Patent ID: 7507565

Claim:
A method to determine efficacy of a test compound in modulating the interaction of a first protein with a second protein in an animal, comprising: (a) providing a first vector, comprising a first polynucleotide that encodes a first fusion protein, wherein the first fusion protein includes a first target domain and a bioluminescence donor domain, wherein the bioluminescence donor domain is a mutated Renilla Luciferase polypeptide according to SEQ ID:No 3, wherein the mutated luciferase polypeptide in the presecence of coelanterazine 400a emits bioluminescence having a wavelength between about 360 to 460 nm and at an enhanced level compared to a non-mutated Renilla luciferase in the prescence of coelanterzine; (b) providing a second vector, comprising a second polynucleotide that encodes a second fusion protein that includes a second target domain and a fluorescent acceptor domain; (c) administering the first vector and the second vector to the animal, thereby generating the first fusion protein and the second fusion protein in the animal; (d) administering coelanterazine 400a to the animal, whereupon a bioluminescence energy having a wavelength of about 360 to about 460 nm is emitted from the bioluminescence donor domain upon interaction with the coelanterazine 400a, and whereupon, when the first fusion protein and the second fusion protein are within about 50 to about 100 Angstroms of each other, a fluoresence energy having a wavelength of about 500 to about 570 nm is emitted from the fluorescent acceptor domain upon accepting the bioluminescence energy; (e) detecting a first bioluminescence energy emission and a first fluoresence energy emission in the animal, and calculating the radiance value for each first energy emission; (f) administering to the animal a test compound; (g) detecting a second bioluminescence energy emission and a second fluoresence energy emission in the animal, and calculating the radiance value of each second energy emission; and (h) comparing the radiance value of each of the first emissions with the corresponding radiance value of the second energy emissions, wherein a change in the radiance values of the first and second fluoresence energy emissions indicates that the test compound modulated the interaction of the first target domain and the second target domain.