Construction and expression of renilla luciferase and green fluo

Details Construction and expression of renilla luciferase and green fluo Construction and expression of renilla luciferase and green fluo

1996-12-23

1999-11-02

Duffy, Patricia A.

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving nucleic acid

435 8, 435 691, 435 697, 435183, 436548, 530350, 536 234, C12Q 166, C12P 2104, G01N 3353, C07H 2104

Patent

active

059767960

ABSTRACT:
A fusion gene is provided comprising the cDNA of Renilla luciferase and the cDNA of the "humanized" Aequorea green fluorescent protein. The fusion gene was used to produce a novel protein, the "Renilla-GFP fusion protein," which displayed both the luciferase activity of Renilla luciferase, and the green fluorescence of GFP. The Renilla-GFP fusion gene is useful as a double marker for monitoring gene expression quantitatively in UV light and by enzyme activity.

REFERENCES:
Kobatake et al. Bioluminescent immunoassay with a protein A-luciferase fusion protein, Anal. Biochem. 208:300-305, 1993.
Ward et al. An energy transfer protein in Coelenterate bioluminescence, J. Biol. Chem. 254:781-788, 1979.
Mitra et al. Fluorescence resonance energy transfer between blue-emitting and red-shifted excitation derivatives of the green fluorescent protein, Gene 173:13-17, 1996.
Mayerhofer, R. et al., "Expression of recombinant Renilla luciferase in transgenic plants results in high levels of light emission," The Plant Journal 7(6): 1031-1038 (1995).
Zolotukhin, S. et al., "A `Humanized Green` Fluorescent Protein cDNA Adapted for High-Level Expression in Mammalian Cells," Journal of Virology 70:4646-4654 (1996).

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