Expression of the infectious bursal disease virus polyprotein in Vero cells using attenuated Salmonella typhimurium as transgenic carrier.
- Author:
Long LI
1
;
Wei-Huan FANG
;
Yong-Jun FAN
;
Jian XU
;
Li FANG
;
Jian-Rong LI
;
Lian YU
Author Information
1. Institute of Preventive Veterinary Medicine, Zhejiang University, Hangzhou, 310029, China.
- Publication Type:Journal Article
- MeSH:
Animals;
Cercopithecus aethiops;
Electroporation;
Genetic Vectors;
genetics;
Infectious bursal disease virus;
genetics;
metabolism;
Polyproteins;
genetics;
Reverse Transcriptase Polymerase Chain Reaction;
Salmonella typhimurium;
genetics;
metabolism;
Transfection;
Vero Cells;
Viral Proteins;
genetics
- From:
Chinese Journal of Biotechnology
2004;20(3):437-440
- CountryChina
- Language: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.
