This proposal focuses on three classes of protein which control olfactory function: olfactory receptor proteins, odorant binding proteins, and odorant degrading enzymes. The last few years have seen the biochemical characterization of each of these components in a variety of animals ranging from rats and frogs to insects, lobsters and snails. We have ourselves amassed a substantial amount of biochemical and molecular biological information of olfactory processing in an insect model system: sex-pheromone reception/transduction in large moths. In previous work we have characterized candidate odorant receptor proteins; isolated, characterized and sequenced the cDNA of odorant binding proteins; and isolated and characterized odorant degrading enzymes. We now propose to isolate and characterize the nucleic acid sequences which encode these proteins, as well as those 5' transcription regulatory sequences which determine and modulate their expression. Nucleic acid and antibody probes will be generated to each of the previously identified proteins. These probes will be used to screen cDNA libraries which have been made from RNA isolated from olfactory tissue. In some cases enhanced cDNA libraries will be constructed using size fractionation and tissue specific subtraction techniques. Selected cDNA inserts will be screened by tissue specific RNA hybridization techniques and by physiological expression assays. Complementary DNA inserts thus selected will be used to isolate and characterize genomic DNA with special attention to the sequences of the 5' transcription regulatory sequences. By comparing the nucleic acid sequences encoding different species of related proteins from different species of related animals patterns will emerge enabling us to understand how tissue and sex specific coordination is accomplished in the nervous system.
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