This volume covers some of the most widely used protocols on nanocanonical amino acids, providing details and advice for users to get each method up and running for their chosen application. ?Chapters have been divided into three parts describing methods for protein production in the test tube, in prokaryotes, and in eukaryotes. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and practical, Noncanonical Amino Acids: Methods and Protocols aims to provide readers with techniques that enable them to design new experiments and create new areas of research.
Part I: Engeneering in the Test Tube
1. Leveraging Formylglycine-Generating Enzyme for Production of Site-Specifically Modified Bioconjugates
Robyn M. Barfield and David Rabuka
2. Artificial Division of Codon Boxes for Expansion of the Amino Acid Repertoire of Ribosomal Polypeptide Synthesis
Yoshihiko Iwane, Takayuki Katoh, Yuki Goto, and Hiroaki Suga
3. Cell-free Protein Synthesis for Multiple Site-Specific Incorporation of Noncanonical Amino Acids Using Cell Extracts from RF-1 deletion E. coli Strains
Eiko Seki, Tatsuo Yanagisawa, and Shigeyuki Yokoyama
4. Tub-tag Labeling; Chemoenzymatic Incorporation of Unnatural Amino Acids
Jonas Helma, Heinrich Leonhardt, Christian P.R. Hackenberger, and Dominik Schumacher
Part II: Engeering in Prokaryotes
5. Directed Evolution of Orthogonal Pyrrolysyl-tRNA Synthetases ilÓ+
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