Cloning and expression of region VP2 gene of Aleutian mink disease parvovirus and analysis of immunogenicity of the recombinant protein.

The primers were designed according to the complete genome sequences of Aleutian mink disease parvovirus(ADV) published in GenBank.The part domains of VP2 gene of ADV strain were amplified by PCR and cloned into pET-28a vector.PCR,restriction endonuclease digestion and sequence analysis proved the insertion position. It was indicated that the prokaryotic expression vectors Pet-28a-VP2 were constructed.The positive recombinant vectors were transformed into recipient germs BL21 for expression by IPTG inducing.The expressed proteins were measured by SDSPAGE and Western blot.Results showed that the proteins could be expressed successfully.The molecular weights of the expressed proteins were separately 21 kD which agreed with the theory presumed.Induced by IPTG at a final concentration of 1 mmol/L,the expressed protein reached the highest quantity after 5 hours of induction.The Western blot results indicated that the expressed antigen proteins could be recognized by the rabbitanti-Mink antiserum and had some immunoreactivity,and could be efficiently expressed in the prokaryotic system with immunological activities.