Discodermolide Biosynthesis Genes

Discodermolide 生物合成基因

• 批准号: 6549229
• 负责人: CHARLES R HUTCHINSON
• 金额: $ 10万
• 依托单位: KOSAN BIOSCIENCES, INC.
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-07-01 至 2004-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6549229
• 关键词: Porifera; antineoplastics; biological products; carbamoylphosphate synthase; drug design /synthesis /production; fermentation; gene expression; immunosuppressive; lactones; molecular cloning; nucleic acid sequence; polyketide synthase; polymerase chain reaction; protein biosynthesis; technology /technique development

DESCRIPTION (provided by applicant): The long-range goal of our research is to develop an economical method for producing the experimental anti-tumor drug, (+) -discodermolide, by microbiological fermentation. Pre-clinical investigation of discodermolide has been hampered by inadequate supply of this marine polyketide. It currently is available by isolation or total synthesis, but the cost of the drug is very high. To enable development of a fermentation based process, the discodermolide biosynthetic genes will be cloned from the marine sponge from which discodermolide is isolated, followed by expression of these genes in a bacterial host. The immediate goal of Phase I research is to identify and clone many of the discodermolide biosynthesis genes by pursuing the following specific aims: (1) Detect modular polyketide synthase (PKS) and carbamoyl phosphate transferase (CPT) gene homologs by PCR analysis of cells or cell extracts of the marine sponge that produces discodermolide. (2) Prepare a fosmid library using DNA isolated from the sponge or its putative microbial symbiont and by PCR analysis identify clones that contain these PKS and CPT genes. (3) Sequence the PKS and CPT genes to the extent required to justify their involvement in discodermolide biosynthesis. Our research will also establish the technological base for a general approach to making many other currently scarce marine polyketides with anti-tumor activity available in large amounts for drug development.

描述（由申请人提供）：我们研究的长期目标是开发一种通过微生物发酵生产实验性抗肿瘤药物（+）-discodermolide的经济方法。由于海洋聚酮化合物供应不足，discodermolide 的临床前研究受到阻碍。目前可通过分离或全合成获得，但该药物的成本非常高。为了开发基于发酵的工艺，将从分离 discodermolide 的海绵中克隆 discodermolide 生物合成基因，然后在细菌宿主中表达这些基因。第一阶段研究的直接目标是通过追求以下具体目标来鉴定和克隆许多 discodermolide 生物合成基因：（1）通过细胞或细胞的 PCR 分析检测模块化聚酮合酶（PKS）和氨基甲酰磷酸转移酶（CPT）基因同源物。产生 discodermolide 的海洋海绵的细胞提取物。 (2) 使用从海绵或其假定的微生物共生体中分离的 DNA 制备 fosmid 文库，并通过 PCR 分析鉴定包含这些 PKS 和 CPT 基因的克隆。 (3) 对 PKS 和 CPT 基因进行测序，以证明它们参与 discodermolide 生物合成的合理性。我们的研究还将为通用方法奠定技术基础，以大量生产许多其他目前稀缺的具有抗肿瘤活性的海洋聚酮化合物，用于药物开发。