Mutated genes for the catalytic protein of Oplophorus luciferase and use thereof
AssigneeJnc Corporation
InventorsSatoshi Inouye, Yuiko Miura, Junichi Sato
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Abstract
A novel luciferase that is distinct from conventional luciferase has been desired. A luciferase mutant comprising an amino acid sequence in which glutamic acid at position 4, arginine at position 11, leucine at position 18 and valine at position 27 are substituted with other amino acids, in the amino acid sequence of SEQ ID NO: 2.
Claims12
A method for producing a recombinant luciferase mutant, which comprises the steps of culturing a transformant transformed with a recombinant vector comprising a polynucleotide comprising a nucleotide sequence encoding the recombinant luciferase mutant and separating/purifying the recombinant luciferase mutant, wherein the recombinant luciferase mutant is selected from the group consisting of:
- (a) a recombinant luciferase mutant comprising the amino acid sequence of SEQ ID NO:
- 2 substituted at the amino acid positions consisting of:
- glutamic acid at position 4;
- arginine at position 11;
- leucine at position 18;
- and
- valine at position 27;
- and,
- (b) a recombinant luciferase mutant comprising the amino acid sequence of SEQ ID NO:
- 2 substituted at the positions consisting of:
- glutamic acid at position 4;
- arginine at position 11;
- leucine at position 18;
- valine at position 27;
- and
- one or more amino acid(s) at position(s) other than positions 4, 11, 18, 27, 33, 43, 44, 54, 68, 72, 75, 90, 115, 124, 138 and 166 of the amino acid sequence of SEQ ID NO:
- 2, wherein the recombinant luciferase mutant has luciferase activity.
A method for assaying the activity of a target promoter, which comprises
- providing a polynucleotide encoding a luciferase mutant as a report gene and fusing the polynucleotide to the target promoter to construct a vector;
- transforming the vector into a host cell;
- contacting the host cell with a luciferin;
- and
- detecting luminescence, thereby assaying the activity of the target promoter,
- wherein the luciferase mutant is selected from the group consisting of:
- (a) a recombinant luciferase mutant comprising the amino acid sequence of SEQ ID NO:
- 2 substituted at the amino acid positions consisting of:
- glutamic acid at position 4;
- arginine at position 11;
- leucine at position 18;
- and
- valine at position 27;
- and,
- (b) a recombinant luciferase mutant comprising the amino acid sequence of SEQ ID NO:
- 2 substituted at the positions consisting of:
- glutamic acid at position 4;
- arginine at position 11;
- leucine at position 18;
- valine at position 27;
- and
- one or more amino acid(s) at position(s) other than positions 4, 11, 18, 27, 33, 43, 44, 54, 68, 72, 75, 90, 115, 124, 138 and 166 of the amino acid sequence of SEQ ID NO:
- 2, wherein the recombinant luciferase mutant has luciferase activity.
Dependencies are inferred from the claim text.