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J Gen Virol 71 (1990), 1339-1345; DOI 10.1099/0022-1317-71-6-1339
© 1990 Society for General Microbiology
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Stable expression of hepatitis delta virus antigen in a eukaryotic cell line

T. B. Macnaughton1, E. J. Gowans1, B. Reinboth2, A. R. Jilbert1 and C. J. Burrell1

1 Division of Medical Virology, Institute of Medical and Veterinary Science, Box 14, Rundle Mall Post Office, Adelaide 5000
and2 Department of Pathology, University of Adelaide, GPO Box 498, Adelaide 5000, South Australia

The gene encoding the hepatitis delta virus structural antigen (HDAg) was linked to a neomycin resistance gene in a retrovirus expression vector, and human HepG2 cells were transfected with the recombinant plasmid. A stable cell line was cloned that expressed HDAg in the nuclei of 100% of cells, in a pattern indicating a close relationship with cell nucleoli. Analysis of partially purified recombinant HDAg by HPLC showed an Mr in the range of 7 x 105 to 2 x 106, which appeared to contain conformation-dependent epitopes, whereas the density of the antigen was 1.19 g/ml by equilibrium centrifugation in caesium chloride, and in rate zonal centrifugation it sedimented with a value of 50S, close to that of particulate hepatitis B virus surface antigen. Immunoblotting demonstrated a single polypeptide with an Mr of 24K which corresponded to the smaller of the two HDAg-specific polypeptides present in infected sera. The recombinant HDAg polypeptide was shown to be a RNA-binding protein with specificity for both genomic and anti-genomic species of hepatitis delta virus RNA.

Received 13 November 1989; accepted 9 February 1990.


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