Abstract
The nucleocapsid (N) protein of the Toscana (TOS) virus was expressed in Escherichia coli by using a pET15b vector. The recombinant protein was purified by affinity chromatography and was characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, immunoblotting, and enzyme immunoassay (EIA). The recombinant antigen was reactive with positive human sera, and the reactivity correlated very well (r = 0.9) with that of a whole-virus antigen when tested by EIA with 30 TOS virus-positive and 30 TOS virus-negative serum samples. The results demonstrate that the recombinant N protein can be easily produced in a procaryotic system and used for diagnostic assays for TOS virus immunity.
MeSH terms
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Antibodies, Viral / blood
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Antigens, Viral / genetics
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Antigens, Viral / isolation & purification
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Base Sequence
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Bunyaviridae / genetics
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Bunyaviridae / immunology*
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Bunyaviridae Infections / diagnosis*
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Bunyaviridae Infections / immunology
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Bunyaviridae Infections / virology
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DNA Primers / genetics
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Escherichia coli / genetics
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Humans
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Immunoenzyme Techniques
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Immunoglobulin G / blood
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Immunoglobulin M / blood
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Nucleocapsid Proteins / genetics
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Nucleocapsid Proteins / immunology*
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Nucleocapsid Proteins / isolation & purification
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Recombinant Proteins / genetics
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Recombinant Proteins / immunology
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Recombinant Proteins / isolation & purification
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Reverse Transcriptase Polymerase Chain Reaction
Substances
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Antibodies, Viral
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Antigens, Viral
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DNA Primers
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Immunoglobulin G
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Immunoglobulin M
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Nucleocapsid Proteins
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Recombinant Proteins