CORE--TOXIN PROBES

核心--毒素探针

• 批准号: 6587614
• 负责人: KATHLEEN S REIN
• 金额: $ 7.35万
• 依托单位: UNIVERSITY OF MIAMI CORAL GABLES
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-04-22 至 2007-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6587614
• 关键词: Dinoflagellates; biomedical facility; gel electrophoresis; high performance liquid chromatography; marine toxins; microorganism culture; microorganism resource /registry; nuclear magnetic resonance spectroscopy; nucleic acid quantitation /detection; photosynthetic bacteria; protein purification; thin layer chromatography; toxicology registry /referral center

Description (provided by applicant): From its inception in 1992, the Center has housed a Toxin Probes Facility Core. The Core provided purified toxins and toxin antibodies to Center Investigators. This Core was temporarily discontinued in 1998 with the departure of the Core Leader and Center Director, Daniel Baden. The Core will be re-established at Florida International University and will be led by Dr. Kathleen Rein, beginning with the April 1, 2001 budget year. The new Core will provide similar services as the old Core. However, the new Core is envisioned to be a significant expansion of the previous Core at the University of Miami MFBS Center with an emphasis on DNA technology. In addition to Center support, funding is being sought from NIEHS via the ARCH (Advanced Research Cooperation in Environmental Health Sciences) mechanism that pairs a minority institution (FIU) with a research institution (UM). In addition, this facility will provide the basis for ongoing collaborations between institutions. Cultures will be grown in 4L carboys to maximum density. In addition to dinoflagellates the investigators will grow toxic prymesiophytes and cyanobacteria. The cyanobacteria have been isolated from the Florida Everglades by Dr. Miro Gantar and Evelyn Gaiser at FIU and will be made available to Center members. The volume of each individual strain at any given time will be dictated by the needs of the Center scientists. Cells will be harvested either by centrifugation prior to toxin purification or DNA/RNA isolation. Alternatively, depending on the specific circumstance, cultures will be extracted directly for toxin. Toxins will be purified by a combination of solvent extraction, column chromatography, preparative TLC and HPLC. Purified material will be analyzed by 400 MHz NMR and stored in sealed ampoules in a locked freezer. DNA and RNA will be isolated by standard protocols, quantitated spectrophotometrically, analyzed by gel electrophoresis for quality and stored at 80 C. This core will interact with the analytical chemistry and electron microscopy core for quality control of purified toxins.

描述（由申请人提供）：自1992年成立以来，中心 拥有毒素探针设施核心。核心提供纯化的毒素 和毒素抗体给中心研究人员。该核心暂时 1998年随着核心领导人和中心的离开而终止 导演丹尼尔·巴登。核心将在佛罗里达州重建 国际大学，将由 Kathleen Rein 博士领导，首先 2001年4月1日的预算年度。新的核心将提供类似的服务 旧的核心。然而，新的核心预计将是一个重要的 迈阿密大学 MFBS 中心原有核心区的扩建 强调DNA技术。除了中心的支持外，资金还正在 通过 ARCH（环境高级研究合作）向 NIEHS 寻求 健康科学）机制，将少数派机构 (FIU) 与 研究机构（UM）。此外，该设施还将提供基础 促进机构之间的持续合作。 培养物将在 4L 大瓶中生长至最大密度。此外 甲藻 调查人员将种植有毒的原中生植物 蓝细菌。蓝藻已从佛罗里达分离出来 大沼泽地由佛罗里达国际大学的 Miro Gantar 博士和 Evelyn Gaiser 博士制作，将制作 可供中心会员使用。任意应变下每个单独应变的体积 给定的时间将由中心科学家的需求决定。细胞会 毒素纯化前通过离心或 DNA/RNA 收获 隔离。或者，根据具体情况、文化 直接提取毒素。毒素将被净化 溶剂萃取、柱色谱、制备薄层色谱和 高效液相色谱法。纯化的材料将通过 400 MHz NMR 进行分析并储存在密封的容器中 安瓿瓶放在上锁的冰箱中。 DNA和RNA将按标准分离 方案，分光光度法定量，凝胶电泳分析 为保证质量并在 80°C 下保存。 该核心将与分析化学和电子显微镜相互作用 纯化毒素质量控制的核心。