QUANTIFYING ACTIVATION DYNAMICS OF SYNTHETIC MAP KINASE CASCADE BY 2P-FLIM

通过 2P-FLIM 量化合成 MAP 激酶级联的激活动力学

• 批准号: 8362578
• 负责人: Casim Sarkar
• 金额: $ 0.07万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-06-01 至 2012-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8362578
• 关键词: Fluorescence; Fluorescence Resonance Energy Transfer; Funding; Grant; Image; Laboratories; MAP Kinase Modules; Methods; National Center for Research Resources; Optics; Phosphotransferases; Photons; Principal Investigator; Process; Publishing; Research; Research Infrastructure; Resources; Saccharomyces cerevisiae; Source; Time; United States National Institutes of Health; Venus; base; cost; novel; sensor

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. We wish to carry out quantitative real-time imaging of the kinase ERK in synthetic mammalian MAP kinase cascade that we have constructed in saccharomyces cerevisiae. Using novel genetically encoded FRET sensors that are specific for ERK, we propose to use 2-photon fluorescence lifetime imaging to quantify its activation dynamics, both temporarily and spatially. The two FRET sensors that we have are based on the fluorescent pairs EGFP-mRFP1 and Cerulean-Venus. We intend to follow the method that was published recently by the group that developed the sensors.

该副本是利用资源的众多研究子项目之一 由NIH/NCRR资助的中心赠款提供。对该子弹的主要支持 而且，副投影的主要研究员可能是其他来源提供的 包括其他NIH来源。 列出的总费用可能 代表subproject使用的中心基础架构的估计量， NCRR赠款不直接向子弹或副本人员提供的直接资金。 我们希望在我们在酿酒酵母中构建的合成哺乳动物MAP激酶级联反应中对激酶ERK进行定量实时成像。使用针对ERK的新型遗传编码的FRET传感器，我们建议使用2光子荧光寿命成像来临时和空间上量化其激活动力学。我们拥有的两个FRET传感器基于荧光对EGFP-MRFP1和Cerulean-Venus。我们打算遵循开发传感器的小组最近发布的方法。