CORE--ANALYTICAL AND BIOMOLECULAR

核心——分析和生物分子

• 批准号: 6564254
• 负责人: CAPDEVILA JORGE H
• 金额: $ 16.26万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-12-01 至 2002-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6564254
• 关键词: arachidonate; biochemistry; biomedical facility; eicosanoid metabolism; gas chromatography mass spectrometry; genetic library; high performance liquid chromatography; immunochemistry; protein purification; radioimmunoassay; recombinant DNA; tissue /cell culture

Intense studies of the chemistry, biochemistry, and physiological significance of the cytochrome P450 branch of the arachidonate cascade has produced significant advances in: a) the number and the importance of biological functions attributed to its metabolites, b) an increased knowledge of metabolite structural diversity, c) the documentation of regio- and stereochemical determinants of product biosynthesis and biological activity, and d) the generation, characterization, and utilization of biospecific probes and recombinant DNA methods for the molecular and functional characterization of these enzymes. In support of Projects 1-6 and, to optimize productive interactions and resources utilization, Core B will apply standard and established methods of eicosanoid extraction, purification, HPLC analysis, GC/MS, LS/MS, protein purification, and recombinant DNA manipulation, to: a) the detection and quantification of eicosanoids in biological samples, b) the storage, and documentation of synthetic standards, c) the storage, and documentation of immunospecific probes, d) the partial purification of recombinant enzymes, and e) the amplification, purification and documentation of cloned cDNAs. The centralization of these routine tasks in Core B will eliminate unnecessary and costly duplications and will provide projects 1-6 with efficient and timely access to synthetic standards, biospecific probes and modern bioanalytical techniques.

对化学、生物化学和生理学的深入研究 花生四烯酸级联的细胞色素 P450 分支的重要性 在以下方面取得了重大进展： a) 生物功能归因于其代谢物，b）增加 代谢物结构多样性的知识，c) 的记录 产品生物合成的区域和立体化学决定因素和 生物活性，以及​​ d) 产生、表征和 利用生物特异性探针和重组DNA方法 这些酶的分子和功能表征。支持 项目 1-6，优化生产性互动和资源 利用，核心 B 将应用标准和既定方法 类二十烷酸提取、纯化、HPLC 分析、GC/MS、LS/MS、蛋白质 纯化和重组 DNA 操作，以：a) 检测和 生物样品中类二十烷酸的定量，b) 储存，以及 合成标准的记录，c) 的存储和记录 免疫特异性探针，d)重组酶的部分纯化， e) 克隆 cDNA 的扩增、纯化和记录。 将这些日常任务集中在核心 B 中将消除 不必要且成本高昂的重复，将为项目 1-6 提供 高效及时地获取合成标准品、生物特异性探针和 现代生物分析技术。