运输应激影响弓形虫速殖子与缓殖子转化的分子机制

弓形虫感染的特点是机会性致病，其生物学基础是速殖子和缓殖子的相互转换。这正是目前弓形虫病研究的热点之一，但相关研究大多从宿主本身免疫状态出发，对应激之后虫体转化过程中蛋白本身的变化研究则很少，而弄清这些不明之处是阻止缓殖子向速殖子转化的基础，更是寻找和开发转化阻断剂和抗弓形虫药物的前提。运输应激是动物弓形虫病诱发的主要原因之一。为此本研究拟在本室建立的弓形虫速殖子与缓殖子转化的运输应激动物模型的基础上，利用双向电泳技术分析在不同应激时间速殖子和缓殖子蛋白的时空表达情况，筛选出转化过程中的差异表达蛋白并进行质谱分析；针对这些蛋白应用TALENs靶向基因敲除技术构建基因缺失和互补虫株，研究相关蛋白在虫体生长、入侵、速殖子与缓殖子转化时的功能；利用免疫共沉淀技术寻找与差异蛋白互作的上下游蛋白，推测其可能的作用途径，从而为阐明速殖子与缓殖子转化的分子机理和抗弓形虫药物的筛选和开发提供理论依据。

Toxoplasma gondii is an opportunistic pathogen. The conversion between tachyzoite and bradyzoite is the character of toxoplasmosis and one of the research hotspot. Although many efforts have been made in the host innate immunity, the study on parasite proteins during conversion stage caused by stress is seriously lacked. To identify such proteins and clarify their functions is the basis of preventing T. gondii from conversion and precondition for the exploration of the blocking conversion agents and drugs against T. gondii. The aim of this study is to screen the toxoplasma proteins expressed under transport stress using two-dimensional electrophoresis in the animal model of transport stress which was established by our laboratory. The knockout and recovered toxoplasma strains will be constructed using TALENs target knockout technology and the functions of the identified proteins in the parasite invasion, replication and conversion between tachyzoite and bradyzoite will be evaluated. The study also attempts to find the parasite proteins interacted with the identified proteins using co-immunoprecipitation and to predict the potential functions. The results of this study will provide foundation for exploring drugs against T. gondii and elucidating the mechanism of conversion between tachyzoite and bradyzoite.