| Literature DB >> 23425213 |
M Hannah Degeling1, M Sarah S Bovenberg, Grant K Lewandrowski, Mark C de Gooijer, Carmen L A Vleggeert-Lankamp, Marie Tannous, Casey A Maguire, Bakhos A Tannous.
Abstract
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, suited for high-throughput applications, was also identified. These results indicate that molecular evolution could be used to modulate Gluc bioluminescence reaction characteristics.Entities:
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Year: 2013 PMID： 23425213 PMCID： PMC3617556 DOI： 10.1021/ac4003134
Source DB: PubMed Journal: Anal Chem ISSN： 0003-2700 Impact factor: 6.986