title: Focusing mutations within random libraries to distinct areas: protein domain library generation by overlap extension.
abstract: Directed evolution is an often used approach toward new proteins with tailor-made properties. It consists of random variation of the coding sequence of a protein followed by an appropriate selection procedure or a suitable type of property read out. In many, if not all cases, it is of significant advantage to constrain the randomly mutagenized DNA sequence to that encoding a particular part of the protein or a distinct domain, and not to mutate the entire gene of the target protein. For this purpose, a three-step, polymerase-based method was developed, which is independent of two flanking restriction sites adjacent to the nucleotide sequence supposed to be mutagenized, and named protein library generation by overlap extension (PDLGO).
authors.affiliation: Institute of Pharmaceutical and Medicinal Chemistry, Heinrich-Heine-University, Düsseldorf, Germany.
authors.forename: Andreas
authors.initials: A
authors.lastname: Gratz
authors.forename: Joachim
authors.initials: J
authors.lastname: Jose
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D019143:Evolution, Molecular; D008969:Molecular Sequence Data; D016296:Mutagenesis; D009154:Mutation; D019151:Peptide Library; D016133:Polymerase Chain Reaction / Q000379:methods; D017434:Protein Structure, Tertiary
chemical_list: D019151:Peptide Library