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Measurement of In Vitro Integration Activity of HIV-1 Preintegration Complexes
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1Center for AIDS Health Disparities Research,Meharry Medical College, 2Department of Biochemistry and Cancer Biology,Meharry Medical College, 3School of Graduate Studies and Research,Meharry Medical College, 4Department of Microbiology and Immunology,Meharry Medical College, 5Tennessee Center for AIDS Research (TN-CFAR),Meharry Medical College
Chapters
- 00:05Title
- 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
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.
