title: A recombinant human G-CSF/GM-CSF fusion protein from E. coli showing colony stimulating activity on human bone marrow cells.
abstract: Granulocyte-Macrophage colony stimulating factor (GM-CSF) and Granulocyte colony stimulating factor (G-CSF) are cytokines involved in the differentiation of bone marrow progenitor cells into myeloid cells. They also activate mature myeloid cells to mediate a variety of antimicrobial activities and inflammatory responses. Recombinant GM-CSF and G-CSF proteins have been used to treat various diseases including cancer and hematopoietic diseases and to isolate peripheral blood progenitor cells for bone marrow transplantation. A plasmid construct expressing recombinant human G-CSF/GM-CSF fusion protein has now been prepared by linking the human G-CSF and GM-CSF coding regions and the recombinant fusion protein has been successfully expressed in E. coli. The recombinant human G-CSF/GM-CSF fusion protein was extracted and purified from the cellular inclusion and refolded into the biologically active form to show colony stimulating activity. The recombinant fusion protein exhibited colony stimulating activity on human bone marrow cell cultures, indicating that the linkage of GM-CSF and G-CSF by a linker peptide may not interrupt activities of the cytokines in the fusion protein. The colony forming unit of the fusion protein was also higher than those of the cultures treated with the same molar numbers of the recombinant human GM-CSF and G-CSF separately, which suggests that the fusion protein presumably retains both G-CSF and GM-CSF activities.
authors.affiliation: Department of Microbiology and School of Biotechnology & Biomedical Science, College of Biomedical Science and Engineering, Inje University, Gimhae 621-749, Korea.
authors.forename: Ah Young
authors.initials: AY
authors.lastname: Lee
authors.forename: Hye Kyung
authors.initials: HK
authors.lastname: Chung
authors.forename: Eun Kyong
authors.initials: EK
authors.lastname: Bae
authors.forename: Jung Suk
authors.initials: JS
authors.lastname: Hwang
authors.forename: Bong Whan
authors.initials: BW
authors.lastname: Sung
authors.forename: Chung Won
authors.initials: CW
authors.lastname: Cho
authors.forename: Jin-Kyoo
authors.initials: JK
authors.lastname: Kim
authors.forename: Kyung
authors.initials: K
authors.lastname: Lee
authors.forename: Jae Yong
authors.initials: JY
authors.lastname: Han
authors.forename: Choon-Taek
authors.initials: CT
authors.lastname: Lee
authors.forename: Hyun Joo
authors.initials: HJ
authors.lastname: Youn
mesh_terms: D001483:Base Sequence; D001854:Bone Marrow Cells / Q000187:drug effects / Q000502:physiology; D002455:Cell Division / Q000187:drug effects; D004926:Escherichia coli / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial; D015971:Gene Expression Regulation, Enzymologic; D016179:Granulocyte Colony-Stimulating Factor / Q000096:biosynthesis* / Q000737:chemistry* / Q000235:genetics / Q000494:pharmacology; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000096:biosynthesis* / Q000737:chemistry* / Q000235:genetics / Q000494:pharmacology; D006801:Humans; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000235:genetics; D011994:Recombinant Proteins
chemical_list: D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D016179:Granulocyte Colony-Stimulating Factor; D016178:Granulocyte-Macrophage Colony-Stimulating Factor