title: Transposon-based approaches for generating novel molecular diversity during directed evolution.
abstract: This chapter introduces a set of transposon-based methods that were developed to sample trinucleotide deletion, trinucleotide replacement, and domain insertion. Each approach has a common initial step that utilizes an engineered version of the Mu transposon called MuDel. The inherent low sequence specificity of MuDel results in its random insertion into target DNA during in vitro transposition. Removal of the transposon using a type IIS restriction endonuclease generates blunt-end random breaks at a frequency of one per target gene and the concomitant loss of 3 bp. Self-ligation or insertion of another DNA cassette results in the sampling of trinucleotide deletion or trinucleotide substitution/domain insertion, respectively.
authors: School of Biosciences, Cardiff University, Museum Avenue, Cardiff, CF10 3AT, UK, jonesdd@cardiff.ac.uk.
authors: D Dafydd
authors: DD
authors: Jones
authors: James A J
authors: JA
authors: Arpino
authors: Amy J
authors: AJ
authors: Baldwin
authors: Matthew C
authors: MC
authors: Edmundson
mesh_terms: D004251:DNA Transposable Elements / Q000235:genetics*; D019020:Directed Molecular Evolution / Q000379:methods*
chemical_list: D004251:DNA Transposable Elements