Journal
/
Bioquímica
/
Toeprinting Analysis of Translation Initiation Complex Formation on Mammalian mRNAs
JoVE Revista
Bioquímica
Se requiere una suscripción a JoVE para ver este contenido.  Inicie sesión o comience su prueba gratuita.
JoVE Revista Bioquímica
Toeprinting Analysis of Translation Initiation Complex Formation on Mammalian mRNAs

Toeprinting Analysis of Translation Initiation Complex Formation on Mammalian mRNAs

DOI:
10.3791/57519-v

•

10:37 min

•

May 10, 2018

•

,
1Department of Chemistry and Biochemistry, Alberta RNA Research and Training Institute,University of Lethbridge, 2Department of Neuroscience and the Canadian Centre for Behavioral Neuroscience (CCBN),University of Lethbridge, 3Arnie Charbonneau Cancer Institute,University of Calgary

Capítulos

  • 00:04Título
  • 00:34Preparation of mRNA
  • 02:59Toeprinting Assay
  • 06:32Sequencing Reactions and Gel Electrophoresis
  • 07:53Results: Importance of IRES Secondary Structure in the Formation of Translation Initiation Complexes on an Uncapped XIAP IRES RNA
  • 09:35Conclusion

Summary

Traducción Automática

Traducción Automática

Toeprinting aims to measure the ability of in vitro transcribed RNA to form translation initiation complexes with ribosomes under a variety of conditions. This protocol describes a method for toeprinting mammalian RNA and can be used to study both cap-dependent and IRES-driven translation.

Tags

Keywords: Toeprinting AnalysisTranslation Initiation ComplexMammalian MRNAsIRESRNA PurificationRabbit Reticulocyte LysateRNase InhibitorATPGMP-PNPDTTSpermidine

Article

Embed

Añadir a la lista de reproducción

Usage Statistics

Videos relacionados

check_url/es/57519?article_type=v

Protein Complex Affinity Capture from Cryomilled Mammalian Cells
check_url/es/57519?article_type=v

Quantitative Immunofluorescence to Measure Global Localized Translation
check_url/es/57519?article_type=v

Genome-wide Quantification of Translation in Budding Yeast by Ribosome Profiling
check_url/es/57519?article_type=v

An In Vitro Single-Molecule Imaging Assay for the Analysis of Cap-Dependent Translation Kinetics
check_url/es/57519?article_type=v

Complementation of Splicing Activity by a Galectin-3 – U1 snRNP Complex on Beads
check_url/es/57519?article_type=v

Optical Tweezers to Study RNA-Protein Interactions in Translation Regulation
check_url/es/57519?article_type=v

Measuring Composition of CD95 Death-Inducing Signaling Complex and Processing of Procaspase-8 in this Complex
check_url/es/57519?article_type=v

Routine Collection of High-Resolution cryo-EM Datasets Using 200 KV Transmission Electron Microscope
check_url/es/57519?article_type=v

Evaluating Biophysical Assays for Characterizing PROTACS Ternary Complexes (Video) | JoVE
check_url/es/57519?article_type=v

Author Spotlight: Characterization of Low-Affinity Protein Interactions in Solution Using MassFluidix Technology

Read Article