Codon-optimized gene for mutated shrimp luciferase and method for use thereof
AssigneeJnc Corporation
InventorsSatoshi Inouye, Junichi Sato
Life of this patent
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Abstract
There has been a demand for a codon-optimized gene for the mutated catalytic domain of Oplophorus luciferase, which is capable of efficiently expressing a protein both in a cultured animal cell and Escherichia coli. There has also been a demand for a substrate coelenterazine analogue showing a higher activity than that of native 19 kDa protein. The invention provides a polynucleotide comprising a polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 2. According to the invention, bis-coelenterazine is used as a substrate coelenterazine analogue suitable for the photoprotein encoded by the polynucleotide comprising the polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 2.
Claims11
An isolated polynucleotide selected from the group consisting of (a) to (d) below:
- (a) a polynucleotide comprising a polynucleotide consisting of the nucleotide sequence of SEQ ID NO:
- 2;
- (b) polynucleotide comprising a polynucleotide encoding a protein consisting of a nucleotide sequence in which 1 to 20 nucleotides are deleted, substituted, inserted and/or added in the nucleotide sequence of SEQ ID NO:
- 2, and having a luminescent catalyst activity by using luciferin as a substrate;
- (c) a polynucleotide comprising a polynucleotide encoding a protein consisting of a nucleotide sequence having at least 90% identity to the nucleotide sequence of SEQ ID NO:
- 2, and having a luminescent catalyst activity by using lucifeuin as a substrate;
- and,
- (d) a polynucleotide comprising a polynucleotide encoding a protein which hybridizes to a polynucleotide consisting of a nucleotide sequence complementary to the nucleotide sequence of SEQ ID NO:
- 2 under high stringency conditions, and has a luminescent catalyst activity by using luciferin as a substrate, wherein the high stringency conditions are 5×SSC, 5×Denhardt's solution, 0.5% (w/v) SDS, 50% (v/v) formamide, and 50° C.
An isolated polynucleotide comprising a polynucleotide consisting of the nucleotide sequence of SEQ ID NO:
- 2.
An isolated polynucleotide consisting of any one of the nucleotide sequences of SEQ ID NO:
- 5, SEQ ID NO:
- 8, SEQ ID NO:
- 10 and SEQ ID NO:
- 14.
Dependencies are inferred from the claim text.