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）：从1992年开始，该中心 已经拥有毒素探针设施核心。提供的核心纯净毒素 和毒素抗体的中心研究者。这个核心是暂时的 随着核心领导和中心的离开，1998年停产 导演丹尼尔·巴登（Daniel Baden）。核心将在佛罗里达重新建立 国际大学，并将由凯瑟琳·雷恩（Kathleen Rein）博士领导 2001年4月1日预算年。新核心将提供类似的服务 旧核心。但是，新核心被认为是重要的 在迈阿密大学MFBS中心扩展以前的核心 强调DNA技术。除了中心支持之外，资金还在 通过拱门寻求NIEHS（环境的高级研究合作 健康科学）机制，将少数族裔机构（FIU）与 研究机构（UM）。此外，该设施将提供基础 进行机构之间的持续合作。 培养物将在4升carboys中生长至最大密度。此外 鞭毛鞭毛研究人员将种植有毒的prysiophytes和 蓝细菌。蓝细菌已从佛罗里达州隔离 Miro Gantar博士和FIU的Evelyn Gaiser博士将被制作 可用于中心成员。每个分数的体积 给定的时间将由中心科学家的需求决定。细胞会 在毒素纯化之前通过离心或DNA/RNA收获 隔离。或者，根据特定情况，文化 将直接用于毒素。毒素将通过 溶剂提取，色谱柱色谱法，制备TLC和 HPLC。纯化的材料将通过400 MHz NMR分析并存储在密封中 安培在锁定的冰箱中。 DNA和RNA将通过标准分离 通过凝胶电泳分析的分光光度法规程，分光光度法 质量并存储在80 C中。 该核心将与分析化学和电子显微镜相互作用 纯化毒素的质量控制的核心。