title: Cloning, expression, and characterization of a DNA ligase from a hyperthermophilic archaeon Thermococcus sp.
abstract: Genomic analysis of a hyperthermophilic archaeon, Thermococcus sp. NA1, revealed an ORF of 1689 bases encoding 562 amino acids that showed a high similarity to DNA ligases from other hyperthermophilic archaea. The ligase, which was designated TNA1_lig (Thermococcus sp. NA1 ligase), was cloned and expressed in Escherichia coli. The recombinant TNA1_lig was purified by metal affinity chromatography. The optimum ligase activity of the recombinant TNA1_lig occurred at 80 degrees C and pH 7.5. The enzyme was activated by MgCl2 and ZnCl2 but was inhibited by MnCl2 and NiCl2. Additionally, the enzyme was activated by either ATP or NAD+.
authors.affiliation: Korean Ocean Research & Development Institute, Ansan P.O. Box 29, Seoul, 425-600, Korea.
authors.forename: Yun Jae
authors.initials: YJ
authors.lastname: Kim
authors.forename: Hyun Sook
authors.initials: HS
authors.lastname: Lee
authors.forename: Seung Seob
authors.initials: SS
authors.lastname: Bae
authors.forename: Jung Ho
authors.initials: JH
authors.lastname: Jeon
authors.forename: Sung Hyun
authors.initials: SH
authors.lastname: Yang
authors.forename: Jae Kyu
authors.initials: JK
authors.lastname: Lim
authors.forename: Sung Gyun
authors.initials: SG
authors.lastname: Kang
authors.forename: Suk-Tae
authors.initials: ST
authors.lastname: Kwon
authors.forename: Jung-Hyun
authors.initials: JH
authors.lastname: Lee
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D000595:Amino Acid Sequence; D001431:Bacteriological Techniques / Q000379:methods; D015870:Gene Expression / Q000235:genetics; D008969:Molecular Sequence Data; D009243:NAD / Q000378:metabolism; D016366:Open Reading Frames / Q000235:genetics; D019712:Thermococcus / Q000201:enzymology* / Q000235:genetics
chemical_list: D009243:NAD; D000255:Adenosine Triphosphate