Acquisition of a 200 kV Cryo-Electron Microscope

描述（由申请人提供）：本提案要求支持购买和安装200 kV低温电子显微镜（Cryo-EM）。Cryo-EM是医学院战略研究计划的关键组成部分，涉及微生物学，医学，艾滋病研究中心和生物防御中心的教师。Cryo-EM允许分析复杂的结构，而没有传统EM中使用的染色方案所引入的伪影。使用图像重建软件，cryo-EM产生复杂结构的3D渲染，包括完整的病毒结构，揭示病毒组装和成熟中涉及的蛋白质/核酸相互作用。几个正在进行的NIH资助的研究项目将通过在UAB校园内使用200 kV cryo-EM而得到显着增强。这些包括确定噬菌体P22成熟的潜在机制（P.Prevelige），分析RNA病毒基因组和病毒衣壳之间的相互作用（A. Ball）、蓝舌病病毒核心的组装（P，Roy）和VSV RNA核衣壳复合物的包装（M. Luo）。噬菌体P2和P4组装的额外研究将由新招募的T.多克兰所有这些项目都集中在多蛋白质复合物的组装机制和在这些复合物中建立核酸-蛋白质相互作用。获得200 kV显微镜是谢尔比研究大楼内开发综合电子显微镜设施计划的关键一步，该设施还将包括300 kV冷冻EM，常规I20 kV透射EM和扫描EM。Cryo-EM设施的负责人将是Terje Dokland博士。该仪器的获得将大大提高UAB科学家的结构生物学能力，他们准备将该技术应用于细胞器和大分子复合物的分析。使用该技术收集的数据将支持结构生物学、病毒学、跨膜通道、生物防御和疫苗开发方面正在进行的高优先级计划。

DESCRIPTION (provided by applicant): This proposal requests support to purchase and install a 200 kV cryo-electron microscope (Cryo-EM). The Cryo-EM is a key component of the strategic research plan of the School of Medicine, involving faculty within the Departments of Microbiology, Medicine, the Center for AIDS Research, and the Center for Bio-defense. Cryo-EM permits analysis of complex structures without the artifacts that are introduced with staining protocols used in conventional EM. Using image reconstruction software, cryo-EM produces 3-D renderings of complex structures, including complete virus structures revealing protein/nucleic acid interactions involved in viral assembly and maturation. Several ongoing NIH-funded research projects will be dramatically enhanced by access to a 200 kV cryo-EM on the UAB campus. These include determination of the mechanisms underlying maturation of the bacteriophage P22 (P. Prevelige), analysis of interactions between an RNA viral genome and the viral capsid (A. Ball), assembly of the bluetongue virus core (P, Roy), and packaging of the VSV RNA nucleocapsid complex (M. Luo). Additional studies of assembly of bacteriophages P2 and P4 will be performed by new recruit T. Dokland. All these projects focus on mechanisms of assembly of multi-protein complexes and establishment of nucleic acid-protein interactions in these complexes. Acquiring a 200 kV microscope is a key step in plans to develop a consolidated electron microscopy facility within the Shelby Research Building that will also include a 300 kV cryo-EM, conventional I20 kV transmission EM and scanning EM. The director of the Cryo-EM facility will be Dr. Terje Dokland. Acquisition of this instrument will dramatically enhance the structural biology capabilities of UAB scientists who are poised to apply this technology to analysis of cellular organelles and macromolecular complexes. Data collected using this technology will support ongoing and high-priority initiatives in structural biology, virology, trans-membrane channels, bio-defense, and vaccine development.