[Expression of the infectious bursal disease virus polyprotein in Vero cells using attenuated Salmonella typhimurium as transgenic carrier]

Sheng Wu Gong Cheng Xue Bao. 2004 May;20(3):437-40.
[Article in Chinese]

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Chlorocebus aethiops
  • Electroporation
  • Genetic Vectors / genetics
  • Infectious bursal disease virus / genetics*
  • Infectious bursal disease virus / metabolism
  • Polyproteins / genetics*
  • Reverse Transcriptase Polymerase Chain Reaction
  • Salmonella typhimurium / genetics*
  • Salmonella typhimurium / metabolism
  • Transfection
  • Vero Cells
  • Viral Proteins / genetics*

Substances

  • Polyproteins
  • Viral Proteins