Mutant luciferases
AssigneePromega Corporation
InventorsKeith V. Wood, Monika G. Gruber
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.
Claims30
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, 232, 236, 237, 242, 244, 245, 248, 282, 283 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.
An isolated DNA molecule comprising a segment having a sequence which encodes a synthetic mutant beetle luciferase having 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 position 282 in the amino acid sequence of LucPplGR of SEQ ID NO:2, 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.
An isolated DNA molecule comprising a segment having a sequence which encodes a synthetic mutant beetle luciferase having 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 position 283 in the amino acid sequence of LucPplGR of SEQ ID NO:2, 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.
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-223L, -R223Q, -R223M, -R223H, -L238R, -L238M, -L238Q, -L238S, -S247H, -S247T, -S247Y -S247F 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.