RRC ID 67134
著者 Kaku T, Sugiura K, Entani T, Osabe K, Nagai T.
タイトル Enhanced brightness of bacterial luciferase by bioluminescence resonance energy transfer.
ジャーナル Sci Rep
Abstract Using 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.
巻・号 11(1)
ページ 14994
公開日 2021-7-22
DOI 10.1038/s41598-021-94551-4
PII 10.1038/s41598-021-94551-4
PMID 34294849
PMC PMC8298465
MeSH Bacterial Proteins / genetics* Bacterial Proteins / metabolism Bioluminescence Resonance Energy Transfer Techniques Cloning, Molecular HEK293 Cells Humans Luciferases, Bacterial / genetics* Luciferases, Bacterial / metabolism Luminescent Proteins / genetics* Luminescent Proteins / metabolism Plants, Genetically Modified Protein Engineering Recombinant Fusion Proteins / metabolism* Tobacco / genetics Tobacco / growth & development Tobacco / metabolism
IF 3.998
リソース情報
ヒト・動物細胞 293T(RCB2202)