Expression and identification of Peste des petits ruminants virus N gene in Escherichia coli.

A pair of primers added EcoRⅠand NotⅠsites was designed and synthesized based on the sequence of the nucleocapsid(N)protein gene of Peste des petits ruminants virus (PPRV) for amplifying the full-length N gene fragment of pCI-neo -PPRN plasmid from Austria. The fragment was cloned into pGM-T vector and transformed into Escherichia coli TOP10. Then the cloned fragment was subcloned into prokaryotic expression vector pET-32a (+), and the recombinant expression plasmid pET-PPRN was expressed in E.coli BL21 (DE3) with IPTG. The size of the recombinant N fusion protein was 80 kD. The recombinant PPRV N fusion protein could be detected with Histidine monoclonal antibody or goat anti-PPRV sera by Western blot.