title: Expression, purification and preliminary X-ray analysis of the BRCT domain from Rhp9/Crb2.
abstract: The BRCT domain from Rhp9 (a Schizosaccharomyces pombe DNA-damage checkpoint protein) has been expressed, purified and crystallized. Overexpression in bacterial cells was achieved by minimizing aeration during host cell growth. A robotic screen was used to determine the solubility parameters; concentration of the protein was achieved by exploiting this information. Single crystals suitable for X-ray analysis were obtained in two forms by vapour diffusion (trigonal, unit-cell parameters a = b = 228.04, c = 70.42 A, and tetragonal, P4/m Laue group symmetry, unit-cell parameters a = b = 72.3, c = 91.1 A).
authors.affiliation: CR-UK DNA Repair Enzyme Group, Institute of Cancer Research, 237 Fulham Road, South Kensington, London SW3 6JB, England.
authors.forename: J A
authors.initials: JA
authors.lastname: Hinks
authors.forename: M
authors.initials: M
authors.lastname: Roe
authors.forename: J C Y
authors.initials: JC
authors.lastname: Ho
authors.forename: F Z
authors.initials: FZ
authors.lastname: Watts
authors.forename: J
authors.initials: J
authors.lastname: Phelan
authors.forename: M
authors.initials: M
authors.lastname: McAllister
authors.forename: L H
authors.initials: LH
authors.lastname: Pearl
mesh_terms: D001331:Automation; D018797:Cell Cycle Proteins / Q000737:chemistry*; D003001:Cloning, Molecular / Q000379:methods; D003460:Crystallization / Q000379:methods; D018360:Crystallography, X-Ray; D009687:Nuclear Proteins / Q000737:chemistry*; D017434:Protein Structure, Tertiary; D029702:Schizosaccharomyces pombe Proteins / Q000737:chemistry*; D012965:Sodium Chloride; D012995:Solubility
chemical_list: D018797:Cell Cycle Proteins; C109867:Crb2 protein, S pombe; D009687:Nuclear Proteins; D029702:Schizosaccharomyces pombe Proteins; D012965:Sodium Chloride