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：（申请人提供）：弓形虫是一种普遍存在的 据估计，这种病原体感染了美国三分之一的人口， 全球个人的百分比（取决于国家，各种 社会学和行为因素等）。这种寄生虫复制 在细胞内广泛存在于各种细胞类型中，并且可以持续数年， 潜伏（组织）囊肿形式。除了弓形虫，原生动物门 顶复门包括临床和/或兽医学上的许多其它寄生虫。 重要性虽然这些生物体引起的疾病在 自然（比较疟疾，例如，与弓形虫病或球虫病）， 所有顶复门寄生虫的致病性影响最终归因于 繁殖，这使得了解寄生虫复制变得重要 目标.所有顶复虫都通过一种独特的过程进行复制， 子细胞在母细胞内同时聚集（称为“配子体”）。 本申请提出探索寄生虫组装的动力学， 弓形虫，因为（1）T.刚地通常一次只形成两种寄生虫， 使复制形态学的研究比 疟原虫或艾美耳球虫物种，和（2）广泛的细胞生物学和 分子遗传学工具现在可用于T.刚地。特别是， 荧光蛋白报告分子现在允许几乎所有已知的亚细胞 在活的寄生虫中可视化的结构，以及 瞬时转染允许快速评估重组质粒功能 (even用于致死转基因）。成像技术允许分析 各种亚细胞器之间的关系随着时间的推移，使用 定量时间推移视频显微镜和图像解卷积，激光 扫描共聚焦显微镜，荧光光漂白和恢复，和 激光烧蚀分子遗传学方法允许基本上 任何寄生虫基因通过随机或有针对性的方法，并确定 造成的损伤我们的目标是确定时间顺序 关键事件T.弓形虫复制，因果关系 与这些过程相关的，以及所涉及的分子机制。