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Expressed Protein Ligation: General Experimental Protocols

  • Joshua Diaz
  • Jean-Philippe PelloisEmail author
Protocol
  • 162 Downloads
Part of the Methods in Molecular Biology book series (MIMB, volume 2133)

Abstract

Expressed protein ligation allows for the attachment of a chemically labeled peptide to the N- or C-terminus of a recombinant protein. In this book chapter, the practical considerations involved in using this protein engineering technology are described. In particular, approaches used to design optimal ligation sites are discussed. In addition, several methods used to generate the reactive fragments required for EPL are highlighted in practical details. Finally, strategies that one can implement to achieve efficient ligation reactions are presented.

Key words

Expressed protein ligation N-terminal cysteine C-terminal thioester Solid-phase peptide synthesis Ligation site Protein engineering Protein chemistry Protein labeling 

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© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  1. 1.Department of Biochemistry and BiophysicsTexas A&M UniversityCollege StationUSA
  2. 2.Department of ChemistryTexas A&M UniversityCollege StationUSA

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