Elsevier

Gene

Volume 106, Issue 2, 15 October 1991, Pages 143-149
Gene

Production of hepatitis B virion-like particles in yeast

https://doi.org/10.1016/0378-1119(91)90193-FGet rights and content

Abstract

We constructed a yeast strain that simultaneously expresses four genes encoding the major S, middle S, large S hepatitis B viral envelope proteins and the core protein under the control of the yeast glyceraldehyde-3-phosphate dehydrogenase promoter and terminator. The lysate from this cell line, examined by immunological, physicochemical methods and electron microscopy, was found to contain spherical particles with a diameter of about 40 nm and a density of 1.25g/ml. These particles reacted with anti-envelope antibodies, but not with anti-core antibodies. However, core antigenicity appeared upon treatment with 3 % Nonidet P-40 that eliminates an outer envelope. These observations suggest production of a virion-like complex structure, or at least its DNA-less analog, consisting of core particle enveloped by antibody-reactive envelope. Such a structure was made only when all the four gene products were synthesized in a yeast cell. This system may be useful for the study of virus structure and assembly, and for improved vaccine development.

References (22)

  • K. Araki et al.

    The essential region for assembly and particle formation in hepatitis B virus surface antigen produced in yeast cells

    Gene

    (1990)
  • R.A. Hitzeman et al.

    Expression of hepatitis B virus surface antigen in yeast

    Nucleic Acids Res.

    (1983)
  • M. Imai et al.

    A receptor for polymerized human and chimpanzee albumins on hepatitis B virus particles co-occurring with HBeAg

    Gastroenterology

    (1979)
  • T. Imamura et al.

    Expression of hepatitis B virus middle and large surface antigen genes in Saccharomyces cerevisiae

    J. Virol.

    (1987)
  • T. Imamura et al.

    Purification and characterization of the hepatitis B virus core antigen produced in the yeast Saccharomyces cerevisiae

    J. Biotechnol.

    (1988)
  • H. Ito et al.

    Transformation of intact yeast cells treated with alkali cations

    J. Bacteriol.

    (1983)
  • E. Jacobs et al.

    Simultaneous synthesis and assembly of various hepatitis B surface proteins in Saccharomyces cerevisiae

    Gene

    (1989)
  • A. Jimenez et al.

    Expression of a transposable antibiotic resistance element in Saccharomyces

    Nature

    (1980)
  • M. Kobayashi et al.

    Recombinant hepatitis B virus surface antigen carrying the pre-S2 region derived from yeast: purification and characterization

    J. Biotechnol.

    (1988)
  • A. Machida et al.

    A polypeptide containing 55 amino acid residues coded by the pre-S region of hepatitis B virus deoxyribonucleic acid bears the receptor for polymerized human as well as chimpanzee albumins

    Gastroenterology

    (1984)
  • D.R. Milich et al.

    Antibody production to the nucleocapsid and envelope of the hepatitis B virus primed by a single synthetic T cell site

    Nature

    (1987)
  • Cited by (10)

    • Effects of Trx2p and Sec23p expression on stable production of hepatitis B surface antigen S domain in recombinant Saccharomyces cerevisiae

      2012, Journal of Biotechnology
      Citation Excerpt :

      These proteins share a common carboxyl terminus; the L protein harbors pre-S1, pre-S2 and S domains, and the M protein possesses pre-S2 and S domains. The S protein equal to the S domain is called small HBV surface antigen (sHBsAg) and consists of 226 amino acids, which is antigenically most significant from vaccine development point of view (Bruss and Ganem, 1991; Shiosaki et al., 1991). The proper orientation of the N-terminus of sHBsAg is assigned to be important to allow lateral interactions between the S proteins in their assembly pathway (Bruss and Ganem, 1991).

    View all citing articles on Scopus
    View full text