CLONING AND TRANSIENT EXPRESSION OF GENES ENCODING THE HUMAN ALPHA-4 AND BETA-2 NEURONAL NICOTINIC ACETYLCHOLINE-RECEPTOR (NACHR) SUBUNITS

Partial cDNA clones generated by RT-PCR were used as probes to clone t he cDNAs encoding the human alpha 4 and beta 2 neuronal nicotinic acet ylcholine receptor (nAChR) subunits. The 2.1-kb alpha 4 cDNA shows 84 and 76% identity to the rat and chicken cDNA sequences, respectively, The deduced amino-acid sequence shares 89 and 84% similarity, respecti vely, with the corresponding rat and chicken proteins, with most of th e divergence occurring in the cytoplasmic domain, The 1721-nucleotide beta 2 sequence was identical to the human beta 2 sequence previously reported. Transfection of the alpha 4 and beta 2 clones into HEK293 ce lls resulted in the formation of binding sites that display high affin ity towards [H-3] cytisine, a characteristic of the alpha 4 beta 2 sub type produced in vivo.