DNA Replication System from Pseudomonas aeruginosa

铜绿假单胞菌 DNA 复制系统

• 批准号: 6444461
• 负责人: Nebojsa JANJIC
• 金额: $ 10万
• 依托单位: REPLIDYNE, INC.
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-02-01 至 2003-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6444461
• 关键词: DNA replication; Pseudomonas aeruginosa; biotechnology; drug discovery /isolation; drug resistance; enzyme complex; gene expression; high throughput technology; molecular cloning; peptide chemical synthesis; protein engineering; protein purification; replicase

DESCRIPTION (provided by applicant): The ongoing emergence of drug-resistant bacteria is rapidly reducing the efficacy of existing antibiotics leading to increased mortality, morbidity and public health burden due to bacterial infections. Since most drugs in late-stage clinical development are analogs of existing drugs to which resistance is expected to develop rapidly, there is an acute need for novel, mechanistically distinct antibacterials. To date, none of the antibacterials target any of the central components of the DNA replication system of bacteria. The overall aim of this project is to assemble a minimal DNA replication system from Pseudomonas aeruginosa, a clinically important pathogen in which the emergence of multidrug resistance is beginning to limit treatment options. We anticipate that 5 subunits will be minimally required to reconstitute the replicative polymerase capable of rapid and processive DNA synthesis: DnaE (DNA polymerase III subunit), DnaN (sliding clamp processivity factor), DnaX (clamp loader ATPase), HoIA and HoIB (accessory subunits of the clamp loader complex). All of these genes are apparent in the recently completed Pseudomonas aeruginosa genome. We plan to clone, express and purity these five proteins and use them to reconstitute a functional DNA replicase. This multiprotein assembly will then be used in high-throughput screens to identify new drug candidates that inhibit this and other bacterial replication systems. PROPOSED COMMERCIAL APPLICATION: NOT AVAILABLE

描述（由申请人提供）：耐药性的不断出现 细菌正在迅速降低现有抗生素的功效，导致 细菌引起的死亡率、发病率和公共卫生负担增加 感染。由于大多数处于后期临床开发的药物都是类似物 预计耐药性会迅速发展的现有药物 迫切需要新颖的、机制独特的抗菌药物。迄今为止，没有一个 抗菌药物针对 DNA 复制的任何核心成分 细菌系统。该项目的总体目标是组装一个最小的 来自铜绿假单胞菌的 DNA 复制系统，一种临床上重要的 多药耐药性的出现开始受到限制的病原体 治疗方案。我们预计至少需要 5 个子单元 重建能够快速持续进行 DNA 的复制聚合酶 合成：DnaE（DNA 聚合酶 III 亚基）、DnaN（滑动钳持续合成能力） 因子）、DnaX（夹钳装载机 ATP 酶）、HoIA 和 HoIB（ 夹具装载机复合体）。所有这些基因在最近的研究中都很明显 完成铜绿假单胞菌基因组。我们计划克隆、表达和纯化 这五种蛋白质并用它们来重建功能性 DNA 复制酶。 然后，这种多蛋白组装体将用于高通量筛选 确定抑制这种细菌和其他细菌复制的新候选药物 系统。 拟议的商业应用：不可用