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
Dans Zh
| WPRIM
| ID: wpr-249967
Responsable en Bibliothèque :
WPRO
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.
Texte intégral:
1
Indice:
WPRIM
Sujet Principal:
Salmonella typhimurium
/
Protéines virales
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Cellules Vero
/
Transfection
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Chlorocebus aethiops
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Électroporation
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Virus de la bursite infectieuse
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RT-PCR
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Polyprotéines
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Vecteurs génétiques
Limites du sujet:
Animals
langue:
Zh
Texte intégral:
Chinese Journal of Biotechnology
Année:
2004
Type:
Article