Actinomycetes from deep sea invertebrates: source of novel biosynthetic genes

来自深海无脊椎动物的放线菌：新型生物合成基因的来源

• 批准号: 6300679
• 负责人: AMY Elizabeth WRIGHT
• 金额: $ 11.56万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-03-10 至 2001-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6300679
• 关键词: Actinomycetales; Streptomyces; antineoplastics; bacterial genetics; biological products; combinatorial chemistry; drug discovery /isolation; genetic library; genetic strain; growth media; marine biology; microorganism culture

Cancer remains the second leading cause of death in the United States. Advances in our understanding of neoplastic progression have provided new targets for intervention which can be utilized as smart assays for drug discovery. Natural products or compounds modeled on them have found significant utility in the treatment of cancer. In this proposal we seek to substantiate our hypothesis that actinomycetes which live in association with deep-sea invertebrates have undergone a unique evolution which in turn provided them with unique biosynthetic capabilities. This genetic information can be extracted and recombinant clones which express both the native genome and novel pathways derived from combinatorial biology can be produced. These clones will produce a diverse library of compounds for cancer drug discovery using target based assays. The project will be carried out in collaboration with the Sherman group from the University of Minnesota (Project 2) who will conduct combinatorial biology studies using DNA from HBOI originated isolates and Novartis Pharmaceutical Corporation (Project 3) who will carry out assays on extracts from clones which incorporate actinomycete biosynthetic gene sequences. Active actinomycete derived clones generated by Minnesota will be chemically deconvoluted at HBOI. The specific goals of the HBOI research project will be to: 1. Provide a diverse set of actinomycetes to Project 2 (U. of Minnesota) for cosmid preparation of native genome sequences and combinatorial biology studies. [Years 1-5 respectively: 60, 80, 60, 40, 40 isolates/year] 2. Using selective media/conditions, isolate additional actinomycete strains from deep water marine invertebrates for use in years 2-5 of the project. 3. Ferment active Streptomyces venezuelae clones derived from actinomycete DNA and carry out bioassay guided purification and structure elucidation of the active components.

癌症仍然是美国第二大死因。我们对肿瘤进展的理解的进步为干预提供了新的目标，可以用作药物发现的智能分析。天然产物或以它们为模型的化合物在治疗癌症方面已经发现了重要的效用。在这一建议中，我们试图证实我们的假设，即与深海无脊椎动物生活在一起的放线菌经历了独特的进化，这反过来又为它们提供了独特的生物合成能力。这种遗传信息可以被提取出来，并且可以产生既表达原生基因组又表达来自组合生物学的新途径的重组克隆。这些克隆将产生一个多样化的化合物文库，用于使用基于靶标的检测发现癌症药物。该项目将与明尼苏达大学的Sherman小组（项目2）和诺华制药公司（项目3）合作进行，前者将使用来自HBOI的分离株的DNA进行组合生物学研究，后者将对包含放线菌生物合成基因序列的克隆提取物进行分析。明尼苏达产生的活性放线菌克隆将在HBOI进行化学反卷积。HBOI研究项目的具体目标将是：1。为项目2（美国明尼苏达大学）提供多种放线菌，用于天然基因组序列的合成和组合生物学研究。[1-5年分别为：60、80、60、40、40株/年]使用选择性培养基/条件，从深水海洋无脊椎动物中分离出额外的放线菌菌株，供项目第2-5年使用。3. 从放线菌DNA中提取发酵活性委内瑞拉链霉菌克隆，并进行生物测定指导下的活性成分纯化和结构解析。