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Monitoring Protein Synthesis in Living Cells With Fluorescent Labeled tRNA FRET Pairs
EP20540
Poster Title: Monitoring Protein Synthesis in Living Cells With Fluorescent Labeled tRNA FRET Pairs
Submitted on 20 Dec 2013
Author(s): Zeev Smilansky, Sima Barhoom, Ian Farrel, Dvir Dahary, Andrew Leask, Peter Vanderklish, Marcelo Ehrlich, Barry S. Cooperman and Orna Elroy-Stein
Affiliations: Anima Cell Metrology, Inc
Poster Views: 1,842
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Poster Information
Abstract: In humans there are 48 distinct isoacceptor tRNAs yielding 1176 distinct tRNA pairs. The E-factor of a protein is defined as the maximal ratio of the frequency of appearance of a specific di-tRNA in the synthesis sequence of the protein of interest relative to its appearance in all other proteins in the relevant cell or tissue under the given experimental conditions.Summary: We transfect cells with tRNAs labeled as FRET donors and acceptors. A FRET signal is generated only when a donor labeled tRNA and an acceptor-labeled tRNA come in close contact (< 7 nM), as they do on the ribosome during the elongation cycle. The intensity of the FRET signal correlates with the number of ribosomes engaged in protein synthesis, providing a real-time, live-cell assay for measuring rates of protein synthesis.Report abuse »
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