Antigenicity of hantavirus nucleocapsid proteins expressed in E. coli.
DNA clones representing the small genomic segment of Nephropathia epidemica virus strain Hällnäs B1 (NEV) and Hantaan virus strain 76-118 (HTV) encoding their nucleocapsid proteins were inserted into the E. coli vector pIN-III-ompA for secretion of proteins into the periplasmic space. The complete HTV and NEV nucleocapsid proteins and two truncated versions of the NEV nucleocapsid proteins were expressed as fusion proteins. Unexpectedly, all products accumulated as insoluble aggregates. Most of the ompA signal peptide remained uncleaved. However, nucleocapsid fusion proteins could be purified from the insoluble fraction by extraction with 8 M urea followed by separation on SDS-PAGE and electroelution. Rabbits were immunized with the eluted proteins and the resulting antibodies reacted specifically with authentic viral nucleocapsid proteins of HTV and NEV. The recombinant nucleocapsid proteins were found to react specifically with various hantavirus-immune sera, but not with human control sera, indicating their suitability as potential diagnostic antigens. This is the first report on the expression of a protein of a NEV serotype strain of hantaviruses by use of recombinant DNA techniques.
Gött
P
P
Zentrum für Molekulare Biologie, Universität Heidelberg, F.R.G.
Zöller
L
L
Yang
S
S
Stohwasser
R
R
Bautz
E K
EK
Darai
G
G
D000956:Antigens, Viral / Q000276:immunology*; D001425:Bacterial Outer Membrane Proteins / Q000276:immunology / Q000378:metabolism; D001483:Base Sequence; D002213:Capsid / Q000235:genetics / Q000276:immunology*; D002478:Cells, Cultured; D003001:Cloning, Molecular; D000939:Epitopes; D006476:Orthohantavirus / Q000276:immunology*; D006480:Hemorrhagic Fever with Renal Syndrome / Q000175:diagnosis; D006801:Humans; D008969:Molecular Sequence Data; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000276:immunology; D014758:Viral Core Proteins / Q000235:genetics / Q000276:immunology*
D000956:Antigens, Viral; D001425:Bacterial Outer Membrane Proteins; D000939:Epitopes; D011993:Recombinant Fusion Proteins; D014758:Viral Core Proteins