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 对于经受住传统抗生素的几种病理。这 已经引起了更大的需求，以提高有效性 抗生素和抗塑性剂。巧合的是，这是 越来越多的证据积累表明广泛的时间 沉默基因链霉菌的分布，用于继发代谢产物， 以及利用这种基因生产新颖的潜力 抗生素。这样的一组沉默基因由这些基因组成 苯氧唑合酶（PHS），放线菌素途径中的关键酶 d静音链霉菌抗生素中的生物合成。这个申请人 意识到放线霉素D作为抗肿瘤的临床使用有限 代理，并建议检查pHS基因的无声同源物 链霉菌作为遗传修饰的潜在候选者 放线霉素的生物合成途径，产生更多的临床用途和L类似物 这种抗生素。 因此，拟议研究的具体目的将包括：（i） 克隆和表征酶的无声基因 来自链霉菌的苯氧唑合酶（PHS），并 链霉菌的青光眼； （ii）阐明 基因的致密化和激活； （iii）隔离 推定途径的整个基因簇； （iv）探索， 通过分子干预，这些沉默的潜在作用 放线霉素或其他抗肿瘤剂的等位基因。 预计拟议的实验将产生重要的 信息量将在继续检查中有用 该细菌属的成员用于合成的无声途径 新颖的抗生素。该项目还将提供急需的 在申请人的本科生培训的机会 机构，在分子生物学和微生物遗传学领域。