Characterization of two membrane transporter genes in the genome of Plasmodium yoeliiGomboc, Pika (Avtor) Keše, Darja (Mentor) Franke-Fayard, Blandine (Komentor) vaccinevaccine developmentinfectious diseasesmalariaPlasmodiumgenetically attenuated parasitesmembrane transportmfs6ctr2biomolecular methodsfluorescent taggingMalaria, an infectious disease caused by parasites from the Plasmodium genus, is one of the leading causes of death and morbidity worldwide and puts almost half of the world population at risk. Due to the emergence and rise of resistance to the insecticides and antimalarial drugs, there is a great need for a vaccine. So far whole sporozoite based immunization has been shown to be the most effective in experimental settings. Genetically attenuated parasites (GAP) have demonstrated significant protection in rodent models of malaria. The ideal GAP would arrest late in the liver stage of the malaria life cycle and provide sterile immunity in the host. However, the route and method of sporozoite administration have not been optimized yet. Intravenous injection (IV) is currently the only effective route of administration, that has been able to induce strong protective immune response. The preferred route is intradermal injection (ID), but several studies have shown that it induces lower immunity compared to IV injection. In order to improve the ID route of administration and determine the suitability of chosen genes as vaccine candidate genes, we aimed to generate mutant parasite lines of two promising membrane transporter genes, mfs6 and ctr2 of P. yoelii. We generated fluorescently tagged and deletion mutants of candidate genes with different biomolecular methods for functional analysis. We also characterized the expression and localization of fluorescently tagged proteins in the blood stage. These lines can be further used to determine expression and localization of tagged genes in the liver stage by infecting hepatocytes in vitro. Deletion mutant parasite lines need to be fully confirmed or further genetically characterized before use for route of administration experiments. Generated transgenic lines can then be used for studying route of administration and generation of GAP vaccine candidates in murine models of malaria.[P. Gomboc]20172017-03-19 07:15:19Magistrsko delo/naloga91091UDK: 616-022.1:616.936:543.9VisID: 130572COBISS_ID: 4765304sl