| Summary: | With sequencing projects identifying novel genes at a great rate, innovative techniques are required to confer information onto these genes in a high throughput manner. Several such techniques are at present being applied to this challenge. The eukaryotic cell is highly compartmentalized and structured, because of this the localisation of a protein is fundamentally linked to its function. Knowledge of the subcellular localisation of a protein can yield information on a protein’s function and on possible protein-protein interactions. In this thesis I describe the development of a multi-system screening methodology for the identification of the subcellular localisation of many individual proteins in a high throughput manner in <i>Drosophila melanogaster.</i> Genes are cloned using a recombinase based cloning system into fluorescently tagged expression vectors for use in cell culture. The candidates are then further analysed in whole animals by generating transgenic organisms which express a fluorescently tagged version of the protein and by <i>in situ </i>hybridization. The localisation data is then combined with data from other sources, such as bioinformatics and RNAi screens, to ascribe functional information onto the gene. In this thesis I concentrated my efforts on screening cytoskeletal proteins for a role in cellular polarity. More specifically proteins were screened for a role in cellular polarity within asymmetrically dividing neural precursors of the developing CNS. Several genes were implicated in this process, for example a possible role for pUf68 in regulating cellular proliferation within the CNS is discussed. In addition, information was discovered for several other genes in processes outside of CNS development. The methodology I describe is amenable to automation and as such is capable of being scaled up to a whole genome level.
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