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Mesure de<em> In vitro</em> Activité d'intégration du VIH-1 préintégration Complexes
JoVE Journal
Immunologia e infezioni
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JoVE Journal Immunologia e infezioni
Measurement of In Vitro Integration Activity of HIV-1 Preintegration Complexes
DOI:

10:34 min

February 22, 2017

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Capitoli

  • 00:05Titolo
  • 01:36Virus Infection of Sup-T1 Cells
  • 03:22Harvesting and Lysis of Infected Cells
  • 04:22Preparation of Cytoplasmic Preintegration Complexes
  • 05:32In Vitro Integration Assay Using Preintegration Complexes
  • 07:40PCR Assays for Quantifying Preintegration Complex Activity
  • 08:29Results: HIV-1 Preintegration Complex-associated vDNA Integration Activity
  • 09:47Conclusion

Summary

Traduzione automatica

This report describes an in vitro assay for measuring the human immunodeficiency virus type 1 (HIV-1) preintegration complex (PIC)-associated integration activity in the cytoplasmic extracts of acutely infected cells. The integration of PIC-associated HIV-1 DNA into heterologous target DNA is quantified by using a nested real-time polymerase chain reaction strategy.

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