期刊
JOURNAL OF BIOMEDICAL OPTICS
卷 18, 期 12, 页码 -出版社
SPIE-SOC PHOTO-OPTICAL INSTRUMENTATION ENGINEERS
DOI: 10.1117/1.JBO.18.12.126013
关键词
mPlum-infrared fluorescent protein 1.4 fusion protein; Forster resonance energy transfer; near-infrared fluorescence; in vivo imaging
资金
- National Science Council of Taiwan [NSC 100-2627-E-010-001]
- Ministry of Education, Aim for the Top University Plan, National Yang-Ming University
- National Research Program for Biopharmaceuticals (NRPB) at the National Science Council (NSC) of Taiwan
Bacteriophytochrome infrared fluorescent protein (IFP) has a long emission wavelength that is appropriate for detecting pathophysiological effects via near-infrared (NIR) based imaging. However, the brightness and photostability of IFP are suboptimal, although an exogenous supply of biliverdin (BV) IX alpha is able to enhance these properties. In this study, we fused a far red mPlum fluorescent protein to IFP 1.4 via a linker deoxyribonucleic acid (DNA) sequence encoding eight amino acids. The brightness of mPlum-IFP 1.4 fusion protein at the IFP emission channel was comparable to that of native IFP 1.4 protein when fusion protein and IFP 1.4 were excited by 543 and 633 nm using confocal microscopy, respectively. Visualization of IFP 1.4 fluorescence by excitation of mPlum in mPlum-IFP 1.4 fusion protein is likely to be associated with Forster resonance energy transfer (FRET). The FRET phenomenon was also predicted by acceptor photobleaching using confocal microscopy. Furthermore, the expression of mPlum-IFP 1.4 fusion protein could be detected in cell culture and in xenograft tumors in the absence of BV using in vivo imaging system, although the BV was still essential for detecting native IFP 1.4. Therefore, this innovative-fluorescent fusion protein would be useful for NIR-based imaging in vitro and in vivo. (C) 2013 Society of Photo-Optical Instrumentation Engineers (SPIE)
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