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Produção de conjuntos de motor de dynein e de Kinesin em nanostructures do ADN origami para a única observação da molécula
JoVE Journal
Biochemistry
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JoVE Journal Biochemistry
Production of Dynein and Kinesin Motor Ensembles on DNA Origami Nanostructures for Single Molecule Observation
DOI:

08:09 min

October 15, 2019

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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

Automatic Translation

O objetivo deste protocolo é formar conjuntos de motores moleculares em nanoestruturas de DNA origami e observar a motilidade do conjunto usando microscopia de fluorescência de reflexão interna total.

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