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。discodermolide的临床前研究受到海洋聚酮供应不足的阻碍。它目前可通过分离或全合成获得，但药物的成本非常高。为了能够开发基于发酵的方法，将从从中分离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。I期研究的近期目标是通过追求以下特定目标来鉴定和克隆许多discodermolide生物合成基因：（1）通过PCR分析产生discodermolide的海绵细胞或细胞提取物来检测模块聚酮合酶（PKS）和氨基甲酰基磷酸转移酶（CPT）基因同源物。(2)使用从海绵或其推定的微生物共生体分离的DNA制备fosmid文库，并通过PCR分析鉴定含有这些PKS和CPT基因的克隆。(3)对PKS和CPT基因进行测序，以证明其参与discodermolide生物合成的合理性。我们的研究还将建立一个通用的方法，使许多其他目前稀缺的海洋聚酮具有抗肿瘤活性的药物开发提供大量的技术基础。