Expression, Purification of Nucleoprotein of IBV and Its Application in Monitoring

doi:10.13523/j.cb.20140411

China Biotechnology

2014, Vol. 34

Issue (4): 65-70 DOI: 10.13523/j.cb.20140411

Expression, Purification of Nucleoprotein of IBV and Its Application in Monitoring

LI Zhong-hua¹, XIAO Yun-cai², BI Ding-ren², HU Si-shun², WU Ren-wei²

1. Fujian Province Animal Disease Control Center, Fuzhou 350003, China;
2. College of Veterinary Medicine, Huazhong Agriculture University, Wuhan 430070, China

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Abstract

In order to detect antibodies against infectious bronchitis virus(IBV) in the serum, the N gene of IBV was cloned, expressed and then utilized in the enzyme linked immunosorbent assay(ELISA). N gene(1230bp) of IBV was amplified by RT-PCR from a strain of IBV H52,and confirmed by sequencing and blast analysis. The N gene was then subcloned into prokaryotic expression vector pGEX-KG in the form of vector pGEX-NP. The fusion protein N-GST which was expressed in E. coli BL21(DE3) was characterized by SDS-PAGE and Western blotting analysis as 80kDa with immunity and dissolubility, and was purified with GST affinity column, then to establish an indirect ELISA for the detection of antibodies against IBV. The results indicate the assay has excellent reduplication, high sensitivity and specificity. It could be applied to detect the antibodies against IBV fast, to provide technical support for sero-epidemiologic survey of IBV infection and to understand how IBV infect animals.

Key words： Infectious bronchitis virus Nucleocapsid protein Prokaryotic expression ELISA

Received: 11 March 2014 Published: 25 April 2014

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Cite this article:

LI Zhong-hua, XIAO Yun-cai, BI Ding-ren, HU Si-shun, WU Ren-wei. Expression, Purification of Nucleoprotein of IBV and Its Application in Monitoring. China Biotechnology, 2014, 34(4): 65-70.

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https://manu60.magtech.com.cn/biotech/10.13523/j.cb.20140411 OR https://manu60.magtech.com.cn/biotech/Y2014/V34/I4/65

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