title: An improved dual-tube megaprimer approach for multi-site saturation mutagenesis.
abstract: Saturation mutagenesis is a powerful tool in protein engineering. Even though QuikChange site-directed mutagenesis method is dominantly used in laboratories, it could not be successfully applied to the generation of a focused mutant library of human glutathione transferase A2-2. In the present study, we further developed an improved versatile dual-tube approach of randomizing difficult-to-amplify targets, exhibiting significant improvement towards equal distribution of nucleotides at randomized sites compared to other published methods.
authors.affiliation: Department of Chemistry, BMC, Uppsala University, Box 576, 75123 Uppsala, Sweden.
authors.forename: Wei
authors.initials: W
authors.lastname: Zhang
authors.forename: Bengt
authors.initials: B
authors.lastname: Mannervik
mesh_terms: D005982:Glutathione Transferase / Q000235:genetics; D006801:Humans; D007527:Isoenzymes / Q000235:genetics; D008967:Molecular Biology / Q000379:methods*; D016296:Mutagenesis; D015202:Protein Engineering / Q000379:methods*
chemical_list: D007527:Isoenzymes; D005982:Glutathione Transferase; C119995:glutathione S-transferase alpha