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Possibilities of synthesis of human glycoproteins in legumes seeds
Ploužková, Tereza ; Ryšlavá, Helena (advisor) ; Vaněk, Ondřej (referee)
The expression of recombinant proteins in seeds is especially advantageous due to the high concentration of storage proteins in a small volume. Recombinant proteins accumulate in seeds at high levels, remain stable for many years, and because of the low content of alkaloids and other substances in seed environment they can be easily extracted. There is also a minimal risk of contamination. This bachelor thesis deals with the testing of transient expression of gene constructs directly in maturing soybean seeds. In this work, we have have prepared gene constructs for strong transient expression of the uidA gene in plant tissues. The impact of the acetosyringone and dithiothreitol additives and the impact of their combinations on the transient expression and activity of the uidA gene have been studied as well. Furthermore, the optimal length and environment of sonication have been studied. Finally, the effectiveness of transient expression by in vitro and in vivo has also been compared. It has been confirmed that acetosyringone has a positive effect only on seeds that had not been previously sonicated. With prolonged sonication, the relative activity of the uidA gene decreases. Furthermore, the positive effect of dithiothreitol on seeds that had not been sonicated has been found. Higher values of the...
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The use of plants for the expression of Human papillomavirus vaccine
Dlabalová, Lucie ; Moravec, Tomáš (advisor) ; Fischer, Lukáš (referee)
Papillomaviruses are causing various diseases from skin warts to the lesions leading to malignant tumours and are widespread among people. For this reason, the current research is trying to develop methods for the production of inexpensive and effective vaccines against both Papillomaviruses and against all other infectious diseases. Currently animal and microbial expression systems are most frequently used for the production of biopharmaceuticals which have several drawbacks and their capacity is limited. This opens up the doors for plants - potentially very efficient producers of biopharmaceuticals. Currently there is rapid development towards the optimization and improvement of the results of plant expression systems and establishing the best and safest methods of their use. This paper summarizes and compares the advantages and disadvantages of different methods of plant transformation, leading either to stable production of the protein of interest in transgenic plants or to transient expression of recombinant virus infecting non-transgenic plants. Furthermore it analyzes the most appropriate plant species, which provide high yields combined with a transformation method and ease of cultivation, describes few basic ways of optimizing expression levels and outlines the future of plant expression systems.
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A comparison of efficacy of direct and indirect methods of genetic transformation of potato (Solanum tuberosum L.)
PŘIBYLOVÁ, Marie
Potato is one of the main targets for genetic improvement by gene transfer. The aim of this study was to compare the efficacy of genetic transformation of potato, cultivar Bintje, using two methods: Agrobacterium tumefaciens mediated transformation and microprojectile bombardment. The same plasmid p35SGUSint, which cosists of 35S CaMV promoter, gus and nptII genes, was used for both transformations of internodal potato explants. Kamamycin selection, transient and stable expressions of {$\beta$}-glucuronidase and PCR amplification of gus and nptII transgenes were used for transgenic plant selection, identification and analysis.
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