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Publication . Part of book or chapter of book . 2014

Transposon-Based Approaches for Generating Novel Molecular Diversity During Directed Evolution

D. Dafydd Jones; James A. J. Arpino; Amy Joy Baldwin; Matthew C. Edmundson;
Open Access
Published: 27 Jun 2014
Publisher: Springer New York

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.

Subjects by Vocabulary

Microsoft Academic Graph classification: Directed evolution Restriction enzyme DNA chemistry.chemical_compound chemistry Composite transposon Transposable element Biology Genetics Transposition (music) Sequence (medicine) Simple transposon

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Part of book or chapter of book . 2014