Plant extensions have been identified as possible substitutes for bovine collagen which could be dangerous because of BSE development in EC. Extension and related genes play an important role in plant development similar to collagen in mammals. Furthermore, proteins coming from plants are well faced by potetntial customers in food industries, pharmaceutical and cosmetic companies. Molecular pharming is probably one of the most innovative technology to produce molecules of interest around the 21st century. Use and transfer of genes has opened a new and exciting field for generating in mass new bioactive molecules. The most important bottlenek in development and commercialization of natural products coming from plant is to acquire necessary large quantities of one molecule. Genetic engineering can offer this goal. Genetic transformation provides a means of introducing gentically engineered gene, preliminary isolated and cloned from various crops, into plant or yeast, and increases the quantity of extension gene pool expression. Methods for plant transformation (Agrobacterium, protoplast, biolistic gun, virus vector system) have to be evaluated in function of plant hosts preliminary chosen in accordance with the goal to over express a molecule of approximatelu 33 kDa. The other possiblity to over express a protein is to clone the cDNA of interest in a yeast vector and to tranform the host yeast. When a transformant has been chosen, optimization of expression is realized under controlled conditions in fermentor (up to 3000litres). Isolated extension cDNAs will be chosen in function of their homologies with bovine and human collagen cDNAs. These steps of isolation and cloning of cDNA will be the start of the project. The second step will be gentic engineering and selection of best transformants (transformation of plants or yeasts, regeneration of plant, selection of transformants). Next step will be optimization of expression and development of techniques to extract and to purify extension. This ambitious project needs competence in molecular biolgy, plant transformation, fermentation and purification methods.
Funding SchemeEAW - Exploratory awards