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  • Expression of the Infectious Bursal Disease Virus Polyprotein in Vero Cells Using Attenuated Salmonella typhimurium as Transgenic Carrier

    Author(s)
    Long, Li
    Wei-Huan, Fang
    Yong-Jun, Fan
    Jian, Xu
    Li, Fang
    Jian-Rong, Li
    Lian, Yu
    Griffith University Author(s)
    Fan, Yongjun
    Year published
    2004
    Metadata
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    Abstract
    To examine if polyprotein gene (VP2/VP4/VP3) of Infectious Bursal Disease Virus (IBDV) could be delivered into mammalian cells and expressed using attenuated Salmonella typhimurium as vector. The IBDV polyprotein gene was amplified by RT-PCR and inserted in to pCI, an eukaryotic expression plasmid. The resulting recombinant pCI-VP2/VP4/VP3 was transformed by electroporation into attenuated Salmonella typhimurium strain ZJ111 (dam- and phoP-), which was then use to transfect the Vero cells. Gene specific RT-PCR revealed that VP2/VP4/VP3 was transcribed into mRNA in the Vero cells. Indirect immunofluorscence assay, SDS-PAGE ...
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    To examine if polyprotein gene (VP2/VP4/VP3) of Infectious Bursal Disease Virus (IBDV) could be delivered into mammalian cells and expressed using attenuated Salmonella typhimurium as vector. The IBDV polyprotein gene was amplified by RT-PCR and inserted in to pCI, an eukaryotic expression plasmid. The resulting recombinant pCI-VP2/VP4/VP3 was transformed by electroporation into attenuated Salmonella typhimurium strain ZJ111 (dam- and phoP-), which was then use to transfect the Vero cells. Gene specific RT-PCR revealed that VP2/VP4/VP3 was transcribed into mRNA in the Vero cells. Indirect immunofluorscence assay, SDS-PAGE and Western-blot analysis showed that VP2/VP4/VP3 was expressed and the product was immuno-reactive with anti-IBDV serum. This work provides essential precondition for developing a new oral DNA vaccine against IBDV.
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    Journal Title
    Chinese Journal of Biotechnology
    Volume
    20
    Issue
    3
    Publisher URI
    http://www.ncbi.nlm.nih.gov/pubmed/15971620
    Subject
    Biomedical Engineering not elsewhere classified
    Biological Sciences
    Technology
    Publication URI
    http://hdl.handle.net/10072/27288
    Collection
    • Journal articles

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