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Design and production of TBEV specific cleavage reporter for in vitro and in cell studies
KOTOUNOVÁ, Eliška
Tick-borne encephalitis virus (TBEV) is an important human pathogen and as of now, there are no specific therapeutics to combat this disease. In vitro and in cell models are essential in studying the disease and in the pursuit of novel therapeutics. This thesis deals with the design and production of two genetically encoded cleavage reporters of TBEV NS2B-NS3 protease activity. The reporters were produced in E. coli and tested with recombinant NS2B-NS3 protease, as well as produced in mammalian cell lines and analyzed for fluorescence upon infection with TBEV.
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Mechanism of dsRNA virus replication: cloning, production and structural characterization of N-terminal domain of sigmaNS
KOTOUNOVÁ, Eliška
Avian reovirus is an important pathogen, which causes substantial losses in poultry industry. NS protein is essential for the virus replication, however, the mechanism and assortment of genomic segments during virus replication is yet not well understood. The structure of NS is not known as previous attempts at crystallization yielded non-diffracting crystals due to the flexible regions in the molecule. This thesis deals with the production and purification of the N-terminal domain of NS, which lacks the exposed flexible region.
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