Inventors:
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
Kimberly RISMA - Cincinnati OH, US
Gediminas VIDUGIRIS - Fitchburg WI, US
Susan WIGDAL - Belleville WI, US
Kristopher ZIMMERMAN - Madison WI, US
Assignee:
PROMEGA CORPORATION - Madison WI
International Classification:
C12Q 1/66, C12N 9/02
US Classification:
435 613, 536 232, 4353201, 435 8, 435 618, 435325
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.