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.
Article
in Chinese
| 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.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Bombyx
/
Transformation, Genetic
/
Recombinant Fusion Proteins
/
Animals, Genetically Modified
/
Cell Line
/
Granulocyte-Macrophage Colony-Stimulating Factor
/
Cell Biology
/
Fibroins
/
Genetic Vectors
/
Genetics
Limits:
Animals
/
Humans
Language:
Chinese
Journal:
Chinese Journal of Biotechnology
Year:
2010
Type:
Article
Similar
MEDLINE
...
LILACS
LIS