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Virologica Sinica, 18 (4) : 371-375, 2003
Research Article
M olecular Cloning of the Infectious Genomic DNA of Porcine circovirus Type 2
Key laboratory of Animal Disease Diagnostic and Immunology,Ministry of Agriculture,Nanjing Agricultural University,Nanjing 210095,China
2.Colle·. P ofAgriculture,Shanghai Jiaotong University,Shanghai 200030,China
3. Anima l Science Department ofAgricultural College。Ningxia University,Hnchuan 750105,China
The complete genome of type 2 Porcine circovirus(pcv一2)was amplified by polymerase
chain reaction(PCR and cloned directly into me EcoRI sites of plasmid pcDNA3,and recominant
plasmid carrying the complete genome was constructed,designated pcDNApcv2.The entire genome of
PCV一2 was purified and recycled from pcDNApcv2 with the digestion of EcoRI enzyme an d then
circular genomic DNA was generated by self-ligating with T4 DNA ligase in vitro.Th e non—infected
PK一1 5 cells were transfected with the PCV一2 circular genome using Lipofectin Reagent.After four
continuous passages,PCV一2 virus and specific antigens were visualized by electron microscopy an d
IFA,respectively.Th us,the cloned circular PC V一2 genomi c DNA generated in this study was infectious
in vitro.
  Published online: 5 Aug 2003
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