Expressed Protein Ligation: General Experimental Protocols.

Joshua Diaz, Jean-Philippe Pellois

Journal: Methods in molecular biology (Clifton, N.J.) 2021;2133():75-117

PMID: 32144664

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.

Address: Department of Biochemistry and Biophysics, Texas A&M University, College Station, TX, USA.; Department of Biochemistry and Biophysics, Texas A&M University, College Station, TX, USA. [email protected].; Department of Chemistry, Texas A&M University, College Station, TX, USA. [email protected].

DOI: 10.1007/978-1-0716-0434-2_5

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https://doi.org/10.1007/978-1-0716-0434-2_5

MeSH Terms: Amino Acids, Cloning, Molecular, Electrophoresis, Polyacrylamide Gel, Escherichia coli, Esters, Gene Expression, Inteins, Peptides, Protein Engineering, Recombinant Fusion Proteins, Solid-Phase Synthesis Techniques, Sulfhydryl Compounds