[Isolation and characteristics of new thermostable DNA ligase from archaea of the genus Thermococcus]

Prikl Biokhim Mikrobiol. 2008 Sep-Oct;44(5):523-8.
[Article in Russian]

Abstract

The DNA ligase gene from thermophilic archaea of the genus Thermococcus (strain 1519) was identified and sequenced in the polymerase chain reaction. The recombinant enzyme LigTh1519 was expressed in Escherichia coli, purified, and characterized. LigTh1519 was capable of ligating the cohesive ends and single-strand breaks in double-stranded DNA (ATP as a cofactor). The optimum conditions for the ligase reaction appeared as follows: 100 mM NaCl, 50 mM MgCl2, pH 7.0-10.5, and temperature 70 degrees C. More than 50% Lig1519 activity were preserved after incubation of the enzyme at 80 degrees C for 30 min. New thermostable DNA ligase LihTh1519 may be used for basic and applied researches in molecular biology and genetic engineering.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Archaeal Proteins / chemistry*
  • Archaeal Proteins / genetics
  • Archaeal Proteins / isolation & purification*
  • DNA Breaks, Single-Stranded
  • DNA Ligase ATP
  • DNA Ligases / chemistry*
  • DNA Ligases / genetics
  • DNA Ligases / isolation & purification*
  • Gene Expression
  • Hot Temperature
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / isolation & purification
  • Thermococcus / enzymology*
  • Thermococcus / genetics

Substances

  • Archaeal Proteins
  • Recombinant Proteins
  • DNA Ligases
  • DNA Ligase ATP