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Title: A Mira-GFP screen : looking for missing genes in bicoid mRNA localisation
Author: Chang, C-W.
Awarding Body: University of Cambridge
Current Institution: University of Cambridge
Date of Award: 2007
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Abstract:
During Drosophila oogenesis, bicoid mRNA is localised to the anterior of the oocyte, where it is translated after egg deposition to pattern the head. However, only a few genes involved in the bcd mRNA localisation have been identified in classic genetic screens. In this thesis, I describe a genetic screen for suppressors of Miranda-GFP, which was designed to identify missing genes required for the bcd mRNA localisation. Mira is not endogenously expressed during oogenesis. However, when Mira-GFP is expressed in the female germ line, the resulting embryos show a maternal-effect lethal bicaudal phenotype. oskar mRNA is localised to both anterior and posterior ends of these embryos and co-localises with Staufen to both ends of the oocyte. Stau is double-strand RNA binding protein and is required for osk and bcd mRNA localisation during oogenesis and asymmetrical prospero mRNA localisation in neuroblasts. Mira functions as an adaptor to couple Stau/pros mRNA to the basal side of neuroblasts. Mutants in genes required for bcd mRNA localisation, such as exuperantia and swallow, suppress the Mira-GFP bicaudal phenotype. Osk mRNA diffuses from the anterior of these oocytes in a similar way to bcd mRNA, indicating that Mira-GFP couples Stau/osk mRNA to the anterior localisation pathway. I further confirmed this by showing that the anterior localisation of Mira-GFP is microtubule-dependent, like bcd mRNA. In order to find genes required for bcd mRNA localisation, I performed a screen for dominant suppressors of the Mira-GFP bicaudal phenotype. I have identified and characterised a number of interesting mutants from this screen, and have shown that several of them cause defects in bcd mRNA localisation.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID: uk.bl.ethos.597452  DOI: Not available
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