STRUCTURE AND EXPRESSION OF A RAT KAPPA OPIOID RECEPTOR GENE

Abstract A gene encoding the rat κ opioid receptor (KOR) was cloned and characterized. Results of rat genomic library screening and genomic Southern blot analysis show the gene represented at one copy/haploid genome. Three introns are present within the gene; however, polymerase chain reaction using different sets of primers specifying neighboring exons indicates that alternative splicing does not occur. Using reverse transcriptionpolymerase chain reaction and primer extension techniques, we are able to demonstrate that two species of transcripts are differentially produced from the KOR gene in a tissue-specific manner. The first transcript that we designate as KOR1 is equivalent to the cDNA sequence reported by other groups and is believed to correspond to KOR subtype 1. KOR1 begins with exon 1 just downstream of two TATA boxes, whereas the second transcript, which we refer to as KORx, begins in intron 1 and thereby retains this intronic sequence in the mature mRNA. Within this intronic sequence there are two potential translation initiation codons that are in-frame with the proposed initiation codon of KOR. The potential open reading frame that starts further upstream in KORx may lead to the translation of a variant KOR protein having a novel peptide sequence at its amino terminus.