title: Expression and purification of non-tagged recombinant mouse SPP1 in E. coli and its biological significance.
abstract: Secreted phosphoprotein 1 (SPP1) is a multifunctional protein expressed by cells from a large variety of tissues. It is involved in many physiological and pathological processes, including bone metabolism, inflammation progress, tumor metastasis, injury repair, and hyperoxia-induced injury. Native SPP1 from multiple species have been isolated from the milk and urine, and recombinant SPP1 with different tags have been expressed and purified from bacteria. In our study, DNA fragments corresponding to mouse SPP1 without signal peptide were built into the pET28a(+) vector, and non-tagged recombinant mouse SPP1 (rmSPP1) was expressed in Escherichia coli BL21(DE3). rmSPP1 was purified using a novel tri-step procedure, and the product features high purity and low endotoxin level. rmSPP1 can effectively increase hepatocellular carcinoma cell (HCC) proliferation in vitro, demonstrating its biological activity.
authors.affiliation: a Shanghai Key Laboratory of Veterinary Biotechnology; College of Agriculture and Biology ; Shanghai Jiao Tong University ; Shanghai , P.R. China.
authors.forename: Shunyan
authors.initials: S
authors.lastname: Weng
authors.forename: Liang
authors.initials: L
authors.lastname: Zhou
authors.forename: Lei
authors.initials: L
authors.lastname: Han
authors.forename: Yunsheng
authors.initials: Y
authors.lastname: Yuan
mesh_terms: D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D005822:Genetic Vectors; D006801:Humans; D053495:Osteopontin / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology*
chemical_list: D011993:Recombinant Fusion Proteins; D053495:Osteopontin
keywords: HCC; biological activity; cell proliferation; purity; recombinant SPP1