Inventors:
Brock BINKOWSKI - Sauk City WI, US
Braeden BUTLER - Madison WI, US
Lance P. ENCELL - Fitchburg WI, US
Frank FAN - Verona WI, US
Brad HOOK - Baraboo WI, US
Paul OTTO - Madison WI, US
Gediminas VIDUGIRIS - Fitchburg WI, US
Susan WIGDAL - Belleville WI, US
Kristopher ZIMMERMAN - Madison WI, US
International Classification:
A01K 67/00
C07H 21/00
C12N 1/00
C12N 15/63
C12N 5/10
US Classification:
800 13, 4353201, 435325, 435243, 435419, 536 232
Abstract:
A polynucleotide encoding a biosensor polypeptide comprising a modified circularly-permuted thermostable luciferase and a linker linking the C-terminal portion of the thermostable luciferase to the N-terminal portion of the thermostable luciferase. The modified circularly-permuted thermostable luciferase is modified relative to a parental circularly-permuted thermostable luciferase. The linker contains a sensor region capable of interacting with a target molecule in a cell. The modified circularly-permuted thermostable luciferase has an enhanced response after interaction of the biosensor with the target molecule relative to the parental circularly-permuted thermostable luciferase in the presence of the target molecule. Alternatively, the modified circularly-permuted thermostable luciferase has an enhanced response after interaction of the biosensor with the target molecule relative to the modified circularly-permuted thermostable luciferase in the absence of the target molecule.