ARACHNOCAMPA LUCIFERASES
First Claim
1. An isolated peptide that displays luciferase activity, whereina. the luciferase activity comprises catalysis of a luminescence reaction dependent upon ATP;
- andb. the luminescence reaction produces an emission spectrum with a maximum emission intensity at wavelengths less than or equal to 530±
5 nm; and
c. the peptide has an amino acid sequence that, when aligned with that of SEQ ID NO;
4, differs from that of SEQ ID NO;
4 by at least one change in amino acid sequence selected from the group consisting of deletion of R218, H245N, G315S, L342S, and T343S.
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Abstract
Nucleotide and amino acid sequences of luciferase peptides that are encoded by genes within the genome of Arachnocampa (Diptera) are disclosed. Specifically provided are functional ATP-dependent luciferases that catalyze luminescence reactions with emission spectra within the blue portion of the spectrum. The present invention specifically provides isolated peptide and nucleic acid molecules, methods of identifying orthologs and active subsequences of the enzyme peptides, and methods of identifying modulators and substrates of the luciferase peptides. Methods of assays, including multiple reporter assays utilizing at least two ATP-dependent luciferases are provided.
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Citations
20 Claims
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1. An isolated peptide that displays luciferase activity, wherein
a. the luciferase activity comprises catalysis of a luminescence reaction dependent upon ATP; - and
b. the luminescence reaction produces an emission spectrum with a maximum emission intensity at wavelengths less than or equal to 530±
5 nm; andc. the peptide has an amino acid sequence that, when aligned with that of SEQ ID NO;
4, differs from that of SEQ ID NO;
4 by at least one change in amino acid sequence selected from the group consisting of deletion of R218, H245N, G315S, L342S, and T343S. - View Dependent Claims (2, 3, 4, 5, 16)
- and
- 6. An isolated peptide having an amino acid sequence that shares at least 70 percent identity with the amino acid sequence shown in SEQ ID NO:
- 14. A method of measuring the activity of at least two reporter enzymes in an aliquot of a sample comprising assaying for activity of a first reporter enzyme by measuring the light signal produced by catalysis of a luminescence reaction by the first reporter enzyme, and assaying for activity of at least a second reporter enzyme by measuring the light signal produced by catalysis of a luminescence reaction by at least a second reporter enzyme, wherein at least one of the first or second reporter enzymes displays luciferase activity comprising catalysis of a luminescence reaction dependent upon ATP and that produces an emission spectrum with a maximum emission intensity at wavelengths less than or equal to 530±
Specification