Expression of hGM-CSF in transformed silkworm BmN cells mediated by non-transposon vector / 生物工程学报
Chinese Journal of Biotechnology
;
(12): 830-836, 2010.
Artigo
em Chinês
| WPRIM
| ID: wpr-292201
ABSTRACT
To develop the stable transformants of the silkworm (Bombyx mori) BmN cells that could continuously express the exogenous gene based on a non-transposon vector, an expression cassette containing human granucyto-macrophage colony-stimulating factor (hGM-CSF) gene driven by ie-1 promoter from B. mori nucleopolyhedrovirus was inserted into pIZT-V5-His to form a recombinant vector pIZT-IE-hGM-CSF, followed by transfecting the constructant into BmN cells, the stable ie-hGM-CSF cell lines were obtained after being selected with Zeocin. PCR result using the genomic DNA of the transformed BmN cells as template illustrated a specific fragment of ie-hGM-CSF, and Western blotting analysis using an antibody against hGM-CSF demonstrated a specific band with a molecular weight of 22 kDa in the transformed cells, meanwhile, the expression level of hGM-CSF determined by ELISA was about 2 814.7 pg in 10(6) transformed BmN cells.
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Bombyx
/
Transformação Genética
/
Proteínas Recombinantes de Fusão
/
Animais Geneticamente Modificados
/
Linhagem Celular
/
Fator Estimulador de Colônias de Granulócitos e Macrófagos
/
Biologia Celular
/
Fibroínas
/
Vetores Genéticos
/
Genética
Limite:
Animais
/
Humanos
Idioma:
Chinês
Revista:
Chinese Journal of Biotechnology
Ano de publicação:
2010
Tipo de documento:
Artigo
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