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http://dx.doi.org/10.5352/JLS.2010.20.11.1577

Expression and Production of Human Granulocyte Colony Stimulating Factor (G-CSF) in Silkworm Cell Line  

Park, Jeong-Hae (Department of Clinical Laboratory Science, Dong Eui University)
Jang, Ho-Jung (Department of Clinical Laboratory Science, Dong Eui University)
Kang, Seok-Woo (Department of Agricultural Biology, National Academy of Agricultural Science, RDA)
Goo, Tae-Won (Department of Agricultural Biology, National Academy of Agricultural Science, RDA)
Chung, Kyung-Tae (Department of Clinical Laboratory Science, Dong Eui University)
Publication Information
Journal of Life Science / v.20, no.11, 2010 , pp. 1577-1581 More about this Journal
Abstract
Granulocyte colony stimulating factor (G-CSF) is a hematopoietic cytokine that stimulates bone marrow cells to proliferate and differentiate into granulocytes. G-CSF is approved and used for therapeutic purposes. The endoplasmic reticulum (ER) signal peptide of hG-CSF was replaced with silkworm-specific signal peptides to express and efficiently secrete recombinant hG-CSF by silkworm cells. Plasmids that contain cDNAs for hG-CSF and hG-CSF fused with silkworm- specific signal peptides of prophenoloxidase activating enzyme (PPAE), protein disulfide isomerase (PDI), and bombyxin (BX) were constructed. The G-CSF protein was expressed in insect cell line BM5 and was detected by western blot analysis. The cells transfected with plasmids containing rhG-CSF genes with silkworm-specific signal sequences released mature rhG-CSF protein more efficiently than the cells transfected with pG-CSF, the plasmid containing human G-CSF gene, including its own signal sequence. The production of hG-CSF reached maximal level at four days post-transfection and remained at a high level until 7 days post-transfection. These data demonstrate that the modification of the human G-CSF mimic to insect proteins synthesized in ER greatly improves the production of the protein.
Keywords
G-CSF; insect cells; ER signal sequence; recombinant protein;
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