(19)
(11)EP 2 284 271 A3

(12)EUROPEAN PATENT APPLICATION

(88)Date of publication A3:
07.03.2012 Bulletin 2012/10

(43)Date of publication A2:
16.02.2011 Bulletin 2011/07

(21)Application number: 10178711.7

(22)Date of filing:  26.10.2000
(51)Int. Cl.: 
C12N 15/53  (2006.01)
C12Q 1/66  (2006.01)
C12N 9/02  (2006.01)
(84)Designated Contracting States:
AT BE CH CY DE DK ES FI FR GB GR IE IT LI LU MC NL PT SE

(30)Priority: 26.10.1999 GB 9925161

(62)Application number of the earlier application in accordance with Art. 76 EPC:
07017621.9 / 1935980
00971589.7 / 1224294

(71)Applicant: Promega Corporation
Madison, WI 53711-5399 (US)

(72)Inventors:
  • Squirrell, David James
    Salisbury Wiltshire SP4 0JQ (GB)
  • Willey, Tara Louise
    Salisbury Wiltshire SP4 0JQ (GB)
  • Murphy, Melanie Jane
    Salisbury Wiltshire SP4 0JQ (GB)
  • White, Peter John
    Salisbury Wiltshire SP4 0JQ (GB)
  • Price, Rachel Louise
    Salisbury Wiltshire SP4 0JQ (GB)

(74)Representative: HOFFMANN EITLE 
Patent- und Rechtsanwälte Arabellastraße 4
81925 München
81925 München (DE)

  


(54)Mutant luciferase


(57) A recombinant protein having luciferase activity and at least 60% similarity to a wild-type luciferase wherein in the sequence of the enzyme, the amino acid residue corresponding to residue 357 in Photinus pyralis luciferase is mutated as compared to the corresponding wild-type luciferase, such that the luciferase enzyme is able to emit light at a different wavelength as compared to the corresponding wild-type luciferase and/or has enhanced thermostability as compared to the corresponding wild-type luciferase. In general, the residue corresponding to 357 in Photinus pyralis luciferase is changed from an acidic amino acid to a non-acidic amino acid and preferably an uncharged polar amino acid such as tyrosine.
Mutant luciferases in accordance with the invention can produce a large (50nm) wavelength shift in emitted light and have good thermostability. The resultant colour shift can be reversed by addition of coenzyme A.
These properties make the mutant particularly useful in a variety of assays.