title: Soluble expression and one-step purification of recombinant mouse interferon-λ3 in Escherichia coli.
abstract: Interferon (IFN)-λ3, a member of the type III IFN family, is a pleiotropic cytokine that exhibits potent antiproliferative, antiviral, and immunoregulatory activities. For further functional study of IFN-λ3, we developed an efficient procedure that includes cloning, expression, and purification to obtain relatively large quantity of mouse IFN-λ3 fusion protein. The mature IFN-λ3 protein-coding region was cloned into the prokaryotic expression vector pET-44. IFN-λ3 contains a hexahistidine tag at its C-terminus. We used Ni(2+)-nitrilotriacetic acid agarose-affinity chromatography to purify the expressed soluble protein. The purified IFN-λ3 inhibited significantly IL-13 production in stimulated RAW264.7 macrophages. Our findings show that the production of soluble IFN-λ3 proteins by the pET-44 vector in Escherichia coli is a good alternative for the production of native IFN-λ3 and could be useful for the production of other IFN proteins.
authors.affiliation: Zhejiang Provincial Key Laboratory of Pathophysiology, Department of Immunology, Ningbo University School of Medicine, Ningbo, 315211, China. yanli319@yahoo.com.
authors.forename: Y Q
authors.initials: YQ
authors.lastname: Wang
authors.forename: M
authors.initials: M
authors.lastname: Zhou
authors.forename: L M
authors.initials: LM
authors.lastname: Zeng
authors.forename: Q Y
authors.initials: QY
authors.lastname: Gao
authors.forename: X L
authors.initials: XL
authors.lastname: Yuan
authors.forename: Y
authors.initials: Y
authors.lastname: Li
authors.forename: M C
authors.initials: MC
authors.lastname: Li
mesh_terms: D000818:Animals; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics*; D005822:Genetic Vectors; D006639:Histidine / Q000235:genetics; D007372:Interferons / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D051379:Mice; D009842:Oligopeptides / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification
chemical_list: C471213:His-His-His-His-His-His; D009842:Oligopeptides; D011993:Recombinant Fusion Proteins; C000602567:interferon-lambda3, mouse; D006639:Histidine; D007372:Interferons