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PCR Amplification and Cloning of Virtually Full-Length HIV-1 Provirus

  • Mika Salminen
Part of the Methods in Molecular Medicine™ book series (MIMM, volume 17)

Abstract

Molecular analyses of HIV-1 variation, and its impact on the biological properties of the virus have by necessity been restricted by the scarcity of cloned proviruses. Only a handful of full-length clones exist (1), and of these, even a smaller fraction are infectious upon transfection into cell lines. Furthermore, the available infectious clones only represent subtypes B and D (1). The main reasons for the lack of infectious clones are related to the relatively low copy number of proviral DNA especially in primary cultures (8).

Keywords

Microfuge Tube Infectious Clone Polymerase Chain Reaction Tube Triethylammonium Salt Disposable Glove 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

References

  1. 1.
    Myers, G., Korber, B., Hanhn, B. K., Jeang, K.-T., Mellors, J. W., McCutchan, F. E., et al. (eds.) (1995) Human Retroviruses and AIDS Theoretical Biology and Biophysics, Group T-10, MS K710, Los Alamos National Laboratory, Los Alamos, NM.Google Scholar
  2. 2.
    Edmonson, P. F. and Mullins, J. I. (1992) Efficient amplification of HIV half-genomes from tissue DNA. Nucleic Acids Res. 20, 4933.PubMedCrossRefGoogle Scholar
  3. 3.
    Hahn, B. H., Gonda, M. A., Shaw, G. M., Popovic, M., Hoxie, J. A., Gallo, R. C., and Wong-Staal, F. (1985) Genomic diversity of the acquired immune deficiency syndrome virus HTLV-III: different viruses exhibit greatest divergence in their envelope genes. Proc. Natl. Acad. Sci. USA 82, 4813–4817.PubMedCrossRefGoogle Scholar
  4. 4.
    Hahn, B. H., Shaw, G. M., Arya, S. K., Popovic, M., Gallo, R. C., and Wong-Staal, F. (1984) Molecular cloning and characterization of the HTLV-III virus associated with AIDS. Nature 312, 166–169.PubMedCrossRefGoogle Scholar
  5. 5.
    Li, Y., Hui, H., Burgess, C. J., Price, R. W., Sharp, P. M., Hahn, B. H., and Shaw, G. M. (1992) Complete nucleotide sequence, genome organization, and biological properties of human immunodeficiency virus type 1 in vivo: evidence for limited defectiveness and complementation. J. Virol. 66, 6587–6600.PubMedGoogle Scholar
  6. 6.
    Li, Y., Kappes, J. C., Conway, J. A., Price, R. W., Shaw, G. M., and Hahn, B. H. (1991) Molecular characterization of human immunodeficiency virus type 1 cloned directly from uncultured human brain tissue: identification of replication-competent and-defective viral genomes. J. Virol. 65, 3973–3985.PubMedGoogle Scholar
  7. 7.
    Salminen, M. O., Koch, C., Sanders-Buell, E., Ehrenberg, P. K., Michael, N. L., Carr, J. K., et al. (1995) Recovery of virtually full-length HIV-1 provirus of diverse subtypes from primary virus cultures using the polymerase chain reaction. Virology 213, 80–86.PubMedCrossRefGoogle Scholar
  8. 8.
    Simmonds, P., Balfe, P., Peutherer, J. F., Ludlam, C. A., Bishop, J. O., and Brown, A. J. (1990) Human immunodeficiency virus-infected individuals contain provirus in small numbers of peripheral mononuclear cells and at low copy numbers. J. Virol. 64, 864–872.PubMedGoogle Scholar
  9. 9.
    Isolation of DNA from mammalian cells: Protocol I (1989) in, Molecular Cloning: A Laboratory Manual (Sambrook, J., Fritch, E. F., and Maniatis, T., eds.), 2nd ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.Google Scholar

Copyright information

© Humana Press Inc., Totowa, NJ 1999

Authors and Affiliations

  • Mika Salminen
    • 1
  1. 1.Department of Chronic Virus DiseasesNational Public Health InstituteHelsinkiFinland

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