A method for introducing site-specific mutations using oligonucleotide primers and its application to site-saturation mutagenesis.
Oligonucleotide primer-directed mutagenesis is a useful molecular biological tool, which is invaluable 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 site-saturation mutagenesis.
O'Donohue
M J
MJ
Laboratoire de Médiateurs Protéiques, INRA Centre de Jouy-en-Josas, France.
Kneale
G G
GG
D017931:DNA Primers; D005821:Genetic Techniques; D016297:Mutagenesis, Site-Directed; D013873:Thionucleotides / Q000737:chemistry
D017931:DNA Primers; D013873:Thionucleotides