CORE--TOXIN PROBES

描述(申请人提供)：该中心自1992年成立以来， 已经安置了一个毒素探测器设施核心。核心提供提纯的毒素 和毒素抗体给中心的调查人员。这个核心暂时是 1998年，随着核心领导和中心的离开而停产 导演丹尼尔·巴登。核心将在佛罗里达州重新建立 并将由凯瑟琳·雷小山博士领导，从 2001年4月1日预算年度。新的核心将提供类似的服务 旧的核心。然而，新的核心被设想为一个重要的 迈阿密大学MFBS中心以前的核心扩展 强调DNA技术。除了中心的支持外，资金正在 通过ARCH(环境高级研究合作)向NIEHS寻求 健康科学)将少数族裔机构(FIU)与 研究机构(UM)。此外，这一设施将提供基础 用于机构之间持续的合作。 培养物将在4L香石竹中生长到最大密度。除了……之外 甲藻研究人员将种植有毒的苔藓植物和 蓝藻。蓝藻是从佛罗里达州分离出来的 Miro Gantar博士和Evelyn Gaiser在FIU的《大沼泽地》，将制作 可供中心成员使用。每一株菌株在任何时候的体积 给出的时间将由中心科学家的需求决定。单元格 通过毒素纯化前的离心法或DNA/RNA法获得 与世隔绝。或者，根据具体情况，文化 将被直接提取为毒素。毒素将通过一种 溶剂萃取、柱层析、制备性TLC和 高效液相色谱柱。提纯后的材料将用400 MHz核磁共振进行分析，并密封储存 安瓶放在上了锁的冰箱里。DNA和RNA将按标准分离 凝胶电泳法定量分光光度法分析方案 为了质量，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。 用于精制毒素质量控制的核心。

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.