Molecular Tools Core

分子工具核心（MTC）的目标是为bioMT教师提供生物分子工具， 更好地实现自己的研究目标。在第一个资助期内，MTC协助了200多名研究人员。 在更大的生物医学界，并在bioMT初级的进步发挥了核心作用 研究项目负责人（RPL）到外部资助的独立调查人员。我们的成功基于 集中设备和专业知识，提供高质量的纯化蛋白质和其他生物分子， 工具，由我们的专家工作人员进行专门的实验室工作，以及对用户进行利用最新技术的培训， 艺术仪器保持在核心内。为了补充我们的教师主任的专业知识，我们招募了 一位经验丰富的首席科学家，负责监督MTC的运作，并提供专家蛋白质 生化支持和咨询。她在蛋白质生产方面的经验对她很有帮助， 顾问，提供从序列分析到表达系统到纯化方案的建议， 有经验的和新手用户。我们已经在E.杆菌和 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），我们已经制定了生化和生物物理表征和QA/QC协议 的最终产品。此外，在第1阶段，MTC创建了一个bioMT服务中心，用于退款 从2020年10月1日起，与真核细胞蛋白质生产相关的用品、培养基和试剂的成本。在 在下一个资助期内，我们会继续为目标1所述的所有四个注册牌照提供宝贵资源。 一个重要的特点是增加了使用昆虫细胞系（Sf 9，Hi5）生产蛋白质的标准方法。 目标二是提高核心业务的效率，精简日常运作， 以更好地服务于研究社区。我们将利用我们收集的资源，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， 在构建体设计、克隆、表达筛选方面的能力，以快速鉴定蛋白质靶点， 减轻bioMT研究人员在蛋白质生产方面的负担。随着MTC的用户数量的增加， 扩大，这是当务之急，我们投资的运作效率的核心，并保持一个强大的用户 在我们向成本分担中心过渡以保证可持续性的过程中，我们第二阶段的另一个关键要素 战略是可持续性。如目标3所述，我们将开发成本回收模式。教师不会被指控 在第二阶段，但我们将与预计成本分摊季度使用量，因此他们可以将成本计入赠款 在第三阶段之前提交。展望未来，MTC还将支持合作试点项目 旨在促成多项目赠款。总体而言，MTC的第二阶段计划将继续促进成功 RPLs和其他bioMT教师，吸引我们的教师在一起，奠定了研究所的长期基础- 过渡到COBRE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.