A high efficiency method for site-directed mutagenesis with any plasmid.
A procedure is described which allows for the site-directed mutagenesis of DNA segments in any double-stranded plasmid with high efficiency. There are no limitations as to the position of the mutation. The protocol involves only simple enzymatic manipulations and no difficult to control operations, such as partial digestions, are required. The method was developed and used to mutagenize two different genes (encoding human interferon-beta and interleukin-2) cloned in a eukaryotic expression vector. For ten mutageneses with different oligodeoxyribonucleotides the average yield of mutants was 60%.
Hofer
B
B
Department of Genetics, Gesellschaft für Biotechnologische Forschung, Braunschweig, F.R.G.
Kühlein
B
B
D001483:Base Sequence; D005796:Genes; D005818:Genetic Engineering / Q000379:methods*; D006801:Humans; D007370:Interferon Type I / Q000235:genetics*; D007376:Interleukin-2 / Q000235:genetics*; D008969:Molecular Sequence Data; D009154:Mutation; D010957:Plasmids
D007370:Interferon Type I; D007376:Interleukin-2