CLONING AND EXPRESSION OF OPIOID RECEPTOR GENES

阿片类药物引起多种生理效应。 几种不同的阿片类药物 受体已经被描述，但它们的基因仍然未知。 我们提出 鉴定并克隆阿片受体基因， 测序并将基因导入哺乳动物细胞。 我们已经确定 人神经母细胞瘤细胞系SK-N-SH-SY 5 Y，其表达丰富的μ- ，而且， δ-受体。 基因分离的策略包括1）使用 从μ受体的肽序列推导的寡核苷酸探针， 2)直接哺乳动物表达克隆使用cDNA文库从 神经母细胞瘤细胞系; 3）聚合酶链反应的应用 利用G蛋白偶联受体之间的序列同一性; 4）筛选 具有来自任何已知阿片样物质的探针的基因组文库的cDNA 受体基因 扩展搜索将包括三种主要的阿片类药物 受体类别及其推定亚型。 不同的知识 阿片受体基因，它们推导的分子结构，它们的组织 特异性表达和稳定转染细胞系的可用性 将允许开发明确的阿片受体亚型测定， 从而为开发高度特异性的 毒品

Opioids cause multiple physiological effects. Several distinct opioid receptors have been described, but their genes remain unknown. We propose to identify and clone opioid receptor genes to deduce their amino acid sequences and transfect the genes into mammalian cells. We have identified a human neuroblastoma cell line, SK-N-SH-SY5Y, which expresses abundant mu- , and also delta-, receptors. Strategies for gene isolation include 1) use of oligonucleotide probes deduced from peptide sequences of the mu receptor, 2) direct mammalian expression cloning using cDNA libraries from neuroblastoma cell lines; 3) application of the polymerase chain reaction using sequence identities among G protein coupled receptors; 4) screening cDNA of genomic libraries with probes derived from any known opioid receptor gene. The extended search will include the three main opioid receptor classes, and their putative subtypes. Knowledge of distinct opioid receptor genes, their deduced molecular structure, their tissue specific expression, and the availability of stably transfected cell lines will permit the development of unequivocal opioid receptor subtype assays, and thereby, provide a powerful tools for development of highly specific drugs.