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Expression and purification of Tick-borne encephalitis virus Capsid protein
SCHMELZER, Jakob
Tick-borne encephalitis virus (TBEV) is an important pathogen of the family Flaviviridae with an emerging health concern. Virus causes severe neurological disorders to human population in Europa and Asia. Despite effective vaccine, there is no specific antiviral treatment till now. The capsid protein encapsules the viral positive-sense single-stranded RNA and is essential in packaging and assembling of new virions within the host. A deeper understanding of these interactions and their implications should lead to new antiviral strategies and drug design. This thesis deals with the expression and purification of the capsid protein without membrane anchor using Escherichia coli expression system.
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Exprese proteinu NS5 viru klíšťové encefalitidy v lidských neurálních buňkách
JAKLOVÁ, Kateřina
This study focuses on the detection of tick-borne encephalitis virus (TBEV) NS5 protein in infected and NS5-transfected DAOY HTB-186 human neural cells. TBEV NS5 protein was shown to localize mainly on the membranes of the endoplasmic reticulum. An interesting finding was also nuclear localization, which is supported by the obtained data from both, confocal microscopy and subcellular fractionation.
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Recombinant production and characterization of protease domain from TBEV nonstructural protein NS3
VEJVODOVÁ, Kateřina
The aim of this thesis was to produce and characterize recombinant protease domain of TBEV nonstructural protein 3 (NS3). The NS3 protein of TBEV is essential in viral replication and plays an indispensable role in cleaving viral polyprotein into structural and nonstructural viral proteins, thus making it an attractive target for antiviral treatment. This thesis summarized the cloning of TBEV NS3 protease,production and purification of recombinant enzyme, biochemical and kinetic characterazion of the protease domain from TBEV NS3.
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Molecular Factors of Cell Antiviral Immunity
SELINGER, Martin
The proposed thesis focuses on the description of flavivirus-host interactions in case of tick-borne encephalitis virus (TBEV) and Zika virus (ZIKV). In more detail, the TBEV-induced host responses in human cells of neural origin and interferon-mediated protection were described together with the identification of a new phenomenon of TBEV-induced host transcriptional and translational shut-off. In addition, virus-derived molecules with hypothetical immunomodulatory characteristics, TuORF and ZIKV sfRNA, were analysed for their presence and possible function during the infection.
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Interaction of tick-borne encephalitis virus with host and vector cells
TYKALOVÁ, Hana
The proposed thesis deals with the various aspects of tick-borne encephalitis virus infection in the host and the vector on the cellular level. It uncovers transcriptomic and proteomic responses in infected cells in the human neurons and astrocytes, and vector cells. It identifies the subgenomic flaviviral RNA as an important pathogenesis effector that can interfere with the vector RNAi pathway, and at the same time denotes the components of this pathway. It also describes the phenomenon of impairment of host protein and rRNA synthesis upon TBEV infection. Moreover, it uncovers the importance of quasispecies in the adaptation to vector and host cells.
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Synthesis of new nucleosides as potential inhibitors of flaviviral replication
Horkelová, Simona ; Nencka, Radim (advisor) ; Janeba, Zlatko (referee)
Viruses of the Flaviviridae family are the causative agents of many dangerous diseases for which we currently have no known cure, and research into new drugs against them therefore represents one of the major challenges for modern medicinal chemistry. Targeting the proteins encoded by viruses is the most common approach to combat them. For flaviviruses, the non- structural protein NS5 exhibiting methyltransferase (MTase) and RNA-dependent-RNA polymerase (RdRp) enzyme activity appears to be one of the most suitable molecular targets. This bachelor thesis deals with the synthesis of new potential drugs capable of inhibition skill of flaviviral RdRp. C-nucleoside analogues were prepared, containing 2 types of heterocyclical base: 3-fluoropicolamide modified in positions 5 or 6 and pyrido[3,2-d]pyrimidine-4-amine modified in positions 6 or 7 using aryl or heteroaromatic substituent Key words: C-nucleosides, polymerase, flaviviruses, Tick-borne encephalitis virus, Suzuki reaction, Grignard reaction
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Antiviral vaccination - tick borne encephalitis virus
Solarová, Pavlína ; Drda Morávková, Alena (advisor) ; Janštová, Vanda (referee)
Tick borne encephalitis is by one of the most severe disease of central nervous system. Agent of this disease is tick-borne encephalitis virus, which is transmitted mainly by tick. Clinical picture of TBE is considerably different and varied in individuals - from quite infection without symptoms up to severe cerebro-spinal meningitis, in such cases attacked persons by this virus leave hospital with whole-life handicap. We can prevent this disease by inoculation. We can see it in the neighboring Austria, where TBE almost became extinct because 90% of population is vaccinated. Vaccines are commercially available from 1979. Nevertheless only 17% population is vaccinated against TBEV. The reason for it could be fact that insurance companies do not provide the vaccines as well as not very effective health education. One of possibilities how we can this tendency reverse would be higher awareness of pupils at schools. In school educational programs there is contained prevention of viral disease, especially in general biology, biology of viruses and education to health.
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