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Proceedings Paper

A new red bimolecular fluorescence complementation based on TagRFP
Author(s): Lingsong Qin; Jun Chu; Ying Zheng; Qingming Luo; Shaoqun Zeng; Zhihong Zhang
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Paper Abstract

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/ΔbFos 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.

Paper Details

Date Published: 16 February 2009
PDF: 6 pages
Proc. SPIE 7191, Fluorescence In Vivo Imaging Based on Genetically Engineered Probes: From Living Cells to Whole Body Imaging IV, 71910G (16 February 2009); doi: 10.1117/12.811375
Show Author Affiliations
Lingsong Qin, Huazhong Univ. of Science and Technology (China)
Jun Chu, Huazhong Univ. of Science and Technology (China)
Ying Zheng, Huazhong Univ. of Science and Technology (China)
Qingming Luo, Huazhong Univ. of Science and Technology (China)
Shaoqun Zeng, Huazhong Univ. of Science and Technology (China)
Zhihong Zhang, Huazhong Univ. of Science and Technology (China)


Published in SPIE Proceedings Vol. 7191:
Fluorescence In Vivo Imaging Based on Genetically Engineered Probes: From Living Cells to Whole Body Imaging IV
Alexander P. Savitsky; Yingxiao Wang, Editor(s)

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