Molecular Biotechnology

, Volume 34, Issue 1, pp 55–68

Mutant library construction in directed molecular evolution

Casting a wider net
  • Tian-Wen Wang
  • Hu Zhu
  • Xing-Yuan Ma
  • Ting Zhang
  • Yu-Shu Ma
  • Dong-Zhi Wei
Review
  • 892 Downloads

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 

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Copyright information

© Humana Press Inc 2006

Authors and Affiliations

  • Tian-Wen Wang
    • 1
  • Hu Zhu
    • 1
  • Xing-Yuan Ma
    • 1
  • Ting Zhang
    • 1
  • Yu-Shu Ma
    • 1
  • Dong-Zhi Wei
    • 1
  1. 1.State Key Laboratory of Bioreactor Engineering, New World Institute of BiotechnologyEast China University of Science and TechnologyShanghaiP. R. China

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