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A Rapid Microtiter Assay for Duck Hepatitis Virus Reverse Transcriptase

  • Michael J. Otto
  • Richard A. Whitaker
Part of the Methods in Molecular Medicine™ book series (MIMM, volume 24)

Abstract

The duck hepatitis B virus (DHBV) genome contains a pol gene that codes for the viral polymerase protein. This enzyme, which is essential for the replication of the virus, has multiple activities including an RNA directed DNA polymerase or reverse transcriptase (RT) activity, an RNase H activity, and a DNA-directed DNA polymerase activity. The assay described in this chapter is designed to measure the ability of test compounds to inhibit the RNA-directed DNA polymerase activity of the DHBV pol gene product. The assay is based on research performed in the laboratory of Dr. Christoph Seeger (1) and on the observation that the e stem loop structure in the pregenomic RNA is required for initiation of DNA synthesis (2, 3, 4).

Keywords

Reaction Plate Rabbit Reticulocyte Lysate Duck Hepatitis Reverse Transcriptase Assay Duck Hepatitis Virus 
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.
    Wang, G. H., Zoulim, F., Leber, E. H., Kitson, J., and Seeger, C. (1994) Role of RNA in enzymatic activity of the reverse transcriptase of hepatitis B viruses. J. Virol. 68, 8437–8442.PubMedGoogle Scholar
  2. 2.
    Wang, G. H. and Seeger, C. (1992) The reverse transcriptase of hepatitis B virus acts as a protein primer for viral DNA synthesis. Cell 71, 663–670.CrossRefPubMedGoogle Scholar
  3. 3.
    Wang, G. H. and Seeger, C. (1993) Novel mechanism for reverse transcription in hepatitis B viruses. J. Virol. 67, 6507–6512.PubMedGoogle Scholar
  4. 4.
    Tavis, J. E., Perri, S., and Ganem, D. (1994) Hepadnavirus reverse transcriptase initiates within the stem-loop of the RNA packaging signal and employs a novel strand transfer. J. Virol. 68, 3536–3543.PubMedGoogle Scholar
  5. 5.
    Zoulim, F. and Seeger, C. (1994) Reverse transcription in hepatitis B viruses is primed by a tyrosine residue of the polymerase. J. Virol. 68, 6–13.PubMedGoogle Scholar

Copyright information

© Humana Press Inc. 2000

Authors and Affiliations

  • Michael J. Otto
    • 1
  • Richard A. Whitaker
    • 1
  1. 1.Department of Clinical ResearchRhone-Poulenc RorerCollegeville

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