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FLIM-FRET Målinger af protein-protein interaktioner i levende bakterier.
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JoVE Journal Biology
FLIM-FRET Measurements of Protein-Protein Interactions in Live Bacteria.

FLIM-FRET Målinger af protein-protein interaktioner i levende bakterier.

DOI:
10.3791/61602-v

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09:26 min

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August 25, 2020

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, , , , , , , , ,
1Université de Strasbourg,Laboratoire de Bioimagerie et Pathologies, UMR CNRS 7021, 2Université de Strasbourg, UMR 7242, ESBS, 3CNRS, UMR 7242, ESBS, 4Groupe Méthode Recherche Clinique,Hôpitaux Universitaires de Strasbourg

Chapters

  • 00:04Introduction
  • 00:41Fluorescent Tag Insertion into the Chromosomal Genome of P. aeruginosa
  • 03:02Preparation of Agarose Pad
  • 03:57Imaging with a Two-photon Microscopy Setup
  • 06:44Results: FLIM-FRET Data Visualization and Analysis
  • 08:57Conclusion

Summary

Automatic Translation

Automatic Translation

Vi beskriver her en protokol til at karakterisere protein-protein interaktioner mellem to meget forskelligt udtrykt proteiner i levende Pseudomonas aeruginosa ved hjælp af FLIM-FRET målinger. Protokollen omfatter bakterier stamme konstruktioner, bakterier immobilisering, billeddannelse og post-imaging dataanalyse rutiner.

Tags

FLIM-FRETProtein-protein InteractionsLive BacteriaLive CellsP.aeruginosaE.coliFluorescent ProteinsMicroscopyAgaroseGentamycin

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