NOVEL DRUG TARGETS FOR TOXOPLASMA GONDII

弓形虫的新药物靶点

• 批准号: 2672253
• 负责人: JOSEPH D SCHWARTZMAN
• 金额: $ 22.27万
• 依托单位: DARTMOUTH COLLEGE
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-08-01 至 2000-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2672253
• 关键词: Toxoplasma gondii; actins; cell motility; gene expression; gene mutation; gene targeting; molecular cloning; myosins; protein structure function; transfection

DESCRIPTION: (Adapted from Applicant's Abstract) The long term goal of this proposal is to develop a strategy to interrupt both acute and reactivation toxoplasmosis. This will be done by taking advantage of gliding motility which is a unique mechanism of parasite mobility. Development of pharmacological agents that interfere with T. gondii invasion and intracellular survival will be facilitated by an understanding of the molecular mechanisms effecting gliding motility of zoites. T. gondii myosins will be the major focus of this work as they are the likely primary motor proteins. Cloning will by PCR amplification of myosin from cDNA and genomic libraries using degenerate oligonucleotide primers designed from highly conserved sequences in heterologous myosin genes. Characterization of such myosin genes may add considerable information regarding the functional and regulatory domains of the parasite myosins. Expression of parasite myosin genes will be studied in motile and immotile pathogenic life stages. Parasite-specific anti-myosin antibodies will be produced for morphologic and immunochemical analysis of myosin distribution. Manipulation of T. gondii myosin expression by mechanisms of transfection, insertional mutagenesis, or gene knockout will allow study of myosin regulation and its role in motility and invasion directly. The mechanism of agents affecting motility will be studied by observing changes in the actomyosin cytoskeleton and the resultant deficits in invasion, secretion or signal transduction. Selection of resistance mutants will allow reverse genetics approaches for discovery of the targets of agents affecting motility.

描述：（改编自申请人的摘要） 建议是制定一种急性和重新激活的策略 弓形虫病。 这将通过利用滑行运动来完成 这是寄生虫迁移率的独特机制。 发展 干扰T. gondii入侵和的药理学剂 细胞内生存将通过对 分子机制影响Zoites滑行运动。 T. Gondii 肌球蛋白将成为这项工作的主要重点，因为它们可能是主要的 运动蛋白。 克隆将通过从cDNA和 基因组文库使用由脱名寡核苷酸引物设计的 异源肌球蛋白基因中的高度保守序列。 表征 这样的肌球蛋白基因可能会添加有关 寄生虫肌球蛋白的功能和调节域。 表达 寄生虫肌球蛋白基因将在运动和侵入性致病寿命中进行研究 阶段。 寄生虫特异性抗肌球蛋白抗体将用于 肌球蛋白分布的形态和免疫化学分析。 通过转染机制操纵弓形虫肌球蛋白的表达， 插入诱变或基因敲除将允许研究肌球蛋白 调节及其在运动和入侵中的作用。 机制 影响运动性的代理人将通过观察变化来研究 肌动蛋白细胞骨架和由此产生的入侵，分泌或 信号转导。 选择阻力突变体将允许反向 遗传学方法是发现影响力的靶标的目标 运动。