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Motility of Single Molecules and Clusters of Bi-Directional Kinesin-5 Cin8 Purified from S. cerevisiae Cells
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Bioquímica
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JoVE Journal Bioquímica
Motility of Single Molecules and Clusters of Bi-Directional Kinesin-5 Cin8 Purified from S. cerevisiae Cells

Motility of Single Molecules and Clusters of Bi-Directional Kinesin-5 Cin8 Purified from S. cerevisiae Cells

DOI:
10.3791/63425-v

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10:46 min

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February 02, 2022

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, , , , , , , , , , ,
1Department of Chemistry,Ben-Gurion University of the Negev, Israel, 2Department of Biotechnology Engineering,Ben-Gurion University of the Negev, Israel

Capítulos

  • 00:05Introduction
  • 01:51Cin8 Overexpression and Purification from Saccharomyces cerevisiae Cells
  • 04:06Single-Molecule Motility Assay with the Purified Cin8
  • 06:29Motility Analysis
  • 09:13Results: Motility Characteristics of Different Cluster Sizes of Cin8 on Single Microtubules
  • 09:58Conclusion

Summary

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Tadução automática

The bi-directional mitotic kinesin-5 Cin8 accumulates in clusters that split and merge during their motility. Accumulation in clusters also changes the velocity and directionality of Cin8. Here, a protocol for motility assays with purified Cin8-GFP and analysis of motile properties of single molecules and clusters of Cin8 is described.

Tags

Kinesin-5 Cin8Bidirectional Motor ProteinSingle-molecule Motility AssaySaccharomyces CerevisiaeGFP-type Full-length Cin8Protein PurificationNuclear ProteinsFluorescent Particles

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