The studies on the transfection of EGFP-fused porcine circovirus Type 2 genome.

This study described construction and transfection of an EGFP-fused Porcine Circovirus Type 2 (PCV2) genome and the recovery of the virus. Posttransfection, PCV2 (ORF1)-EGFP/pSK, PCV2 (ORF3)-EGFP/pSK, PCV2 (ORF4)-EGFP/pSK and PCV2(ORF5)-EGFP/pSK showed no fluorescent signals in transfected cells, while green fluorescent signals were observed in the nuclei of PK-15 cells after PCV2 (ORF2)-EGFP/pSK transfection. The presence of ORF2-EGFP fusion protein was demonstrated by dual signals of green fluorescence and anti-PCV2 antibodies conjugated with rhodamine in an immunofluorescence assay (IFA). Furthermore, the released EGFP-fused PCV2 genome was demonstrated by real-time PCR.