Construction of Eukaryotic Expression Vector and Production of Antibody to Open Reading Frame 4 of Porcine Circovirus Type 2

doi:10.11924/j.issn.1000-6850.2013-2065

Chinese Agricultural Science Bulletin ›› 2014, Vol. 30 ›› Issue (2): 60-64.doi: 10.11924/j.issn.1000-6850.2013-2065

Special Issue: 畜牧兽医

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Construction of Eukaryotic Expression Vector and Production of Antibody to Open Reading Frame 4 of Porcine Circovirus Type 2

Received:2013-07-31 Revised:2013-10-10 Online:2014-01-15 Published:2014-01-15

Abstract

Abstract: The objective of this study was to construct the eukaryotic expression vector of open reading frame 4 (ORF4) of porcine circovirus type 2 (PCV2) and produce the antibody against ORF4. The PCV2 ORF4 gene was amplified by PCR method and cloned into the eukaryotic expression vector pCDNA3.1 (+) or pEGFP-C1, then the recombinant plasmid pCDNA-ORF4 and pEGFP-ORF4 was constructed successfully. After transfection of the recombinant plasmid pEGFP-ORF4 in PK-15 cells, the expression of pEGFP-ORF4 protein was obvious, and the fluorescence signal was mainly localized in the nuclei. After inoculation of the recombinant plasmid pCDNA-ORF4 in BALB/c mice, the specific immune reaction of the inoculated mice sera with PCV2 was observed, and the antibodies titer were all above 1:26 after the fourth inoculation, was indicative of the induction of the antibodies to PCV2 ORF4. The results indicated that two kinds of eukaryotic expression vector of PCV2 ORF4 were constructed and the antibodies against PCV2 ORF4 were prepared, which laid the foundation for researches on PCV2 ORF4 gene functions.

References

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Construction of Eukaryotic Expression Vector and Production of Antibody to Open Reading Frame 4 of Porcine Circovirus Type 2

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