Center for Structural Biology of HIV RNA

核心2.结晶学 摘要 CRNA晶体核心将与CRNA研究人员合作，解决RNA的晶体结构-- 蛋白质复合体、RNA和蛋白质。核心的一项关键活动是提供分子生物学方面的专业知识 为RNA和RNA-蛋白质复合体量身定做，包括结构设计、表达测试和开发 纯化方案，以及纯化的大分子和络合物的生物物理表征。这些 分子生物学和生物物理学方面的贡献不仅对核心的结晶学焦点至关重要， 但也可用于CRNA项目，用于蛋白质或RNA-蛋白质的生产和表征 复合体。核心还将提供构建设计、表达质粒和方案以及纯化 蛋白质，RNA和RNA-蛋白质复合体到CRNA实验室。密歇根大学的核心实验室将为 作为材料和协议的储存库。核心团队由PIS珍妮特·史密斯(密歇根大学)共同领导 和张劲伟(NIDDK，NIH)，他带来了设计RNA和RNA-蛋白质复合体的专业知识 结晶。核心团队将专注于三个目标。核心将开发或调整尖端方法，以 RNA设计促进表达，RNA-蛋白质共表达，RNA的结晶伴侣，以及 体外RNA合成能力的建立。为了支持其结晶学使命，核心将 开发先进的方法来帮助RNA结晶，提高衍射质量和解决物相 含RNA晶体的问题，提高含RNA结构的结晶学精确度，以及 以促进结晶学和电子显微镜的结合。

Core 2. Crystallography Summary The CRNA Crystallography Core will collaborate with CRNA investigators to solve crystal structures of RNA- protein complexes, RNAs and proteins. A key activity of the Core is to provide expertise in molecular biology tailored for RNAs and RNA-protein complexes, including construct design, expression testing, and development of purification protocols, and in biophysical characterization of purified macromolecules and complexes. These contributions in molecular biology and biophysics are not only critical to the crystallography focus of the Core, but are also available to CRNA Projects as needed for production and characterization of proteins or RNA-protein complexes. The Core will also provide construct designs, expression plasmids and protocols as well as purified proteins, RNAs and RNA-protein complexes to CRNA labs. The Core lab at the University of Michigan will serve as a repository for materials and protocols. The Core team is co-led by PIs Janet Smith (University of Michigan) and Jinwei Zhang (NIDDK, NIH), who brings expertise in the design of RNAs and RNA-protein complexes for crystallization. The Core team will focus on three aims. The Core will develop or adapt cutting-edge methods for RNA designs to facilitate expression, RNA-protein co-expression, crystallization chaperones for RNA, and establishment of an in vitro RNA synthesis capability. In support of its crystallography mission, the Core will develop cutting-edge methods to aid RNA crystallization, to improve the diffraction quality and solve the phase problem for RNA-containing crystals, to improve crystallographic refinement for RNA-containing structures, and to facilitate the combination of crystallography and electron microscopy.