FLUORESCENCE RESONANCE ENERGY TRANSFER DETECTION OF cAMP IN LIVING CELLS USING GFP VARIANTS

摘要

The present invention establishes a technology to monitor cAMP changes in living cells using two GFP variants (ECFP and EYFP). The present invention describes a construct in which the gene encoding the R subunit is essentially a linker between the genes encoding ECFP and EYFP. Following cAMP binding to both cAMP binding domains (CAB) of the recombinantly expressed R subunit, the R subunit undergoes a conformational change, thereby reducing the distance between ECFP and EYFP, which is subsequently detected by fluorescence resonance energy transfer.