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KMID : 0613820080180070912
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Journal of Life Science
2008 Volume.18 No. 7 p.912 ~ p.917
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Biological Activity of Recombinant Human Granulocyte Colony-Stimulating Factor and Isolation of the Somatic Cell Transfected EGFP-hG-CSF Gene
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Park Jong-Ju
Min Kwan-Sik
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Abstract
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To investigate the biological activity of recombinant human granulocyte colony-stimulating factor (rec-hG-CSF) in mammalian cells, hG-CSF gene was cloned using the cDNA extracted from the human squamous carcinoma cell lines and rec-hG-CSF was produced in CHO cell lines. To analyze the biological activity in vivo, the rec-hG-CSF protein was injected into mice subcutaneously on days 0 and 2. Blood was withdrawn for white blood cell (WBC) determination 5 days after the first injection. WBC values were found to have increased significantly. A pEGFP-mUII-hG-CSF vector was transfected into somatic cell lines isolated from bovine fetal cells. The colony expressing EGFP signals was observed with a confocal microscope. These data suggest that the rec-hG-CSF produced in this study has potent activity in vivo. Thus, the results of this biological activity show that rec-hG-CSF can be enhanced considerably by genetic engineering that affects potential activity, including mutations, which add the oligosaccharide chain and construct double-fusion proteins. A pEGFP-mUII-hG-CSF vector can be utilized for the production of cloned transgenic livestock.
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KEYWORD
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G-CSF, recombinant, biologicalactivity, pEGFP-mUII-hG-CSFvector
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