Last Updated: 18/06/2024
Elucidation and application of biological molecular basis that characterizes malaria parasite
Objectives
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This study will select gene groups that are highly expressed during the erythroid stage from among 40 candidates, narrow down the list to those that are considered to be particularly important based on information such as location of expression and whether they are essential or non-essential, and conduct detailed functional analysis. Drugs that inhibit the action of antibodies against proteins and the functions of proteins will also be selected. This will help elucidate the biological and molecular basis that characterizes malaria parasites, which will be useful in countermeasures aimed at eradicating malaria. |
It is a protein molecule that is conserved only among Plasmodium species, has a highly conserved amino acid sequence, does not have a characteristic domain from which its function can be inferred, and has not been analyzed in detail so far. Conserved proteins (CPs) are considered to be important molecules involved in the molecular basis that characterizes the malaria parasite. In the first year, the research target was narrowed down to molecule X by selecting molecules whose expression was observed during the erythrocytic stage of malaria parasites from 41 candidate CPs. The possibility that this molecule might localize to the apicoplast was also discovered. In 2021, the second year, 10 candidate molecules were looked for that are predicted to interact with this molecule X, and attempted to construct vectors and express His-Tagged proteins for each of them. Since the malaria parasite gene has a very high AT content and is difficult to express in E. coli, an artificial gene with codons optimized for E. coli was synthesized, cloned into an expression vector (pET15b), and used. However, it is difficult to express the protein in the soluble fraction, and selection of Escherichia coli strains and examination of culture conditions have been repeated. In addition, in order to simplify the analysis of protein interactions, the NanoBit system (Promega) could be used, which can detect interactions by luminescence, and have started constructing the vectors necessary for this. In addition, in order to confirm the localization of molecule X in the protozoa, we started to produce recombinant protozoa by fusing molecule X with a fluorescent protein.
Apr 2020 — Mar 2023
$38,109
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