Titlebook: Genetically Incorporated Non-Canonical Amino Acids; Methods and Protocol Yu-Hsuan Tsai,Simon J. Els?sser Book 2023 The Editor(s) (if applic

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Genetic Code Expansion in Mammalian Cellsch a unique handle into the protein of interest (POI), bioorthogonal reactions can be utilized in live cells to monitor or manipulate the interaction, translocation, function, and modification of the POI. Here, we describe a basic protocol outlining the necessary steps to incorporate a ncAA into a P

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Generation of Amber Suppression Cell Lines Using CRISPR-Cas9ing cell. The archaeal pyrrolysine-tRNA/pyrrolysine-tRNA synthetase (PylT/RS) pair from . (.) has been established for incorporation of a wide range of noncanonical amino acids (ncAAs) in mammalian cells. Once integrated in an engineered protein, ncAAs allow for simple click-chemistry derivatization

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Genetically Encoded 1,2-Aminothiol for Site-Specific Modification of a Cellular Membrane Protein viaion on a target protein is through a reaction between bioorthogonal functionalities. Indeed, various bioorthogonal reactions have been developed, including a recently reported reaction between 1,2-aminothiol and ((alkylthio)(aryl)methylene)malononitrile (TAMM). Here, we describe the procedure that c

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Site-Specific Incorporation of Sulfotyrosine into Proteins in Mammalian Cellsl processes and the development of human diseases, including AIDS and cancer. To facilitate the study of PTS in live mammalian cells, an approach for the site-specific synthesis of tyrosine-sulfated proteins (sulfoproteins) was developed. This approach takes advantage of an evolved . tyrosyl-tRNA sy

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Book 2023ion 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 Mole

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