Thermostable luciferases and methods of production
AssigneePromega Corporation
InventorsKeith V. Wood, Monika G. Wood, Mary P. Hall
Life of this patent
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Abstract
Luciferase enzymes with greatly increased thermostability, e.g., at least half lives of 2 hours at 50° C., cDNAs encoding the novel luciferases, and hosts transformed to express the luciferases, are disclosed. Methods of producing the luciferases include recursive mutagenesis. The luciferases are used in conventional methods, some employing kits.
Claims11
A thermostable beetle luciferase variant of the luciferase of SEQ ID NO:37, wherein said luciferase variant retains at least 50% activity in aqueous solution for at least 2 hours at a temperature of 50° C. and has up to 42 amino acids modified from SEQ ID NO:37, wherein said modifications comprise a substitution of residue 2 with A, a substitution of residue 91 with I or V, a substitution of residue 183 with Y, a substitution of residue 220 with I or L, a substitution of residue 221 with A, a substitution of residue 249 with M, a substitution of residue 262 with V, a substitution of residue 294 with L, a substitution of residue 353 with N or K, a substitution of residue 354 with N or G, a substitution of residue 386 with P, a substitution of residue 399 with G, a substitution of residue 542 with T, a substitution of residue 543 with N, a substitution of residue 544 with G, and a deletion of residue 545 of SEQ ID NO:37.
Dependencies are inferred from the claim text.