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A method for introducing site-specific mutations using oligonucleotide primers and its application to site-saturation mutagenesis

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Abstract

Oligonucleotide primer-directed mutagenesis is a useful molecular biological tool, which is invalubable for the study of the structure/function relationships in proteins and for the creation of mutant proteins possessing modified or novel biological activities. Mutagenesis studies in which a site-saturation approach is employed require a high-efficiency mutagenesis procedure, which will generate a population of mutated molecules containing an even distribution of all possible amino acid changes, or a subset thereof. This article describes such a mutagenesis technique and discusses the adaptations that are necessary to perform sitesaturation mutagenesis.

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Authors and Affiliations

  1. Laboratoire de Médiateurs Protéiques, INRA Centre de Jouy-en-Josas, Domaine de Vilvert, 78352, Jouy-en-Josas Cedex, France

    Michael J. O’Donohue

  2. Biophysics Laboratories, University of Portsmouth, England

    G. Geoff Kneale

Authors
  1. Michael J. O’Donohue
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  2. G. Geoff Kneale
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O’Donohue, M.J., Geoff Kneale, G. A method for introducing site-specific mutations using oligonucleotide primers and its application to site-saturation mutagenesis. Mol Biotechnol 6, 179–189 (1996). https://doi.org/10.1007/BF02740772

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  • DOI: https://doi.org/10.1007/BF02740772

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