Cloning and characterization of luciferase from an Asian firefly Pygoluciola qingyu and its comparison with other beetle luciferases
The bioluminescence system of luminescent beetles has extensive applications in biological imaging, protein labeling and drug screening. To explore wild luciferases with excellent catalytic activity and thermal stability, we cloned the luciferase of Pygoluciola qingyu, one species living in areas of high temperature and with strong bioluminescence, by combining transcriptomic sequencing and reverse transcription polymerase chain reaction (RT-PCR). The total length of luciferase gene is 1638 bp and the luciferase consists 544 amino acids. The recombinant P. qingyu luciferase was produced in vitro and its characteristics were compared with those of eight luciferases from China firefly species and two commercial luciferases. Compared with these luciferases, the P. qingyu luciferase shows the highest luminescence activity at room temperature (about 25–28 ℃) with similar KM value for d-luciferin and ATP to the Photinus pyralis luciferase. The P. qingyu luciferase activity was highest at 35 ℃ and can keep high activity at 30–40 ℃, which suggests the potential of P. qingyu luciferase for in vivo and cell application. Our results provide new insights into P. qingyu luciferase and give a new resource for the application of luciferases. Graphical abstract.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2024 |
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| Erschienen: |
2024 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:23 |
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| Enthalten in: |
Photochemical & photobiological sciences - 23(2024), 4 vom: 05. März, Seite 719-729 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Li, Jun [VerfasserIn] |
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| Links: |
Volltext [lizenzpflichtig] |
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| Anmerkungen: |
© The Author(s), under exclusive licence to European Photochemistry Association, European Society for Photobiology 2024. Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law. |
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| doi: |
10.1007/s43630-024-00547-0 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| 520 | |a The bioluminescence system of luminescent beetles has extensive applications in biological imaging, protein labeling and drug screening. To explore wild luciferases with excellent catalytic activity and thermal stability, we cloned the luciferase of Pygoluciola qingyu, one species living in areas of high temperature and with strong bioluminescence, by combining transcriptomic sequencing and reverse transcription polymerase chain reaction (RT-PCR). The total length of luciferase gene is 1638 bp and the luciferase consists 544 amino acids. The recombinant P. qingyu luciferase was produced in vitro and its characteristics were compared with those of eight luciferases from China firefly species and two commercial luciferases. Compared with these luciferases, the P. qingyu luciferase shows the highest luminescence activity at room temperature (about 25–28 ℃) with similar KM value for d-luciferin and ATP to the Photinus pyralis luciferase. The P. qingyu luciferase activity was highest at 35 ℃ and can keep high activity at 30–40 ℃, which suggests the potential of P. qingyu luciferase for in vivo and cell application. Our results provide new insights into P. qingyu luciferase and give a new resource for the application of luciferases. Graphical abstract | ||
| 700 | 1 | |a Liu, Wei |e verfasserin |4 aut | |
| 700 | 1 | |a Liu, Guichun |e verfasserin |4 aut | |
| 700 | 1 | |a Dong, Zhiwei |e verfasserin |4 aut | |
| 700 | 1 | |a He, Jinwu |e verfasserin |4 aut | |
| 700 | 1 | |a Zhao, Ruoping |e verfasserin |4 aut | |
| 700 | 1 | |a Wang, Wen |e verfasserin |4 aut | |
| 700 | 1 | |a Li, Xueyan |e verfasserin |0 (orcid)0000-0003-0457-7846 |4 aut | |
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