Expression of Recombinant Cellulase Cel5A from Trichoderma reesei in Tobacco Plants

The overall goal of the following experiment is to express target proteins transiently in tobacco to gain an idea about the feasibility of plant-based enzyme expression in a short timeframe. This transient expression is achieved by infiltrating tobacco leaves with a strain of agrobacterium tumor fain, which contains a vector incorporating the gene of interest as a second step After incubation infiltrated plant leaves are harvested and protein extracts are partially purified to obtain a working enzyme solution. Next protein analysis is performed in order to assess the effects of plant-based expression on the enzyme and to verify its enzymatic activity.

Results are obtained that show protein size possible glycosylation effects and enzymatic activity based on western blotting, zy and substrate degradation assays. This method can help answer key questions about plant biomass conversion and can also provide insight into the activity of enzymes after being transiently expressed in tobacco. Visual demonstration of this method is critical as the infiltration steps are difficult to learn due to the leaves being easily damaged by the syringe herger clothes and I will be performing the experiments Under sterile conditions.