Molecular Tools Core

分子工具核心(MTC)的目标是向生物移植教师提供生物分子工具，以允许他们 以更好地实现他们的研究目标。在第一个资助期内，MTC帮助了200多名研究人员 在更大的生物医学社区内，并在生物医学初级培训的发展中发挥了核心作用 研究项目负责人(RPL)到外部资助的独立调查人员。我们的成功是建立在 提供高质量纯化蛋白质和其他生物分子的设备和专业知识的集中化 工具，由我们的专家工作人员专心致志的实验室工作，以及培训用户使用最新的 保持在核心内的艺术仪器。为了补充我们教务主任的专业知识，我们聘请了 一位经验丰富的首席科学家监督MTC的运作并提供专家蛋白质 生物化学支持和咨询。她在蛋白质生产方面的经验对她很有帮助，特别是 顾问，提供从序列分析到表达系统再到纯化方案的建议 有经验和新手的用户。我们已经建立了高效的大肠杆菌和蛋白质生产管道 哺乳动物细胞和优化的纯化方案。与分子相互作用和 成像核心(MIIC)，我们已经开发了用于生化和生物物理表征以及QA/QC的协议 最终产品的价格。此外，在第一阶段，MTC创建了一个按存储容量使用计费的BioMT服务中心 从2020年10月1日起，与真核细胞蛋白生产相关的用品、介质和试剂的成本。在……里面 在下一个资助期内，我们会继续向目标1所详述的所有四个注册助产士提供宝贵的资源。 一个重要的特征将是增加使用昆虫细胞系(Sf9，Hi5)生产蛋白质的标准方法。 目标二是提高核心业务的效率，精简日常业务和整体 吞吐量，以更好地服务于研究界。我们将在我们的资源集合的基础上，包括菌株， 载体、内部生产的酶和树脂。作为我们第二阶段战略的一部分，我们将扩大吞吐量 在构建设计、克隆、表达筛选以快速识别蛋白质靶标方面的能力， 减轻生物MT研究人员对蛋白质生产的负担。由于MTC的用户数量已经 扩大后，我们必须投资于提高核心业务的效率，并保持强大的用户 随着我们过渡到成本分担中心以保证可持续性，我们将建立一个可持续的基础。我们第二阶段的另一个关键要素 战略是可持续发展。正如目标3所述，我们将发展成本回收模式。教职员工不会被起诉 在第二阶段，但我们将与预计成本共享季度使用量，以便它们可以将成本包括在赠款中 在第三阶段之前提交。展望未来，MTC还将支持合作试点项目 旨在形成多项目赠款的核心。总体而言，MTC第二阶段计划将继续促进成功。 将我们的教职员工聚集在一起，为研究所的长期- 过渡到科布雷独立。

The goal of the Molecular Tools Core (MTC) is to provide biomolecular tools to the bioMT faculty to allow them to better achieve their research goals. During the first funding period the MTC assisted over 200 researchers within the greater biomedical community and played a central role in the advancement of the bioMT junior research project leaders (RPL) to externally funded independent investigators. Our success is based on the centralization of the equipment and expertise for providing high-quality purified proteins and other biomolecular tools, dedicated laboratory work by our expert staff, as well as training of users in the utilization of state-of-the- art instrumentation maintained within the core. To complement the expertise of our faculty Director, we recruited a highly experienced Lead Scientist to oversee the operation of the MTC and to provide expert protein biochemistry support and consultation. Her experience in protein production has served her well particularly as a consultant, providing advice from sequence analysis to expression systems to purification protocols to both experienced and novice users. We have established efficient pipelines for protein production in E. coli and mammalian cells and optimized purification schemes. In collaboration with the Molecular Interactions and Imaging Core (MIIC), we have developed protocols for biochemical and biophysical characterization and QA/QC of the final products. Additionally, during phase 1, the MTC created a bioMT Service Center for chargeback costs of supplies, media and reagents related to eukaryotic cell protein production starting October 1, 2020. In the next funding period, we will continue to provide valuable resources to all four RPLs as detailed in Aim 1. One vital feature will be the addition of standard methods for protein production using insect cell lines (Sf9, Hi5). Aim 2 is to enhance the efficient operation of the core, streamlining the day-to-day operations and overall throughput to better serve the research community. We will build on our collection of resources, including strains, vectors, in-house produced enzymes and resins. As part of our phase II strategy, we will expand our throughput capabilities with respect with construct design, cloning, expression screenings to rapidly identify protein targets, reducing the burden of the bioMT investigators on protein production. As the number of users of the MTC has expanded, it is imperative that we invest in the efficiency of the operation of the core and maintain a robust user base as we transition to a cost-share center to guarantee sustainability. Another key element of our phase II strategy is sustainability. As outlined in Aim 3, we will develop cost-recovery models. Faculty will not be charged during phase II, but we will share quarterly usage with projected costs, so they can include costs in grant submissions prior to phase III. Looking towards the future, the MTC will also support collaborative pilot projects designed to nucleate multi-project grants. Overall, the phase II plan for MTC will continue to catalyze the success of RPLs and other bioMT faculty, drawing our faculty together and laying the foundation for the Institute’s long- term transition to COBRE independence.