Expression, purification, crystallization and preliminary crystallographic analysis of a thermostable DNA ligase from the archaeon Thermococcus sibiricus.

T E Petrova, E Y Bezsudnova, B D Dorokhov, E S Slutskaya, K M Polyakov, P V Dorovatovskiy, N V Ravin, K G Skryabin, M V Kovalchuk, V O Popov

Bach Institute of Biochemistry, RAS, Moscow, Russian Federation. petrova@impb.psn.ru

Acta crystallographica. Section F, Structural biology and crystallization communications 2012 Feb 1

filter terms:

DNA ligases join single-strand breaks in double-stranded DNA by catalyzing the formation of a phosphodiester bond between adjacent 5'-phosphate and 3'-hydroxyl termini. Their function is essential to maintain the integrity of the genome in DNA replication, recombination and repair. A recombinant ATP-dependent DNA</a> ligase from the hyperthermophilic anaerobic archaeon Thermococcus sibiricus was expressed in Escherichia coli and purified. Crystals were grown by vapour diffusion using the hanging-drop method with 17%(w/v) PEG 4000 and 8.5%(v/v) 2-propanol as precipitants. A diffraction experiment was performed with a single crystal, which diffracted X-rays to 3.0 Å resolution. The crystal belonged to space group P2(1)2(1)2(1), with unit-cell parameters a = 58.590, b = 87.540, c = 126.300 Å.

Citation

T E Petrova, E Y Bezsudnova, B D Dorokhov, E S Slutskaya, K M Polyakov, P V Dorovatovskiy, N V Ravin, K G Skryabin, M V Kovalchuk, V O Popov. Expression, purification, crystallization and preliminary crystallographic analysis of a thermostable DNA ligase from the archaeon Thermococcus sibiricus. Acta crystallographica. Section F, Structural biology and crystallization communications. 2012 Feb 1;68(Pt 2):163-5