Review

Molecular Biotechnology

, Volume 34, Issue 1, pp 55-68

First online:

Mutant library construction in directed molecular evolution

Casting a wider net
  • Tian-Wen WangAffiliated withState Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology
  • , Hu ZhuAffiliated withState Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology
  • , Xing-Yuan MaAffiliated withState Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology
  • , Ting ZhangAffiliated withState Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology
  • , Yu-Shu MaAffiliated withState Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology
  • , Dong-Zhi WeiAffiliated withState Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology Email author 

Rent the article at a discount

Rent now

* Final gross prices may vary according to local VAT.

Get Access

Abstract

Directed molecular evolution imitates the natural selection process in the laboratory to find mutant proteins with improved properties in the expected aspects by exploring the encoding sequence space. The success of directed molecular evolution experiment depends on the quality of artificially prepared mutant libraries and the availability of convenient high-throughput screening methods. Well-prepared libraries promise the possibility of obtaining desired mutants by screening a library containing a relatively small number of mutants. This article summarizes and reviews the currently available methodologies widely used in directed evolution practices in the hope of providing a general reference for library construction. These methods include error-prone polymerase chain reaction (epPCR), oligonucleotide-based mutagenesis, and genetic recombination exemplified by DNA shuffling and its derivatives. Another designed method is also discussed, in which B-lymphocytes are fooled to mutate nonantibody foreign proteins through somatic hypermutation (SHM).

Index Entries

Directed molecular evolution error-prone PCR oligonucleotide-based mutagenesis DNA shuffling mutator strain somatic hypermutation