BIOSYNTHESIS OF ISOFLAVONOID AND FLAVONOID NUTRIENTS

异黄酮和类黄酮营养素的生物合成

• 批准号: 7610258
• 负责人: Nancy Paiva
• 金额: $ 7.79万
• 依托单位: UNIVERSITY OF OKLAHOMA HLTH SCIENCES CTR
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-05-01 至 2008-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7610258
• 关键词: Alfalfa; Anabolism; Animal Feed; Animals; Anthocyanins; Area; Biological; Biological Availability; Biological Factors; Biomedical Research; Catechin; Chemicals; Computer Retrieval of Information on Scientific Projects Database; Condition; Copper Sulfate; Diet; Environment; Enzymes; Escherichia coli; Factor Analysis; Flavonoids; Funding; Future; Gene Expression; Genes; Glucosides; Goals; Grant; Institution; Measures; Medicago truncatula; Nutrient; Oligonucleotides; Pathway interactions; Pichia; Plants; Proanthocyanidins; RNA; Reagent; Relative (related person); Research; Research Personnel; Resources; Reverse Transcriptase Polymerase Chain Reaction; Sampling; Seedling; Seeds; Source; Testing; Tissues; United States National Institutes of Health; Variant; base; beta Glucosidases; beta-Glucosidase; enzyme pathway; expression cloning; gene discovery; improved; laboratory facility; microbial; mutant; nutrition; research study; vector

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The long term goal of this research is to identify and characterize clones encoding biosynthetic enzymes essential to the accumulation of beneficial natural products in plants. These clones may later be used to analyze factors which effect the accumulation of these compounds in plants, to improve their availability in crop species, or to facilitate biomedical nutrition and bioavailability studies. The main natural product areas are anthocyanins, catechins/condensed tannins, and isoflavonoids. Initially, much effort involved acquiring and propagating necessary biological reagents, including mutant plant lines accumulating visibly different levels of natural products, and re-organizing laboratory facilities from strictly chemical research to a more biomedical research environment. Vectors for microbial over-expression of cloned Medicago truncatula beta-glucosidases active on isoflavonoid glucosides have been constructed, and are being tested in E. coli and Pichia pastoris, with the goal of producing and purifying sufficient enzyme to test in animal feeding studies. Condensed tannins from seeds from alfalfa mutants will be analyzed, for comparison with gene expression analysis. Conditions for induction of isoflavonoid pathway enzymes in M. truncatula seedlings using copper sulfate will be investigated. RT-PCR primers will be used to measure the relative expression levels of known flavonoid and isoflavonoid pathway genes in induced tissues or in tissues from mutant and wild-type plant lines. Satisfactory RNA samples will be probed using 70-mer oligo arrays developed for M. truncatula. In addition to revealing the gene-expression differences conferring the natural product variations in mutant lines, these experiments should facilitate future microarray-based gene discovery or animal diet analysis.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 这项研究的长期目标是识别和表征编码对植物中有益天然产物必不可少的生物合成酶的克隆。 这些克隆后来可用于分析影响这些化合物在植物中积累的因素，以提高其在农作物物种中的可用性或促进生物医学营养和生物利用度研究。 主要的天然产物区域是花青素，儿茶素/凝结的单宁和异黄酮。 最初，大量精力涉及获取和繁殖必要的生物试剂，包括突变的植物线积累明显不同的天然产品，以及将实验室设施从严格的化学研究中重新组织到更生物医学研究环境。 已经构建了活性在异类葡萄糖糖苷上的克隆美食丁香β-葡萄糖苷酶微生物过表达的载体，并正在大肠杆菌和肉芽素食中进行测试，目的是在动物喂养研究中产生和净化足够的酶以产生和净化足够的酶来测试。 将分析来自苜蓿突变体的种子的凝结单宁，以与基因表达分析进行比较。 将研究使用硫酸铜的截骨幼苗中诱导异黄素途径酶的条件。 RT-PCR引物将用于测量诱导组织或突变体和野生型植物系的组织中已知类黄酮和异黄素途径基因的相对表达水平。 将使用为截短开发的70-mer寡核阵列探测令人满意的RNA样品。 除了揭示了赋予突变系的自然产物变化的基因表达差异外，这些实验还应促进未来的基于微阵列的基因发现或动物饮食分析。