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J Virol. 1974 August; 14(2): 384-391
Copyright © 1974 American Society for Microbiology. All Rights Reserved.

DNA of a Human Hepatitis B Virus Candidate

William S. Robinson, David A. Clayton and Richard L. Greenman

Departments of Medicine and Pathology, Stanford University School of Medicine, Stanford, California 94305

ABSTRACT

Particles containing DNA polymerase (Dane particles) were purified from the plasma of chronic carriers of hepatitis B antigen. After a DNA polymerase reaction with purified Dane particle preparations treated with Nonidet P-40 detergent, Dane particle core structures containing radioactive DNA product were isolated by sedimentation in a sucrose density gradient. The radioactive DNA was extracted with sodium dodecyl sulfate and isolated by band sedimentation in a preformed CsCl gradient. Examination of the radioactive DNA band by electron microscopy revealed exclusively circular double-stranded DNA molecules approximately 0.78 µm in length. Identical circular molecules were observed when DNA was isolated by a similar procedure from particles that had not undergone a DNA polymerase reaction. The molecules were completely degraded by DNase 1. When Dane particle core structures were treated with DNase 1 before DNA extraction, only 0.78-µm circular DNA molecules were detected. Without DNase treatment of core structures, linear molecules with lengths between 0.5 and 12 µm, in addition to the 0.78-µm circles were found. These results suggest that the 0.78-µm circular molecules were in a protected position within Dane particle cores and the linear molecules were not within core structures. Length measurements on 225 circular molecules revealed a mean length of 0.78 ± 0.09 µm which would correspond to a molecular weight of around 1.6 x 106. The circular molecules probably serve as primer-template for the DNA polymerase reaction carried out by Dane particle cores. Thermal denaturation and buoyant density measurements on the Dane particle DNA polymerase reaction product revealed a guanosine plus cytosine content of 48 to 49%.

J Virol. 1974 August; 14(2): 384-391
Copyright © 1974 American Society for Microbiology. All Rights Reserved.



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Copyright © 1974 by the American Society for Microbiology. All rights reserved.