Polynucleotide encoding luciferase
View Patent ↗Briefly described, embodiments of this disclosure include polynucleotides that encode mutant Cnidarian luciferases that exhibit modulated properties as compared to the corresponding wild-type luciferases, and the modulated properties include at least one of: modulated stability; enhanced light output; and modulated emission maximum. Embodiments of the present disclosure also include polypeptides or fragments thereof encoded by the polynucleotides, constructs including the polynucleotide, expression cassettes, cells, methods of producing the polynucleotides and polypeptides, antibodies, transgenic cells and/or animals, kits, and the like.
1. An isolated polynucleotide encoding a mutant Cnidarian luciferase, wherein the mutant luciferase has the sequence of SEQ ID NO:27.
2. The polynucleotide of claim 1 , wherein the polynucleotide encodes a fusion protein comprising the mutant Cnidarian luciferase and at least one fusion partner.
3. The polynucleotide of claim 2 , wherein the at least one fusion partner comprises an epitope tag.
4. The polynucleotide of claim 2 , wherein the at least one fusion partner comprises a targeting moiety.
5. The polynucleotide of claim 4 , wherein the targeting moiety is selected from the group consisting of: an antibody, a receptor, a ligand, and binding fragments or mimetics thereof.
6. The polynucleotide of claim 5 , wherein the targeting moiety is a ligand that specifically binds to VEGF receptor 2.
7. The polynucleotide of claim 6 , wherein the targeting moiety is VEGF 121.
8. A construct comprising: a vector and a polynucleotide according to claim 1 .
9. An expression cassette comprising:
a polynucleotide according to claim 1 .
10. A cell, or the progeny thereof, comprising an expression cassette according to claim 9 as part of an extrachromosomal element or integrated into the genome of a host cell as a result of introduction of the expression cassette into the host cell.
11. A method of producing a luciferase, the method comprising:
growing a cell according to claim 10 , whereby the protein is expressed; and
isolating the protein substantially free of other proteins.
12. A kit comprising: a polynucleotide according to claim 1 .
13. The kit of claim 12 , wherein the kit further comprises a substrate for the luciferase.
14. The kit of claim 13 , wherein the substrate is a coelenterazine.