SILENT GENES FOR STREPTOMYCIN SYNTHESIS IN STREPTOMYCES

链霉菌中链霉素合成的沉默基因

• 批准号: 2612756
• 负责人: ANTHONY C MADU
• 金额: $ 0.3万
• 依托单位: VIRGINIA UNION UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-09-30 至 1998-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2612756
• 关键词: Escherichia coli; SDS polyacrylamide gel electrophoresis; Streptomyces; actinomycin; alleles; antineoplastic antibiotics; bacterial DNA; bacterial genetics; drug design /synthesis /production; enzyme biosynthesis; gene deletion mutation; genetic manipulation; genetic transcription; genetic translation; microorganism metabolism; molecular cloning; northern blottings; nucleic acid sequence; southern blotting

Recently, there has been a growing urgency to develop lasting therapies for several pathologies that have withstood traditional antibiotics. This has given rise to, among others, a greater need to develop more effective antibiotics and anti-neoplastic agents. Coincidentally, this comes at a time when a mounting evidence is accumulating indicating the widespread distribution in Streptomycetes of silent genes for secondary metabolites, and the potential to harness such genes for the production of novel antibiotics. One such group of silent genes consists of those for phenoxazinone synthase (PHS), a key enzyme in the pathway of actinomycin D biosynthesis in Streptomyces antibiotics of silent. This applicant realizes the limited clinical use of actinomycin D as an antineoplastic agent, and proposes to examine the silent homologs of the PHS gene in Streptomycetes, as potential candidates for a genetic modification of the actinomycin biosynthetic pathway to produce clinically more use&l analogs of this antibiotic. The specific aims of the proposed research will therefore include: (i) cloning and characterizing the silent gene(s) for the enzyme phenoxazinone synthase (PHS) from Streptomyces coelicolor, and Streptomyces glaucasence; (ii) elucidating the mechanism of cryptification, and activation, of the gene(s); (iii) isolating the entire cluster of genes for the putative pathway; and (iv) exploring, through molecular intervention, the potential role of these silent alleles in the production of actinomycin or other antineoplastic agents. It is anticipated that the proposed experiments will yield a significant amount of information which will be useful in continued examination of members of this bacterial genus for silent pathways for the synthesis of novel antibiotics. This project will also provide a much-needed opportunity for the training of undergraduate students at the applicant institution, in the area of molecular biology and microbial genetics.

最近，开发持久疗法的紧迫性越来越大 几种能抵抗传统抗生素的疾病这 因此，除其他外，更需要制定更有效的 抗生素和抗肿瘤剂。巧合的是， 当越来越多的证据积累表明广泛的 次级代谢产物沉默基因在链霉菌中的分布， 以及利用这些基因产生新的 抗生素一组这样的沉默基因包括那些 放线菌素代谢途径中的关键酶吩恶嗪酮合酶 D链霉菌生物合成中沉默的抗生素。此申请人 实现了放线菌素D作为抗生素的有限临床应用， 代理，并建议检查沉默的同源物的PHS基因在 链霉菌，作为基因修饰的潜在候选者， 放线菌素生物合成途径产生临床上更多的用途&L类似物 这种抗生素。 因此，拟议研究的具体目标将包括： 克隆和表征酶的沉默基因 来自天蓝色链霉菌的吩恶嗪酮合酶（PHS），和 灰绿链霉菌;（ii）阐明 （iii）分离所述基因的密码子化和激活; 推定途径的整个基因簇;和（iv）探索， 通过分子干预，这些沉默的潜在作用 在放线菌素或其他抗生素生产中的等位基因。 预计拟议的实验将产生重大影响。 这将是有用的，在继续审查的信息量 这种细菌属的成员沉默途径的合成 新型抗生素该项目还将提供急需的 在申请人的本科生的培训机会 在分子生物学和微生物遗传学领域，