Korean Journal of Veterinary Research 2009;49(1):39-44.
Production of recombinant nucleocapsid protein of Newcastle disease virus in Escherichia coli for a diagnostic ELISA
Hyun-Il Kim1, Kyoung-Phil Park1, Chan-Hee Park1, Hyun-Ah Cho1, Ho-Suk Yang1, Tae-Wook Hahn2
1Lab of Clinical Pathology, Institute of Cheilbio
2School of Veterinary Medicine, Kangwon National University
Transmission of avian viruses both bird-to-bird and from birds to non-avian species is a major health concern. Newcastle disease virus (NDV) is an economically important avian virus that poses substantial risks to the poultry industry. Rapid and sensitive diagnostic methods, such as the enzymelinked immunosorbent assay (ELISA), are required to track such infections. To develop an ELISA for detecting anti-NDV antibody in avian sera, the nucleocapsid protein (NCP) gene of the NDV La Sota strain was cloned and expressed in Escherichia coli and the 513-amino acid recombinant NCP was purified by Ni-NTA affinity chromatography. To evaluate its ability to replace NDV whole virus antigen as a coating antigen, NCP-coated and whole NDV-coated ELISAs were tested and compared using a panel of NDV positive antisera from chickens. Results using purified NCP were highly correlated with those obtained using whole NDV (r= 0.927), demonstrating that recombinant NCP expressed in Escherichia coli is a suitable substitute antigen for whole NDV in a diagnostic ELISA.
Key Words: E. coli expression, ELISA, Newcastle disease virus, nucleocapsid protein

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