弓形虫能从各类宿主细胞中获取包括鞘脂类脂质在内的各类营养物质。鞘脂类的转运主要受宿主Rab蛋白的调控，尤其是Rab14。抑制宿主Rab14的蛋白表达后，检测到纳虫空泡中的鞘脂类显著减少，提示弓形虫可能通过募集Rab14来获取鞘脂类用于胞内增殖。本研究拟通过基因编辑技术构建rab14四环素诱导条件性缺失Vero细胞系，动态、可调节地研究Rab14对弓形虫获取细胞鞘脂类的作用；通过间接免疫荧光、免疫印迹等方法探究弓形虫感染对Rab14蛋白表达量及时空定位分布的影响；通过过表达或沉默Rab14蛋白，验证Rab14在鞘脂类转运中的功能，并明确其对弓形虫胞内增殖的影响；探索不同种源Rab14蛋白对鞘脂类运输作用的异同，以期解释Rab14蛋白参与弓形虫胞内增殖的相关作用机制，最终为揭示弓形虫摄取营养物质信号转导的分子机制提供新的理论依据，为控制弓形虫的胞内增殖及为弓形虫病的防治提供新的药物靶点和思路.

Toxoplasma gondii is able to salvage various nutritions including sphingolipid from host cells for development. The transportation of sphingolipid inside cells is mainly regulated by the Rab family of proteins, especially the Rab14 protein. Dominant-negative expression of Rab14 protein of host cell results in the significantly decreased sphingolipid transported into parasitophorous vacuole, suggesting T. gondii salvaging sphingolipid for cellular proliferation probably through recruitment Rab14 by parasitophorous vacuole membrane. In order to dynamically study the effect of Rab14 on T. gondii salvaging sphingolipid, we screening the conditional Rab14 knockout Vero cells via gene editing technology. Indirect immunofluorescence and western blot will be used to study the location and expression of Rab14. By overexpressed or negative expressed of Rab14 protein to validation its specific role in sphingolipid transportation, and to explicate the effect of Rab14 on cellular proliferation of T. gondii. In addition, different host cells will be used to explore whether the function of Rab14 isspecies-dependent. Therefore, we may explain the mechanism of Rab14 participating in the cellular proliferation of T. gondii. Thus the development in the discovery of new drug targets against T. gondii, in innovative medicines and in effective vaccines will be made.