Expression of Porcine Circovirus Type 2ORF2 Gene in Myocardial Cells of Detached Cherry Valley Duck

摘要

The objective of this study was to construct the recombinant eukaryotic expression plasmid of ORF2 gene harboring enhanced green fluorescent protein(EGFP) report gene. ORF2 gene of porcine circovirus type2 cloned by PCR was ligated to the expression vector pEGFP-N1,which contains enhanced green fluorescent protein(EGFP) report gene,the recombinant eukaryotic expression plasmid pEGFP-N1-ORF2 was constructed successfully and was transfected into prepared duck myocardial cells(DMCs) by lipofectin. According to the result,the fluorescence expression was directly detected with fluorescence microscope,and the expression of ORF2 were analyzed by RT-PCR and indirect immunofluorescence assay(IFA) respectively. About 48 h after transfection,green fluorescent can be observed on transfected cells: T-PCR and IFA were positive. This indicated that ORF2 gene of PCV2 was expressed efficiently in transfected duck myocardial cells.