| Abstract [eng] |
Tissue engineering’s main goal is to restore and improve the functionality of damaged tissues or organs. New advances in this field have created a possibility to grow artificial organs in a lab using artificial extracellular matrix, cells and bioactive compounds. The method still has some issues, the main one being the biological efficiency of the artificial scaffold. The scaffold mimics the native tissue architecture and environment as well as participates in many cellular processes. To fulfil this function labs use extracellular compounds, specifically recombinant proteins. Extracellular proteins have high molecular weight, so their production using commonly used bacterial expression systems is problematic. As an alternative for this method, plant based expression systems are used. Compared to bacterial expression systems, plant based ones have many advantages and so this method is more appealing to use for mass production. The production of extracellular matrix mimicking protein in Nicotiana tabacum and Daucus carota plant expression systems was investigated. Both of these plants are widely used for the production of recombinant proteins. Binary system vector, carrying the gene for the chimeric protein, was constructed and a transgenic Agrobacterium tumefaciens line was created. The bacteria were later used for plant transformation. The transformation of tobacco protoplasts and leaf discs as well as carrot cell suspension and hypocotyls was attempted. The transformation success of the leaf discs, cell suspension and hypocotyls was assessed by using selective growth mediums. The transformation success of carrot hypocotyls was also assessed by doing capillary electrophoresis on their DNA. |
