Overexpression, purification and crystallization of a thermostable DNA ligase from the archaeon Thermococcus sp. 1519

Acta Crystallogr Sect F Struct Biol Cryst Commun. 2009 Apr 1;65(Pt 4):368-71. doi: 10.1107/S1744309109007799. Epub 2009 Mar 25.

Abstract

DNA ligases catalyze the sealing of 5'-phosphate and 3'-hydroxyl termini at single-strand breaks in double-stranded DNA and their function is essential to maintain the integrity of the genome in DNA metabolism. An ATP-dependent DNA ligase from the archaeon Thermococcus sp. 1519 was overexpressed, purified and crystallized. Crystals were obtained using the hanging-drop vapour-diffusion method employing 35%(v/v) Tacsimate pH 7.0 as a precipitant and diffracted X-rays to 3.09 A resolution. They belonged to space group P4(1)2(1)2, with unit-cell parameters a = b = 79.7, c = 182.6 A.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Crystallization
  • Crystallography, X-Ray
  • DNA Ligases / chemistry*
  • DNA Ligases / isolation & purification*
  • Electrophoresis, Polyacrylamide Gel
  • Enzyme Stability
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / isolation & purification
  • Temperature*
  • Thermococcus / enzymology*

Substances

  • Recombinant Proteins
  • DNA Ligases