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Single‐Plasmid‐Based System for Efficient Noncanonical Amino Acid Mutagenesis in Cultured Mammalian Cells
Author(s) -
Cohen Sarit,
Arbely Eyal
Publication year - 2016
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.201500681
Subject(s) - transfection , amino acid , mutagenesis , plasmid , expression vector , biology , cell culture , biochemistry , transfer rna , chemistry , microbiology and biotechnology , mutation , recombinant dna , gene , genetics , rna
We describe a new expression system for efficient non‐canonical amino acid mutagenesis in cultured mammalian cells by using the pyrrolysine tRNA synthetase/tRNA CUA Pyl pair. A significant improvement in the incorporation of non‐canonical amino acids into proteins was obtained by combining all the required genetic components into a single and compact vector that can be efficiently delivered to different mammalian cell lines by conventional transfection reagents.