Oplophorus-derived luciferases, novel coelenterazine substrates, and methods of use
AssigneePromega Corporation
InventorsBrock Binkowski, Lance P. Encell, Mary Hall, Matthew B. Robers, Michael R. Slater, Keith V. Wood, Monika G. Wood
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Abstract
An isolated polynucleotide encoding a modified luciferase polypeptide and substrates. The OgLuc variant polypeptide has at least 60% amino acid sequence identity to SEQ ID NO: 1 and at least one amino acid substitution at a position corresponding to an amino acid in SEQ ID NO: 1. The OgLuc variant polypeptide has at least one of enhanced luminescence, enhanced signal stability, and enhanced protein stability relative to the corresponding polypeptide of the wild-type Oplophorus luciferase.
Claims5
A composition comprising a circularly permuted polypeptide having an N-terminal segment linked to a C-terminal segment, wherein the N-terminal segment of the circularly permuted polypeptide has at least 95% sequence identity to a portion of SEQ ID NO:
- 89 from a circular permutation site to the C-terminus of SEQ ID NO:
- 89, wherein the C-terminal segment of the circularly permuted polypeptide has at least 95% sequence identity to a portion of SEQ ID NO:
- 89 from the N-terminus to the circular permutation site of SEQ ID NO:
- 89, wherein the circular permutation site is within a transition between predicted protein structural motifs of a polypeptide of SEQ ID NO:
- 89, and wherein the circularly permuted polypeptide exhibits luciferase activity in the presence of a coelenterazine substrate.
Dependencies are inferred from the claim text.