title: Expression, refolding, and characterization of a novel recombinant dual human stem cell factor.
abstract: A novel recombinant dual human stem cell factor (rdhSCF) gene which consisted of a full-length hSCF(1-165aa) cDNA and a truncated hSCF (1-145aa) cDNA, linked by a peptide (GGGGSGGGGSGG) coding region, was constructed and cloned into Escherichia coli expression vector pET-22b. The rdhSCF was expressed at high level in E. coli BL21(DE3) and existed mainly as inclusion bodies. The inclusion bodies were solubilized in urea and refolded by ion-exchange chromatography. After renaturation, the purity of the yielded rdhSCF was up to 90%. Cell proliferation assay showed that the specific activity of the rdhSCF was 2.86x10(5) U/mg, about 1.66 times as high as that of monomer rhSCF expressed in E. coli.
authors.affiliation: School of Life Science and State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, Nanjing 210093, PR China.
authors.forename: Haiqin
authors.initials: H
authors.lastname: Lu
authors.forename: Yuhui
authors.initials: Y
authors.lastname: Zang
authors.forename: Yuguan
authors.initials: Y
authors.lastname: Ze
authors.forename: Jie
authors.initials: J
authors.lastname: Zhu
authors.forename: Tao
authors.initials: T
authors.lastname: Chen
authors.forename: Junhai
authors.initials: J
authors.lastname: Han
authors.forename: Junchuan
authors.initials: J
authors.lastname: Qin
mesh_terms: D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D005822:Genetic Vectors; D016298:Hematopoietic Cell Growth Factors / Q000096:biosynthesis* / Q000737:chemistry* / Q000235:genetics; D006801:Humans; D002479:Inclusion Bodies / Q000737:chemistry / Q000235:genetics; D017510:Protein Folding; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000737:chemistry* / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000737:chemistry* / Q000235:genetics
chemical_list: D016298:Hematopoietic Cell Growth Factors; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; C505956:dual stem cell factor, human