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Gene cloning, expression, crystallization and preliminary X-ray analysis of Thermus thermophilus arginyl-tRNA synthetase (2001)

Shimada, Atsushi ; Nureki, Osamu ; Dohmae, Naoshi ; [et al.]

Copenhagen : International Union of Crystallography (IUCr)

Acta crystallographica 57 (2001), S. 272-275

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ISSN: 1399-0047

Source: Crystallography Journals Online : IUCR Backfile Archive 1948-2001

Topics: Chemistry and Pharmacology , Geosciences , Physics

Notes: The gene encoding the highly thermostable arginyl-tRNA synthetase (ArgRS) from Thermus thermophilus was cloned and overexpressed in Escherichia coli under the control of the T7 promoter. The recombinant ArgRS was purified by two chromatographic steps and was crystallized by the hanging-drop vapour-diffusion method using PEG 8000 and ethylene glycol as precipitants. The crystals belong to the hexagonal space group P65, with unit-cell parameters a = b = 156.04 (7), c = 87.17 (4) Å. X-ray data to 2.8 Å resolution were collected at room temperature from a native crystal using an in-house X-ray source. Uranium, platinum and selenomethionine derivatives were found to be useful for phasing by the multiple isomorphous replacement method with anomalous scattering. The flash-frozen crystals diffracted beyond 2.3 Å resolution using synchrotron radiation from the beamline 41XU at SPring-8 (Harima).

Type of Medium: Electronic Resource

URL: http://dx.doi.org/10.1107/S0907444900016255

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