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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

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

DOI:
10.3791/4026-v

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10:15 min

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July 06, 2012

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1Center for Gene Regulation in Health and Disease, Department of Biological, Geological and Environmental Sciences,Cleveland State University

Chapters

  • 00:05Title
  • 03:55DNA Template Preparation, In vitro Transcription, and In vitro Translation
  • 05:30Isolation and Resolution of Nascent Polypeptides
  • 06:54Representative Translational Pause Results
  • 08:35Conclusion

Summary

Automatic Translation

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

RibosomePolypeptidesTranslational Pause SitesMRNACodon UsageSynonymous CodonsNon-uniform ElongationMRNA Secondary StructureRibosome MovementTranslational Elongation RatesCodon BiasTRNAsRare CodonsPause SitesProtein ExpressionMRNA StabilityProtein Folding

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