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BK-多瘤病毒非编码控制区域驱动转录活性通过流细胞测定的测量
JoVE Journal
Genética
Author Produced
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JoVE Journal Genética
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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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

Summary

Tadução automática

在本手稿中,提出了使用 HEK293T 细胞转染的基于 FACS 的 BK-多瘤病毒转录活性的测量方案,该细胞与表达 tdTomato 和 eGFP 的双向报告质粒转染。该方法进一步允许定量确定新化合物对病毒转录的影响。

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