How do you build a parasite?

如何构建寄生虫？

• 批准号: 6497397
• 负责人: JOHN M. MURRAY
• 金额: $ 27.18万
• 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/6497397
• 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Gondii无处不在 病原体感染了大约三分之一的美国人口和10-90 全球个人中的百分比（取决于国家，各种 社会学和行为因素等）。该寄生虫复制 细胞内各种细胞类型，并且可以持续多年 潜在（组织）囊肿形式。除了弓形虫，原生动物门 Apicomplexa包括许多其他临床和/或兽医寄生虫 重要性。尽管这些生物引起的疾病有很大差异 自然（例如，将疟疾与弓形虫病或球虫病进行比较）， 所有Apicomplexan寄生虫的致病影响最终都可以归因于 扩散，这使得理解寄生虫复制是重要的 目标。所有apicomplexana通过一个独特的过程复制，其中多个 女儿在母细胞中同时组装（称为“精神分裂症”）。 该应用程序建议探索寄生虫组装的动态 弓形虫，因为（1）T。gondii通常一次仅形成两个寄生虫 对复制形态进行研究比在 疟原虫或碘植物，以及（2）广泛的细胞生物学和 分子遗传工具现在可用于T. gondii。尤其， 现在，荧光蛋白报道允许几乎所有已知的亚细胞 在活寄生虫中可视化的结构，效率 瞬态转染允许快速评估重组质粒功能 （即使是致命的转基因）。成像技术允许分析 随着时间的流逝，各种亚细胞细胞器之间的关系，使用 定量的延时视频显微镜和图像反卷积，激光器 扫描共聚焦显微镜，荧光光漂白和恢复，以及 激光消融。分子遗传方法允许基本上突变 通过随机或靶向方法的任何寄生虫基因，并鉴定 负责的病变。我们旨在威慑 T. gondii复制中的关键事件，因果关系 与这些过程以及所涉及的分子机甲相关。