Abstract: To study the antiviralactivity of swine viperin in pigs,we extracted and cloned the swine Viperin genes from PK-15 cells. PK-15 cells were seeded onto a 24-well plate and treaded with IFN-α. After 24 h,the cells were harvested to extract the total RNA,and to amplify the Viperin gene by RT-PCR. Then the gene was cloned into the pEASY-blunt simple plasmid for sequencing. The antigenicity and hydrophobicity of swine Viperin were analyzed by DNA Star software,and the fragment with good immunogenicity were chosen and amplified,which was inserted into the prokaryotic expression vector pET-28a⁺. E.coli BL21 containing recombinant plasmid pET-sVIP was induced by IPTG. SDS-PAGE analysis demonstrated that the recombinant protein was expressed in the form of inclusion bodies in E.coli. Then inclusion body protein was purified. The BALB/c mice (9 day old) were immunized two times using the purified recombinants Viperin protein emulsified with equal amounts of Freund's complete adjuvant. Two weeks after final immunization,the serums were collected and the antibody specificity was identified by Western Blot and IFA assay. The result showed that the swine viperin polyclonal antibody could react with the sViperin protein expressed by eukaryotic expression vector pVAX-sVIP. Preparation of polyclonal antibody laid the foundation for further studying the antiviral activity of swine Viperin.

Key words: Swine Viperin protein, Prokaryotic expression, Polyclonal antibody, E.coli

CLC Number: 

• Q78

• S828