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| Phylogenetic analysis of Porcine Circovirus 2 isolated from Henan Province and Expression of the ORF2 Gene |
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Abstract Two pairs of primers were designed according to the published ORF2 gene sequence of PCV2, and ORF2 genes was amplified from PCV2 strain isolated from Henan province .The PCR products was cloned into pMD18-T easy vector and the recombinant plasmids named pMD18- ORF2 were obtained. The sequence of the cloned PCR product was analyzed and compared with other PCV2 isolate in GenBank. The results indicated that the homology of their nucleotide sequence was 92.4% to 99.6% ,and ORF2 gene displayed 90.6％to 97.9％identity based on amino acids with other PCV2 strains. Meanwhile, a fragment of ORF2 gene with a size of 587bp was amplified by PCR from pMD18-T-ORF2 of PCV2 and cloned into pET-32a vector .The recombinant plasmid was successfully expressed in Escherichia Coli BL21 by induction of IPTG. The molecular weight of fused recombinant protein was 40KD by SDS-PAGE and could be recognized by antiserum against PCV2 by Western-Blot.
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Received: 17 November 2008
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