Site‐directed Mutagenesis of Double‐stranded Plasmids


Site‐directed mutagenesis can be used to mutate DNA fragments inserted into plasmid vectors using one oligonucleotide and three restriction enzymes.

Keywords: mutagens

Figure 1.

Site‐directed mutagenesis of double‐stranded plasmids.



Cordell B, Bell G, Tisher E et al. (1979) Isolation and characterization of a cloned rat insulin gene. Cell 18: 533–543.

De Chiara TM, Erlitz F and Tarnowski SJ (1986) Procedures for in vitro DNA mutagenesis of human‐leukocyte interferon sequences. Methods in Enzymology 199: 403–415.

Derbyshire KM, Salvo JJ and Grindley NDF (1986) A simple and efficient procedure for saturation mutagenesis using mixed oligodeoxynucleotides. Gene 46: 145–152.

Lai D, Zhu X and Pestka S (1993) A simple and efficient method for site‐directed mutagenesis with double‐stranded plasmid DNA. Nucleic Acids Research 21: 3977–3980.

Li B, Langer JA, Schwartz B and Pestka S (1989) Creation of phosphorylation sites in proteins – construction of a phosphorylatable human interferon‐alpha. Proceedings of the National Academy of Sciences of the USA 86: 558–562.

Morinaga Y, Franceschini T, Inouye S and Inouye M (1984) Improvement of oligonucleotide‐directed site‐specific mutagenesis using double‐stranded plasmid DNA. Bio/Technology 2: 636–639.

Vlasuk GP and Inouye S (1983) Site specific mutagenesis using synthetic oligodeoxynucleotides as mutagens. In: Inouye M (ed.) Experimental Manipulation of Gene Expression, appendix 2, pp. 291–303. New York: Academic Press.

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Jones, P(Jul 2003) Site‐directed Mutagenesis of Double‐stranded Plasmids. In: eLS. John Wiley & Sons Ltd, Chichester. [doi: 10.1038/npg.els.0003798]