Method of detecting biologically active substances by using gree

Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving antigen-antibody binding – specific binding protein...

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435 71, 435 77, 435 691, 435183, 4353201, 435325, 4352523, 43525411, 536 231, 536 235, 536 234, C12Q 100, C12N 510, C12N 1512, C07K 14435

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059587130

ABSTRACT:
The present invention provides a method for detecting a biologically active substance that affects intracellular processes mediated through a protein kinase or second messenger, which comprises incubating a cell with a test substance and measuring any change caused by the test substance in the fluorescence of (i) green fluorescent protein (GFP) having a protein kinase recognition site or (ii) GFP that has been modified to contain a second messenger binding domain.

REFERENCES:
patent: 4220450 (1980-09-01), Maggio
patent: 5683888 (1997-11-01), Campbell
Martin Chalfie et al., "Green Fluorescent Protein As A Marker For Gene Expression", Science vol. 263, Feb. 11, 1994, pp. 802-805.
Sala-Newby et al. Engineering a bioluminescent indicator for cyclic AMP-dependent protein kinase. Biochemical Journal. vol. 279, pp. 727-732, 1991.
Sala-Newby et al. Engineering firefly luciferase as an indicator of cyclic AMP-dependent protein kinase in living cells. FEBS Letters. vol. 307, No. 2, pp. 241-244, Jul. 1992.
Sala-Newby et al. Engineering protein kinase recognition sites into firefly luciferase and aequorin. In Biolumin. Chemilumin. Proc. Int. Symp., Stanely et al., eds. Wiley, UK, pp. 23-26, 1991.
Campbell et al. Engineering bioluminescent proteins: a new experimental strategy for measuring intracellular signalling in live cells. Methodol. Surv. Biochem. Anal. vol. 21, pp. 311-316, 1991.

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