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Mutants of firefly luciferase with large changes in wavelengths of emitted light
TL Willey1; DJ Squirrell2; Peter J White*1
(1) Babraham Institute, Babraham, Cambridge CB2 4AT, UK; (2) DERA, Porton Down, Salisbury, Wiltshire SP4 0JQ, UK *(email@example.com)
Combinatorial cassette mutagenesis was used to create a library of mutants of the North American firefly luciferase. Mutant luciferases were expressed in Escherichia coli and selected, using an in vivo assay, for changes in spectral distribution of emitted light and improved thermostability. Combinations of mutations at two positions on the surface of the enzyme were found to have very large effects on both wavelength and thermostability in in vivo and in vitro assays. A possible alternative mechanism for the molecular basis for the colour of bioluminescence is proposed.
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