Advertisement

Assays for Measuring Hepadnaviral Supercoiled DNA

  • Stephen A. Locamini
  • Gilda M. Civitico
Part of the Methods in Molecular Medicine™ book series (MIMM, volume 24)

Abstract

Amajor reason for treatment failure during antiviral therapy of chronic hepatitis B infection is thought to be the persistence of the key replicative intermediate, the viral covalently closed circular (CCC) or supercoiled DNA (1,2). Investigators studying the structure and function of hepadnaviral CCC DNA (3) have provided evidence that suggests that this structure exists in the nucleus of infected hepatocytes as a heterogeneous population of viral minichromosomes, which range from half to fully chromatinized, thought to be owing to their association with variable numbers of nucleosomes.

Keywords

Southern Blot Hybridization Microfuge Tube Replicative Intermediate Infected Hepatocyte Southern Blot Hybridization Analysis 
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.
    Hirota, K., Sherker, A. H., Omata, M, Yokosuka, O., and Okuda, K. (1987) Effects of adenine arabinoside on scrum and i n trail c pad c implicative forms of duck hepatitis B virus in chronic infection. Hepatology 7, 24–28.CrossRefPubMedGoogle Scholar
  2. 2.
    Mason, W. S., Cullen, J., Saputelli, J., Wu, T.-T., Liu, C, London, W, T., Lustbader, E., Schaffcr, P., O’Connell, A. P., Fourel, I, Aldrich, C. E., and Jilbert, A. R. (1994) Characterization of the antiviral effects of 2′carbodeoxyguanosine in ducks chronically infected with duck hepatitis B virus. Hepatology 19, 398–411.CrossRefPubMedGoogle Scholar
  3. 3.
    Newbold, J. E., Xin, H., Tencza, M., Sherman, G., Dean, J., Bowden, S., and Locarnini, S. (1995) The covalently closed duplex form of hepadnaviral genome exists in situ as a heterogeneous population of viral minichromoxomes. J. Virol. 69, 3350–3357.PubMedGoogle Scholar
  4. 4.
    Tuttleman, J., Pugh, J., and Summers, J. (1986) In vitro experimental infection of primary duck hepatocyte cultures with duck hepatitis B virus. J. Virol. 58, 17–25.PubMedGoogle Scholar
  5. 5.
    Wu, T.-T., Coates, L., Aldrich, C, Summers, J., and Mason, W. S. (1990) In hepatocytes infected with duck hepatitis B virus, the template for viral RNA synthesis is amplified by an intracellular pathway. Virology 175, 255–261.CrossRefPubMedGoogle Scholar
  6. 6.
    Hirt, B. (1967) Selective extraction of polyoma DNA from infected mouse cell cultures. J. MoL Biol. 119, 487–506.Google Scholar
  7. 7.
    Luscombe, C., Pederson, J., Bowden, S., and Locarnini, S. (1994) Alterations in intrahepatic expression of duck hepatitis B viral markers with ganciclovir therapy. Liver 14, 182–192.PubMedGoogle Scholar
  8. 8.
    Wang, Y., Luscombe, C., Bowden, S., Shaw, T., and Locarnini, S. (1995) Inhibition of duck hepatitis B virus DNA replication by antiviral chemotherapy with ganciclovir-nalidixic acid. Antimicrob. Agents Chemother. 39, 556–558.PubMedGoogle Scholar
  9. 9.
    Lin, E., Luscombe, C., Wang, Y-Y., Shaw, T., and Locarnini, S. (1996) The guanine nucleoside analogue penciclovir is active against chronic duck hepatitis B virus infection in vivo. Antimicrob. Agents Chemother. 40, 413–418.PubMedGoogle Scholar
  10. 10.
    Bishop, N., Civitico, G., Wang, Y., Guo, K., Gust, I. D., and Locarnini, S. (1990) Antiviral strategies in chronic HBV infection: I. Rstablishment of a suitable in vitro system using the duck hepatitis B virus model. J. Med. Virol. 31, 82–89.CrossRefPubMedGoogle Scholar
  11. 11.
    Moore, D. (1995) Preparation and analysis of DNA, in Current Protocols in Molecular Biology, vol. 1 (Ausubel, F. M., Brent, R., Kingston R. E., Moore, D. D. Siedman, J. G., Smith, J. A., and Struhl, K., eds.), Wiley, New York.Google Scholar
  12. 12.
    Jilbert, A., Freiman, J., Burrell, C, Holmes, M., Gowans, E., Roweand, R., Hall, P., and Cossart, Y. (1988) Virus-liver cell interactions in duck hepatitis B virus infection: A study of virus dissemination within the liver. Gastroenterology 95, 1375–1382.PubMedGoogle Scholar

Copyright information

© Humana Press Inc. 2000

Authors and Affiliations

  • Stephen A. Locamini
    • 1
  • Gilda M. Civitico
    • 2
  1. 1.Research and Molecular DevelopmentVictorian Infectious Diseases Reference LaboratoryNorth MelbourneAustralia
  2. 2.Victorian Infectious Diseases Reference LaboratoryFairfield HospitalFairfieldUK

Personalised recommendations