Cloning and expression of human stem cell factor fused with thrombopoietin mimetic peptide in Escherichia coli.
Thrombopoietin (TPO) acts synergistically with stem cell factor (SCF) in hematopoiesis and megakaryopoiesis. In this work, we designed the expression of SCF fused with the monomer or the dimer of TPO mimetic peptide through a flexible peptide linker. The recombinant fusion proteins were produced in E. coli DH5alpha at up to 25% of total cell proteins. The resultant inclusion bodies were refolded by dilution and purified by ion-exchange chromatography. Subsequent biological activity assays showed that the fusion proteins exhibited higher potency than recombinant human SCF, indicating that they have a potential role for pharmaceutical applications.
Su
Lin
L
National Laboratory of Medical Molecular Biology, Department of Biochemistry and Molecular Biology, Peking Union Medical College, Beijing, China.
Chen
Songsen
S
Yang
Kegong
K
Liu
Changzheng
C
Liang
Zhiquan
Z
D019677:Artificial Gene Fusion / Q000379:methods*; D002852:Chromatography, Ion Exchange; D004926:Escherichia coli / Q000235:genetics; D005312:Fetal Blood / Q000187:drug effects*; D005786:Gene Expression Regulation / Q000235:genetics*; D006801:Humans; D002479:Inclusion Bodies; D046911:Macromolecular Substances; D010455:Peptides / Q000737:chemistry / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D010957:Plasmids / Q000235:genetics*; D053628:Receptors, Thrombopoietin / Q000819:agonists; D019089:Stem Cell Factor / Q000737:chemistry / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology
D046911:Macromolecular Substances; D010455:Peptides; D053628:Receptors, Thrombopoietin; D019089:Stem Cell Factor; C515728:thrombopoietin mimetic peptide