Development of live cell imaging experimental system in babesia parasite

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The system for stable expression of green fluorescent protein (GFP) in babesia parasite was developed and the GFP-expressing parasite was observed through time-lapse video microscopy. (1) Stable expression of GFP in B. bovis using the WR99210/hDHFR selection system was achieved. The expression cassette was applied for the development of gene targeting (knockout) technique and the knockout parasite was established for the first time. (2) The gliding motility of B. bovis merozoites was revealed for the first time by in vitro time-lapse video microscopy of the GFP-expressing parasite. In vivo infection model of the parasite with SCID mice, with which a part of the circulating erythrocyte had been replaced with bovine cells, was also developed to investigate the gliding motility of the parasite in detail through time-lapse video microscopy.