Cloning, expression, and characterization of a DNA ligase from a hyperthermophilic archaeon Thermococcus sp.
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+.
Kim
Yun Jae
YJ
Korean Ocean Research & Development Institute, Ansan P.O. Box 29, Seoul, 425-600, Korea.
Lee
Hyun Sook
HS
Bae
Seung Seob
SS
Jeon
Jung Ho
JH
Yang
Sung Hyun
SH
Lim
Jae Kyu
JK
Kang
Sung Gyun
SG
Kwon
Suk-Tae
ST
Lee
Jung-Hyun
JH
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
D009243:NAD; D000255:Adenosine Triphosphate