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Title: Shaw potassium channel genes in Drosophila
Author: Hodge, J. J. L.
Awarding Body: University of Cambridge
Current Institution: University of Cambridge
Date of Award: 2001
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Potassium (K+) channels shape the electrical activities of cells by changing the frequency and duration of action potentials and by setting the resting potential. The huge diversity of K+ channels is conserved across species and is thought to be required to uniquely customise both the active excitable and passive properties of different cell types. When this study began, Drosophila voltage-gated K+ currents were thought to be encoded by the four cloned members of this family Shaker, Shab, Shaw and Shal. Mutant analysis had only been performed with Shaker. In order to investigate the in vivo function of another member of the family, a number of methods of making Shaw mutants were used. A more accurate genomic structure and location of Shaw was determined, and I attempted to identify P-element inserts in Shaw. However all the candidate inserts identified were located too far from Shaw to be of practical use for generating mutations affecting Shaw. A dominant negative strategy to remove Shaw function was therefore performed. PCR mutagenesis was used to generate both epitope-tagged full-length and truncated mutant versions of Shaw. Additional control constructs were also made: a full-length wildtype Shaw and a dominant negative version of Shaker. Transgenic lines were generated containing the constructs whose expression was GAL4 inducible. Expression of mutant Shaw, using a number of GAL4 lines, caused an ether-sensitive leg-shaking phenotype.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available