Mutant luciferases
AssigneePromega Corporation
InventorsKeith V. Wood, Monika G. Wood
Life of this patent
Estimating the term…
Abstract
The invention provides active, non-naturally occurring mutants of beetle luciferases and DNAs which encode such mutants. A mutant luciferase of the invention differs from the corresponding wild-type luciferase by producing bioluminescence with a wavelength of peak intensity that differs by at least 1 nm from the wavelength of peak intensity of the bioluminescence produced by the wild-type enzyme. The mutant luciferases and DNAs of the invention are employed in various biosensing applications.
Claims15
An isolated DNA molecule comprising a segment having a sequence which encodes a synthetic mutant beetle luciferase comprising an amino acid sequence that differs from that of the corresponding wild-type luciferase by at least one amino acid substitution, the position of the amino acid substitution corresponding to a position in the amino acid sequence of LucPplGR of SEQ ID NO:2 selected from the group consisting of position 215, 224, 238, 242, 247 and 348, wherein the mutant luciferase produces bioluminescence having a shift in wavelength of peak intensity of at least 1 nanometer relative to the bioluminescence produced by the wild-type luciferase, wherein for the encoded amino acid sequence, the corresponding wild-type luciferase is LucPplGR of SEQ ID NO:2, and wherein the encoded synthetic mutant luciferase includes an amino acid substitution selected from the group consisting of LucPplGR-R215Q, -R215S, -R215Y, -R215K, -R215C, -R215E, -R215F, -H242S, -H348A, -V224F/L238M, and -V224F/S247Y.
An isolated DNA molecule comprising a segment having a sequence which encodes a mutant beetle luciferase having an amino acid sequence that differs from that of the corresponding wild-type luciferase LucPplGR by at least one amino acid substitution, wherein the encoded mutant luciferase is selected from the group consisting of LucPplGR-R215Q, -R215S, -R215Y, -R215K, -R215C, -R215E, -R215F, -H242S, -H238A, -V224F/L238M, and -V224F/S247Y and the encoded mutant luciferase produces bioluminescence having a shift in wavelength of peak intensity of at least 1 nanometer relative to the bioluminescence produced by the wild-type luciferase.
Dependencies are inferred from the claim text.