Mutant luciferases
Described are mutant luciferases, nucleic acids that encode them, cells and animals expressing them, methods of use thereof, and kits.
1. A polypeptide comprising a mutant luciferase,
wherein said mutant luciferase is at least 95% identical to the full length of SEQ ID NO: 5 and comprises a lysine at position 218 and an alanine at position 342, and
wherein, compared to the luciferase of SEQ ID NO: 5, said mutant luciferase has improved bioluminescence using CycLuc2 as the substrate.
2. A kit comprising the polypeptide of claim 1 and a cognate substrate.
3. A method of producing bioluminescence in a sample, the method comprising:
providing a sample comprising the polypeptide of claim 1 ; and
contacting the sample with a cognate substrate.
4. The method of claim 3 , further comprising detecting bioluminescence in the sample.
5. The method of claim 3 , wherein the sample comprises a living cell, a living animal, a tissue or organ, or a cell or tissue extract.
6. A method of producing bioluminescence in a sample, the method comprising:
providing a sample comprising the polypeptide of claim 1 , and
contacting the sample with a substrate, wherein
the substrate is CycLuc1-8, MeNH-LH2, iBuNH-LH2, (3-hydroxypropyl)NH-LH2, or Me2N-LH2.
7. The method of claim 6 , further comprising detecting bioluminescence in the sample.
8. The method of claim 6 , further comprising obtaining an image of the bioluminescence in the sample.
9. The method of claim 6 , wherein the substrate is CycLuc1, CycLuc4, CycLuc2, or CycLuc7.
10. A kit for use in a method of producing bioluminescence in a sample, the kit comprising the polypeptide of claim 1 , and a substrate, wherein
the substrate is CycLuc1-8, MeNH-LH2, iBuNH-LH2, (3-hydroxypropyl)NH-LH2, or Me2N-LH2.
11. The kit of claim 10 , wherein the substrate is CycLuc1, CycLuc4, CycLuc2, or CycLuc7.
12. The mutant luciferase of claim 1 , wherein the cognate substrate is CycLuc2.
13. The kit of claim 2 , wherein the cognate substrate is CycLuc2.
14. The method of claim 3 , wherein the cognate substrate is CycLuc2.
15. The method of claim 6 , wherein the substrate is CycLuc2.
16. The kit of claim 10 , wherein the substrate is CycLuc2.