Citation: Cap protein, . Construction and Identification of Recombinant Adenoviruses Containingthe Cap Protein of Porcine Circovirus 2 .VIROLOGICA SINICA, 2006, 21(1) : 29-33.

Construction and Identification of Recombinant Adenoviruses Containingthe Cap Protein of Porcine Circovirus 2

  • Available online: 20 January 2006
  • Porcine circovirus 2 (PCV2) has been implicated as the etiological agent of a new disease, named postweaning multisystemic wasting syndrome (PMWS). PCV2 contains two major open reading frames (ORFs): ORF1 and ORF2, encoding replication-associated (Rep) protein and capsid (Cap) protein, respectively. The cap protein is a candidate 2 for recombinant PCV vaccine, because it possesses several potential antigen epitopes. In the study, the cap gene was amplified by PCR, and cloned into the transfer vector pShuttle-CMV subsequentially. The recombinant plasmid and bone vector pAdEasy-1 were cotransformed into BJ5183 to generate the recombinant plasmid rAd-ORF2 pAd-Cap. The recombinant adenovirus (rAd-Cap) was generated by transfection and plaque purification in in 293 cells. The transcription and expression of Cap protein in rAd-Cap infected 293 cells were confirmed by RT-PCR、indirect-ELISA, Western blot and IPMA. The study laid the foundation for development of the recombinant vaccine against PCV2.

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    Construction and Identification of Recombinant Adenoviruses Containingthe Cap Protein of Porcine Circovirus 2

    • 1. Key Laboratory of Animal Diseases Diagnostic and Immunology, Ministry of Agriculture, Nanjing Agricultural University, Nanjing 210095,China

    Abstract: Porcine circovirus 2 (PCV2) has been implicated as the etiological agent of a new disease, named postweaning multisystemic wasting syndrome (PMWS). PCV2 contains two major open reading frames (ORFs): ORF1 and ORF2, encoding replication-associated (Rep) protein and capsid (Cap) protein, respectively. The cap protein is a candidate 2 for recombinant PCV vaccine, because it possesses several potential antigen epitopes. In the study, the cap gene was amplified by PCR, and cloned into the transfer vector pShuttle-CMV subsequentially. The recombinant plasmid and bone vector pAdEasy-1 were cotransformed into BJ5183 to generate the recombinant plasmid rAd-ORF2 pAd-Cap. The recombinant adenovirus (rAd-Cap) was generated by transfection and plaque purification in in 293 cells. The transcription and expression of Cap protein in rAd-Cap infected 293 cells were confirmed by RT-PCR、indirect-ELISA, Western blot and IPMA. The study laid the foundation for development of the recombinant vaccine against PCV2.

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