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The human olfactory system can recognize anddiscriminate a large number of different odorant molecules. The detection ofchemically distinct odorants starts with the binding of an odorant ligand to aspecific receptor protein on the olfactory neuron cell surface. To address theproblem of olfactory perception at a molecular level, we have expressed andcharacterized different olfactory receptors with several expression systems.Here we provide the first documentation of functional expression of odorantreceptors using the Semliki Forest virus system. The human odorant OR 17-40receptor and the rat I7 receptor were functionally expressed in vertebratekidney cells (HEK293) using recombinant Semliki Forest viruses. Receptors wereexpressed as a fusion protein with the Nterminal membrane import sequence ofthe guinea pig serotonin receptor. Experiments employing the Ca2+sensitive dyefura-2 revealed a fast, transient increase in the [Ca2+] after application ofthe specific agonists helional and octanal to HEK293 cells infected withviruses containing RNA for the human odorant OR 17-40 receptor and the rat I7receptor, respectively.

Functional Expression and Characterization of Odorant Receptors Using the Semliki Forest Virus System