Expression of NSP 3AB gene of Encephalomyocarditis virus (EMCV) in E.coli and development of monoclonal antibodies against 3AB protein

China Biotechnology

2008, Vol. 28

Issue (9): 27-31 DOI:

Expression of NSP 3AB gene of Encephalomyocarditis virus (EMCV) in E.coli and development of monoclonal antibodies against 3AB protein

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Abstract

Abstract Objective: To express the EMCV 3AB gene by prokaryotic expression systerm , and prepare monoclonal antibodies against it. Method: NSP 3AB gene of Encephalomyocarditis virus (EMCV) was amplified and cloned into a prokaryotic expression vector pGEX-6P-1 and a recombinant protein 3AB with high antigenicity was expressed in E. coli. Balb / c mice were immunized by purified recombinant 3AB protein of inclusion-body, and the splenocytes of the immunized mice were fused with murine myeloma cells to produce hybridoma cell line. Results: After subcloning by 3 times, one strain of hybridoma cell line steadily secreting antibodies of 3AB protein was obtained, named 2D12. The McAb belongs to IgG1 /κ. The McAb and was confirmed by indirect immunofluorescent assay (IFA) and Western-blotting. Conclusion: These results can provide a potential value for structural and functional studies of EMCV-3AB and early diagnosis of Encephalomyocarditis virus infection.

Key words： Encephalomyocarditis virus (EMCV) non-structural protein Prokaryotic expression Monoclonal antibody

Received: 28 March 2008 Published: 25 September 2008

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. Expression of NSP 3AB gene of Encephalomyocarditis virus (EMCV) in E.coli and development of monoclonal antibodies against 3AB protein. China Biotechnology, 2008, 28(9): 27-31.

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https://manu60.magtech.com.cn/biotech/ OR https://manu60.magtech.com.cn/biotech/Y2008/V28/I9/27

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