Mining the aspergillus nidulans secondary metabolome

挖掘构巢曲霉次级代谢组

基本信息

批准号：
7813486
负责人：
BERL Ray OAKLEY
金额：
$ 58.96万
依托单位：
UNIVERSITY OF KANSAS LAWRENCE
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-09-30 至 2012-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7813486
关键词：
Acids Aflatoxins Anabolism Antibiotic Resistance Anticholesteremic Agents Appearance Aspergillus Aspergillus nidulans Autonomous Replication Biological Factors Carbon Cell Nucleus Cells Chromatin Chromosomes Cloning Cluster Analysis Collection Communities Complement Complex Cosmid Vector Cosmids DNA DNA Restriction Enzymes Data Decision Making Detection Diagnostic Diagnostic tests Direct Repeats Distant Emericella Engineering Enzymes Equation Escherichia coli Event Excision Fermentation Fibrinogen Figs - dietary Frequencies Funding Gel Gene Cluster Gene Targeting Genes Genetic Genetic Recombination Genome Genomic Library Genomics Goals Grant Growth Homologous Gene Human Individual Institutes Joints Knock-out Learning Length Libraries Ligation Lovastatin Maintenance Maps Mass Spectrum Analysis Methods Mining Molecular Molecular Genetics Molecular Weight Monascus Mutation Natural Products Chemistry Organism Oryza Pathogenicity Pathway interactions Plasmid Cloning Vector Plasmids Probability Procedures Production Program Research Project Grants Property Proteins Publications Publishing Pyrithiamine Reaction Recovery Recycling Regulation Relative (related person)Repression Research Research Personnel Resistance Resources Riboflavin Running Sepharose Side Site Sorting - Cell Movement Source Sterigmatocystin Structure Sucrose System Testing Time Toxic effect United States National Institutes of Health Universities Visual Wisconsin Work Writing base combat density design experience fungal genetics fungus gene replacement genome database genome sequencing homologous recombination improved insight interest liquid chromatography mass spectrometry liquid chromatography mass spectroscopy novel particle pathogen pressure programs promoter public health relevance recombinase research study response sound success tool transcription factor vector volunteer

项目摘要

DESCRIPTION (provided by applicant): This is an application for a competitive revision of Program Project grant (5P01GM084077) entitled "Mining the Aspergillus nidulans Secondary Metabolome," It is written in response to Notice Number NOT-OD-09-058, Notice Title: NIH Announces the Availability of Recovery Act Funds for Competitive Revision Applications. The goal of the Program Project grant as funded is to use a synergistic combination of genomics, molecular genetics and natural products chemistry to mine the Aspergillus nidulans secondary metabolome for useful natural products. Although the grant has been in effect for less than one year, we are making excellent progress and wish to extend this approach to the hundreds of other fungi which likely make medically useful secondary metabolites, but lack the excellent genetic and molecular genetics tools that have been developed for A. nidulans. Since developing molecular genetic systems for these fungi is likely to be difficult and extremely time consuming, we wish, instead, to develop A. nidulans as a system for heterologous expression of secondary metabolite gene clusters from other fungi. In the requested two year period of support, we propose to improve A. nidulans as a host organism for heterologous expression of secondary metabolites by deleting A. nidulans secondary metabolite gene clusters that is expressed at high levels under normal growth conditions. This may allow greater production of target secondary metabolites, and should certainly allow target secondary metabolites to be detected more easily by mass spectroscopy. We will then develop two cosmid-based systems for expression of gene clusters from other fungi. We will develop the system using gene clusters of Aspergillus terreus, an economically and medically important fungus with a sequenced genome, but with a much less well developed set of molecular genetics tools than A. nidulans. We will develop a system for integrating gene clusters from A. terreus into the A. nidulans genome and a system for maintenance of A. terreus gene clusters on autonomously replicating cosmid vectors. We will analyze production of secondary metabolites by liquid chromatography/mass spectroscopy and develop procedures that allow us to obtain production of adequate quantities of secondary metabolites for purification and structural analysis. PUBLIC HEALTH RELEVANCE: Fungal secondary metabolites are a rich source of medically valuable compounds. Molecular genetics approaches in Aspergillus nidulans are providing a very efficient system for identifying new secondary metabolites and deducing the steps of the biosynthetic pathways that produce them. This application will allow us to apply the same approach to other fungi that lack the molecular tools available for A. nidulans.

描述（由申请人提供）：这是对程序项目赠款进行竞争性修订的申请（5P01GM084077），题为“挖掘“挖掘Aspergillus nidulans Secondary Ansabolome”，它是响应通知号而不是09-09-058的通知号，通知标题：NIH通知标题：NIH宣布恢复行动型的竞争力富有竞争力的适用性。作为资助的计划项目赠款的目标是将基因组学，分子遗传学和天然产品化学的协同组合用于有用的天然产物，以挖掘asspergillus nidulans次要代谢组。尽管该赠款的生效不到一年，但我们正在取得良好的进步，并希望将这种方法扩展到可能使医学上有用的二次代谢物的其他真菌扩展到其他真菌，但缺乏为nidulans开发的出色遗传和分子遗传学工具。由于这些真菌为这些真菌开发分子遗传系统可能很困难且耗时，因此我们希望开发A. nidulans作为来自其他真菌的继发代谢物基因簇异源表达的系统。在要求的两年支持期间，我们建议通过删除A. nidulans二级代谢产物基因簇来改善A. nidulans作为宿主生物，以异源表达次级代谢产物。这在正常生长条件下以高水平表达。这可以允许更多的靶标代谢产物产生，并且应允许通过质谱法更容易检测目标二级代谢产物。然后，我们将开发两个基于宇宙的系统，用于表达其他真菌的基因簇。我们将使用Aspergillus terreus的基因簇开发系统，这是一种具有测序基因组的经济和医学上重要的真菌，但与A. nidulans相比，发达的一组分子遗传学工具集要差得多。我们将开发一个系统，用于将基因簇从曲霉的基因组整合到A. nidulans基因组中，并在自主复制宇宙载体上维持曲霉基因簇维护的系统。我们将通过液相色谱/质谱分析二级代谢产物的生产，并开发程序，使我们能够获得足够数量的二级代谢产物进行纯化和结构分析。公共卫生相关性：真菌二级代谢产物是医学上有价值的化合物的丰富来源。曲霉中的分子遗传学方法正在提供一个非常有效的系统，用于鉴定新的二级代谢产物并推断产生它们的生物合成途径的步骤。该应用程序将使我们能够将相同的方法应用于缺乏可用于曲霉的分子工具的其他真菌。