An improved fluorescent protein-based expression reporter system that utilizes bioluminescence resonance energy transfer and peptide-assisted complementation

2020 ◽  
Vol 56 (25) ◽  
pp. 3625-3628 ◽  
Author(s):  
Taishi Kakizuka ◽  
Akira Takai ◽  
Keiko Yoshizawa ◽  
Yasushi Okada ◽  
Tomonobu M Watanabe
Keyword(s):  
Gene Expression ◽  
Energy Transfer ◽  
Fluorescent Protein ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Bioluminescence Resonance Energy Transfer ◽  
Reporter System ◽  
Resonant Energy ◽  
Maturation Delay ◽  
Resonant Energy Transfer

A peptide-assisted complementation strategy with screened peptides has succeeded to eliminate maturation delay of a bioluminescence resonant energy transfer reporter for gene expression monitoring.

scholarly journals Enhanced brightness of bacterial luciferase by bioluminescence resonance energy transfer

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Tomomi Kaku ◽  
Kazunori Sugiura ◽  
Tetsuyuki Entani ◽  
Kenji Osabe ◽  
Takeharu Nagai
Keyword(s):  
Energy Transfer ◽  
Mammalian Cells ◽  
Fluorescent Protein ◽  
Color Change ◽  
Yellow Fluorescent Protein ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Bioluminescence Resonance Energy Transfer ◽  
Bacterial Luciferase ◽  
Bacterial Bioluminescence

AbstractUsing the lux operon (luxCDABE) of bacterial bioluminescence system as an autonomous luminous reporter has been demonstrated in bacteria, plant and mammalian cells. However, applications of bacterial bioluminescence-based imaging have been limited because of its low brightness. Here, we engineered the bacterial luciferase (heterodimer of luxA and luxB) by fusion with Venus, a bright variant of yellow fluorescent protein, to induce bioluminescence resonance energy transfer (BRET). By using decanal as an externally added substrate, color change and ten-times enhancement of brightness was achieved in Escherichia coli when circularly permuted Venus was fused to the C-terminus of luxB. Expression of the Venus-fused luciferase in human embryonic kidney cell lines (HEK293T) or in Nicotiana benthamiana leaves together with the substrate biosynthesis-related genes (luxC, luxD and luxE) enhanced the autonomous bioluminescence. We believe the improved luciferase will forge the way towards the potential development of autobioluminescent reporter system allowing spatiotemporal imaging in live cells.

A simple strategy based on upconversion nanoparticles for a fluorescent resonant energy transfer biosensor

2015 ◽  
Vol 3 (3) ◽  
pp. 458-464 ◽  
Author(s):  
Hao Zhu ◽  
Yujie Ding ◽  
Anqi Wang ◽  
Xu Sun ◽  
Xing-Cai Wu ◽  
...  
Keyword(s):  
Energy Transfer ◽  
Fluorescence Resonance Energy Transfer ◽  
Dna Detection ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Upconversion Nanoparticles ◽  
Resonant Energy ◽  
Simple Strategy ◽  
Resonant Energy Transfer ◽  
Fluorescence Resonance

A novel biosensor was fabricated for lysozyme and DNA detection based on fluorescence resonance energy transfer between UCNPs and TAMRA-aptamer.

scholarly journals Unmodified Prolactin (PRL) and S179D PRL-Initiated Bioluminescence Resonance Energy Transfer between Homo- and Hetero-Pairs of Long and Short Human PRL Receptors in Living Human Cells

2005 ◽  
Vol 19 (5) ◽  
pp. 1291-1303 ◽  
Author(s):  
Dunyong Tan ◽  
David A. Johnson ◽  
Wei Wu ◽  
Lingfang Zeng ◽  
Yen Hao Chen ◽  
...  
Keyword(s):  
Energy Transfer ◽  
Fluorescent Protein ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Living Cells ◽  
Bioluminescence Resonance Energy Transfer ◽  
Cdna Sequences ◽  
Receptor Isoforms ◽  
Renilla Reniformis ◽  
Green Fluorescent

Abstract We have used bioluminescence resonance energy transfer (BRET) to examine the interaction between human prolactins (PRLs) and the long (LF) and two short isoforms (SF1a and SF1b) of the human PRL receptor in living cells. cDNA sequences encoding the LF, SF1a, and SF1b were subcloned into codon-humanized vectors containing cDNAs for either Renilla reniformis luciferase (Rluc) or a green fluorescent protein (GFP2) with a 12- or 13-amino acid linker connecting the parts of the fusion proteins. Transfection into human embryonic kidney 293 cells demonstrated maintained function of Rluc and GFP2 when linked to the receptors, and confocal microscopy demonstrated the localization of tagged receptors in the plasma membrane by 48 h after transfection. All three tagged receptors transduced a signal, with the LF and SF1a stimulating, and SF1b inhibiting, promoter activity of an approximately 2.4-kb β-casein-luc construct. Both unmodified PRL (U-PRL) and the molecular mimic of phosphorylated PRL, S179D PRL, induced BRET with all combinations of long and short receptor isoforms except SF1a plus SF1b. No BRET was observed with the site two-inactive mutant, G129R PRL. This is the first demonstration, 1) that species homologous PRL promotes both homo- and hetero-interaction of most long and short PRLR pairs in living cells, 2) that both U-PRL and S179D PRL are active in this regard, and 3) that there is some aspect of SF1a-SF1b structure that prevents this particular hetero-receptor pairing. In addition, we conclude that preferential pairing of different receptor isoforms is not the explanation for the different signaling initiated by U-PRL and S179D PRL.

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