title: Expression, purification and crystallization of the cell-division protein YgfE from Escherichia coli.
abstract: An open reading frame designated b2910 (ygfE) in the Escherichia coli K12-MG1655 genome sequence, identified as a possible homologue to the cell-division protein ZapA, was cloned into the high-expression plasmid pETDuet-1 and overexpressed in E. coli BL21 (DE3)-AI. The protein was purified in three steps to 99% purity. Crystals were obtained by the hanging-drop vapour-diffusion method at 291 K from a wide range of screened conditions, but principally from solutions containing 0.1 M HEPES pH 7.0, 18% PEG 6000, 5 mM CaCl2. Diffraction data to 1.8 A were collected at the European Synchrotron Radiation Facility (ESRF). The crystals belong to space group P6(1)22 or P6(5)22, with unit-cell parameters a = 53.8, b = 53.8, c = 329.7 A, alpha = beta = 90, gamma = 120 degrees.
authors.affiliation: Faculty of Life Sciences, University of Manchester, Oxford Road, Manchester M13 9PT, England.
authors.forename: Stephen G
authors.initials: SG
authors.lastname: Addinall
authors.forename: Kenneth A
authors.initials: KA
authors.lastname: Johnson
authors.forename: Timothy
authors.initials: T
authors.lastname: Dafforn
authors.forename: Corinne
authors.initials: C
authors.lastname: Smith
authors.forename: Alison
authors.initials: A
authors.lastname: Rodger
authors.forename: Raul Paco
authors.initials: RP
authors.lastname: Gomez
authors.forename: Katherine
authors.initials: K
authors.lastname: Sloan
authors.forename: Anne
authors.initials: A
authors.lastname: Blewett
authors.forename: David J
authors.initials: DJ
authors.lastname: Scott
authors.forename: David I
authors.initials: DI
authors.lastname: Roper
mesh_terms: D002352:Carrier Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D002455:Cell Division; D003001:Cloning, Molecular; D003460:Crystallization; D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000166:cytology / Q000201:enzymology; D029968:Escherichia coli Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D005784:Gene Amplification; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification
chemical_list: D002352:Carrier Proteins; D029968:Escherichia coli Proteins; D011994:Recombinant Proteins; C520744:ZapA protein, E coli; C118060:ssnA protein, E coli