Protein Production Core

蛋白质生产核心

• 批准号: 9360236
• 负责人: Philip Charles Bourne
• 金额: $ 23.87万
• 依托单位: UNIVERSITY OF OKLAHOMA
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/9360236
• 关键词: Advisory Committees; Affinity; Bacteria; Biological Sciences; Businesses; Cells; Centers of Research Excellence; Chromatography; Collaborations; Communities; Core Facility; Core Protein; Crystallization; Crystallography; Custom; Cytolysis; Development; Educational workshop; Ensure; Equipment; Escherichia coli; Exclusion; Faculty; Fee-for-Service Plans; Fees; Freezing; Funding; Goals; Grant; Hydrophobicity; Income; Individual; Insecta; Ion Exchange; Laboratories; Laboratory Research; Modernization; Molecular; Oklahoma; Performance; Phase; Positioning Attribute; Postdoctoral Fellow; Process; Production; Productivity; Proteins; Protocols documentation; Publications; Research; Research Personnel; Research Support; Role; Sampling; Scientist; Secure; Structural Protein; Students; System; Techniques; Training; Work; Yeasts; base; biophysical properties; experimental study; graduate student; instrument; instrumentation; light scattering; pressure; programs; protein expression; protein function; protein purification; protein structure; senior faculty; structural biology; student training; training opportunity

PROJECT SUMMARY (Protein Production Core) The Protein Production Core (PPC) serves as research support for the Oklahoma COBRE in Structural Biology (OCSB). It is a state-of the art facility for the expression and purification of protein for OCSB-supported researchers and the wider structural biology community. The PPC produces high quality proteins in a reliable and efficient manner on a fee-for-service basis and provides hands-on-training and expert advice in all aspects of protein expression and purification available in the facility. The PPC staff consists of a Director, Dr. Philip Bourne and a Graduate Student training position. The graduate student typically assists in the PPC for a semester. They are trained by the PPC director to use all of the equipment in the facility and assist in the day- to-day activities of the PPC. The PPC is guided by an advisory committee comprised of one junior and two senior faculty researchers. The PPC facility was established using OCSB Phase I funding. Milestones achieved during this period included: (i) renovation of laboratory space to create a stand-alone secure core facility, (ii) establishing the facility with modern instrumentation for protein expression and purification, and (iii) development of a user base. The user base has grown to twenty two research groups, including over 65 individual PPC users. Progress has been demonstrated by evidence of increased productivity of the user base i.e., publications and grant support. The PPC works closely with the investigators and the OCSB-supported Macromolecular Crystallography Laboratory (MCL) to produce homogeneous, highly purified protein for crystallization trials. The PPC is equipped with instrumentation and expertise frequently not found within individual research laboratories. Equipment housed in the PPC includes two refrigerated medium pressure chromatography systems and a large selection of columns for different purification techniques (affinity, hydrophobic, ion-exchange, and size exclusion). Other core equipment includes: an Emulsiflex-C3 homogenizer for lysis of yeast and bacterial cells; a high performance Avanti centrifuge, two freezers (-80 °C and -20 °C), and a BioRad ChemiDoc imager that detects UV, white and chemiluminescence. During the proposed Phase II of the OCSB, planned expansion of the PPC includes the addition of a low pressure chromatography system to ease the burden on the two medium pressure systems and expand the protein purification throughput of the facility. The addition of a small self-contained Dynamic Light Scattering instrument will allow for assessing the quality of samples prior to crystallization trials. This addition is ideal for a multi-user facility where a quick assessment of sample quality prior to crystallization experiments would be extremely useful. The increased protein production capacity and the expanded biophysical characterization will meet the existing demand and will help expand the user base. The larger user base will lead to more income from user fees and this will move the facility closer to self-sufficiency.

项目总结（蛋白质生产核心） 蛋白质生产核心（PPC）作为俄克拉荷马州COBRE结构生物学的研究支持 （公务员事务局）。这是一个国家的最先进的设施，表达和纯化蛋白质的OCSB支持的， 研究人员和更广泛的结构生物学社区。PPC以可靠的方式生产高质量的蛋白质。 以按服务收费的方式提供高效的方式，并提供各方面的实践培训和专家建议 蛋白质表达和纯化的能力。PPC的工作人员包括一名主任，菲利普博士 伯恩和一个研究生培训职位。研究生通常协助PPC为一个 学期他们接受PPC主任的培训，使用设施中的所有设备，并在白天提供协助- PPC的日常活动。PPC由一个咨询委员会指导，该委员会由一名初级和两名 高级研究员。PPC设施是利用OCSB第一阶段资金建立的。里程碑 在此期间取得的成就包括：㈠翻修实验室空间，以建立一个独立的安全核心 设施，（ii）建立具有用于蛋白质表达和纯化的现代仪器的设施，以及（iii） 发展用户群。用户群已经增长到22个研究小组，其中包括超过65个研究小组。 个人PPC用户。有证据表明，用户群的生产力有所提高，这表明取得了进展 也就是说，出版物和赠款支持。 PPC与研究人员和OCSB支持的高分子晶体学密切合作 实验室（MCL）生产均匀，高度纯化的蛋白质结晶试验。PPC是 配备的仪器和专业知识往往没有发现在个别研究实验室。 PPC中的设备包括两个制冷中压色谱系统和一个 多种色谱柱可供选择，适用于不同的纯化技术（亲和、疏水、离子交换和尺寸 排除）。其他核心设备包括：用于酵母菌和细菌裂解的Emulsiflex-C3均质器 细胞;一台高性能Avanti离心机，两台冷冻机（-80 °C和-20 °C）和一台BioRad ChemiDoc成像仪 检测紫外线、白色和化学发光。 在建议的有组织公务员事务局第II期工程期间，计划中的私营保安公司扩建工程包括增设一个低 压力色谱系统，以减轻两个中压系统的负担，并扩大 该设施的蛋白质纯化吞吐量。增加了一个小型的自包含动态光散射 仪器将允许在结晶试验之前评估样品的质量。该添加物非常适合 多用户设施，在结晶实验之前快速评估样品质量， 非常有用增加的蛋白质生产能力和扩大的生物物理特性 将满足现有的需求，并将有助于扩大用户群。更大的用户群将导致更多 这将使该设施更接近自给自足。