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Polysome Fractionation and Analysis of Mammalian Translatomes on a Genome-wide Scale
JoVE Journal
Biologia
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JoVE Journal Biologia
Polysome Fractionation and Analysis of Mammalian Translatomes on a Genome-wide Scale

Polysome Fractionation and Analysis of Mammalian Translatomes on a Genome-wide Scale

DOI:
10.3791/51455-v

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10:56 min

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May 17, 2014

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, , , , , ,
1Lady Davis Institute and Department of Oncology,McGill University, 2Department of Oncology-Pathology,Karolinska Institutet, 3Goodman Cancer Centre and Department of Biochemistry,McGill University

Capítulos

  • 00:05Título
  • 01:54Preparation of Sucrose Gradient
  • 03:38Isolation and Sedimentation of Polysomes
  • 06:59Polysome Fractionation and RNA Extraction
  • 09:22Results: Analytics of Translational Activity in MCF7 Cells Using Polysome Fractionation
  • 10:24Conclusion

Summary

Tadução automática

Tadução automática

Ribosomes play a central role in protein synthesis. Polyribosome (polysome) fractionation by sucrose density gradient centrifugation allows direct determination of translation efficiencies of individual mRNAs on a genome-wide scale. In addition, this method can be used for biochemical analysis of ribosome- and polysome-associated factors such as chaperones and signaling molecules.

Tags

Keywords: MRNA TranslationTranslatomeRibosomePolysomeSucrose Density Gradient CentrifugationAnota AlgorithmGenome-wide AnalysisProtein FoldingPost-translational Modifications

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