Abstract
For studies of translational regulation, it is often important to determine the distribution of ribosomes along a particular mRNA species. Knowledge of the exact positions of stalled ribosomes may aid in the identification of features of mRNA sequence and structure that result in ribosome stalling. We therefore devised an assay to allow the determination of ribosome distribution on an mRNA with single nucleotide precision (1). This assay can be used to map the positions of ribosomes during in vitro translation of any mRNA for which a full-length cDNA clone is available. Since development of this assay, it has been used to show that ribosomes frequently pause at initiation and termination codons (1,2), to examine the arrest of translation by signal recognition particle (1,3,4) and to demonstrate that ribosomes pause at the site of a pseudoknot required for frameshifting (5). The assay has also been adapted to monitor the positions of paused ribosomes on abundant mRNAs in vivo (6,7).
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Wolin, S.L. (1998). Analyses of Ribosome Distribution During In Vitro Translation. In: Martin, R. (eds) Protein Synthesis. Methods in Molecular Biology, vol 77. Springer, Totowa, NJ. https://doi.org/10.1385/0-89603-397-X:1
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DOI: https://doi.org/10.1385/0-89603-397-X:1
Publisher Name: Springer, Totowa, NJ
Print ISBN: 978-0-89603-397-9
Online ISBN: 978-1-59259-563-1
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