(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 |
(22) | Date of filing: 26.10.2000 |
| (51) | International Patent Classification (IPC): |
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(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
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(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) |
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| |
(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.