University of Groningen
Morphologic analysis of the apicoplast formation in Plasmodium falciparum
Linzke, Marleen
DOI:
10.33612/diss.107482905
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Publication date: 2019
Link to publication in University of Groningen/UMCG research database
Citation for published version (APA):
Linzke, M. (2019). Morphologic analysis of the apicoplast formation in Plasmodium falciparum. University of Groningen. https://doi.org/10.33612/diss.107482905
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Malaria, caused by Plasmodium spp., remains with more than 200 million cases and more than 400.000 deaths per year one of the most severe diseases worldwide. The steady decrease of malaria cases has stalled in recent years, with stable numbers in the past five years. No highly efficient vaccine is available so far and just a few antimalarial drugs are available on the market. Spreading drug resistance against almost all available drugs has rendered them ineffective in combating the disease. New drugs are urgently needed to prevent a new rise of this severe disease. The apicoplast, a relict plastid of secondary endosymbiosis, is a promising source for new drug targets due to its prokaryotic ancestry and its absence in the mammalian host. The plastid has been shown to be essential to the survival of the Plasmodium parasite. Understanding its biology further will lead to the discovery of novel drug targets which can be exploited for drug discovery in the future. This work is focusing on the mode of replication of the apicoplast of P. falciparum during its erythrocytic life cycle. We focus on the Min system utilised during the division of the apicoplast`s ancestry, the chloroplast. We could identify a possible homologue of one of the main components of this system, the division inhibitor MinD, in the plasmodial genome. To further characterise and prove that this protein is a key part of apicoplast division during replication of the Plasmodium parasite this work had the following objectives:
Cloning of the open reading frame of PfMinD into the expression vector pASK-IBA3 and the transfection vector pARL 1a+
Analysis of the activity and oligomerisation of the recombinant protein including substrate profiling
Mutagenic analysis of important domains by site-directed mutagenesis
Generation of transgenic P. falciparum parasite overexpressing MinD or a mutated version of MinD
Localisation of the protein within P. falciparum using GFP chimeras
Evaluation of the overexpression of MinD on the viability of P. falciparum via transgenic parasites
Mutagenic analysis of the protein by co-transfection to GFP-tagged reference proteins in order to study apicoplast morphology
