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In planta production of TMV (Tobacco mosaic virus) nanoparticles of specific length
Dlabalová, Lucie ; Moravec, Tomáš (advisor) ; Šafářová, Dana (referee)
Tobacco mosaic virus (TMV) is one of the most investigated viruses and its attributes and structure are therefore well-known. In this work, we have chosen TMV as a biotemplate for the adjustable-length particles production in plants. The viral RNA and coat protein of TMV self-assemble into particles under physiological conditions. The particle length depends on the length of packaged RNA. The encapsidation signal that is necessary for preferential viral RNA packaging by coat protein disks is known and characterized since the 1980's. In this work, we have proposed a two-component system based on a Nicotiana bentamiana plants infection with packaging competent defective RNA (dRNA) and a helper virus RNA which provides all the components necessary for dRNA replication and packaging. The encapsidation signal in the helper virus sequence was removed to avoid formation of particles of incorrect length. Some of our helper viruses contained a coat protein with modified region of the particle's inner channel. This modification should allow specific binding of metal atoms within the core of the rod shaped particle. Several variants of dRNA and helper viruses were prepared to identify individual areas important for the replication, encapsidation and nanoparticle stability. We focused on the particle formation...
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In planta production of TMV (Tobacco mosaic virus) nanoparticles of specific length
Dlabalová, Lucie ; Moravec, Tomáš (advisor) ; Šafářová, Dana (referee)
Tobacco mosaic virus (TMV) is one of the most investigated viruses and its attributes and structure are therefore well-known. In this work, we have chosen TMV as a biotemplate for the adjustable-length particles production in plants. The viral RNA and coat protein of TMV self-assemble into particles under physiological conditions. The particle length depends on the length of packaged RNA. The encapsidation signal that is necessary for preferential viral RNA packaging by coat protein disks is known and characterized since the 1980's. In this work, we have proposed a two-component system based on a Nicotiana bentamiana plants infection with packaging competent defective RNA (dRNA) and a helper virus RNA which provides all the components necessary for dRNA replication and packaging. The encapsidation signal in the helper virus sequence was removed to avoid formation of particles of incorrect length. Some of our helper viruses contained a coat protein with modified region of the particle's inner channel. This modification should allow specific binding of metal atoms within the core of the rod shaped particle. Several variants of dRNA and helper viruses were prepared to identify individual areas important for the replication, encapsidation and nanoparticle stability. We focused on the particle formation...
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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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