Expression and Bioactivity of Nucleoprotein from Peste des Petits Ruminants Virus

doi:10.11924/j.issn.1000-6850.casb16010125

Abstract

Abstract: To get the specific antigen nucleoprotein (N) and prepare a method for the rapid detection of antibody level of peste des petits ruminants virus (PPRV) in serum, nucleoprotein (N) gene of PPRV was amplified. Based on the published sequence of the N gene of PPRV in GenBank, two pairs of primer were designed for amplifying the N gene. The amplified fragment was inserted into pCold Ⅰ plasmid via two unique restriction sites of NdeⅠ and Hind Ⅲ. The complete gene encoding the protein of PRRV(N) was subcloned into the expression vector pCold Ⅰ. After transformed into E.coli BL21(DE3), a 58 kD fusion protein was expressed. The expressed product was analyzed by SDS-PAGE and Western Blotting, the results revealed that the expressed fusion protein could specifically react with anti-serum against PRRV. The fusion protein was further purified and used as the coating antigen to establish an indirect ELISA method for detecting PRRV in serum. Compared with the imported standard ELISA, the coincidence rate of the two methods was 98.3%. The expressed nucleoprotein could be used for the preparation of a rapid detection of antibody level of PPRV.

CLC Number:

S852.65

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