Valassina M, Soldateschi D, Dal Maso G M, Santini L, Bianchi S, Valensin P E, Cusi M G
Department of Molecular Biology, Section of Microbiology, University of Siena, 53100 Siena, Italy.
J Clin Microbiol. 1998 Nov;36(11):3170-2. doi: 10.1128/JCM.36.11.3170-3172.1998.
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.
通过使用pET15b载体，托斯卡纳（TOS）病毒的核衣壳（N）蛋白在大肠杆菌中得以表达。重组蛋白经亲和层析纯化，并通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳、免疫印迹和酶免疫测定（EIA）进行表征。该重组抗原与人类阳性血清发生反应，在用30份TOS病毒阳性和30份TOS病毒阴性血清样本进行EIA检测时，其反应性与全病毒抗原的反应性相关性非常好（r = 0.9）。结果表明，重组N蛋白能够在原核系统中轻松产生，并用于TOS病毒免疫的诊断检测。
