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Isolation of Ribosome Bound Nascent Polypeptides in vitro to Identify Translational Pause Sites Along mRNA
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Isolation of Ribosome Bound Nascent Polypeptides in vitro to Identify Translational Pause Sites Along mRNA

Article DOI: 10.3791/4026 • 10:15 min • July 6th, 2012
July 6th, 2012 •
1, 1
1Center for Gene Regulation in Health and Disease, Department of Biological, Geological and Environmental Sciences, Cleveland State University

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Summary
Automatic Translation

A technique to identify translational pause sites on mRNA is described. This procedure is based on isolation of nascent polypeptides accumulating on ribosomes during in vitro translation of a target mRNA, followed by the size analysis of the nascent chains using a denaturing gel electrophoresis.

Tags

Ribosome Polypeptides Translational Pause Sites MRNA Codon Usage Synonymous Codons Non-uniform Elongation MRNA Secondary Structure Ribosome Movement Translational Elongation Rates Codon Bias TRNAs Rare Codons Pause Sites Protein Expression MRNA Stability Protein Folding
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