一种具潜力的弓形虫新型蛋白TgDOC2.1功能及作用机制的全面研究

Toxoplasma gondii is widely distributed and infection of humans and animals can have very serious consequence. Currently, we still lack of an ideal drug to combat T. gondii's infection, due to its complex life cycle. In order to clarify the molecular mechanisms of Toxoplasma pathogenicity and to completely prevent T. gondii's infecting as soon as possible, we are going to study a great potential protein of T. gondii, which was named as TgDOC2.1. This kind of protein was first discovered in mutants F-P2 of T. gondii. by Andrew Farrell et al, and and proved to play an important role in Toxoplasma gondii invasion of host cells and exocytosis proliferation, and its homologous protein is also present in P. falciparum.We intend to analyze the gene and predicted the structure and function of the TgDOC2.1 protein of T. gondii by bioinformatics method ,and to use X-ray crystal diffraction, NMR, immunofluorescence, immunoelectron microscopy, Yeast two-hybrid system ,RNAi, GST pull-down, knockout, chromatin immunoprecipitation assay (CHIP) and other technology to study the protein comprehensively, to explore the biological function and manners of the TgDOC2.1 protein in T. gondii, to study the signal transduction pathway it mediated. To provide experimental basis with the molecular mechanisms of Toxoplasma pathogenic and to offer theoretical basis for design and select new prevention and treatment programs for toxoplasmosis.

弓形虫的生活史复杂，目前尚缺乏理想的药物来防治弓形虫感染。为及早阐明弓形虫致病的分子机制，彻底地防治弓形虫感染人畜，我们筛选出一种极具潜力的弓形虫蛋白，称为TgDOC2.1。该蛋白首先由Andrew Farrell等在弓形虫F-P2突变株发现，并被证明在弓形虫入侵宿主细胞及胞吐增殖方面具有重要作用，且其同源体蛋白还存在于恶性疟原虫。我们拟通过生物信息学等方法分析预测该蛋白的结构和功能，并通过X射线晶体衍射、免疫荧光、免疫电镜、酵母双杂交、RNAi、GST pull-down、knockout、染色质免疫沉淀等技术对该蛋白进行全方位的研究，探索其在弓形虫体内具体的生物学功能、发挥作用的方式、介导的信号转导通路等，为阐明弓形虫致病的分子机制提供实验依据，为弓形虫病新防治方案的设计与选择奠定理论基础。

弓形虫的生活史复杂，目前尚缺乏理想的药物来防治弓形虫感染。为及早阐明弓形虫致病的分子机制，彻底地防治弓形虫感染人畜，我们筛选出一种极具潜力的弓形虫蛋白，称为TgDOC2.1。该蛋白首先由Andrew Farrell等在弓形虫F-P2突变株发现，并被证明在弓形虫入侵宿主细胞及胞吐增殖方面具有重要作用，且其同源体蛋白还存在于恶性疟原虫。在本研究中我们通过生物信息学等方法分析预测该蛋白的结构和功能。成功提取弓形虫RH株的总RNA并反转录成cDNA。实验中克隆出TgDOC2.1并成功构建了此基因的真核重组表达质粒。为阐明弓形虫致病的分子机制提供了实验依据，为弓形虫病新防治方案的设计与选择奠定理论基础。