Modified renilla luciferase nucleic acids and methods of use
This invention provides modified nucleotide sequences encoding luciferase that have greater expression than wild type luciferase.
1. A nucleic acid molecule comprising a reporter nucleotide sequence encoding a polypeptide comprising a Renilla luciferase amino acid sequence having at least 99% sequence identity with SEQ ID NO:1, wherein the Renilla luciferase amino acid sequence is encoded by a luciferase nucleotide sequence having at least 95% sequence identity with SEQ ID NO:2.
2. The nucleic acid molecule of claim 1 wherein the Renilla luciferase amino acid sequence is encoded by a luciferase nucleotide sequence having at least 98% sequence identity with SEQ ID NO:2.
3. The nucleic acid molecule of claim 1 wherein the Renilla luciferase amino acid sequence is encoded by a luciferase nucleotide sequence having at least 99% sequence identity with SEQ ID NO:2.
4. The nucleic acid molecule of claim 1 wherein the Renilla luciferase amino acid sequence is encoded by a luciferase nucleotide sequence having at least 99.9% sequence identity with SEQ ID NO:2.
5. The nucleic acid molecule of claim 1 wherein the Renilla luciferase amino acid sequence is encoded by a luciferase nucleotide sequence identical with SEQ ID NO:2.
6. The nucleic acid molecule of claim 1 wherein the polypeptide is Renilla luciferase (SEQ ID NO:1) (RenS).
7. The nucleic acid molecule of claim 6 wherein RenS is encoded by the nucleotide sequence of SEQ ID NO:2.
8. The nucleic acid molecule of claim 1 wherein the polypeptide is a fusion protein comprising the Renilla luciferase amino acid sequence of SEQ ID NO:1.
9. The nucleic acid molecule of claim 8 wherein the Renilla luciferase amino acid sequence is encoded by the nucleotide sequence of SEQ ID NO:2.
10. The nucleic acid molecule of claim 8 wherein the fusion protein is RenSP (SEQ ID NO: 3).
11. The nucleic acid of claim 10 wherein the RenSP is encoded by the nucleotide sequence of SEQ ID NO: 4.
12. The nucleic acid molecule of claim 8 wherein the fusion polypeptide comprises a protein destabilization domain that decreases the half-life of Renilla luciferase.
13. The nucleic acid molecule of claim 1 wherein the luciferase nucleotide sequence contains no more than 10 regulatory motifs selected from the motifs of FIG. 3 .
14. A recombinant nucleic acid comprising a promoter operably linked with a reporter nucleotide sequence, wherein the reporter nucleotide sequence encodes a polypeptide comprising a Renilla luciferase amino acid sequence having at least 99% sequence identity with SEQ ID NO:1, wherein the Renilla luciferase amino acid sequence is encoded by a luciferase nucleotide sequence having at least 95% sequence identity with SEQ ID NO:2.
15. The recombinant nucleic acid of claim 14 wherein the polypeptide comprises Renilla luciferase (SEQ ID NO:1) (RenS) encoded by a luciferase nucleotide sequence having at least 99% sequence identity with SEQ ID NO:2.
16. The recombinant nucleic acid of claim 14 wherein the Renilla Luciferase amino acid sequence is encoded by the luciferase nucleotide sequence of SEQ ID NO:2.
17. The recombinant nucleic acid of claim 14 wherein the polypeptide is RenSP (SEQ ID NO: 3).
18. The recombinant nucleic acid of claim 14 comprised in a plasmid.
19. A recombinant cell comprising a recombinant nucleic acid comprising a promoter operably linked with a reporter nucleotide sequence, wherein the reporter nucleotide sequence encodes a polypeptide comprising a Renilla luciferase amino acid sequence having at least 99% sequence identity with SEQ ID NO:1, wherein the Renilla luciferase amino acid sequence is encoded by a luciferase nucleotide sequence having at least 95% sequence identity with SEQ ID NO:2.
20. The recombinant cell of claim 19 wherein the polypeptide comprises Renilla luciferase (SEQ ID NO:1) (RenS) encoded by a luciferase nucleotide sequence having at least 99% sequence identity with SEQ ID NO:2.
21. The recombinant cell of claim 19 wherein the luciferase nucleotide sequence has at least 98% sequence identity with SEQ ID NO:2.
22. The recombinant cell of claim 19 wherein the luciferase nucleotide sequence is SEQ ID NO:2.
23. The recombinant cell of claim 19 which is a mammalian cell.
24. A kit comprising: (1) a vector containing a nucleic acid molecule comprising a reporter nucleotide sequence encoding a polypeptide comprising a Renilla luciferase amino acid sequence having at least 99% sequence identity with SEQ ID NO:1, wherein the Renilla luciferase amino acid sequence is encoded by a luciferase nucleotide sequence having at least 95% sequence identity with SEQ ID NO:2; and (2) a luciferase substrate.
25. The kit of claim 24 wherein the polypeptide comprises Renilla luciferase (SEQ ID NO:1) (RenS) encoded by a luciferase nucleotide sequence having at least 99% sequence identity with SEQ ID NO:2.
26. The kit of claim 24 wherein the luciferase nucleotide sequence has at least 98% sequence identity with SEQ ID NO:2.
27. The kit of claim 24 wherein the luciferase nucleotide sequence is SEQ ID NO:2.
28. The kit of claim 24 wherein the Luciferase substrate is coelenterazine.
29. The nucleic acid molecule of claim 1 further comprising a cloning site positioned relative to the reporter nucleotide sequence such that an expression control sequence inserted into the cloning site becomes operably linked with the luciferase-encoding nucleotide sequence.
30. The nucleic acid molecule of claim 1 , wherein the Renilla luciferase amino acid sequence is SEQ ID NO:1.
31. The nucleic acid molecule of claim 2 , wherein the Renilla luciferase amino acid sequence is SEQ ID NO:1.
32. The nucleic acid molecule of claim 3 , wherein the Renilla luciferase amino acid sequence is SEQ ID NO:1.
33. The nucleic acid molecule of claim 4 , wherein the Renilla luciferase amino acid sequence is SEQ ID NO:1.
34. The nucleic acid molecule of claim 13 , wherein the Renilla luciferase amino acid sequence is SEQ ID NO:1.
35. The recombinant nucleic acid of claim 14 , wherein the Renilla luciferase amino acid sequence is SEQ ID NO:1.
36. The recombinant cell of claim 19 , wherein the Renilla luciferase amino acid sequence is SEQ ID NO:1.
37. The recombinant cell of claim 21 , wherein the Renilla luciferase amino acid sequence is SEQ ID NO:1.
38. The recombinant cell of claim 23 , wherein the Renilla luciferase amino acid sequence is SEQ ID NO:1.
39. The kit of claim 24 , wherein the Renilla luciferase amino acid sequence is SEQ ID NO:1.
40. The kit of claim 26 , wherein the Renilla luciferase amino acid sequence is SEQ ID NO:1.
41. The kit of claim 28 , wherein the Renilla luciferase amino acid sequence is SEQ ID NO:1.