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Highly chromophoric Cy5-methionine for N-terminal fluorescent tagging of proteins in eukaryotic translation systems.

Sci Rep.. 2017-09; 
Kim JM, Seong BL.
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Gene Synthesis ... was constructed as follows: the HIV-1 Tat gene was selected from the HIV-1 complete genome (GenBank ID: {"type":"entrez-nucleotide","attrs":{"text":"NC_001802","term_id":"9629357","term_text":"NC_001802"}}NC_001802), chemically synthesized (GenScript), and used as a ... Get A Quote

Abstract

Despite significant advances on fluorescent labeling of target proteins to study their structural dynamics and function, there has been need for labeling with high quantum yield ensuring high sensitivity and selectivity without sacrificing the biological function of the protein. Here as a technical advancement over non-canonical amino acid incorporation, we provided a conceptual design of the N-terminal fluorescent tagging of proteins. Cy5-labeled methionine (Cy5-Met) was chemically synthesized, and then the purified Cy5-Met was coupled with synthetic human initiator tRNA by methionine tRNA synthetase. Cy5-Met-initiator tRNA (Cy5-Met-tRNAi) was purified and transfected into HeLa cells with HIV-Tat plasmid, resu... More

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