This detailed volume explores non-canonical amino acids (ncAAs) through their site-specific incorporation by genetic code expansion (GCE). The collection provides a broad resource of methods for implementing GCE in E. coli, mammalian cells, and animals, highlighting specific applications ranging from fluorescence labeling to photocontrol and the study of protein post-translational modification. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step and readily…mehr
This detailed volume explores non-canonical amino acids (ncAAs) through their site-specific incorporation by genetic code expansion (GCE). The collection provides a broad resource of methods for implementing GCE in E. coli, mammalian cells, and animals, highlighting specific applications ranging from fluorescence labeling to photocontrol and the study of protein post-translational modification. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step and readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Genetically Incorporated Non-Canonical Amino Acids: Methods and Protocols serves as an ideal source of methodologies that can be adapted and extended, migrated to different model systems, and combined in new ways to help explore a wide range of biological questions and to augment industrial and pharmaceutical protein engineering.
Focused Engineering of Pyrrolysyl tRNA Synthetase Based Orthogonal Translation Systems for the Incorporation of Various Non Canonical Amino Acids. Engineering Homogeneous Photoactive Antibodies Fragments. Repurposing Photosensitizer Proteins through Genetic Code Expansion to Facilitate Photo Biocatalysis. Genetic Encoding of a Fluorescent Non Canonical Amino Acid as a FRET Donor for Analysis of Deubiquitinase Activities. Creating Selenocysteine Specific Reporters Using Inteins. Protein Expression with Biosynthesized Non Canonical Amino Acids. Reprogramming Initiator and Nonsense Codons to Simultaneously Install Three Distinct Non Canonical Amino Acids into Proteins in E. coli. Encoding Non Canonical Amino Acids into Phage Displayed Proteins. Genetically Encoded Non Canonical Amino Acids in Proteins to Investigate Lysine Benzoylation. Semi Synthesis of Glutamine Methylated Proteins Enabled by Genetic Code Expansion. Genetic Code Expansion in Mammalian Cells. Generation of Amber Suppression Cell Lines Using CRISPR Cas9. Genetic Code Expansion in Mammalian Cells through Quadruplet Codon Decoding. Genetically Encoded 1,2 Aminothiol for Site Specific Modification of a Cellular Membrane Protein via TAMM Condensation. Conformational GPCR BRET Sensors Based on Bioorthogonal Labeling of Non Canonical Amino Acids. Selective Inhibition of Kinase Activity in Mammalian Cells by Bioorthogonal Ligand Tethering. Site Specific Incorporation of Sulfotyrosine into Proteins in Mammalian Cells. Small Molecule Phosphine Activation of Protein Function in Zebrafish Embryos with an Expanded Genetic Code. Non Canonical Amino Acid Incorporation in Mice.
Focused Engineering of Pyrrolysyl tRNA Synthetase Based Orthogonal Translation Systems for the Incorporation of Various Non Canonical Amino Acids. Engineering Homogeneous Photoactive Antibodies Fragments. Repurposing Photosensitizer Proteins through Genetic Code Expansion to Facilitate Photo Biocatalysis. Genetic Encoding of a Fluorescent Non Canonical Amino Acid as a FRET Donor for Analysis of Deubiquitinase Activities. Creating Selenocysteine Specific Reporters Using Inteins. Protein Expression with Biosynthesized Non Canonical Amino Acids. Reprogramming Initiator and Nonsense Codons to Simultaneously Install Three Distinct Non Canonical Amino Acids into Proteins in E. coli. Encoding Non Canonical Amino Acids into Phage Displayed Proteins. Genetically Encoded Non Canonical Amino Acids in Proteins to Investigate Lysine Benzoylation. Semi Synthesis of Glutamine Methylated Proteins Enabled by Genetic Code Expansion. Genetic Code Expansion in Mammalian Cells. Generation of Amber Suppression Cell Lines Using CRISPR Cas9. Genetic Code Expansion in Mammalian Cells through Quadruplet Codon Decoding. Genetically Encoded 1,2 Aminothiol for Site Specific Modification of a Cellular Membrane Protein via TAMM Condensation. Conformational GPCR BRET Sensors Based on Bioorthogonal Labeling of Non Canonical Amino Acids. Selective Inhibition of Kinase Activity in Mammalian Cells by Bioorthogonal Ligand Tethering. Site Specific Incorporation of Sulfotyrosine into Proteins in Mammalian Cells. Small Molecule Phosphine Activation of Protein Function in Zebrafish Embryos with an Expanded Genetic Code. Non Canonical Amino Acid Incorporation in Mice.
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