Overview of Receptor Cloning

Norman Nash1

1 Acadia Pharmaceuticals, San Diego, California
Publication Name:  Current Protocols in Pharmacology
Unit Number:  Unit 6.1
DOI:  10.1002/0471141755.ph0601s00
Online Posting Date:  May, 2001
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Receptors are generally categorized into four major superfamilies according to their structural and genetic similarities: steroid receptors, growth factor receptors, ionotropic receptors (i.e., ligand‐gated ion channels), and metabotropic receptors (i.e., G protein‐coupled receptors). This unit describes the methodology and critical issues for cloning receptors by several methods. Although cloning by protein purification was the first major method, it is now less frequently employed. The early, more tedious procedures broadened knowledge about receptors and established a strong foundation that has enabled the utilization of the newer approaches discussed in this unit, including: homology cloning by screening with oligonucleotides, DNA fragments or the products of direct polymerase chain reaction (PCR); expression cloning using function, ligand binding, and antibody recognition; and differential display techniques.

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Table of Contents

  • Cloning Strategies
  • Critical Parameters
  • Figures
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Literature Cited

Literature Cited
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   Kramer, M.F. and Coen, D.M. 1995. Enzymatic amplification of DNA by PCR: Standard procedures and optimization. In Current Protocols in Molecular Biology (F.M. Ausubel, R. Brent, R.E. Kingston, D.D. Moore, J.G. Seidman, J.A. Smith, and K. Struhl, eds.) pp. 15.1.1‐15.1.9. John Wiley & Sons, New York.
   Liang, P. and Pardee, A.B. 1992. Differential display of eukaryotic messenger RNA by means of the polymerase chain reaction. Science 257:967‐971.
   Masu, Y., Nakayama, K., Tamaki, H., Harada, Y., Kuno, M., and Nakanishi, S. 1987. cDNA cloning of bovine substance‐K receptor through oocyte expression system. Nature (Lond.) 329:836‐838.
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   Moriyoshi, K., Masu, M., Ishii, T., Shigemoto, R., Mizuno, N., and Nakanishi, S. 1991. Molecular cloning and characterization of the rat NMDA receptor. Nature (Lond.) 354:31‐37.
   Nash, N., Heilman, C., Rees, H., and Levey, A. 1997. Cloning and localization of exon 5‐containing isoforms of the NMDAR1 subunit in human and rat brains. J. Neurochem. 69:485‐493.
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Key References
   Ausubel et al., 1998. See above.
  An excellent source of general molecular biology techniques.
   Liang and Pardee, 1995. See above.
  Original description of differential display, showing careful controls and analysis.
   Numa, 1989. See above.
  A good review of ionotropic receptors.
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