Prokaryotic Expression Of The Nucleocapsid Protein Of Peste Des Petits Ruminants Virus

Peste des petits ruminants (PPR) is an acute and highly contagious viral disease of sheep , and goats are particularly serious harm, wild animals can be infected ,which is characterized by pyrexia, ocular and nasaldischarges, erosive stomatitis and diarrhoea,and which is caused by peste des petits ruminants virus. PPR was listed as the A disease by OIE,the morbidity and mortality rates can be as high as 100% and 100%, respectively . According the notice of the ministry agriculture, in July of 2007,the first outbreak of peste des petits ruminants in Tibet was confirmed.The nucleocapsid protein of the peste des petits ruminants virus was the major structure proteins and the protective antigen, it induces important biofunctions and play important roles in the pathogenic process and immune response.It was the good signs for PPRV differential diagnosis.In this paper,a pair of primers were designed and synthesized based on the sequence of the nucleocapsid(N)protein genes of peste des petits ruminants virus(PPRV) ,which are added EcoR I and Not I sites for amplifiting the Full-length fragment of the N gene from recombinant plasmids named pCI-neo-N from Austria using PCR..The fragment was cloned into pGM-T vector and transformed into E.coli TOP10, DNA sequence was determined . Then the cloned fragment was subcloned into prokaryotic expression vector pET-32a(+), and the recombinant expression plasmids were expressed in E.coli BL21 (DE3) with inducement. The size of the recombinant fusion protein was 80 kD (N) analyzed by SDS-PAGE.The recombinant fusion protein showed positive reaction with Histidine monoclonal antibody and goat positive serum by Western blotting.