CORE--GENETIC CROSSES WITH TOXOPLASMA GONDII

核心--与弓形虫的遗传杂交

• 批准号: 6099481
• 负责人: ELMER R PFEFFERKORN
• 金额: --
• 依托单位: PALO ALTO MEDICAL FOUNDATION RES INST
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-12-01 至 1998-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6099481
• 关键词: Toxoplasma gondii; cats; drug resistance; microorganism genetics; molecular cloning; mutant; protozoal cyst; recombinant DNA

Toxoplasmic encephalitis is a major opportunistic infection in AIDS patients. Many of these patients cannot tolerate prolonged administration of pyrimethamine-sulfadiazine. Thus new drugs are needed to treat to prevent this disease. The rational search for improved drug would profit from knowledge as to the mechanism of action of current and future candidate drugs. A useful clue as to mechanism of action can be obtained by studying drug-resistant mutants of Toxoplasma gondii. One way to take advantage of these mutants is to use genetic analysis to identify the gene that confers resistance. Now that is a formal genetic map of T. gondii based on RFLP markers has been established, it is possible to localize drug-resistance genes. This Core Facility will isolate drug-resistant mutants of the PLK strain of T. gondii with the aid of chemical mutagenesis. These mutants will be crossed with a CEP strains of T. gondii that has three unlinked drug-resistance markets. These crosses will be carried out in the definitive host, the cat. Oocysts from the cat feces will be purified and allowed to sporulate. The bradyzoites will be released from sporulated oocysts and grown as tachyzoites in cultured human fibroblasts. Recombinant parasites that inherited drug resistance markers from both parents will be cloned and characterized. The recombinant clones will be sent to Dr. John Boothroyd's laboratory or RFLP analysis.

弓形虫脑炎是艾滋病的主要机会感染 患者。 这些患者中有许多无法忍受延长 丙烯胺 - 磺胺嗪的给药。 因此需要新药 治疗以防止这种疾病。 合理搜索改进的药物 将从知识中获利到当前和当前的作用机理 未来的候选药物。 关于行动机制的有用线索可能是 通过研究弓形虫弓形虫的耐药突变体获得。 一 利用这些突变体的方法是使用遗传分析 确定赋予抗药性的基因。 现在这是一个正式的遗传 已经建立了基于RFLP标记的T. gondii的地图，这是 可以定位耐药基因。 这个核心设施将 分离甲虫的PLK菌株的抗药性突变体与 化学诱变的帮助。 这些突变体将与CEP交叉 T. gondii的菌株具有三个未连接的药物抗药性市场。 这些十字架将在权威的主机猫中进行。 猫粪的卵囊将被纯化并允许孢子散发。 Bradyzoites将从孢子的卵囊中释放出来，并成长为 培养的人成纤维细胞中的tachyzoites。 重组寄生虫 父母双方的遗传抗药性标志物将被克隆，并且 特征。 重组克隆将发送给约翰博士 Boothroyd的实验室或RFLP分析。