Chemistry: molecular biology and microbiology – Enzyme – proenzyme; compositions thereof; process for... – Oxidoreductase
Patent
1995-06-02
1997-12-23
LeGuyader, John L.
Chemistry: molecular biology and microbiology
Enzyme , proenzyme; compositions thereof; process for...
Oxidoreductase
435 697, C12N 902
Patent
active
057006733
ABSTRACT:
A method is disclosed for producing a protein which expresses bioluminescence activity which involves combining two polydeoxyribonucleotides, one containing a continuous sequence of codons encoding a polypeptide which comprises a single covalently bonded molecular structure and which catalyzes the oxidation of insect luciferin to yield light and the other which causes DNA transcription, and obtaining the polypeptide by transcription and subsequent translation. The insect luciferin is derived from bioluminescent insect, preferably Diptera and Coleoptera (fireflies and beetles). Hybrid proteins are similarly formed by inclusion of an additional polydeoxyribonucleotide encoding for a second polypeptide such that their respective polypeptide-encoding reading frames form a continuous reading frame. Also disclosed is a method for quantitatively assaying a fluid for the presence of an unknown quantity of antigen using the hybrid proteins bonded to antibody directed against the antigen, determining luminescence after reaction of the antibody and antigen, and determining the amount of the antigens by comparison of the determined luminescence with the luminescence previously measured under equivalent conditions for equivalent hybrid proteins reacted under equivalent conditions with known amounts of the antigen.
REFERENCES:
Wood J. Bioluminescence and Chemiluminescence, 5:107-114 (1990).
Gruber, "Investigations on redevelopment of multiplexed living cell biosensors using beetle luciterase", Masters Thesis, Univ. of Wisconson-Madison, 1993.
De Wet Jeffrey R.
Helinski Donald R.
Howell Stephen H.
McElroy, deceased Marlene D.
Ow David W.
LeGuyader John L.
The Regents of the University of California
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