Expression of hGM-CSF in transformed silkworm BmN cells mediated by non-transposon vector.
- Author:
Huimei CHEN
1
;
Guangli CAO
;
Renyu XUE
;
Chengliang GONG
Author Information
1. College of Pre-clinical Medical and Biological Science, Soochow University, Suzhou 215123, China.
- Publication Type:Journal Article
- MeSH:
Animals;
Animals, Genetically Modified;
Bombyx;
cytology;
genetics;
metabolism;
Cell Line;
Fibroins;
genetics;
Genetic Vectors;
genetics;
Granulocyte-Macrophage Colony-Stimulating Factor;
biosynthesis;
genetics;
Humans;
Recombinant Fusion Proteins;
biosynthesis;
genetics;
Transformation, Genetic
- From:
Chinese Journal of Biotechnology
2010;26(6):830-836
- CountryChina
- Language:Chinese
-
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.
