CORE--ANALYTICAL AND BIOMOLECULAR

核心——分析和生物分子

• 批准号: 6105364
• 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: 1999
• 资助国家: 美国
• 起止时间: 1999-02-01 至 1999-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6105364
• 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.

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） 归因于其代谢物的生物学功能，B）增加的 代谢物结构多样性的知识，c） 产物生物合成的区域和立体化学决定因素， 生物活性，和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和，以优化生产性互动和资源 核心B将采用标准和既定的方法， 类花生酸提取，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 定量生物样品中的类二十烷酸，B）储存，和 合成标准品的文件，c）储存和文件 免疫特异性探针，d）重组酶的部分纯化， 和e）克隆cDNA的扩增、纯化和记录。 将这些例行任务集中到核心B中， 不必要的和昂贵的重复，并将提供项目1-6， 有效和及时地获得合成标准品、生物特异性探针， 现代生物分析技术。