The present invention provides ligand detection means capable of
exhibiting two-dimensional information (wavelength and intensity of
luminescent signal) responding to multiple signals triggered by a ligand
via a target protein, while taking advantage of the merit of the
single-molecule-format bioluminescent probe.
The present invention could provide a luminescent probe set which is
comprised of a fusion protein comprising a ligand recognition protein and
a molecular recognition domain to which the ligand recognition protein
bind upon conformational change, wherein the fusion protein is sandwiched
between split Lighting Enzyme fragments, the probe set can emit multiple
luminescence by utilizing Lighting Enzymes emitting lights with multiple
wavelength, wherein these multiple components can be tandemly arranged by
using C-terminal fragment of the Lighting Enzyme.
By using a living cell line transfected with gene of multicolor
luminescent probe set or single-molecule-format multicolor bioluminescent
probe, it becomes possible to distinguish and detect bioactivity level of
a target ligand in a complex context of the living cell two-dimensionally
(wavelength versus intensity) in multi colors, and to quantitatively
evaluate multiple effects (anticancer and carcinogenesis actions, agonist
and antagonist) of a ligand represented by a drug at once by
two-dimensional information of different colors in short time.