2011•Chinese Veterinary ScienceRequires access
Study on the construction of eukaryotic expression plasmid pEGFP N1 ORF2 and its expression in duck myocardial cells
Zhiqiang Shen
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Abstract
In this study,ORF2 gene of PCV2 was cloned via PCR method,and the eukaryotic expression plasmid of ORF2 gene with enhanced green fluorescent protein(EGFP) report gene was constructed.The recombinant plasmid was transfected with lipofectin method prepared duck myocardial cells(DMCs).The fluorescence expression was directly detected with fluorescence microscope,and the expression of ORF2 were tested by RT-PCR and indirect immunofluorescence assay(IFA),respectively.About 48h after transfected,green fluorescent can be detected in transfected cells,and RT-PCR and IFA were positive.The results indicated that ORF2 gene was expressed efficiently in transfected duck myocardial cells.