Publication: The fusion Vibrio campbellii luciferase as a eukaryotic gene reporter
Issued Date
1970-01-01
Resource Type
ISSN
18734863
01681656
01681656
Other identifier(s)
2-s2.0-84868448986
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Mahidol University
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SCOPUS
Bibliographic Citation
Journal of Biotechnology. Vol.162, No.2-3 (1970), 346-353
Suggested Citation
Ruchanok Tinikul, Kittisak Thotsaporn, Wichit Thaveekarn, Sarawut Jitrapakdee, Pimchai Chaiyen The fusion Vibrio campbellii luciferase as a eukaryotic gene reporter. Journal of Biotechnology. Vol.162, No.2-3 (1970), 346-353. doi:10.1016/j.jbiotec.2012.08.018 Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/10568
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Title
The fusion Vibrio campbellii luciferase as a eukaryotic gene reporter
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Abstract
© 2012 Elsevier B.V. Bacterial luciferase from Vibrio campbellii is a thermostable enzyme with an in vitro thermal inactivation half-life of ∼1020 min at 37 °C. The enzyme also binds tightly to reduced FMN. In this study, a V. campbellii fusion luciferase construct in which the α and β subunits are linked with a decapeptide was made and characterized. In general, the overall enzymatic properties of the two enzymes are similar. Expression of the enzymes in Escherichia coli demonstrated that the V. campbellii fusion luciferase emits less light than the native luciferase, but still emits a much greater amount of light than native luciferase from Vibrio harveyi and Photobacterium leiognathi TH1. The intensity of light emitted by the V. campbellii fusion luciferase was more than 80-fold greater than that from the V. harveyi native luciferase when expressed at 37 °C. Biochemical characterization has shown that the V. campbellii fusion luciferase also retains a high binding affinity for reduced flavin mononucleotide and high thermostability. The levels of bioluminescence emitted by the V. campbellii fusion luciferase expressed in HEK293T cells reached ∼1 × 10 6 Relative Light Units/mg total protein. These findings suggest that the V. campbellii fusion luciferase is a promising candidate for further development as a luciferase-based reporter for eukaryotic systems.