Entwicklung eines plant-based Impfstoffs gegen den Lungenwurm des Rindes

Masterarbeit aus dem Jahr 2013 im Fachbereich Biologie - Genetik / Gentechnologie, Note: 1,0, Gottfried Wilhelm Leibniz Universität Hannover (Pflanzen Genetik), Sprache: Deutsch, Abstract: Genetically modified plants, which are used in the production of industrial materials or§pharmaceuticals instead of feed and food production, are nowadays referred to as§pharmaceutical plants. Due to genetic modification, these pharmaceutical plants§produce important proteins (vaccines or antibodies) for pharmaceutical use as§enzymes for different applications. In order to produce these proteins/enzymes the§coding DNA are incorporated into the genome of plant where the plants will express§the protein to high level producing low cost molecules compared with other§expression systems like cell culture, hypridoma, fermentas. As well as, when the§target proteins has a lethal effect on E.coli hindering the overexpression. In temperate§climates such as Northern Germany the lungworm (Dictyocaulus viviparous) is one of§the economically most important parasites of cattle. This parasite causes parasitic§bronchitis disease, which occurs in varying intensity and in some cases can lead to§death of the infected animals.§It was possible to control a lung worm infection with anthelmintics and even in some§countries with a live vaccine, which consists of X-ray attenuated larvae and pasturage§technical measures. This vaccine can only induce a robust immunity in cattle for a§few months. In addition, the production of these vaccines is very costly and suffers§from limited durability. Therefore, an attempt was made in this work to produce§recombinant proteins in high purity and in large amounts by using tobacco plants and§to use these proteins instead of the previously available immunization as a vaccine§with high efficiency.§First, the vectors were constructed harbouring SOD (superoxide dismutase§pGIIMH35S-SOD vector size approximately 6138 bp). and NMT (N-methyl)§(pGIIMH35S NMT-HIS vector size approximately 6322 bp) since expression of SOD§and NMT showed a toxic effect in E. coli, alternative system is needed like plantAbstract§IV§based expression. Therefore, they were genetically transfered into tobacco plants§using Agrobacterium tumefaciens mediated transformation. Different molecular§biology methods were used to confirm the integration of the genes into gDNA of§tobacco like PCR and RT-PCR using different sets of primers at DNA and RNA level§and functional assays using Basta herbicide and enzymatic assys at protein level.§The desired proteins were detected by western blot and their enzyme activity was§measured. [...]