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首页> 外文会议>Fluorescence in vivo imaging based on genetically engineered probes: From living cells to whole body imaging IV >A new red bimolecular fluorescence complementation based on TagRFP
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A new red bimolecular fluorescence complementation based on TagRFP

机译：基于TagRFP的新型红色双分子荧光互补

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摘要

Fluorescent proteins have become extremely popular tools for in vivo imaging as well as the study of localization, motility and interaction of proteins in living cells. Bimolecular fluorescence complementation (BiFC) analysis based on fluorescent proteins enables direct and high throughput visualization of protein-protein interactions in living cells. Two red Bimolecular Fluorescent Complementation (BiFC) systems based on mRFP variants have been reported. However, some physical-chemical characteristics of mRFP limited their applications, such as low pH-stability, relative low brightness and low maturation rate. We have developed a new red BiFC system based on TagRFP, a novel monomeric red fluorescent protein with high brightness, complete chromophore maturation, prolonged fluorescence lifetime and high pH-stability. In this study, bFos and bJun were used as the positive protein-protein interaction pair, a mutant of bFos (bFos) and bJun were used as the negative protein-protein interaction pair. bFos/△bFos was fused to N-terminal fragment of TagRFP, and bJun was fused to C-terminal fragment of TagRFP. The BiFC systems based on TagRFP was confirmed in living mammalian cells. Furthermore, the BiFC based on TagRFP allow analyzing multi-protein interactions when combined with other BiFC systems. Thus, the BiFC based on TagRFP is very useful for investigating the complicated and significant molecular mechanisms of multi-protein complex in living cells.
机译：荧光蛋白已成为用于体内成像以及研究活细胞中蛋白质的定位，运动性和相互作用的极为流行的工具。基于荧光蛋白质的双分子荧光互补（BiFC）分析可实现活细胞中蛋白质与蛋白质相互作用的直接和高通量可视化。已有两个基于mRFP变体的红色双分子荧光互补（BiFC）系统。但是，mRFP的某些物理化学特性限制了它们的应用，例如低pH稳定性，相对较低的亮度和较低的成熟率。我们已经开发了基于TagRFP的新型红色BiFC系统，该系统是一种新型的单体红色荧光蛋白，具有高亮度，完全的发色团成熟，延长的荧光寿命和高pH稳定性。在这项研究中，bFos和bJun被用作正蛋白-蛋白相互作用对，bFos（bFos）和bJun的突变体被用作负蛋白-蛋白相互作用对。将bFos /△bFos与TagRFP的N末端片段融合，将bJun与TagRFP的C末端片段融合。在活的哺乳动物细胞中证实了基于TagRFP的BiFC系统。此外，基于TagRFP的BiFC与其他BiFC系统结合使用时，可以分析多种蛋白质的相互作用。因此，基于TagRFP的BiFC对于研究活细胞中多蛋白复合物的复杂而重要的分子机制非常有用。

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  • 来源
  • 会议地点 San Jose CA(US)
  • 作者

    Lingsong Qin; Jun Chu; Ying Zheng; Qingming Luo; Shaoqun Zeng; Zhihong Zhang;

  • 作者单位

    Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics, Huazhong University of Science and Technology, P.R.China;

    Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics, Huazhong University of Science and Technology, P.R.China;

    Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics, Huazhong University of Science and Technology, P.R.China;

    Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics, Huazhong University of Science and Technology, P.R.China;

    Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics, Huazhong University of Science and Technology, P.R.China;

    Britton Chance Center for Biomedical Pho;

  • 会议组织
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

    bimolecular fluorescent complementation (BiFC); TagRFP; protein interactions;

    机译：双分子荧光互补（BiFC）; TagRFP;蛋白质相互作用;

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