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Research Information Expression of ORF2 and ORF3 genes of porcine circovirus typ...  

Update Date: [2013-10-30]

Plan Year: 2004
 
    Porcine circovirus type2 represents the causative agent of a new porcine disease which spread in form of an epidemic all over the world in the past years.  In the previous studies demonstrated that the ORF2-encoded structure protein of PCV2 was associated with neutralizing epitopes.  But the functions of ORF3-encoded protein of PCV2 is unknow now.  The purpose of the this study is to investigate the ability of ORF2- and OFR3-encoded protein of PCV2 to elict the neutralizing antibodies in host by means of expressed protein immunization and DNA immunization.  The viral DNA was extracted from Taiwanese PCV2, ORF2- and ORF3-encoding region was amplified by PCR.  The PCR product was inserted into the pRSET vector.  The BL21 (DE3) cell was transfected with pRSET-ORF2 and pRSET-ORF3 plasmid.  The ORF2 and ORF3 recombinant protein was expressed after IPTG inducing and detected with anti-PCV2 polyclonal antibody by Western blotting.  Then, the intraperitoneal injection of recombinant protein into BALB/c mice was performed. Serum samples were collected and checked for ORF2- and ORF3-specific neutralizing antibodies by virus neutralization test.