Abstract
This chapter describes three different strategies of receptor mutagenesis with their advantages, disadvantages, and limitations. Oligonucleotide-directed mutagenesis using either the Altered Sites® II in vitro mutagenesis system or the GeneTailor® site-directed mutagenesis system can generate base substitutions/deletions/insertions that yield single/multiple amino acid substitutions/deletions/insertions and/or N- or C-terminal truncations in GPCRs. Polymerase chain reaction-based mutagenesis strategies allow substitutions/deletions/insertions of larger domains within GPCRs, creating truncated receptors or receptor chimeras.
In addition, some guidelines are given and examples are provided to facilitate design and interpretation of mutational experiments.
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Blomenröhr, M., Vischer, H.F., Bogerd, J. (2004). Receptor Mutagenesis Strategies for Examination of Structure-Function Relationships. In: Willars, G.B., Challiss, R.A.J. (eds) Receptor Signal Transduction Protocols. Methods in Molecular Biology, vol 259. Humana Press. https://doi.org/10.1385/1-59259-754-8:307
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DOI: https://doi.org/10.1385/1-59259-754-8:307
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