É necessária uma assinatura da JoVE para visualizar este conteúdo. Faça login ou comece sua avaliação gratuita.
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
Capítulos
- 00:00Título
- 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
En este manuscrito, se presenta un protocolo para realizar la medición basada en FACS de la actividad transcripcional BK-polioomavirus mediante el uso de células HEK293T transcritas con un plásmido reportero bidireccional que expresa tdTomato y eGFP. Este método permite además determinar cuantitativamente la influencia de nuevos compuestos en la transcripción viral.
