How do you build a parasite?

如何构建寄生虫？

• 批准号: 6628090
• 负责人: JOHN M. MURRAY
• 金额: $ 27.2万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-02-15 至 2004-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6628090
• 关键词: Toxoplasma gondii; fluorescence microscopy; green fluorescent proteins; intracellular parasitism; microorganism reproduction; video microscopy

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.

描述：（由申请人提供）：弓形虫是一种普遍存在的弓形虫。 估计感染三分之一美国人口和 10-90 人的病原体 全世界个人的百分比（取决于国家、各种 社会学和行为因素等）。这种寄生虫会复制 细胞内存在于多种细胞类型中，并且可以在其中持续数年 潜伏（组织）囊肿形式。除弓形虫外，原生动物门 顶复门包括许多其他临床和/或兽医寄生虫 重要性。尽管这些生物体引起的疾病差异很大 性质（例如，将疟疾与弓形虫病或球虫病进行比较） 所有顶复门寄生虫的致病影响最终可归因于 增殖，这使得了解寄生虫复制变得重要 目标。所有 Apicomplexana 都通过独特的过程进行复制，其中多个 女儿们在母细胞内同时聚集（称为“分裂”）。 该应用旨在探索寄生虫组装的动态 弓形虫，因为 (1) 弓形虫通常一次仅形成两种寄生虫， 使复制形态的研究比复制形态的研究更容易处理 疟原虫或艾美耳球虫属物种，以及 (2) 广泛的细胞生物和 现在可用于弓形虫的分子遗传学工具。尤其， 荧光蛋白报告基因现在允许几乎所有已知的亚细胞 活体寄生虫中可视化的结构以及效率 瞬时转染可快速评估重组质粒功能 （即使是致命的转基因）。成像技术可以分析 随着时间的推移，各种亚细胞器之间的关系，使用 定量延时视频显微镜和图像反卷积、激光 扫描共焦显微镜、荧光光漂白和恢复，以及 激光烧蚀。分子遗传学方法允许本质上的突变 通过随机或靶向方法的任何寄生虫基因，以及鉴定 负责的病变。我们的目标是确定时间顺序 弓形虫复制中的关键事件，因果关系 与这些过程相关的以及所涉及的分子机制。