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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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- Authors
- Issue Date
- 2002
- Publisher
- Springer
- Citation
- Journal of Microbiology, vol.40 no.1, pp. 77-81
- Keywords
- 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.
- ISSN
- 1225-8873
- Language
- English
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