[Expression of the infectious bursal disease virus polyprotein in Vero cells using attenuated Salmonella typhimurium as transgenic carrier].
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.
Li
Long
L
Institute of Preventive Veterinary Medicine, Zhejiang University, Hangzhou, 310029, China.
Fang
Wei-Huan
WH
Fan
Yong-Jun
YJ
Xu
Jian
J
Fang
Li
L
Li
Jian-Rong
JR
Yu
Lian
L
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*
D020815:Polyproteins; D014764:Viral Proteins