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EC Number Crystallization (Commentary)
Display the word mapDisplay the reaction diagram Show all sequences 3.1.11.5crystallisation of a Rec-B like nuclease by the sitting-drop vapour-diffusion method using polyethylene glycol 8000 as the precipitant. The crystals belong to the orthorhombic space group C222(1), with unit-cell parameters a = 81.5, b= 159.8, c = 100.8 A. There is a dimer in the asymmetric unit with its local twofold axis running parallel to the crystallographic twofold screw axis. The crystals diffract to about 2 A and a complete native data set is collected to 2.65 A resolution
Display the word mapDisplay the reaction diagram Show all sequences 3.1.11.5purified recombinant enzyme complex RecBCD bound with a DNA substrate (a 350 kDa complex) or ATP analogue ADPNP, mixing of 0.0015 ml of 5 mg/ml protein, and a 1.75fold excess of DNA, 2 mM ADPNP, and 4 mM MgCl2, in 20 mM Tris-HCl, pH 7.5, 50 mM NaCl, and 1 mM TCEP, with 0.03 ml of 0.1% detergent n-dodecyl beta-D-maltoside on ice, incubation at 4°C overnight, X-ray diffraction structure determination and analysis at 3.8 A resolution, molecular replacement and modelling using the crystal structure of RecBCD in complex with an extended DNA fork (PDB ID 3K70) as a template
Display the word mapDisplay the reaction diagram Show all sequences 3.1.11.5vapour-diffusion hanging drop method, crystal structure of a complex of Escherichia coli RecBCD enzyme bound to ablunt-ended DNA hairpin
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