Skip to main content
PMC home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1980 Apr;77(4):1842–1846. doi: 10.1073/pnas.77.4.1842

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

I W Cummings, J K Browne, W A Salser, G V Tyler, R L Snyder, J M Smolec, J Summers
PMCID: PMC348604  PMID: 6246507

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 lambda gtWES and subcloned into the kanamycin resistance plasmid pA01. 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 small amount of nucleic acid homology previously reported between the HBV and WHV DNAs could be demonstrated between the cloned viral DNAs.

Full text

PDF
1845

Images in this article

1844Image
on p.1844
1844Image
on p.1844
1844Image
on p.1844
1845Image
on p.1845
1845Image
on p.1845

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bauer W., Vinograd J. The interaction of closed circular DNA with intercalative dyes. I. The superhelix density of SV40 DNA in the presence and absence of dye. J Mol Biol. 1968 Apr 14;33(1):141–171. doi: 10.1016/0022-2836(68)90286-6. [DOI] [PubMed] [Google Scholar]
  2. Chan H. W., Israel M. A., Garon C. F., Rowe W. P., Martin M. A. Molecular cloning of polyoma virus DNA in Escherichia coli: lambda phage vector system. Science. 1979 Mar 2;203(4383):887–892. doi: 10.1126/science.217088. [DOI] [PubMed] [Google Scholar]
  3. Charnay P., Mandart E., Hampe A., Fitoussi F., Tiollais P., Galibert F. Localization on the viral genome and nucleotide sequence of the gene coding for the two major polypeptides of the hepatitis B surface antigen (HBs Ag). Nucleic Acids Res. 1979 Sep 25;7(2):335–346. doi: 10.1093/nar/7.2.335. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Charnay P., Pourcel C., Louise A., Fritsch A., Tiollais P. Cloning in Escherichia coli and physical structure of hepatitis B virion DNA. Proc Natl Acad Sci U S A. 1979 May;76(5):2222–2226. doi: 10.1073/pnas.76.5.2222. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cohen S. N., Chang A. C. A method for selective cloning of eukaryotic DNA fragments in Escherichia coli by repeated transformation. Mol Gen Genet. 1974;134(2):133–141. doi: 10.1007/BF00268415. [DOI] [PubMed] [Google Scholar]
  6. Denhardt D. T. A membrane-filter technique for the detection of complementary DNA. Biochem Biophys Res Commun. 1966 Jun 13;23(5):641–646. doi: 10.1016/0006-291x(66)90447-5. [DOI] [PubMed] [Google Scholar]
  7. Enquist L. W., Madden M. J., Schiop-Stanley P., Vande Woude G. F. Cloning of herpes simplex type 1 DNA fragments in a bacteriophage lambda vector. Science. 1979 Feb 9;203(4380):541–544. doi: 10.1126/science.216076. [DOI] [PubMed] [Google Scholar]
  8. Fritsch A., Pourcel C., Charnay P., Tiollais P. Clonage du génome du virus de l'hépatite B dans Escherichia coli. C R Acad Sci Hebd Seances Acad Sci D. 1978 Dec;287(16):1453–1456. [PubMed] [Google Scholar]
  9. Galibert F., Mandart E., Fitoussi F., Tiollais P., Charnay P. Nucleotide sequence of the hepatitis B virus genome (subtype ayw) cloned in E. coli. Nature. 1979 Oct 25;281(5733):646–650. doi: 10.1038/281646a0. [DOI] [PubMed] [Google Scholar]
  10. Gillespie D., Spiegelman S. A quantitative assay for DNA-RNA hybrids with DNA immobilized on a membrane. J Mol Biol. 1965 Jul;12(3):829–842. doi: 10.1016/s0022-2836(65)80331-x. [DOI] [PubMed] [Google Scholar]
  11. Grunstein M., Hogness D. S. Colony hybridization: a method for the isolation of cloned DNAs that contain a specific gene. Proc Natl Acad Sci U S A. 1975 Oct;72(10):3961–3965. doi: 10.1073/pnas.72.10.3961. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Katz L., Kingsbury D. T., Helinski D. R. Stimulation by cyclic adenosine monophosphate of plasmid deoxyribonucleic acid replication and catabolite repression of the plasmid deoxyribonucleic acid-protein relaxation complex. J Bacteriol. 1973 May;114(2):577–591. doi: 10.1128/jb.114.2.577-591.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Smithies O., Blechl A. E., Denniston-Thompson K., Newell N., Richards J. E., Slightom J. L., Tucker P. W., Blattner F. R. Cloning human fetal gamma globin and mouse alpha-type globin DNA: characterization and partial sequencing. Science. 1978 Dec 22;202(4374):1284–1289. doi: 10.1126/science.725604. [DOI] [PubMed] [Google Scholar]
  14. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  15. Summers J., O'Connell A., Millman I. Genome of hepatitis B virus: restriction enzyme cleavage and structure of DNA extracted from Dane particles. Proc Natl Acad Sci U S A. 1975 Nov;72(11):4597–4601. doi: 10.1073/pnas.72.11.4597. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Summers J. Physical map of polyoma viral DNA fragments produced by cleavage with a restriction enzyme from Haemophilus aegyptius, endonuclease R-HaeIII. J Virol. 1975 Apr;15(4):946–953. doi: 10.1128/jvi.15.4.946-953.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Summers J., Smolec J. M., Snyder R. A virus similar to human hepatitis B virus associated with hepatitis and hepatoma in woodchucks. Proc Natl Acad Sci U S A. 1978 Sep;75(9):4533–4537. doi: 10.1073/pnas.75.9.4533. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Werner B. G., Smolec J. M., Snyder R., Summers J. Serological relationship of woodchuck hepatitis virus to human hepatitis B virus. J Virol. 1979 Oct;32(1):314–322. doi: 10.1128/jvi.32.1.314-322.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES