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Mesure de bk-polyomavirus Non-Coding Control Region Driven Transcriptional Activity Via Flow Cytometry
JoVE Journal
Genetics
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JoVE Journal Genetics
Measurement of BK-polyomavirus Non-Coding Control Region Driven Transcriptional Activity Via Flow Cytometry
DOI:

11:54 min

July 13, 2019

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Chapters

  • 00:00Title
  • 01:15Collection of Blood and or Urine Samples and Isolation of BKPyV-DNA
  • 02:11Amplification and Sequencing of the Non-coding Control Region (NCCR)
  • 03:37Cloning of the Non-coding Control Region (NCCR) into the Dual Fluorescence Reporter
  • 05:26Transient Transfection of HEK293T Cells with the Dual Fluorescence Reporter Plasmid and Treatment with Potential Antiviral Agents
  • 07:23Fluorescence Microscopy and Flow Cytometry
  • 09:22Results and Data Interpretation
  • 11:26Conclusion

Summary

Automatic Translation

Dans ce manuscrit, un protocole est présenté pour effectuer la mesure FACS-basée de bk-polyomavirus activité transcriptionnelle en utilisant des cellules HEK293T transfecté avec un plasmide journaliste bidirectionnel exprimant tdTomato et eGFP. Cette méthode permet en outre de déterminer quantitativement l'influence de nouveaux composés sur la transcription virale.

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