Molecular Biology Core

项目摘要/摘要 在第一和第二阶段，微生物致病和宿主炎症反应中心(CMPHIR) 建立和维护研究和技术进步核心，以填补 由UAMS医学院维持的机构核心中可用的资源。其中之一就是DNA 测序核心。然而，这个核心的能力仅限于单基因测序。虽然这是一次 重要和必要的服务，但这是不够的，特别是在基因组和下一代的时代 测序技术。因此，该中心还投入巨资，以增加 现有的DNA测序核心。我们完成了这项任务，在第三阶段，我们建议将这些 在分子生物学核心(核心B)的共同保护伞下提供的服务，能够提供CMPHIR和 其他UAMS调查人员提供他们需要的资源，特别是关于UAMS的病原体方面 寄主-病原体方程。核心B的活动包括提高获得高质量RNA的能力， 用于分析的DNA和蛋白质样品.使用常规Sanger的高通量单基因测序 方法；微生物基因组和基因组文库的下一代测序；蛋白质表达和 纯化；和抗体生产。拟议的核心B将通过CMPHIR的组合得到支持 资金和持续的机构支持。日常运作将由主任乔恩·S·布莱文斯博士监督， 谁将与2名全职技术人员一起工作。其中一名技术人员将对DNA负有主要责任 另一个人将主要负责核心B部分的蛋白质部分。为了确保 一致性和促进自我可持续性，核心B仪器将由这些技术人员代表操作 CMPHIR和UAMS调查人员按服务收费。这将使我们能够恢复一个重要的 成本的比例，从而确保升级和维护该核心设备的能力 在我们进入第三阶段及以后的过程中需要这样做。

PROJECT SUMMARY/ABSTRACT In Phases I and II, the Center for Microbial Pathogenesis and Host Inflammatory Responses (CMPHIR) established and maintained a Research and Technical Advancement Core to fill experimental gaps in the resources available in institutional cores maintained by the UAMS College of Medicine. One of these was a DNA Sequencing Core. However, the capability of this core was limited to single-gene sequencing. While this was an important and necessary service, it was not sufficient, particularly in an era of genomic and next-generation sequencing technology. Thus, the CMPHIR also invested heavily in augmenting the resources available in the existing DNA Sequencing Core. We accomplished this task, and, in Phase III, we propose to integrate these services under the common umbrella of a Molecular Biology Core (Core B) capable of providing CMPHIR and other UAMS investigators with the resources they need, particularly with respect to the pathogen side of the host–pathogen equation. The activities of Core B include enhancing the capability of obtaining high-quality RNA, DNA, and protein samples for analysis; high-throughput single-gene sequencing using conventional Sanger methods; next-generation sequencing of microbial genomes and genomic libraries; protein expression and purification; and antibody production. The proposed Core B will be supported through a combination of CMPHIR funding and continued institutional support. Daily operations will be supervised by Dr. Jon S. Blevins, Director, who will work with 2 full-time technicians. One of these technicians will have primary responsibility for the DNA component of Core B, while the other will have primary responsibility for the protein component. To ensure consistency and promote self-sustainability, Core B instruments will be operated by these technicians on behalf of CMPHIR and UAMS investigators on a fee-for-service basis. This will allow us to recover a significant proportion of the costs and thereby ensure the ability to upgrade and maintain the equipment in this core as needed as we move forward into Phase III and beyond.