title: A rapid purification procedure of recombinant integration host factor from Escherichia coli.
abstract: A rapid procedure for the large-scale isolation of recombinant integration host factor (IHF) protein from Escherichia coli is presented. The protein was overproduced in the E. coli K5746 strain, whose construction has already been described. The procedure consists of a mild extraction of protein and fractionation by ammonium sulfate. A single-step affinity chromatography on heparin-Sepharose provided very pure IHF protein. A Mono-S FPLC column was used to highly concentrate the pure IHF for crystallization trials. Attempts to crystallize IHF produced small stable crystals that have a large number of molecules in the asymmetric unit and to date diffract poorly. Further attempts to crystallize IHF under other conditions as well as in a complex with the putative DNA binding site are underway.
authors.affiliation: European Molecular Biology Laboratory, c/o DESY, Hamburg, Germany.
authors.forename: C E
authors.initials: CE
authors.lastname: Vorgias
authors.forename: K S
authors.initials: KS
authors.lastname: Wilson
mesh_terms: D000645:Ammonium Sulfate; D001426:Bacterial Proteins / Q000235:genetics / Q000302:isolation & purification*; D005591:Chemical Fractionation; D002846:Chromatography, Affinity; D002852:Chromatography, Ion Exchange; D003460:Crystallization; D004268:DNA-Binding Proteins / Q000235:genetics / Q000302:isolation & purification; D004926:Escherichia coli / Q000737:chemistry* / Q000235:genetics; D034982:Integration Host Factors; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification
chemical_list: D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; D034982:Integration Host Factors; D011994:Recombinant Proteins; D000645:Ammonium Sulfate