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., Soldateschi, D., Dal Maso, G.M., Santini, L., Bianchi, S., Valensin, P.E., et al. (1998). Diagnostic potential of Toscana virus N protein expressed in Escherichia coli. JOURNAL OF CLINICAL MICROBIOLOGY, 36(11), 3170-3172 [10.1128/jcm.36.11.3170-3172.1998].
Diagnostic potential of Toscana virus N protein expressed in Escherichia coli
Cusi M. G.
1998-01-01
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.
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https://hdl.handle.net/11365/30946
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