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1Department of Nephrology,University of Duisburg-Essen, University Hospital Essen, 2Institute for Virology,University of Duisburg-Essen, University Hospital Essen, 3Center for Translational Medicine, Medical Department I, Marien Hospital Herne,University Hospital of the Ruhr-University of Bochum, 4Department of Infectious Diseases, University of Duisburg-Essen,University Hospital Essen Hufelandstr
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
In this manuscript, a protocol is presented to perform FACS-based measurement of BK-polyomavirus transcriptional activity by using HEK293T cells transfected with a bidirectional reporter plasmid expressing tdTomato and eGFP. This method further allows to quantitatively determine the influence of novel compounds on viral transcription.
