Luciferase enzymes for use with thermostable luciferins in bioluminescent assays
AssigneePromega Corporation
InventorsMichael Killoran, Ce Shi, Mary Hall, Lance P. Encell, Thomas Kirkland
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Abstract
Provided herein are enhanced luciferase enzymes for use with thermostable luciferin analogs for bioluminescent assays. In particular, the present disclosure provides compositions, assays, and methods for performing a bioluminescent assay using enhanced, high-activity luciferase enzymes compatible with thermostable luciferins, such as 5,5-disubstituted luciferin analogs.
Claims21
-76. (canceled)
A luciferase polypeptide capable of producing an increased bioluminescent signal in the presence of a 5,5-disubstituted luciferin compared to a bioluminescent signal produced by a luciferase polypeptide of SEQ ID NO:
- 1 in the presence of the 5,5-disubstituted luciferin, wherein the luciferase polypeptide is not naturally occurring and comprises at least 70% sequence identity with the amino acid sequence of SEQ ID NO:
- 2, and less than 100% sequence identity with the amino acid sequence of SEQ ID NO:
- 1, wherein the polypeptide comprises at least one amino acid substitution at position 244, 249, 337, 339, 240, 254, 344, 300, 396, 245, 285, 315, 109, 193, 214, 218, 228, 234, 262, 287, 294, 305, 306, 309, 316, 335, 533, 133, 233, 236, 503, 107, and/or 121 relative to the amino acid sequence of SEQ ID NO:
- 1.
An assay system for detecting or quantifying ATP in a sample, the system comprising:
- a reagent composition comprising a luciferase polypeptide of claim 1, a 5,5-disubstituted luciferin;
- and a sample comprising or suspected of comprising ATP.
Dependencies are inferred from the claim text.