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
em Zh
| WPRIM
| ID: wpr-249967
Biblioteca responsável:
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.
Texto completo:
1
Índice:
WPRIM
Assunto principal:
Salmonella typhimurium
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Proteínas Virais
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Células Vero
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Transfecção
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Chlorocebus aethiops
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Eletroporação
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Vírus da Doença Infecciosa da Bursa
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Reação em Cadeia da Polimerase Via Transcriptase Reversa
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Poliproteínas
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Vetores Genéticos
Limite:
Animals
Idioma:
Zh
Revista:
Chinese Journal of Biotechnology
Ano de publicação:
2004
Tipo de documento:
Article