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| Fully Funded Studentship:Enhancing the production of omega-3 polyunsaturates in marine microbes | |||||||||||||
The project will comprise of a number of independent workpackages as follows. The primary focus will be the manipulation of the diatom Thalassiosira pseudonana, which is rich in EPA but contains only low levels of DHA. The Napier group has already published a full characterisation of the T. pseudonana genes required for LC-PUFA biosynthesis (Tonon et al., FEBS J., 2005). WP1. Enhancing the levels of DHA in T. pseudonana. Two additional enzyme activities required for the conversion of EPA to DHA will be introduced in the diatom by biolistic transformation under the control of a constitutive promoter. Transgenic cells will be recovered via selectable markers and evaluated for the presence of altered fatty acid profile. In the case of this and subsequent WPs, the lipid profile will be determined by a state-of-the-art lipidomic analysis present in the Napier group at RRES – this will be performed by the student, who will thus gain valuable experience in mass spectrometry and also the handling of large datasets. WP2. As a reciprocal experiment to WP1, antisense RNA will be used to knockdown the endogenous levels of specific transcripts for components of LC-PUFA biosynthesis in T. pseudonana. Specifically, antisense will be used to silence either the 6-desaturase (which will block the synthesis of EPA and DHA) or the 5-elongase (which will block the synthesis of only DHA. These proposed experiments would define (for the first time) the role of PUFA s in the lifecycle of the diatom. The significance of LC-PUFAs to membrane function and cell survival will be examined by studying the growth of the transgenic cells under different conditions (temperature, CO2, nutrient status) and will be further evaluated by microarray analysis using a custom T. psuedonana chip. WP3. Using a more global approach, we will seek to identify genetic components, which control the synthesis and composition of lipids in T. pseudonana. Specifically, we aim to identify transcription factors (TFs) which modulate omega-3 LC-PUFA biosynthesis and the synthesis of neutral lipids. The endogenous transcript levels of these T. pseudonana TFs will be modulated by transgenic over-expression or antisense knockdown to determine their role in regulation of lipid metabolism, using the approaches outlined above. The focus of this fully funded IBTI Club Studentship will also give the student an excellent introduction into the reality of “Excellence with impact” and how biosciences can deliver benefits to the public. Funding Notes Funding is available for UK/EU students. Funding awarded for this project will cover tuition fees and stipend for UK students. EU students may be eligible for full funding, or tuition fees only, depending on the funding source. International students will not be eligible for this funding however they are still welcome to apply for the project but would have to find alternative funding. Tonon T, Sayanova O, Michaelson LV, Qing R, Harvey D, Larson TR, Li Y, Napier JA, Graham IA (2005) Fatty acid desaturases from the microalga Thalassiosira pseudonana. FEBS J. 2005 Jul;272(13):3401-12. |
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