| Title | A Recombinant Human GM-CSF Protein Expressed as an Inclusion form in Escherichia coli Stimulates Colony Formation and Cell Proliferation in vitro |
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| Author | Ah Young Lee, Jin-Kyoo Kim 1 , Hye Kyung Chung, Eun Kyong Bae, Jung Suk Hwang, Chung Won Cho, Dong Seok Lee, Jae Yong Han 2 , Choon-Taek Lee 3 , Soon- |
| Address | Department of Microbiology and School of Biotechnology and Biomedical Science, College of Biomedical Engineering, Inje University, Kimhae 621-749, Kor |
| Bibliography | Journal of Microbiology, 40(1),77-81, 2002, |
| DOI | |
| Key Words | GM-CSF, recombinant protein, expression construct, colony stimulating activity |
| Abstract | Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a pleiotropic hematopoietic growth factor involved in the development of myeloid cells from bone marrow, and an activator of mature myeloid cells functioning in a variety of antimicrobial and inflammatory responses. Recently, recombinant GM-CSF is increasingly under clinical study for treatment of various diseases including cancer, infectious diseases and hematopoietic diseases as well as for an immune response modulator. In this study, we constructed a recombinant human GM-CSF (rhGM-CSF) expression plasmid with a pelB leader sequence and His.Tag under T7 promoter control. The expression construct was shown to produce a recombinant protein of 20 kDa in the 8M urea preparation, indicating the rhGM-CSF may be expressed as an insoluble inclusion form. The 20 kDa recombinant protein in 8M urea was transformed into the water-soluble form by dialysis against PBS buffer (phosphate buffered saline). The soluble rhGM-CSF protein was shown to stimulate colony formation and cell proliferation in vitro, indicating that the rhGM-CSF could be refolded into its native form to show colony stimulating activity. |
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