Cloning, Expression, and Characterization of a DNA Ligase from a Hyperthermophilic Archaeon Thermococcus Sp. |
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| Authors: | Yun Jae Kim Hyun Sook Lee Seung Seob Bae Jung Ho Jeon Sung Hyun Yang Jae Kyu Lim Sung Gyun Kang Suk-Tae Kwon Jung-Hyun Lee |
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| Institution: | (1) Korean Ocean Research & Development Institute, Ansan P.O. Box 29, Seoul, 425-600, Korea;(2) Department of Genetic Engineering, Sungkyunkwan University, 300 Chunchon-dong, Jangan-gu, Suwon, 440-746, Korea |
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| 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 °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+.
Revisions requested 27 October 2005; Revisions received 14 December 2005 |
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| Keywords: | Archaea DNA ligase Thermococcus sp |
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