[5] Cloning of G protein-coupled opioid receptors using degenerate PCR and low-stringency homology screening

David K. Grandy, Qun Yong Zhou, Claudia Bouvier, Carmen Saez, James R. Bunzow

    Research output: Research - peer-reviewArticle

    • 2 Citations

    Abstract

    In summary we have described methods that we have used to clone the classical μ, δ, and κ opioid receptors and a new receptor, LC132, that based on structural arguments we predict is also a member of the opioid receptor gene family. The combination of low-stringency homology screening and degenerate PCR provides a powerful strategy for the cloning of members of this G protein-coupled receptor family. When these cloning techniques are combined with transient and/or stable expression and classical pharmacological analyses it is often possible to determine which receptor subtype is encoded by the cloned cDNA. Given the sensitivity of the molecular approaches, in particular PCR, and what has been observed for other G protein-coupled receptors that bind the same, or similar ligands, we expect that more receptor genes will be identified than were orginally predicted from earlier pharmacological data. The cloning of LC132 is a perfect example of the situation that molecular neuropharmacologists now face; that is the ability to clone and express a new receptor type whose sequence allows its inclusion within an established gene family yet whose endogenous ligand is unknown. We are confident, however, that in time the endogenous ligand for orphan receptors such as LC132 will be determined by combining expression in tissue culture with pharmacological and physiological analyses.

    LanguageEnglish (US)
    Pages90-104
    Number of pages15
    JournalMethods in Neurosciences
    Volume25
    Issue numberC
    DOIs
    StatePublished - 1995

    Fingerprint

    Opioid Receptors
    G-Protein-Coupled Receptors
    Organism Cloning
    Pharmacology
    Ligands
    Polymerase Chain Reaction
    Genes
    Clone Cells
    kappa Opioid Receptor
    delta Opioid Receptor
    mu Opioid Receptor
    Complementary DNA

    ASJC Scopus subject areas

    • Neuroscience(all)

    Cite this

    [5] Cloning of G protein-coupled opioid receptors using degenerate PCR and low-stringency homology screening. / Grandy, David K.; Zhou, Qun Yong; Bouvier, Claudia; Saez, Carmen; Bunzow, James R.

    In: Methods in Neurosciences, Vol. 25, No. C, 1995, p. 90-104.

    Research output: Research - peer-reviewArticle

    Grandy, David K. ; Zhou, Qun Yong ; Bouvier, Claudia ; Saez, Carmen ; Bunzow, James R./ [5] Cloning of G protein-coupled opioid receptors using degenerate PCR and low-stringency homology screening. In: Methods in Neurosciences. 1995 ; Vol. 25, No. C. pp. 90-104
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