title: Production of bioactive human hemangiopoietin in Escherichia coli.
abstract: To devise an efficient approach for production of human hemangiopoietin (hHAPO), the gene of hHAPO was synthesized and subcloned into the pSUMO vector with a SUMO tag at the N-terminus. The expression construct was then transformed into the expression strain E. coli BL21(DE3). The fusion protein was expressed in soluble form and identified by SDS-PAGE and Western blotting. The fusion protein was purified to 90% purity by metal chelate chromatography with a yield of 45 mg per liter fermentation culture. The SUMO tag was removed by cleavage with SUMO protease at room temperature for 1 h, and the hHAPO was then re-purified by the metal chelate chromatography. Finally, about 21 mg hHAPO was obtained from 1 liter of fermentation culture with no less than 95% purity. The recombinant hHAPO significantly stimulated the proliferation of human umbilical vein endothelial cells.
authors.affiliation: Jiangsu Province Key Laboratory for Molecular and Medical Biotechnology, Life Science College, Nanjing Normal University, Nanjing, Jiangsu, 210046, PR China.
authors.forename: Jie
authors.initials: J
authors.lastname: Zhang
authors.forename: Jian-Feng
authors.initials: JF
authors.lastname: Li
authors.forename: Shuang-Quan
authors.initials: SQ
authors.lastname: Zhang
mesh_terms: D015153:Blotting, Western; D002460:Cell Line; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000378:metabolism*; D006801:Humans; D011509:Proteoglycans / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D000099158:Proteoglycan 4
chemical_list: D011509:Proteoglycans; D011993:Recombinant Fusion Proteins; C419197:PRG4 protein, human; D000099158:Proteoglycan 4