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Applying Fluorescence Resonance Energy Transfer (FRET) to Examine Effector Translocation Efficiency by Coxiella burnetii during siRNA Silencing
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Immunologia e infezioni
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JoVE Journal Immunologia e infezioni
Applying Fluorescence Resonance Energy Transfer (FRET) to Examine Effector Translocation Efficiency by Coxiella burnetii during siRNA Silencing

Applying Fluorescence Resonance Energy Transfer (FRET) to Examine Effector Translocation Efficiency by Coxiella burnetii during siRNA Silencing

DOI:
10.3791/54210-v

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10:29 min

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July 06, 2016

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, ,
1Department of Microbiology and Immunology, Peter Doherty Institute for Infection and Immunity,University of Melbourne

Capitoli

  • 00:05Titolo
  • 01:02Reverse Transfection of siRNA and Seeding of HeLa 229 Cells
  • 04:25Infection of Reverse Transfected Cells
  • 05:39Addition of BlaM Substrate to Determine Level of Translocation
  • 08:23Results: FRET Measurements of Coxiella burnetti Effector Translocation Efficiency in siRNA Silenced HeLa Cells
  • 09:37Conclusion

Summary

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Investigating the interactions between bacterial pathogens and their hosts is an important area of biological research. Here, we describe the necessary techniques to measure effector translocation by Coxiella burnetii during siRNA gene silencing using BlaM substrate.

Tags

FRETEffector TranslocationSiRNA SilencingCoxiella BurnetiiCellular MicrobiologyHost-pathogen InteractionsChlamydiaSalmonellaE. ColiBeta-lactamaseACCM2HeLa CellsTransfection96-well Plate

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