A new red bimolecular fluorescence complementation based on TagRFP

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

doi:10.1117/12.811375

16 February 2009 A new red bimolecular fluorescence complementation based on TagRFP

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

Proceedings Volume 7191, Fluorescence In Vivo Imaging Based on Genetically Engineered Probes: From Living Cells to Whole Body Imaging IV; 71910G (2009) https://doi.org/10.1117/12.811375
Event: SPIE BiOS, 2009, San Jose, California, United States

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.

Citation Download Citation

Lingsong Qin, Jun Chu, Ying Zheng, Qingming Luo, Shaoqun Zeng, and Zhihong Zhang "A new red bimolecular fluorescence complementation based on TagRFP", Proc. SPIE 7191, Fluorescence In Vivo Imaging Based on Genetically Engineered Probes: From Living Cells to Whole Body Imaging IV, 71910G (16 February 2009); https://doi.org/10.1117/12.811375

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