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Production of Dynein and Kinesin Motor Ensembles on DNA Origami Nanostructures for Single Molecule Observation
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生物化学
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JoVE 杂志 生物化学
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

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08:09 min

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October 15, 2019

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

Chapters

  • 00:04Title
  • 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

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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

DNA OrigamiCytoskeletal Motor ProteinsDyneinKinesinMyosinCargo TransportSingle Molecule ObservationYeast ExpressionProtein PurificationIgG Affinity Purification

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