BIOSYNTHESIS OF ANTHRACYCLINE ANTIBIOTICS

蒽环类抗生素的生物合成

• 批准号: 3172998
• 负责人: CHARLES R HUTCHINSON
• 金额: $ 14.22万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 1986
• 资助国家: 美国
• 起止时间: 1986-09-01 至 1991-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3172998
• 关键词: Escherichia coli; Streptomyces; anthracyclines; antineoplastic antibiotics; bacterial genetics; biotechnology; biotransformation; endonuclease; enzyme structure; gene expression; gene mutation; genetic recombination; genetic transcription; genetic translation; mass spectrometry; molecular cloning; nuclear magnetic resonance spectroscopy; plasmids; radiotracer; structural genes; thin layer chromatography; tissue /cell culture; tritium; ultracentrifugation

This proposal requests continuation of our study of the biochemistry and genetics of anthracycline antibiotic biosynthesis for a five year period. During this time we will (i) isolate and study the properties of the socalled " aromatic polyketide synthases" and their structural genes, (ii) define the genetic and biochemical organization of anthracycline antibiotic biosynthetic pathways with special emphasis on their regulation at DNA, RNA and protein levels, and (iii) explore the possibilities for expressing Streptomyces genes in other actinomycetes and creating new anthracycline antibiotics by genetic engineering. The major project during this period will be a study of tetracenomycin C, and antitumor antibiotic produced by Streptomyces glaucescens. We will complete the cloning of the tcm genes then dissect the tcm gene cluster by standard techniques individual tcm genes. Their structures will be studied by nucleotide sequencing, S1 mapping, and other methods to locate the regulatory regions and transcriptional/translational start/stop sites. This information and the results of Northern hybridizations will be used to identify the transcriptional units within the tcm gene cluster. The enzymes catalyzing the formation of the aromatic polyketide intermediates of TcmC biosythesis will be overproduced by inserting their genes in suitable expression vectors to facilitate the study of their physical and biochemical properties. In this way we will establish information about the regulation of TcmC biosynthesis at the DNA, RNA and protein levels. Subsidiary projects will involve cloning the genes for the production of elloramycin, an analog of TcmC; using the tcm genes as probes for genes governing related biochemical pathways in Streptomyces; expressing the tcm genes in other actinomycetes; and exploring the possibilities for the construction of hybrid anthracyclines by genetic engineering.

这项建议要求我们继续研究 蒽环类抗生素生物合成的生物化学和遗传学 为期五年。 在此期间，我们将（i）隔离和 研究所谓的“芳香聚酮”的性质 （ii）定义了遗传和结构基因， 蒽环类抗生素生物合成的生化组织 特别强调它们在DNA，RNA 和蛋白质水平，以及（iii）探索表达的可能性 链霉菌基因在其他放线菌和创造新的 蒽环类抗生素基因工程。 这一期间的主要项目将是研究 四环霉素C和由其生产的抗肿瘤抗生素 灰色链霉菌 我们将完成 然后按照标准对中药基因簇进行解剖， 技术个别中医药基因。 它们的结构将被研究 通过核苷酸测序、S1作图和其他方法， 定位调控区域和转录/翻译 启动/停止站点。 这些信息和北方的结果 杂交将用于鉴定转录单位 在TCM基因簇中。 酶催化 TcmC的芳族聚酮中间体的形成 生物合成将通过将它们的基因插入 合适的表达载体，以促进它们的研究。 物理和生物化学性质。 这样我们就能建立 关于TcmC生物合成调节的信息， DNA、RNA和蛋白质水平。 附属项目将包括克隆基因， 生产埃洛霉素，TcmC的类似物;使用tcm 作为控制相关生化途径基因探针的基因 在链霉菌中表达;在其他 放线菌;并探讨建设的可能性 用基因工程技术培育杂交蒽环类抗生素。