Cloning, expression and characterization of the viral proteins

Over the last twenty years three new coronavirus strains have appeared in human population, that cause dangerous infections of respiratory tract. At this moment no approved drug exists, that could mitigate the infection caused by these viruses. One of the possible targets of these drugs are proteins responsible for viral genome replication. Nsp13 is primarily a helicase, that unwinds double stranded nucleic acids in the direction of 5'-3', but it also increases activity of replication and transcription complex, it participates in the synthesis of 5' cap on viral RNA and it probably mediates the backtracking of replication and transcripton complex during the process of RNA repair. On top of that nsp13 is strongly conserved among all coronaviruses and it is possible target not only for COVID-19 treatment, but also infections caused by potential future coronaviruses. Theoretical part of the thesis briefly describes the life cycle of SARS-CoV-2, functions of individual proteins of this virus, that participate in replication and transcription of RNA and it summarizes current knowledge about function of nsp13 and its potential inhibitors. In experimental part we have prepared the expression vector for nsp13, that was used for the production of nsp13. Nsp13 was then isolated and purified. (In Czech) Key..