Cloning and characterization of wheat peptide transporters by functional complementation of yeast mutants

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Project objective

Objectives and content of the project During seed germination in plants, the nitrogen supplied to the growing embryo is mostly in form of peptides transported from the endosperm. These small peptides are a consequence of proteolytic activities present in the seed endosperm. The objective of this project is to study this peptide transport process starting through the cloning and characterization of wheat peptide transporter by functional complementation in well-characterized yeast mutants. This project will let understand how the wheat embryo obtain the nitrogenous compounds, required for growth, from protein storage reserves in the endosperm of the seed. We plan to carry out: 1. Isolation of wheat cDNA clones by functional complementation of yeast peptide transport mutants. 2. Sequence analysis of the cDNA selected. 3. Characterization of wheat peptide transporter in yeast transformants. 4. Site-specific mutagenesis and analysis in yeast transformants. 5. Expression pattern of wheat transporter in wheat. Training content (objective, benefit and expected impact) Molecular cloning of higher plant genes by functional complementation in yeast mutants is a powerful and original tool which allows us to circumvent the problems associated with traditional techniques. We propose, using this approach, the isolation and characterization of wheat peptide transporter genes. The results obtained in this work can yield substancial information regarding the role that they can play in normal plant growth and development which can open new perspectives in the field of biotechnological applications. On the other hand, this is an ambitious project that will allow me to apply, in Dr. Cejudo's laboratory, all the scientific techniques and skill acquired during my postdoctoral stay in France working on Saccharomyces cerevisiae. Links with industry/ industrial relevance This is an industrially relevant proposal because of the important applications of this crop plant. The host institution, particularly the Dr. Cejudo's lab, has ongoing collaboration in the subject of the project with the french company Rhone- Poulenc. This collaboration is financed by the Spain Programme for Collaboration betwen Industry and Public Research Centers .

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Participants

• Aristotle University of Thessaloniki · ThessalonikiCoordinatorGreece

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