Rivista
/
Biochimica
/
Production of Dynein and Kinesin Motor Ensembles on DNA Origami Nanostructures for Single Molecule Observation
JoVE Journal
Biochimica
È necessario avere un abbonamento a JoVE per visualizzare questo.  Accedi o inizia la tua prova gratuita.
JoVE Journal Biochimica
Production of Dynein and Kinesin Motor Ensembles on DNA Origami Nanostructures for Single Molecule Observation

Production of Dynein and Kinesin Motor Ensembles on DNA Origami Nanostructures for Single Molecule Observation

DOI:
10.3791/60369-v

•

08:09 min

•

October 15, 2019

•

,
1Department of Biological Sciences,Smith College, 2Center for Microscopy and Imaging,Smith College

Capitoli

  • 00:04Titolo
  • 00:46Galactose-Induced Motor Protein Growth, Expression, and Harvesting
  • 01:39Cell Lysis and Soluble Protein Extraction
  • 02:36IgG Affinity Purification
  • 03:17DNA Oligonucleotide Labeling and TEV Cleavage
  • 04:50Chassis Purification
  • 05:56Results: Representative Motor Ensemble Production
  • 07:26Conclusion

Summary

Traduzione automatica

Traduzione automatica

The goal of this protocol is to form ensembles of molecular motors on DNA origami nanostructures and observe the ensemble motility using total internal reflection fluorescence microscopy.

Tags

Keywords: DNA OrigamiCytoskeletal Motor ProteinsDyneinKinesinMyosinCargo TransportSingle Molecule ObservationYeast ExpressionProtein PurificationIgG Affinity Purification

Article

Embed

AGGIUNGI ALLA PLAYLIST

Usage Statistics

Video correlati

check_url/it/60369?article_type=v

High Precision FRET at Single-molecule Level for Biomolecule Structure Determination
check_url/it/60369?article_type=v

A Simple, Robust, and High Throughput Single Molecule Flow Stretching Assay Implementation for Studying Transport of Molecules Along DNA
check_url/it/60369?article_type=v

Single-molecule Manipulation of G-quadruplexes by Magnetic Tweezers
check_url/it/60369?article_type=v

Covalent Immobilization of Proteins for the Single Molecule Force Spectroscopy
check_url/it/60369?article_type=v

Proteome-wide Quantification of Labeling Homogeneity at the Single Molecule Level
check_url/it/60369?article_type=v

OaAEP1-Mediated Enzymatic Synthesis and Immobilization of Polymerized Protein for Single-Molecule Force Spectroscopy
check_url/it/60369?article_type=v

Conventional BODIPY Conjugates for Live-Cell Super-Resolution Microscopy and Single-Molecule Tracking
check_url/it/60369?article_type=v

Single Cell Analysis Of Transcriptionally Active Alleles By Single Molecule FISH
check_url/it/60369?article_type=v

Single-Molecule Dwell-Time Analysis of Restriction Endonuclease-Mediated DNA Cleavage
check_url/it/60369?article_type=v

Making Precise and Accurate Single-Molecule FRET Measurements using the Open-Source smfBox

Read Article