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Double expression systems with respect to parasitic protozoa
Gromyko, Anastasiia ; Verner, Zdeněk (advisor) ; Kraeva, Natalia (referee)
Protozoan parasite infections continue to pose a significant health challenge in developing countries, resulting in hundreds of thousands of deaths each year. These parasites exhibit a complex multi-stage life cycle and possess unique cellular structures. However, many of their biological processes remain poorly understood. Multigene expression is a promising approach to address this knowledge gap, as it enables the expression of functional protein complexes in vivo, the addition of fluorescent protein tags for visualization of protein localization within the cell, and the study of protein-protein interactions. This bachelor's thesis reviews the current knowledge on available systems and approaches for studying key model parasitic protozoan species. Keywords: expression systems, Trypanosoma brucei, Leishmania tarentolae, Entamoeba histolytica, Giardia intestinalis, Trichomonas vaginalis, Toxoplasma gondii, Plasmodium falciparum, pET-Duet, CRISPR-Cas9
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Cellular Protein Transport and Its Role in Patogenesis
Najdrová, Vladimíra ; Doležal, Pavel (advisor) ; Uzlíková, Magdalena (referee)
The main topic of this thesis are the protein secretion processes in several important human parasites - Toxoplasma gondii, Plasmodium falciparum, Trypanosoma cruzi, Leishmania spp. and Giardia intestinalis. Described here are the parasite's and the host proteins which participate in the pathogenic processes involving the protein secretion. As shown here, the protein secretion into the host environment is one of key tools serving the parasite to survive within and manipulate the host organism. Interestingly, different parasitic organisms use functionally and evolutionary distinct strategies to fulfill this aim. Key words secretory pathway, translocon, signal sequence, Toxoplasma gongii, Plasmodium falciparum, Trypanosoma cruzi, Leishmania spp., Giardia intestinalis
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Cellular Protein Transport and Its Role in Patogenesis
Najdrová, Vladimíra ; Doležal, Pavel (advisor) ; Uzlíková, Magdalena (referee)
The main topic of this thesis are the protein secretion processes in several important human parasites - Toxoplasma gondii, Plasmodium falciparum, Trypanosoma cruzi, Leishmania spp. and Giardia intestinalis. Described here are the parasite's and the host proteins which participate in the pathogenic processes involving the protein secretion. As shown here, the protein secretion into the host environment is one of key tools serving the parasite to survive within and manipulate the host organism. Interestingly, different parasitic organisms use functionally and evolutionary distinct strategies to fulfill this aim. Key words secretory pathway, translocon, signal sequence, Toxoplasma gongii, Plasmodium falciparum, Trypanosoma cruzi, Leishmania spp., Giardia intestinalis
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Phenotyping of a glutamate dehydrogenase a null mutant of \kur{Plasmodium falciparum}
PERNER, Jan
Glutamate dehydrogenase a (GDHa) has been suggested as a potential drug target against the malaria parasite Plasmodium falciparum. GDHa knockout cell line was generated and needed a phenotypic description by means of molecular biology and biochemistry. The knockout cell line was tested for higher oxidative stress sensitivity, levels of relevant proteins and gene transcripts were quantified. Furthermore, concentrations of two key molecules enabling redox homeostasis, glutathione and NADPH, were attempted to quantify. Finally, we attempted to disrupt a gene of another glutamate dehydrogenase, gdhb, which did not result in formation of viable parasites. In conclusion, GDHa is not a suitable drug target and GDHb needs to be further elucidated.
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