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Analyse af<em> Yersinia enterocolitica</em> Effector Translokation i værtsceller Brug Beta-lactamase Effector Fusions
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Immunologia e infezioni
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JoVE Journal Immunologia e infezioni
Analysis of Yersinia enterocolitica Effector Translocation into Host Cells Using Beta-lactamase Effector Fusions

Analyse af<em> Yersinia enterocolitica</em> Effector Translokation i værtsceller Brug Beta-lactamase Effector Fusions

DOI:
10.3791/53115-v

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12:23 min

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October 13, 2015

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, , ,
1Institute for Medical Microbiology, Virology and Hygiene,University Medical Center Hamburg-Eppendorf, 2UKE Microscopy Imaging Facility,University Medical Center Hamburg-Eppendorf

Capitoli

  • 00:05Titolo
  • 01:41Preparation of Y. enterocolitica Strains for Cell Culture Infection
  • 03:38Infection of HeLa Cells and Loading with CCF4
  • 05:44Laser Scanning Microscopy
  • 07:17Image Quantification
  • 10:52Results: Analysis of Yersinia enterocolitica Effector Translocation
  • 11:54Conclusion

Summary

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Effector translocation into host cells via a type III secretion system is a common virulence strategy among gram-negative bacteria. A beta-lactamase effector fusion based assay for quantitative analysis of translocation was applied. In Yersinia infected cells, conversion of a FRET reporter by the beta-lactamase is monitored using laser scanning microscopy.

Tags

Keywords: Yersinia EnterocoliticaType III Secretion SystemEffector TranslocationBeta-lactamase FusionFRET ReporterQuantitative AnalysisHeLa CellsLaser Scanning Microscopy

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