CORE--TOXIN PROBES

描述（由申请人提供）：自1992年成立以来，中心 有一个毒素探测器设施核心内核提供纯化的毒素 和毒素抗体给中心研究者这个核心暂时 1998年，随着核心领导人和中心的离开， 导演，丹尼尔巴登。核心将在佛罗里达重建 国际大学，将由凯瑟琳·赖因博士领导，从 2001年4月1日财政年度。新的核心将提供类似的服务， 旧的核心。然而，新的核心设想是一个重要的 扩建了迈阿密大学MFBS中心的前核心， 强调DNA技术。除了中心的支持外， 环境科学高级研究合作组织（Advanced Research Cooperation in Environmental 健康科学）机制，将少数民族机构（金融情报室）与 研究机构（UM）。此外，该设施将提供基础， 机构之间的持续合作。 培养物将在4 L大玻璃瓶中生长至最大密度。除了 调查人员将种植有毒的prymesiophytes， 蓝藻蓝藻已经从佛罗里达分离出来 大沼泽地由米罗·甘塔尔博士和伊夫林·盖泽在FIU，并将作出 提供给中心成员。每个单独菌株在任何温度下的体积 具体时间取决于中心科学家的需要。细胞将 通过在毒素纯化前离心或DNA/RNA 隔离另外，根据具体情况， 会直接提取毒素毒素将被净化， 溶剂萃取、柱色谱法、制备型TLC和 高效液相色谱法将通过400 MHz NMR分析纯化的材料，并将其密封储存在 把安瓿锁在冰箱里将通过标准品分离DNA和RNA 方案，通过凝胶电泳定量，分析 并在80 ℃下储存。 这个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核心。

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.