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Volume 18 Issue 4
August 2003
    Citation: LUYin—hua, HUA Xiu—guo, CUI Li, TAN Guo—lei, XU Li—hua ’, HUANG Wei-jian, CHEN De．sheng ．CHEN Pu—yan. M olecular Cloning of the Infectious Genomic DNA of Porcine circovirus Type 2 .VIROLOGICA SINICA, 2003, 18(4) : 371-375.
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    M olecular Cloning of the Infectious Genomic DNA of Porcine circovirus Type 2

    • 1.

      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

    • Available online: 05 August 2003
    • 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．
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      M olecular Cloning of the Infectious Genomic DNA of Porcine circovirus Type 2

      • 1. 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

      Abstract: 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．

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