Diagnostic potential of Toscana virus N protein expressed in Escherichia coli.
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.
Valassina
M
M
Department of Molecular Biology, Section of Microbiology, University of Siena, 53100 Siena, Italy.
Soldateschi
D
D
Dal Maso
G M
GM
Santini
L
L
Bianchi
S
S
Valensin
P E
PE
Cusi
M G
MG
D000914:Antibodies, Viral / Q000097:blood; D000956:Antigens, Viral / Q000235:genetics / Q000302:isolation & purification; D001483:Base Sequence; D002043:Bunyaviridae / Q000235:genetics / Q000276:immunology*; D002044:Bunyaviridae Infections / Q000175:diagnosis* / Q000276:immunology / Q000821:virology; D017931:DNA Primers / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D006801:Humans; D007124:Immunoenzyme Techniques; D007074:Immunoglobulin G / Q000097:blood; D007075:Immunoglobulin M / Q000097:blood; D019590:Nucleocapsid Proteins / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction
D000914:Antibodies, Viral; D000956:Antigens, Viral; D017931:DNA Primers; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D019590:Nucleocapsid Proteins; D011994:Recombinant Proteins