Skip to main content
NCBI 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
. 1981 Jul;78(7):4510–4514. doi: 10.1073/pnas.78.7.4510

Production of immunologically active surface antigens of hepatitis B virus by Escherichia coli.

P MacKay, M Pasek, M Magazin, R T Kovacic, B Allet, S Stahl, W Gilbert, H Schaller, S A Bruce, K Murray
PMCID: PMC319821  PMID: 6170067

Abstract

Several plasmids have been constructed which direct the synthesis of hepatitis B virus surface antigens in Escherichia coli either as the native polypeptide or fused to other plasmid encoded polypeptides. When injected into rabbits, extracts from bacteria carrying some of these plasmids induced the synthesis of antibodies to the antigens even though the extracts did not give satisfactory positive results in radioimmunoassay for them. Either the NH2-terminal segment or the COOH-terminal segment of the surface antigens alone was sufficient to elicit the immune response, but antibodies against the two segments showed different specificities. The results emphasize the value of an in vivo assay for the presence of antigens in crude cell extracts and illustrate the feasibility of this type of screening with laboratory animals.

Full text

PDF
4510

Selected References

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

  1. Benton W. D., Davis R. W. Screening lambdagt recombinant clones by hybridization to single plaques in situ. Science. 1977 Apr 8;196(4286):180–182. doi: 10.1126/science.322279. [DOI] [PubMed] [Google Scholar]
  2. Bolivar F., Rodriguez R. L., Greene P. J., Betlach M. C., Heyneker H. L., Boyer H. W., Crosa J. H., Falkow S. Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene. 1977;2(2):95–113. [PubMed] [Google Scholar]
  3. Broome S., Gilbert W. Immunological screening method to detect specific translation products. Proc Natl Acad Sci U S A. 1978 Jun;75(6):2746–2749. doi: 10.1073/pnas.75.6.2746. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Burrell C. J., Mackay P., Greenaway P. J., Hofschneider P. H., Murray K. Expression in Escherichia coli of hepatitis B virus DNA sequences cloned in plasmid pBR322. Nature. 1979 May 3;279(5708):43–47. doi: 10.1038/279043a0. [DOI] [PubMed] [Google Scholar]
  5. Burrell C. J., Proudfoot E., Keen G. A., Marmion B. P. Carbohydrates in hepatitis B antigen. Nat New Biol. 1973 Jun 27;243(130):260–262. doi: 10.1038/newbio243260a0. [DOI] [PubMed] [Google Scholar]
  6. Charnay P., Gervais M., Louise A., Galibert F., Tiollais P. Biosynthesis of hepatitis B virus surface antigen in Escherichia coli. Nature. 1980 Aug 28;286(5776):893–895. doi: 10.1038/286893a0. [DOI] [PubMed] [Google Scholar]
  7. 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]
  8. Clewell D. B. Nature of Col E 1 plasmid replication in Escherichia coli in the presence of the chloramphenicol. J Bacteriol. 1972 May;110(2):667–676. doi: 10.1128/jb.110.2.667-676.1972. [DOI] [PMC free article] [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. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  11. Maxam A. M., Gilbert W. A new method for sequencing DNA. Proc Natl Acad Sci U S A. 1977 Feb;74(2):560–564. doi: 10.1073/pnas.74.2.560. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
  13. Murray N. E., Bruce S. A., Murray K. Molecular cloning of the DNA ligase gene from bacteriophage T4. II. Amplification and preparation of the gene product. J Mol Biol. 1979 Aug 15;132(3):493–505. doi: 10.1016/0022-2836(79)90271-7. [DOI] [PubMed] [Google Scholar]
  14. Neurath A. R., Hashimoto N., Prince A. M. Sialyl residues in hepatitis B antigen: their role in determining the life span of the antigen in serum and in eliciting an immunological response. J Gen Virol. 1975 Apr;27(1):81–91. doi: 10.1099/0022-1317-27-1-81. [DOI] [PubMed] [Google Scholar]
  15. Pasek M., Goto T., Gilbert W., Zink B., Schaller H., MacKay P., Leadbetter G., Murray K. Hepatitis B virus genes and their expression in E. coli. Nature. 1979 Dec 6;282(5739):575–579. doi: 10.1038/282575a0. [DOI] [PubMed] [Google Scholar]
  16. Purcell R. H., Wong D. C., Alter H. J., Holland P. V. Microtiter solid-phase radioimmunoassay for hepatitis B antigen. Appl Microbiol. 1973 Oct;26(4):478–484. doi: 10.1128/am.26.4.478-484.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Reeve J. N. Bacteriophage infection of minicells: a general method for identification of "in vivo" bacteriophage directed polypeptide biosynthesis. Mol Gen Genet. 1977 Dec 14;158(1):73–79. doi: 10.1007/BF00455121. [DOI] [PubMed] [Google Scholar]
  18. Reeve J. N. Mucopeptide biosynthesis by minicells of Escherichia coli. J Bacteriol. 1977 Jul;131(1):363–365. doi: 10.1128/jb.131.1.363-365.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
  20. Shih J. W., Gerin J. L. Immunochemistry of hepatitis B surface antigen (HBsAg): preparation and characterization of antibodies to the constituent polypeptides. J Immunol. 1975 Sep;115(3):634–639. [PubMed] [Google Scholar]
  21. Sninsky J. J., Siddiqui A., Robinson W. S., Cohen S. N. Cloning and endonuclease mapping of the hepatitis B viral genome. Nature. 1979 May 24;279(5711):346–348. doi: 10.1038/279346a0. [DOI] [PubMed] [Google Scholar]
  22. Talmadge K., Stahl S., Gilbert W. Eukaryotic signal sequence transports insulin antigen in Escherichia coli. Proc Natl Acad Sci U S A. 1980 Jun;77(6):3369–3373. doi: 10.1073/pnas.77.6.3369. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Valenzuela P., Gray P., Quiroga M., Zaldivar J., Goodman H. M., Rutter W. J. Nucleotide sequence of the gene coding for the major protein of hepatitis B virus surface antigen. Nature. 1979 Aug 30;280(5725):815–819. doi: 10.1038/280815a0. [DOI] [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