SpeedyGenes: Exploiting an Improved Gene Synthesis Method for the Efficient Production of Synthetic Protein Libraries for Directed Evolution

Methods Mol Biol. 2017;1472:63-78. doi: 10.1007/978-1-4939-6343-0_5.

Abstract

Gene synthesis is a fundamental technology underpinning much research in the life sciences. In particular, synthetic biology and biotechnology utilize gene synthesis to assemble any desired DNA sequence, which can then be incorporated into novel parts and pathways. Here, we describe SpeedyGenes, a gene synthesis method that can assemble DNA sequences with greater fidelity (fewer errors) than existing methods, but that can also be used to encode extensive, statistically designed sequence variation at any position in the sequence to create diverse (but accurate) variant libraries. We summarize the integrated use of GeneGenie to design DNA and oligonucleotide sequences, followed by the procedure for assembling these accurately and efficiently using SpeedyGenes.

Keywords: Directed evolution; Error correction; Gene synthesis; Protein libraries; Synthetic biology.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Base Sequence
  • Cloning, Molecular
  • Codon
  • Directed Molecular Evolution*
  • Escherichia coli / genetics
  • Genes, Synthetic*
  • Oligonucleotides / genetics
  • Peptide Library
  • Polymerase Chain Reaction
  • Transformation, Bacterial
  • User-Computer Interface

Substances

  • Codon
  • Oligonucleotides
  • Peptide Library