REGULATION OF ANTIBIOTIC SYNTHESIS IN STREPTOMYCES

链霉菌中抗生素合成的调控

基本信息

批准号：
3137011
负责人：
GEORGE H JONES
金额：
$ 9.76万
依托单位：
UNIVERSITY OF MICHIGAN AT ANN ARBOR
依托单位国家：
美国
项目类别：
财政年份：
1986
资助国家：
美国
起止时间：
1986-12-01 至 1989-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/3137011
关键词：
DNA Streptomyces actinomycin bacterial genetics biotechnology enzyme mechanism gene expression genetic manipulation genetic recombination genetic transcription molecular cloning nucleic acid sequence structural genes

项目摘要

The studies proposed in this application are designed to examine the mechanism and regulation of actinomcyin synthesis in Streptomyces antibioticus. In previous experiments, the structural gene for phenoxazinone synthase (PHS), a key enzyme in the actinomycin biosynthetic pathway was cloned. In the present application it is proposed to: (1) Characterize a recently isolated DNA fragment which overlaps the ends of the PHS gene for sequences reponsible for glucose catabolite repression of PHS synthesis. Techniques of in vitro mutagenesis will be used to identify those sequences: (2) Clone the gene for the enzyme which activates 4-methyl-3-hydroxyanthranilic acid, the precursor of the phenoxazinone chromphore in the actinomycin biosynthetic pathway: (3) Characterize the clones obtained in (2) using molecular biological and biochemical techniques; (4) Examine the possible regulation of gene expression in S. antibioticus by RNA polymerase. For these studies, experiments will be designed to examine the possible specificity of RNA polymerase sigma factors in the transcription of S. antibioticus genes. The proposed studies are significant because they will provide information relating to the regulation of antibiotic synthesis in particular and secondary metabolism in general in an extremely interesting class of organisms. The proposed research will also provide information on gene organization and expression generally in prokaryotes. Finally, the proposed studies may lead to a level of understanding of the mechanisms of antibiotic production which will allow the use of genetic engineering techniques to construct organisms which are capable of hyperproduction of existing antibiotics or the production of new antibiotics. Thus, the proposed studies will have potential applications to industry and to clinical and veterniary medicine.

本应用程序中提出的研究旨在检查阳阳离素合成的机理和调节链霉菌抗生素。在以前的实验中，苯氧唑合酶（PHS）的结构基因，一种关键酶在放线霉素中，生物合成途径被克隆。在目前的应用建议：（1）表征最近孤立的DNA片段与pHS基因的末端重叠对于pHS葡萄糖分解代谢物抑制的序列合成。体外诱变技术将用于识别这些序列：（2）克隆酶的基因激活4-甲基-3-羟基雄性酸（前体）放线菌素生物合成中的苯高氧嗪染色体途径：（3）使用（2）中获得的克隆表征分子生物学和生化技术；（4）检查 RNA可能调节抗生素链球菌中基因表达聚合酶。对于这些研究，实验将设计为检查RNA聚合酶Sigma因子的可能特异性在抗生素基因的转录中。拟议的研究很重要，因为它们将提供与调节抗生素合成有关的信息一般而言有趣的生物类。拟议的研究也将通常提供有关基因组织和表达的信息在原核生物中。最后，拟议的研究可能导致一个水平了解抗生素生产机制这将允许使用基因工程技术构建能够超生产的生物现有的抗生素或新抗生素的产生。因此，拟议的研究将对行业有潜在的应用以及临床和审查医学。