Approaches to receptor cloning

This chapter discusses the approaches to receptor cloning. It has often been observed that when multiple approaches have developed, they are all likely to have limitations. Successful and efficient receptor cloning requires the development of a reasonable overall strategy and the selection of the most appropriate methodologies for the problem. Before a cloning strategy is developed, assessment of the reasons, which the clone is being isolated and of the receptor class, is critical. Representing the two extremes of difficulty is cloning a multi-subunit receptor of unknown structure, such as seminal nicotinic acetylcholine receptor cloning, and cloning a fragment of a receptor cyclic DNA (cDNA) across mammalian species for use in nuclease protection assays or in situ hybridization, such as a monkey dopamine receptor probe with the sequence already characterized in several other mammalian species. In the case of a multiple subunit receptor, expression cloning may be impossible and the arduous task of receptor purification and microsequencing or antibody generation the requisite initial steps.