Enhanced brightness of bacterial luciferase by bioluminescence resonance energy transfer.

Bacterial Proteins / genetics Bioluminescence Resonance Energy Transfer Techniques Cloning, Molecular HEK293 Cells Humans Luciferases, Bacterial / genetics Luminescent Proteins / genetics Plants, Genetically Modified Protein Engineering Recombinant Fusion Proteins / metabolism Nicotiana / genetics

Journal

Scientific reports
ISSN: 2045-2322
Titre abrégé: Sci Rep
Pays: England
ID NLM: 101563288

Informations de publication

Date de publication:
22 07 2021
Historique:
received: 24 02 2021
accepted: 06 07 2021
entrez: 23 7 2021
pubmed: 24 7 2021
medline: 10 11 2021
Statut: epublish

Résumé

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.

Identifiants

DOI: 10.1038/s41598-021-94551-4 PMID: 34294849 PMC: PMC8298465
pubmed: 34294849
doi: 10.1038/s41598-021-94551-4
pii: 10.1038/s41598-021-94551-4
pmc: PMC8298465
doi:

Substances chimiques

Bacterial Proteins 0
Luminescent Proteins 0
Recombinant Fusion Proteins 0
yellow fluorescent protein, Bacteria 0
Luciferases, Bacterial EC 1.14.14.3

Types de publication

Journal Article Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

IM

Pagination

14994

Informations de copyright

© 2021. The Author(s).

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Auteurs

Tomomi Kaku (T)

SANKEN (The Institute of Scientific and Industrial Research), Osaka University, Ibaraki, Osaka, 567-0047, Japan.

Kazunori Sugiura (K)

SANKEN (The Institute of Scientific and Industrial Research), Osaka University, Ibaraki, Osaka, 567-0047, Japan.

Tetsuyuki Entani (T)

SANKEN (The Institute of Scientific and Industrial Research), Osaka University, Ibaraki, Osaka, 567-0047, Japan.

Kenji Osabe (K)

SANKEN (The Institute of Scientific and Industrial Research), Osaka University, Ibaraki, Osaka, 567-0047, Japan.

Takeharu Nagai (T)

SANKEN (The Institute of Scientific and Industrial Research), Osaka University, Ibaraki, Osaka, 567-0047, Japan. ng1@sanken.osaka-u.ac.jp.

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Classifications MeSH

questionsmedicales.fr › Acides aminés, peptides et protéines › Protéines › Protéines bactériennes › Enhanced brightness of bacterial luciferase by...
questionsmedicales.fr › Techniques d'investigation › Techniques de chimie analytique › Photométrie › Mesures de luminescence › Techniques de transfert d'énergie par résonance de bioluminescence › Enhanced brightness of bacterial luciferase by...
questionsmedicales.fr › Techniques d'investigation › Techniques génétiques › Clonage moléculaire › Enhanced brightness of bacterial luciferase by...
questionsmedicales.fr › Cellules › Cellules épithéliales › Cellules HEK293 › Enhanced brightness of bacterial luciferase by...
questionsmedicales.fr › Eucaryotes › Animaux › Chordés › Vertébrés › Mammifères › Eutheria › Primates › Haplorhini › Catarrhini › Hominidae › Humains › Enhanced brightness of bacterial luciferase by...
questionsmedicales.fr › Acides aminés, peptides et protéines › Protéines › Protéines luminescentes › Luciferases › Luciférases des bactéries › Enhanced brightness of bacterial luciferase by...
questionsmedicales.fr › Acides aminés, peptides et protéines › Protéines › Protéines luminescentes › Enhanced brightness of bacterial luciferase by...
questionsmedicales.fr › Stades d'organismes › Organismes génétiquement modifiés › Végétaux génétiquement modifiés › Enhanced brightness of bacterial luciferase by...
questionsmedicales.fr › Techniques d'investigation › Techniques génétiques › Génie génétique › Ingénierie des protéines › Enhanced brightness of bacterial luciferase by...
questionsmedicales.fr › Acides aminés, peptides et protéines › Protéines › Protéines recombinantes › Protéines de fusion recombinantes › Enhanced brightness of bacterial luciferase by...