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…mehr
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.
Leveraging Formylglycine-Generating Enzyme for Production of Site-Specifically Modified Bioconjugates.- Artificial Division of Codon Boxes for Expansion of the Amino Acid Repertoire of Ribosomal Polypeptide Synthesis.- Cell-free Protein Synthesis for Multiple Site-Specific Incorporation of Noncanonical Amino Acids Using Cell Extracts from RF-1 deletion E. coli Strains.- Tub-tag Labeling; Chemoenzymatic Incorporation of Unnatural Amino Acids.- Directed Evolution of Orthogonal Pyrrolysyl-tRNA Synthetases in Escherichia coli for the Genetic Encoding of Noncanonical Amino Acids.- Genetic Code Expansion in Enteric Bacterial Pathogens.- Self-directed in Cell Production of Methionine Analogue Azidohomoalanine by Synthetic Metabolism and its Incorporation into Model Proteins.- Residue-specific Incorporation of no Canonical Amino Acids for Protein Engineering.- Using Amber and Ochre Nonsense Codons to Code Two Different Noncanonical Amino Acids in One Protein Gene.- Genetic Incorporation of Unnatural Amino Acids into Proteins of Interest in Streptomyces venezuelae ATCC 15439.- Expression and Purification of Site-specifically Lysine-acetylated and Natively-folded Proteins for Biophysical Investigations.- Site-specific Incorporation of Sulfotyrosine Using an Expanded Genetic Code.- Site-specific Protein Labeling With Tetrazine Amino Acids.- Mapping of Protein Interfaces in Live Cells Using Genetically Encoded Crosslinkers.- Generation of Stable Amber Suppression Cell Lines.- Trapping Chromatin Interacting Proteins with Genetically Encoded, UV-Activatable Crosslinkers In Vivo.- Genetically Encoding Unnatural Amino Acids in Neurons in vitro and in the Embryonic Mouse Brain for Optical Control of Neuronal Proteins.- Genetic Code Expansion-and Click Chemistry-Based Site--Specific Protein Labelling For Intracellular DNA--PAINT Imaging.- MultiBacTAG- Genetic Code Expansion using the Baculovirus Expression System in Sf21 Cells.- Production and Chemoselective Modification of Adeno-associated Virus Site-specifically Incorporating an Unnatural Amino Acid Residue into its Capsid.- Generation of Intramolecular FRET Probes via Noncanonical Amino Acid Mutagenesis.- Fluorogenic Tetrazine--siliconrhodamine Probe for the Labeling of Non-canonical Amino Acid Tagged Proteins.- Site-specific Protein Labeling Utilizing Lipoic Acid Ligase (LplA) and Bioorthogonal Inverse Electron Demand Diels-Alder Reaction.- Genetic Encoding of Unnatural Amino Acids in C. elegans.
Leveraging Formylglycine-Generating Enzyme for Production of Site-Specifically Modified Bioconjugates.- Artificial Division of Codon Boxes for Expansion of the Amino Acid Repertoire of Ribosomal Polypeptide Synthesis.- Cell-free Protein Synthesis for Multiple Site-Specific Incorporation of Noncanonical Amino Acids Using Cell Extracts from RF-1 deletion E. coli Strains.- Tub-tag Labeling; Chemoenzymatic Incorporation of Unnatural Amino Acids.- Directed Evolution of Orthogonal Pyrrolysyl-tRNA Synthetases in Escherichia coli for the Genetic Encoding of Noncanonical Amino Acids.- Genetic Code Expansion in Enteric Bacterial Pathogens.- Self-directed in Cell Production of Methionine Analogue Azidohomoalanine by Synthetic Metabolism and its Incorporation into Model Proteins.- Residue-specific Incorporation of no Canonical Amino Acids for Protein Engineering.- Using Amber and Ochre Nonsense Codons to Code Two Different Noncanonical Amino Acids in One Protein Gene.- Genetic Incorporation of Unnatural Amino Acids into Proteins of Interest in Streptomyces venezuelae ATCC 15439.- Expression and Purification of Site-specifically Lysine-acetylated and Natively-folded Proteins for Biophysical Investigations.- Site-specific Incorporation of Sulfotyrosine Using an Expanded Genetic Code.- Site-specific Protein Labeling With Tetrazine Amino Acids.- Mapping of Protein Interfaces in Live Cells Using Genetically Encoded Crosslinkers.- Generation of Stable Amber Suppression Cell Lines.- Trapping Chromatin Interacting Proteins with Genetically Encoded, UV-Activatable Crosslinkers In Vivo.- Genetically Encoding Unnatural Amino Acids in Neurons in vitro and in the Embryonic Mouse Brain for Optical Control of Neuronal Proteins.- Genetic Code Expansion-and Click Chemistry-Based Site--Specific Protein Labelling For Intracellular DNA--PAINT Imaging.- MultiBacTAG- Genetic Code Expansion using the Baculovirus Expression System in Sf21 Cells.- Production and Chemoselective Modification of Adeno-associated Virus Site-specifically Incorporating an Unnatural Amino Acid Residue into its Capsid.- Generation of Intramolecular FRET Probes via Noncanonical Amino Acid Mutagenesis.- Fluorogenic Tetrazine--siliconrhodamine Probe for the Labeling of Non-canonical Amino Acid Tagged Proteins.- Site-specific Protein Labeling Utilizing Lipoic Acid Ligase (LplA) and Bioorthogonal Inverse Electron Demand Diels-Alder Reaction.- Genetic Encoding of Unnatural Amino Acids in C. elegans.
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