[Expression of hGM-CSF in transformed silkworm BmN cells mediated by non-transposon vector].
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.
Chen
Huimei
H
College of Pre-clinical Medical and Biological Science, Soochow University, Suzhou 215123, China.
Cao
Guangli
G
Xue
Renyu
R
Gong
Chengliang
C
D000818:Animals; D030801:Animals, Genetically Modified; D012831:Bombyx / Q000166:cytology / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D005349:Fibroins / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics*; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000096:biosynthesis* / Q000235:genetics; D006801:Humans; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D014170:Transformation, Genetic
D011993:Recombinant Fusion Proteins; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D005349:Fibroins