Expression of the infectious bursal disease virus polyprotein in Vero cells using attenuated Salmonella typhimurium as transgenic carrier / 生物工程学报
Chinese Journal of Biotechnology
;
(12): 437-440, 2004.
Article
in Chinese
| WPRIM
| ID: wpr-249967
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.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Salmonella typhimurium
/
Viral Proteins
/
Vero Cells
/
Transfection
/
Chlorocebus aethiops
/
Electroporation
/
Infectious bursal disease virus
/
Reverse Transcriptase Polymerase Chain Reaction
/
Polyproteins
/
Genetic Vectors
Limits:
Animals
Language:
Chinese
Journal:
Chinese Journal of Biotechnology
Year:
2004
Type:
Article
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