Expression, refolding, and characterization of a novel recombinant dual human stem cell factor.
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.
Lu
Haiqin
H
School of Life Science and State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, Nanjing 210093, PR China.
Zang
Yuhui
Y
Ze
Yuguan
Y
Zhu
Jie
J
Chen
Tao
T
Han
Junhai
J
Qin
Junchuan
J
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
D016298:Hematopoietic Cell Growth Factors; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; C505956:dual stem cell factor, human