Skip to main content

In Vitro DNA Recombination by Random Priming

  • Protocol
  • 4542 Accesses

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 231))

Abstract

Variation coupled to selection is the hallmark of natural evolution. Although there is no full agreement concerning the best way to create variation, mutation or recombination (1), computational simulation studies have demonstrated the importance of homologous recombination in the evolution of biological systems (2,3). As compared to random mutagenesis, recombination may be advantageous in combining beneficial mutations that have arisen independently and may be synergistic, while simultaneously removing deleterious mutations.

This is a preview of subscription content, log in via an institution.

Buying options

Protocol
USD   49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD   84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD   109.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD   109.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Learn about institutional subscriptions

Springer Nature is developing a new tool to find and evaluate Protocols. Learn more

References

  1. Bäck, T. (1996) Evolutionary Algorithms in Theory and Practice: Evolution Strategies, Evolutionary Programming, Genetic Algorithms. Oxford University Press, New York, NY.

    Google Scholar 

  2. Holland, J. H. (1992) Genetic algorithms. Sci. Am. 267, 66–72.

    Article  Google Scholar 

  3. Forrest, S. (1993) Genetic algorithms: principles of natural selection applied to computation. Science 261, 872–878.

    Article  PubMed  CAS  Google Scholar 

  4. Schmidt-Dannert, C. (2001) Directed evolution of single proteins, metabolic pathways, and viruses. Biochemistry 40, 13,125–13,136.

    Article  PubMed  CAS  Google Scholar 

  5. Zhao, H. and Zha, W. (2002) Evolutionary methods for protein engineering, in Enzyme Functionality: Design, Engineering and Screening, (Svendsen, A., ed.) Marcel Dekker, New York, NY, in press.

    Google Scholar 

  6. Zhang, Y. X., Perry, K., Vinci, V. A., Powell, K., Stemmer, W. P., and del Cardayre, S. B. (2002) Genome shuffling leads to rapid phenotypic improvement in bacteria. Nature 415, 644–646.

    Article  PubMed  CAS  Google Scholar 

  7. Stemmer, W.P. (1994) Rapid evolution of a protein in vitro by DNA shuffling. Nature 370, 389–391.

    Article  PubMed  CAS  Google Scholar 

  8. Stemmer, W.P. (1994) DNA shuffling by random fragmentation and reassembly: in vitro recombination for molecular evolution. Proc. Natl. Acad. Sci. USA 91, 10,747–10,751.

    Article  PubMed  CAS  Google Scholar 

  9. Zhao, H., Giver, L., Shao, Z., Affholter, J. A., and Arnold, F. H. (1998) Molecular evolution by staggered extension process (StEP) in vitro recombination. Nat. Biotechnol. 16, 258–261.

    Article  PubMed  CAS  Google Scholar 

  10. Shao, Z., Zhao, H., Giver, L., and Arnold, F. H. (1998) Random-priming in vitro recombination: an effective tool for directed evolution. Nucleic Acids Res. 26, 681–683.

    Article  PubMed  CAS  Google Scholar 

  11. Coco, W. M., Levinson, W. E., Crist, M. J., et al. (2001) DNA shuffling method for generating highly recombined genes and evolved enzymes. Nat. Biotechnol. 19, 354–359.

    Article  PubMed  CAS  Google Scholar 

  12. Hodgson, C.P. and Fisk, R. Z. (1987) Hybridization probe size control: optimized ‘oligolabelling’. Nucleic Acids Res. 15, 6295.

    Article  PubMed  CAS  Google Scholar 

  13. Zhao, H. and Arnold, F. H. (1997) Optimization of DNA shuffling for high fidelity recombination. Nucleic Acids Res. 25, 1307–1308.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2003 Humana Press Inc., Totowa, NJ

About this protocol

Cite this protocol

Esteban, O., Woodyer, R.D., Zhao, H. (2003). In Vitro DNA Recombination by Random Priming. In: Arnold, F.H., Georgiou, G. (eds) Directed Evolution Library Creation. Methods in Molecular Biology™, vol 231. Humana Press. https://doi.org/10.1385/1-59259-395-X:99

Download citation

  • DOI: https://doi.org/10.1385/1-59259-395-X:99

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-285-8

  • Online ISBN: 978-1-59259-395-8

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics