Directed Molecular Evolution Reveals Gaussia Luciferase Variants with Enhanced Light Output Stability
| Autor: | M. Hannah Degeling, Casey A. Maguire, Marie Tannous, Bakhos A. Tannous, Mark C. de Gooijer, Grant K. Lewandrowski, M. Sarah S. Bovenberg, Carmen L.A.M. Vleggeert-Lankamp |
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| Přispěvatelé: | Neurosurgery, Other Research |
| Rok vydání: | 2013 |
| Předmět: |
Luciferases
Light biology Chemistry Molecular Sequence Data biology.organism_classification Molecular biology Article Analytical Chemistry Gaussia Molecular evolution Luminescent Measurements Mutation Biophysics Bioluminescence Light emission Luciferase Amino Acid Sequence Emission spectrum Directed Molecular Evolution |
| Zdroj: | Degeling, M H, Bovenberg, M S S, Lewandrowski, G K, de Gooijer, M C, Vleggeert-Lankamp, C L A, Tannous, M, Maguire, C A & Tannous, B A 2013, ' Directed Molecular Evolution Reveals Gaussia Luciferase Variants with Enhanced Light Output Stability ', Analytical Chemistry, vol. 85, no. 5, pp. 3006-3012 . https://doi.org/10.1021/ac4003134 Analytical Chemistry, 85(5), 3006-3012. American Chemical Society Analytical Chemistry, 85(5), 3006-3012 |
| ISSN: | 0003-2700 |
| Popis: | Gaussia Luciferase (Gluc) has proven to be a powerful mammalian cell reporter for monitoring numerous biological processes in immunology, virology, oncology and neuroscience. Current limitations of Gluc as a reporter include its emission of blue light which is absorbed by mammalian tissues, limiting its use in vivo, and a flash-type bioluminescence reaction, making it unsuited for high-throughput applications. To overcome these limitations, a library of Gluc variants was generated using directed molecular evolution and screened for relative light output, a shift in emission spectrum, and glow-type light emission kinetics. Several variants with a 10–15 nm shift in their light emission peak were found. Further, a Gluc variant that catalyzes a glow-type bioluminescence reaction yielding over 10 minutes of stable light output, suited for high-throughput applications, was also identified. These results indicate that molecular evolution could be used to modulate Gluc bioluminescence reaction characteristics. |
| Databáze: | OpenAIRE |
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