STREPTOCOCCAL MULTIENZYME DNA REPLICATION SYSTEM

链球菌多酶 DNA 复制系统

• 批准号: 2715279
• 负责人: VLADIMIR S KERY
• 金额: $ 10.05万
• 依托单位: ENZYCO, INC.
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-08-01 至 1999-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2715279
• 关键词: DNA replication; Streptococcus pyogenes; antibacterial agents; binding proteins; biological products; drug design /synthesis /production; drug screening /evaluation; gene expression; method development; molecular cloning; structural genes

The only carefully studied bacterial replication system, Escherichia. coli, requires 23 proteins for in vitro reconstitution of an authentic replication system. The overall objective of these proposed studies is to establish a complete DNA replication system from an important gram- positive pathogen, Streptococcus pyogenes. We will exploit available genomic information to express the Streptococcus homologs of known replication proteins. we will construct templates containing the Streptococcal replication origin to provide a functional assay for Streptococcal replication factors. Factors not accessible through genomic searches will be isolated by biochemical procedures using their requirement to reconstitute Streptococcus-specific replication. The corresponding structural genes for isolated factors will be identified and expressed. The resulting multicomponent replication system will be developed into a high throughput assay format and, together with a collaborative pharmaceutical partner, existing compound collections will be screened for novel lead compounds that can be developed into anti- bacterials that target the replication apparatus. The goal of this phase I application is to clone four centrally important components: the origin, the origin binding initiator protein, the replicative polymerase and the processivity factor. The protein products will be purified and characterized. The origin will be characterized in terms of its interaction with the origin specific initiator protein. PROPOSED COMMERCIAL APPLICATION: The resulting Streptococcus multiprotein replication system will be used to attract a pharmaceutical company partner to exploit it for high throughput screens for lead compounds for the development of novel anti- bacterials. Given the expanding drug resistance of a number of important gram-positive pathogens, a new antibiotic should achieve commercial viability in the $23 billion antibacterial market.

唯一仔细研究过的细菌复制系统，埃希里希亚。 Coli，需要23个蛋白质才能在体外重建正宗 复制系统。这些拟议研究的总体目标是 从一个重要的基因组建立完整的DNA复制系统- 阳性病原菌，化脓性链球菌。我们将利用现有的资源 表达已知链球菌同源物的基因组信息 复制蛋白。我们将构造包含 链球菌复制来源提供一种功能性检测方法 链球菌复制因子。无法通过访问的因素 基因组搜索将通过生化程序进行分离，使用其 重建链球菌特异性复制的要求。这个 将确定与分离因子相对应的结构基因 并表达了。由此产生的多组件复制系统将是 开发成高通量分析格式，并与 合作制药合作伙伴，现有的化合物收藏将 筛选出可开发成抗癌药物的新型先导化合物 以复制装置为目标的细菌。这一阶段的目标是 I应用程序将克隆四个最重要的组件： 起源，起源结合启动子蛋白，复制聚合酶 以及过程性因素。蛋白质产品将被提纯并 特色化的。起源将从以下几个方面来描述 与来源特定的启动子蛋白相互作用。 建议的商业应用： 由此产生的链球菌多蛋白复制系统将被用于 吸引一家制药公司的合作伙伴利用它来获得高利润 用于开发新型抗氧剂的先导化合物的生产能力筛选 细菌剂。鉴于许多重要的抗药性不断扩大 革兰阳性病原菌，一种新的抗生素应该实现商业化 在230亿美元的抗菌市场上的生存能力。