Journal
/
Biologie
/
Removal of an Internal Translational Start Site from mRNA While Retaining Expression of the Full-Length Protein
Journal JoVE
Biologie
Un abonnement à JoVE est nécessaire pour voir ce contenu.  Connectez-vous ou commencez votre essai gratuit.
Journal JoVE Biologie
Removal of an Internal Translational Start Site from mRNA While Retaining Expression of the Full-Length Protein

Removal of an Internal Translational Start Site from mRNA While Retaining Expression of the Full-Length Protein

DOI:
10.3791/63405-v

•

05:48 min

•

March 16, 2022

•

, , ,
1Nora Eccles Harrison Cardiovascular Research and Training Institute,University of Utah, 2Department of Biomedical Sciences,Cedars-Sinai Medical Center

Chapitres

  • 00:04Introduction
  • 00:42DNA Extraction
  • 01:45DNA Amplification by PCR
  • 02:42Incubation with Restriction Enzyme
  • 03:34Preparation of 1.5% Agarose Gel for Electrophoresis and DNA Band Detection
  • 04:22Results: Genotype Analysis and Recombination Efficiency
  • 05:21Conclusion

Summary

Traduction automatique

Traduction automatique

The present protocol describes a single M213L mutation in Gja1 that retains full-length Connexin43 generation but prevents translation of the smaller GJA1-20k internally translated isoform.

Tags

MRNAProtein ExpressionGenotypingPCRRestriction EnzymeTail SampleThermal CyclerPCR Master MixNlaIIIAgarose Gel Electrophoresis

Article

Embed

AJOUTER À LA PLAYLIST

Usage Statistics

Vidéos Connexes

The Preparation of Primary Hematopoietic Cell Cultures From Murine Bone Marrow for Electroporation

Homemade Site Directed Mutagenesis of Whole Plasmids

Staining of Proteins in Gels with Coomassie G-250 without Organic Solvent and Acetic Acid

Using an Automated Cell Counter to Simplify Gene Expression Studies: siRNA Knockdown of IL-4 Dependent Gene Expression in Namalwa Cells

Budding Yeast Protein Extraction and Purification for the Study of Function, Interactions, and Post-translational Modifications

Identification of Post-translational Modifications of Plant Protein Complexes

siRNA Screening to Identify Ubiquitin and Ubiquitin-like System Regulators of Biological Pathways in Cultured Mammalian Cells

In Vitro Synthesis of Modified mRNA for Induction of Protein Expression in Human Cells

Green Fluorescent Protein-based Expression Screening of Membrane Proteins in Escherichia coli

A Comparative Analysis of Recombinant Protein Expression in Different Biofactories: Bacteria, Insect Cells and Plant Systems

Read Article