Elucidation of survival strategy through parasite membrane in intrahepatic malaria parasite

The original plan was to overexpress EXP1 with the BioID tag in Plasmodium bergii (P. berghei) to comprehensively identify the protozoan parasitic membrane proteins. However, it was decided to change the tag to TurboID and miniTurbo, which can be biotinylated in a shorter time than BioID. This makes it possible to investigate protein-protein interactions over time. Since the protein localized in PVM is considered to change in the prophase, metaphase, and anaphase of infection, it is possible to obtain a more detailed overview of the PVM protein by using TurboID and miniTurbo. First, it was decided to conduct a biotinylation experiment using P. falciparum, which is easier to prepare experimental samples than the endohepatic protozoan. As a result, it is possible to search for the optimum experimental conditions for the biotinification experiment, and the biotinification experiment of the endohepatic protozoan can be performed more smoothly. In the first year, a plasmid was constructed that episomatically overexpresses a fusion protein in which miniTurbo is added to EXP1 of P. falciparum. In addition, since some proteins may show cytotoxicity when overexpressed, the preparation of a construct in which miniTurbo was added to the C-terminal of EXP1 was started at the same time. The plan is to use these constructs to see if the PVM protein identified in previous studies can be identified. In addition, by substituting the EXP1 sequence with another protein coding sequence, it can be used as a new research tool for protein-protein interactions in the red pelvic phase.

Project details

Basic Science

Sep 2020 — Mar 2021

$26,456

Japan