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ניטור העצרת של מופרש בקטריאלי ארסי פקטור שימוש Crosslinking אתר ספציפי
JoVE Journal
Imunologia e Infecção
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JoVE Journal Imunologia e Infecção
Monitoring the Assembly of a Secreted Bacterial Virulence Factor Using Site-specific Crosslinking
DOI:

11:33 min

December 17, 2013

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Capítulos

  • 00:05Título
  • 01:55Culture Preparation
  • 02:59Bpa Incorporation and Preparation for Photocrosslinking
  • 04:51Pulse-chase Labeling and UV Irradiation
  • 07:33Immunoprecipitation and Crosslinking Product Detection
  • 08:27Results: Interaction of the Autotransporter EspP wirh Molecular Chaperones and Components of the Bam Complex
  • 10:54Conclusion

Summary

Tadução automática

This article illustrates the use of pulse-chase radio labeling in combination with site-specific photocrosslinking to monitor interactions between a protein of interest and other factors in E. coli. Unlike traditional chemical cross-linking methods, this approach generates high resolution “snapshots” of an ordered assembly pathway in a living cell.

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