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A quantitative assay for HIV cDNA integration in vivo: analysis of factors modulating early infection.

Butler SL, Hansen MS, Bushman FD; Conference on Retroviruses and Opportunistic Infections.

Program Abstr 8th Conf Retrovir Oppor Infect Conf Retrovir Oppor Infect 8th 2001 Chic Ill. 2001 Feb 4-8; 8: 127 (abstract no. 285).

Salk Inst for Biological Studies, La Jolla, CA.

Background and Methods: We have developed improved methods for quantitating HIV cDNA forms and used them to identify host cell systems that modulate early infection. We used fluorescence- monitored PCR to quantitate HIV cDNAs generated by reverse transcription, circularization, and integration. The integration assay employed a novel quantitative form of Alu-PCR that should be widely applicable to studies of integrating viruses and gene transfer vectors generally. Results and Conclusions: We found that only 1 in 7 to 1 in 20 unintegrated cDNA molecules actually formed integrated proviruses. In different cell types, viral cDNAs were eliminated by either degradation or circularization and loss by dilution during subsequent cell growth. We are using the quantitative framework to examine factors influencing early infection. For example, we find that, as reported previously, action of the proteosome promotes degradation of cDNA forms. Additional studies focus on the roles of small-molecule inhibitors and cellular antiviral systems.

Publication Types:
  • Meeting Abstracts
Keywords:
  • AIDS Vaccines
  • Acquired Immunodeficiency Syndrome
  • Biological Assay
  • Communicable Diseases
  • DNA, Complementary
  • HIV Infections
  • HIV Seropositivity
  • Polymerase Chain Reaction
  • Proviruses
  • Research
  • analysis
Other ID:
  • GWAIDS0006572
UI: 102244068

From Meeting Abstracts




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