Expression of Green Fluorescent Protein in Both Spodoptera frugiperda Cells and Bombyx mori Larvae by Ac-Bm Hybrid Virus
Journal of the Korean Society of Virology
; : 225-232, 1998.
Article
em En
| WPRIM
| ID: wpr-81418
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WPRO
ABSTRACT
We have expressed GFP in Sf9 and Bm5 cells or Bombyx by larvae by using Ac-Bm hybrid virus capable of replicating in both Bm5 and Sf9 cells. Genomic DNA of Ac-Bm hybrid virus expressing P-galactosidase was cotransfected with baculovirus transfer vector containing GFP gene, pBacPAK-GFP in Sf9 cells. The Ac-Bm hybrid virus harboring GFP was named as Ac-Bm hybrid virus-GFP. The Ac-Bm hybrid virus-GFP-infected insect cells were easily selected by detecting the emission of GFP from each well of cell culture dish on the UV illuminator. GFP produced by Ac-Bm hybrid virus-GFP in Sf9 and Bm5 cells or B. mori larvae was confirmed by SDS-PAGE and Western blot analysis using GFP antibody. In addition, B. mori larvae infected with Ac-Bm hybrid virus-GFP was apparently appeared fluorescence from the whole body at 5 days postinoculation. The fluorescence of GFP from the hemolymph and fat body of B. mori larvae infected with Ac-Bm hybrid virus-GFP was also observed by fluorescence microscope. In conclusion, our results demonstrated that in baculovirus expression vector system, use of Ac-Bm hybrid virus have an additional advantage of expanded host range for producing recombinant proteins.
Palavras-chave
Texto completo:
1
Índice:
WPRIM
Assunto principal:
Bombyx
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Proteínas Recombinantes
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DNA
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Corpo Adiposo
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Hemolinfa
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Western Blotting
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Baculoviridae
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Spodoptera
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Técnicas de Cultura de Células
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Eletroforese em Gel de Poliacrilamida
Limite:
Animals
Idioma:
En
Revista:
Journal of the Korean Society of Virology
Ano de publicação:
1998
Tipo de documento:
Article