Related Experiment Videos
Expression, purification, crystallization and preliminary X-ray diffraction analysis of uracil
J Barchue1, J Symersky, S V Narayana
1Center for Macromolecular Crystallography, University of Alabama at Birmingham, 1918 University Boulevard, Birmingham, AL 35294, USA.
Abstract:
Recombinant uracil phosphoribosyltransferase (UPRT) enzyme of Toxoplasma gondii was expressed in Escherichia coli and purified from the cell-free extract by a combination of chromatographic steps. The recombinant protein was enzymatically active when tested in an in vitro UPRT assay. The purified protein was crystallized using the hanging-drop vapor-diffusion technique with ammonium phosphate as precipitant. The crystallized protein also exhibited UPRT activity. Crystals diffract to 2.4 A resolution and belong to space group P3121 or P3221 with unit-cell dimensions a = b = 119.9, c = 70.8 A and two molecules per asymmetric unit.