeless and eyes absent have been associated with mammalian disease. Based on these studies and the discovery of CDS expression in human retina, this gene would appear to be a good candidate for causing a retinal phenotype in mammals. The aim of the work presented here has been to isolate and characterise the murine homologues of eye-cds. To this end both cDNA and genomic clones have been isolated using a combination of traditional molecular biology techniques and a bioinformatic approach. Using the identified clones, the structures of Cdsl and Cds2 have been confirmed. The clones were subsequently used to characterise the genes. In this thesis I have identified the expression pattern of both genes, showing that Cds2 is ubiquitously expressed in both the embryonic and adult mouse whereas Cdsl expression is more specific to the adult eye and brain. In addition, Cdsl has been shown to be expressed only in the rod photoreceptor cells within the retina. Cdsl and Cds2 clones were also used to map the genes to chromosome 5E1 and 2G1 respectively. The remit of this thesis was further expanded to include the potential function of the murine genes. To this end, transgenic constructs of both genes were made for the rescue of the mutant Drosophila phenotype and a construct was prepared as a first step in the generation of a Cds2 null mouse. In summary, the study reported in this thesis has succeeded in isolating and characterising the murine Cdsl and Cds2 genes. The expression pattern of the two genes has been determined both in a large panel of tissues and within specific cell types. The chromosomal location of both genes has been determined and constructs made for use in developing transgenic mice and flies for the study of gene function.

Type: Thesis (Doctoral)
Title: The isolation and characterisation of murine CDP-diacylglycerol synthase genes, Cds1 and Cds2.
Identifier: PQ ETD:602538
Open access status: An open access version is available from UCL Discovery
Language: English
Additional information: Thesis digitised by Proquest Third party copyright material has been removed from the e-thesis.
UCL classification: ?? IOE ??
URI: http://discovery.ucl.ac.uk/id/eprint/1446613
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