Production of hepatitis B virion-like particles in yeast.
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 1.25 g/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.
Institute for Molecular and Cellular Biology, Osaka University, Japan.
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K
Shiosaki
K
K
Takata
S
S
Nishimura
H
H
Mizokami
K
K
Matsubara
D015153:Blotting, Western; D002499:Centrifugation, Density Gradient; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000235:genetics; D006512:Hepatitis B Core Antigens / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism*; D006514:Hepatitis B Surface Antigens / Q000096:biosynthesis / Q000187:drug effects / Q000302:isolation & purification / Q000378:metabolism*; D006515:Hepatitis B virus / Q000302:isolation & purification / Q000378:metabolism* / Q000648:ultrastructure; D008854:Microscopy, Electron; D017830:Octoxynol; D010957:Plasmids / Q000235:genetics; D011092:Polyethylene Glycols / Q000494:pharmacology; D011401:Promoter Regions, Genetic / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D014779:Virus Replication / Q000502:physiology
D006512:Hepatitis B Core Antigens; D006514:Hepatitis B Surface Antigens; D011092:Polyethylene Glycols; D017830:Octoxynol; C010615:Nonidet P-40; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases