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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
I detta manuskript, ett protokoll presenteras för att utföra FACS-baserade mätningar av BK-polyomavirus transkriptionell aktivitet genom att använda HEK293T celler transfekterade med en dubbelriktad reporter plasmid uttrycker tdtomato och egfp. Denna metod gör det möjligt att kvantitativt bestämma påverkan av nya föreningar på viral transkription.
