Bioanalytical systems based on the Bioluminescence Resonance Energy Transfer (BRET)
are widely used in fundamental biochemical studies, as well as for screening and analysis of
biologically active compounds. The Renilla luciferase is the most often used energy donor in this
system despite the fact that it has low bioluminescence quantum yield and demonstrates not so stable
luminescence in time as the firefly luciferase. Moreover, the bioluminescence λmax is observed in the
green region of the spectrum, which complicates signal recording in tissues during in vivo
experiments. The firefly luciferases do not have such drawbacks and show great promise for
applications in BRET systems. Different versions of BRET systems based on firefly luciferases and
the methods for increasing their efficiency are considered in this review; examples of the use of BRET systems based on
the firefly luciferases for highly sensitive determination of proteases and for homogeneous immunoassays are presented.
Keywords: Luciferase, bioluminescence, luciferase-based assay, bioluminescence resonance energy transfer, BRET.
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