Development of live cell imaging experimental system in babesia parasite

建立了绿色荧光蛋白(GFP)在巴贝斯虫中稳定表达的体系，并用时移视频显微镜观察了表达GFP的寄生虫。(1)利用WR99210/hDHFR筛选系统实现了绿色荧光蛋白在牛黄杆菌中的稳定表达。表达盒被应用于基因打靶(敲除)技术的发展，并首次建立了敲除寄生虫。(2)表达绿色荧光蛋白的牛黄杆菌裂殖子的体外时移视频显微镜首次显示了其滑动运动能力。建立了用牛细胞代替部分循环红细胞的SCID小鼠体内感染模型，用时间推移视频显微镜详细观察了寄生虫的滑行运动。

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.