title: The studies on the transfection of EGFP-fused porcine circovirus Type 2 genome.
abstract: 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.
authors.affiliation: School of Veterinary Medicine, National Taiwan University, Taipei 10617, Taiwan.
authors.forename: Hsu-Chung Gabriel
authors.initials: HC
authors.lastname: Chen
authors.forename: Shu-Hsiang
authors.initials: SH
authors.lastname: Yang
authors.forename: Tsun-Yung
authors.initials: TY
authors.lastname: Kuo
authors.forename: Shiow-Suey
authors.initials: SS
authors.lastname: Lai
mesh_terms: D000818:Animals; D002460:Cell Line; D017925:Circovirus / Q000235:genetics* / Q000302:isolation & purification; D003001:Cloning, Molecular; D004279:DNA, Viral; D005455:Fluorescent Antibody Technique / Q000662:veterinary; D005456:Fluorescent Dyes / Q000032:analysis*; D016679:Genome, Viral / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics*; D010957:Plasmids / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction / Q000662:veterinary; D011993:Recombinant Fusion Proteins / Q000032:analysis; D013552:Swine; D014162:Transfection / Q000379:methods / Q000662:veterinary*
chemical_list: D004279:DNA, Viral; D005456:Fluorescent Dyes; D011993:Recombinant Fusion Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins