Detection of Immunogenicity of Bacteria-Expressed PCV2 Genome
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S852.65

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    Abstract:

    Objective To find out whether porcine circovirus type 2 (PCV_2) total genome could be expressed by E.coli and whether the recombinant proteins still be biologically active. MethodsThe N-end fragment of PCV2 ORF2 gene of 737-421nt was PCR amplified, cloned and fusion expressed with the expression vector pGEX-6p-1 in E. coli, resulting in the recombinant clone of pGEX-PCV737-421. Both the C-end fragment of the cap gene and the rep gene of PCV2 virus were expressed by E. coli previously. Then SDS-PAGE assay of the three recombinant proteins was performed and Western-blotting against PCV2-specific antiserum was carried out in turn. Finally both the recombinant proteins of the Cap gene were inoculated into SPF BALB/c mice to prepare anti-mouse polyserum in order to find out if the serum could be used to identify the virus antigens propagated in PK15 cell line through indirect fluorescence assay (IFA) experiment. ResultBoth Western-blotting and IFA assays could identify the expressed protein bands and virus antigens specifically. ConclusionThe immunogen assay demonstrated that E.coli expressed recombiant proteins of PCV2 total genome have immunoreactivity.

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  • Received:
  • Revised:April 12,2005
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