Expression of the Infectious Bursal Disease Virus Polyprotein in Vero Cells Using Attenuated Salmonella typhimurium as Transgenic Carrier
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Wei-Huan, Fang
Yong-Jun, Fan
Jian, Xu
Li, Fang
Jian-Rong, Li
Lian, Yu
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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 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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Chinese Journal of Biotechnology
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20
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3
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Biomedical Engineering not elsewhere classified
Biological Sciences
Technology