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Sequence Homology-Independent Protein Recombination (SHIPREC)

  • Andrew K. Udit
  • Jonathan J. Silberg
  • Volker Sieber
Part of the Methods in Molecular Biology™ book series (MIMB, volume 231)

Abstract

Genomic recombination is widely recognized as the principal mechanism by which proteins evolve new functions (1). Having realized the importance of recombination in evolution, scientists have developed a variety of methods to mimic this phenomenon in the lab to create libraries of gene chimeras. While the ideal technique would allow for the recombination of any parental genes, most available methods are limited to recombining closely related sequences, i.e., those with ≥70% sequence identity, resulting in crossovers biased towards regions of high identity. This occurs because most recombination methods, e.g., Stemmer-shuffling (2), StEP (3), RACHITT (4), and in vivo methods (5), rely on template switching to generate chimeras. However, two methods have recently been described which are capable of generating chimeric libraries independent of sequence identity. Sequence homology-independent protein recombination (SHIPREC) (6) is described in this chapter; another method, incremental truncation for the creation of hybrid enzymes (ITCHY) (7), is also described in this volume. Both methods are capable of generating chimeric libraries containing all possible single crossovers between the two parental genes.

Keywords

Gene Fusion Ethylene Diamine Tetraacetic Acid Ethylene Diamine Tetraacetic Acid Zymo Research Chloramphenicol Acetyl Transferase 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Humana Press Inc., Totowa, NJ 2003

Authors and Affiliations

  • Andrew K. Udit
    • 1
  • Jonathan J. Silberg
    • 1
  • Volker Sieber
    • 2
  1. 1.Division of Chemistry and Chemical EngineeringCalifornia Institute of TechnologyPasadena
  2. 2.Degussa Texturant Systems GmbHFreisingGermany

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