title: Efficient recovery of a functional extracellular domain of bovine IgG2 Fc receptor (boFcgamma2R) from inclusion bodies by a rapid dilution refolding system.
abstract: The extracellular domain of the boFcgamma2R gene was constructed and cloned into the Escherichia coli expression vector pET-28a. The recombinant protein was expressed at high level in E. coli BL21(DE3) and existed mainly as inclusion bodies. The inclusion bodies were solubilized in 6 M guanidine hydrochloride and refolded by rapid dilution. After renaturation, the purity of the recovered recombinant protein was up to 95%. ELISA assay showed that the renatured recombinant protein could inhibit bovine IgG2 binding to expressed boFcgamma2R on the COS-7 cell surface with an IC50 value of 0.68 microM. The overall yield of the active rsbo2R was up to 20 mg/l of culture. Crystals of the rsbo2R were grown at 293 K by the hanging-drop vapour diffusion method showed weak diffraction.
authors.affiliation: Henan Provincial Key Laboratory of Animal Immunology, Henan Academy of Agricultural Sciences, Zhengzhou 450002, China. zhanggaiping2003@yahoo.com.cn
authors.forename: Gaiping
authors.initials: G
authors.lastname: Zhang
authors.forename: Jun
authors.initials: J
authors.lastname: Xi
authors.forename: Xuannian
authors.initials: X
authors.lastname: Wang
authors.forename: Junqing
authors.initials: J
authors.lastname: Guo
authors.forename: Hua
authors.initials: H
authors.lastname: Zhang
authors.forename: Yanyan
authors.initials: Y
authors.lastname: Yang
authors.forename: Songlin
authors.initials: S
authors.lastname: Qiao
authors.forename: Li
authors.initials: L
authors.lastname: Wang
authors.forename: Hong
authors.initials: H
authors.lastname: Zhang
authors.forename: Liyang
authors.initials: L
authors.lastname: He
authors.forename: Yancai
authors.initials: Y
authors.lastname: Zhu
mesh_terms: D000818:Animals; D019556:COS Cells; D002417:Cattle; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D003460:Crystallization / Q000379:methods; D004797:Enzyme-Linked Immunosorbent Assay; D007074:Immunoglobulin G / Q000276:immunology*; D002479:Inclusion Bodies / Q000276:immunology*; D017510:Protein Folding; D020673:Protein Renaturation; D017452:Receptors, IgG / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification* / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D007074:Immunoglobulin G; D017452:Receptors, IgG; D011994:Recombinant Proteins