Isolation, characterization, and comparison of recombinant DNAs derived from genomes of human hepatitis B virus and woodchuck hepatitis virus.

Abstract

The human hepatitis B virus (HBV) and the woodchuck hepatitis virus (WHV) are closely related by several criteria and belong to the same class of DNA viruses. The DNA genomes from these viruses are difficult to obtain in quantities required for biochemical analysis. We have, therefore, cloned these two DNAs in the vector XgtWES and subcloned into the kanamycin resistance plasmid pAOl. Comparison of the recombinant DNAs with authentic viral DNAs by specific hybridization, size, and restriction enzyme analysis suggests that the recombinants contain the complete genome of each virus. The nominal size of the cloned HBV genome was 3150 base pairs, compared to 3200 base pairs for the cloned WHV genome. The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U. S. C. §1734 solely to indicate this fact.

Maintenance of Woodchuck Hepatitis Virus Activity in Woodchuck Hepatocyte Primary Culture

“…Indeed, a very close relationship between human (HBV) and woodchuck (WHV) viruses has been demonstrated by the striking similarity in their genome organization (Cummings et al, 1980;Galibert et al, 1982) and by serological cross-reactions Stannard et al, 1983). Furthermore, chronically infected woodchucks have been found to develop hepatocellular carcinoma even more frequently than infected human patients (London, 1983;Popper et al, 1981).…”

Detection of Hepatitis B Virus-specific DNA in the Genomes of Human Hepatocellular Carcinoma and Liver Cirrhosis Tissues

“…DNA was transferred from the gels to nitrocellulose filters by a modification (Koshy et aL, 1980) of the Southern technique. Filters were hybridized with cloned HBV DNA (pAO 1-HBV) (Cummings et al, 1980), labelled with c~-[32p]dATP (New England Nuclear, 400 Ci/mmol) to a specific activity of 10 s ct/min/pg, by the nick translation procedure of Rigby et al (1977). Reactions contained 50 mM-tris-HC1 pH 7.5, 5 mM-MgC12, 10 mM-2-mercaptoethanol, 50 pg/ml bovine serum albumin, 1.5 mM each of three non-labelled dNTPs and 5 /~M of the fourth dNTP which was uJZP-labelled (400 to 600 Ci/mmol), 1/~g DNA and 100 units DNA polymerase I. Hybridization reactions were done in the presence of 10% dextran sulphate as described by Wahl et al (1979) and also contained 50% formamide, 5 × SSC, 5 × Denhardt's reagent, 20 raM-sodium phosphate pH 6.5, 100/tg/ml sonicated, denatured salmon sperm DNA and 100 ng labelled probe per filter.…”

Chronic hepatitis in chimpanzee carriers of hepatitis B virus: morphologic, immunologic, and viral DNA studies.

“…In a human hepatocellular carcinoma line (PLC/PRF/5) in tissue culture, HBV DNA has recently been found to be integrated into the host genome (7,8). With development of newer techniques in nucleic acid analysis and availability of recombinant cloned HBV DNA probes (9)(10)(11)(12)(13)(14), this question could be addressed in small samples of liver by using very sensitive techniques.…”