title: Mutagen: a random mutagenesis method providing a complementary diversity generated by human error-prone DNA polymerases.
abstract: Random mutagenesis is one of the most effective methodologies to generate variant libraries for directed protein evolution. Indeed, this approach requires no structural or mechanistic information and can uncover unexpected beneficial mutations. Here, we describe a new random mutagenesis method based on the use of human error-prone DNA polymerases (pol beta, pol eta and pol iota). This approach allows the random introduction of mutations through a single replication step followed by a selective PCR amplification of the replicated mutated sequences. The libraries generated using this methodology display different mutation rates and complementary mutational spectra. By taking advantage of the mutation bias of naturally highly error-prone DNA polymerases, MutaGen thus appears as a very useful tool for gene and protein randomization.
authors.affiliation: Antibody Engineering and Molecular Evolution Department, MilleGen SA, Labège, France. philippe.mondon@millegen.com
authors.forename: Philippe
authors.initials: P
authors.lastname: Mondon
authors.forename: David
authors.initials: D
authors.lastname: Grand
authors.forename: Nathalie
authors.initials: N
authors.lastname: Souyris
authors.forename: Stéphane
authors.initials: S
authors.lastname: Emond
authors.forename: Khalil
authors.initials: K
authors.lastname: Bouayadi
authors.forename: Hakim
authors.initials: H
authors.lastname: Kharrat
mesh_terms: D004259:DNA-Directed DNA Polymerase / Q000235:genetics*; D006801:Humans; D016296:Mutagenesis
chemical_list: D004259:DNA-Directed DNA Polymerase