Use of prokaryotically expressed nucleocapsid protein as positive antigen in ELISA.

S S Kumar, R Renji, M Saini, A C Goel, B Sharma
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引用次数: 2

Abstract

A cDNA library of Rinderpest vaccine virus was prepared in Zap Express vector (Stratagene). The Rinderpest 'N' gene specific clones were selected, characterized and thereafter expressed in E. coli XLOLR strain. The expressed protein was found to be immunogenic in western blot with hyperimmune sera. It reacted with rinderpest and 'N' protein specific monoclonal antibodies in Enzyme Linked Immunosorbent Assay (ELISA). Prokaryotically expressed 'N' protein also gave precipitin band in counter immunoelectrophoresis test (CIE). The expression of N protein was sufficient for its utility as positive antigen in CIE and ELISA used for rinderpest diagnosis.

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用原核表达的核衣壳蛋白作为ELISA阳性抗原。
在Zap Express载体(Stratagene)上制备牛瘟疫苗病毒cDNA文库。筛选牛瘟“N”基因特异性克隆,对其进行鉴定,并在大肠杆菌XLOLR中表达。高免疫血清western blot结果显示表达蛋白具有免疫原性。在酶联免疫吸附试验(ELISA)中与牛瘟和“N”蛋白特异性单克隆抗体反应。原核表达的“N”蛋白在反向免疫电泳(CIE)中也出现沉淀带。在CIE和ELISA中,N蛋白的表达足以作为牛瘟诊断的阳性抗原。
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