title: [Expression of the infectious bursal disease virus polyprotein in Vero cells using attenuated Salmonella typhimurium as transgenic carrier].
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.
authors.affiliation: Institute of Preventive Veterinary Medicine, Zhejiang University, Hangzhou, 310029, China.
authors.forename: Long
authors.initials: L
authors.lastname: Li
authors.forename: Wei-Huan
authors.initials: WH
authors.lastname: Fang
authors.forename: Yong-Jun
authors.initials: YJ
authors.lastname: Fan
authors.forename: Jian
authors.initials: J
authors.lastname: Xu
authors.forename: Li
authors.initials: L
authors.lastname: Fang
authors.forename: Jian-Rong
authors.initials: JR
authors.lastname: Li
authors.forename: Lian
authors.initials: L
authors.lastname: Yu
mesh_terms: D000818:Animals; D002522:Chlorocebus aethiops; D018274:Electroporation; D005822:Genetic Vectors / Q000235:genetics; D007243:Infectious bursal disease virus / Q000235:genetics* / Q000378:metabolism; D020815:Polyproteins / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012486:Salmonella typhimurium / Q000235:genetics* / Q000378:metabolism; D014162:Transfection; D014709:Vero Cells; D014764:Viral Proteins / Q000235:genetics*
chemical_list: D020815:Polyproteins; D014764:Viral Proteins