SMALL-MOLECULAR COVALENT LABELING OF RECOMBINANT PROTEINS FOR LIGHT AND EM OBS

用于光和 EM OBS 的重组蛋白的小分子共价标记

• 批准号: 7957599
• 负责人: Mark H Ellisman
• 金额: $ 20.26万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-06-15 至 2010-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7957599
• 关键词: Biological Preservation; Computer Retrieval of Information on Scientific Projects Database; Detection; Eels; Funding; Grant; Hybrids; Image Analysis; Institution; Label; Methods; Molecular; Noise; Nucleic Acids; Performance; Proteins; Recombinant Proteins; Reporter; Reporting; Research; Research Personnel; Resources; Signal Transduction; Source; Spectrum Analysis; System; Time; United States National Institutes of Health; protein aggregation; sample fixation

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. During the previous funding period, we have invested considerable effort and time in merging tetracysteine tags with other reporter systems to generate hybrids with enhanced performance for both LM and EM detection. This report covers the progress we made in: (1) Enhancing the sensitivity and the signal/noise ratio of protein detection with biarsenicals. (2) Detecting nucleic acids and interacting proteins in correlated LM/EM studies. (3) Developing a photo-reversible protein aggregation tag for functional studies. (4) Labeling with biarsenicals after fixation. (5) Developing methods for the simultaneous preservation of fluorescent signals, ultrastructure and antigenicity. (6) Developing multiple labeling strategies using EEL spectroscopy.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目及 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 在之前的资助期间，我们投入了大量的精力和时间将四半胱氨酸标签与其他报告系统合并，以生成具有增强的 LM 和 EM 检测性能的杂交体。本报告涵盖了我们在以下方面取得的进展： (1)提高双砷化合物检测蛋白质的灵敏度和信噪比。 (2) 检测相关 LM/EM 研究中的核酸和相互作用蛋白。 (3) 开发用于功能研究的光可逆蛋白质聚集标签。 (4)固定后用双砷标记。 (5) 开发同时保存荧光信号、超微结构和抗原性的方法。 (6) 利用 EEL 光谱开发多种标记策略。