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《Biotechnology》 2008-01
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Cloning of Nucleocapsid Gene and Its Procaryotic Expression of Porcine Reproductive and Respiratory Syndrome Virus(PRRSV)

ZHAO Xi-hong,WEN Xiao-bo,RAN Xu-hua,ZHOU En-min (Animal Science and Technology College,Heilongjiang August First Land Reclamation University,Daqing 163319,China)  
Objective:A recombinant DNA was constructed to express nucleocapsid protein(N)gene of porcine reproductive and respiratory syndrome virus(PRRSV)VR 2332 strain with gene engineering.Methods: The nucleocapsid protein(N)gene of porcine reproductive and respiratory syndrome virus(PRRSV) VR 2332 strain was amplified by RT-PCR and cloned into the multiple cloning site of pET30a(+)vector.The recombinant rpET30a-PRRSV/N was identified and transformed into E.coli BL21(DE3))pLysS competent cell and induced with IPTG.The results was tested by SDS-PAGE and Western blotting.Results: The SDS-PAGE and Western blotting results showed that the recombinant N protein was 19 kDa in size and specifically reacted with anti-PRRSV /N protein monoclonal antibody,indicating the recombinant protein had high specificity of immunereaction and can be used as antigen of diagnostic assay for detection antibodies of PRRSV in practice.Conclusion: The N protein of PRRSV was expressed in Escherichia coli BL2(DE3))pLysS host cells.
【CateGory Index】: Q78;S852.65
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