How do you build a parasite?

描述：(申请人提供)：弓形虫是一种普遍存在的 据估计，三分之一的美国人口和10-90%的人感染病原体 全球个人百分比(取决于国家/地区，不同 社会和行为因素等)。这种寄生虫可以复制 存在于多种细胞类型中，并可持续多年 潜伏的(组织)囊状。除了弓形虫，原生动物门 尖端复合体包括临床和/或兽医的许多其他寄生虫 重要性。尽管由这些微生物引起的疾病在 自然(例如，将疟疾与弓形虫病或球虫病进行比较)， 所有顶端复合体寄生虫的致病影响最终归因于 增殖，这使得了解寄生虫复制变得重要 进球。所有的Apicomplexana都通过一个独特的过程进行复制，在这个过程中 子代同时聚集在母细胞内(称为“分裂生殖”)。 这个应用程序建议探索寄生虫组装的动力学 弓形虫，因为(1)弓形虫通常一次只形成两种寄生虫， 使对复制形态的研究比在 疟原虫或艾美耳球虫物种，以及(2)广泛的细胞生物学和 弓形虫的分子遗传学工具现已问世。特别是， 荧光蛋白记者现在几乎允许所有已知的亚细胞 在活体寄生虫中可视化的结构，以及 瞬时转染法可快速评估重组质粒的功能 (即使是致命的转基因)。成像技术允许分析 不同亚细胞器之间随时间的关系，使用 激光定量时移视频显微镜和图像去卷积 扫描共聚焦显微镜、荧光光漂白和恢复，以及 激光消融。分子遗传学方法允许基本基因突变 任何寄生虫基因的随机或靶向方法，并鉴定 负责的损伤。我们的目标是阻止按时间顺序 弓形虫复制中的关键事件及其因果关系 与这些过程相关联，以及涉及的分子机制。

DESCRIPTION: (Provided by applicant): Toxoplasma gondii is a ubiquitous pathogen infecting an estimated one-third of the US population and 10-90 percent of individuals worldwide (depending on the country, various sociological and behavioral factors, etc). This parasite replicates intracellularly in a wide range of cell types and can persist for years in latent (tissue) cyst form. In addition to Toxoplasma, the protozoan phylum Apicomplexa includes many other parasites of clinical and/or veterinary importance. Although the diseases caused by these organisms differ greatly in nature (compare malaria, for example, with toxoplasmosis, or coccidiosis), the pathogenic impact of all apicomplexan parasites is ultimately attributable to proliferation, which makes understanding parasite replication an important goal. All Apicomplexana replicate by a distinctive process in which multiple daughters assemble simultaneously within the mother cell (termed 'schizogony'). This application proposes to explore the dynamics of parasite assembly in Toxoplasma, because (1) T. gondii normally forms only two parasites at a time, making studies on the morphology of replication much more tractable than in Plasmodium or Eimeria species, and (2) a wide range of cell biological and molecular genetic tools are now available for T. gondii. In particular, fluorescent protein reporters now permit virtually all known subcellular structures to be visualized in living parasites, and the efficiency of transient transfection permits rapid assessment of recombinant plasmid function (even for lethal transgenes). Imaging techniques permit the analysis of relationships between various subcellular organelles over time, using quantitative time-lapse video microscopy and image deconvolution, laser scanning confocal microscopy, fluorescence photobleaching and recovery, and laser ablation. Molecular genetic approaches permit the mutation of essentially any parasite gene by either random or targeted methods, and identification of the lesions responsible. We aim to deterrnine the chronological order of critical events in T. gondii replication, the cause and effect relationships associated with these processes, and the molecular mechamsms involved.