Biological Reagents

摘要 该核心提供支持艾滋病毒/宿主结构和 猎豹中心内的机械学研究。细菌表达系统将成为研究的主力 生产重组蛋白质，而不需要复杂的折叠/翻译后修饰，而 真核表达系统将被用来生产更复杂的蛋白质。均匀糖基化或 将使用先进的组织培养和酶促方法制造脱糖蛋白。1-3膜 蛋白质将通过真核表达和全面洗涤剂筛选相结合来产生 4肽合成将被用于生产配体、抑制剂和蛋白质片段。 更大的合成肽/蛋白质将使用天然的化学连接产生，从而能够引入 5最后，一种自动化的高通量噬菌体展示筛选和 鉴定平台将用于生产用于结晶学的Fab抗体片段 伴侣、冷冻-EM基准物、生物化学/细胞生物学工具，以及稳定/捕获异类蛋白质 用于结构研究的特定构象的络合物6

ABSTRACT This Core provides peptide, protein, antibody and RNA reagents that support HIV/Host structural and mechanistic studies within the CHEETAH Center. Bacterial expression systems will be the workhorse for producing recombinant proteins without complex folding/post-translational modification requirements, while eukaryotic expression systems will be employed to produce more complex proteins. Uniformly glycosylated or deglycosylated proteins will be made using advanced tissue culture and enzymatic approaches.1-3 Membrane proteins will be produced by coupling eukaryotic expression with comprehensive detergent screening protocols.4 Peptide synthesis will be employed for the production of ligands, inhibitors, and protein fragments. Larger synthetic peptides/proteins will be produced using native chemical ligation to enable the introduction of site-specific labels and modifications.5 Finally, an automated high-throughput phage display screening and characterization platform will be used to produce Fab antibody fragments for use as crystallography chaperones, cryo-EM fiducials, biochemistry/cell biology tools, and to stabilize/trap heterogeneous protein complexes in specific conformations for structural studies.6