초고온성 써모코커스 NA1의 열안정한 DNA ligase 클로닝과 특성

Title
초고온성 써모코커스 NA1의 열안정한 DNA ligase 클로닝과 특성
Alternative Title
Cloning and Characterization of a Thermostable DNA Ligase Gene
Author(s)
김윤재; 임재규; 배승섭; 양성현; 이현숙; 강성균; 김상진; 이정현; 권석태
KIOST Author(s)
Lee, Hyun Sook(이현숙)Kang, Sung Gyun(강성균)Lee, Jung Hyun(이정현)
Publication Year
2005-10-09
Abstract
An extremely thermophilic archaeal strain, NA1 was isolated from a deep-sea hydrothermal vent area at the PACMANUS field in the East Manus Basin. On the basis of 16S rRNA gene sequences, the NA1 was classified to genus, Thermococcus. As a part of genomic research of the NA1, a gene encoding DNA ligase was identified. To characterize the ligase, the gene encoding DNA ligase was cloned into Escherichia coli by PCR. The DNA ligase gene consists of 1,689 bp, corresponding to a polypeptide of 562 amino acids with a predicted molecular mass of 61,820 Da. Sequence comparison with previously reported DNA ligases and the presence of conserved six motifs (Ⅰ, Ⅲ, Ⅲa, Ⅳ, Ⅴ, Ⅵ) suggested that NA1 DNA ligase was an ATP-dependent DNA ligase group. The deduced amino acid sequence of the NA1 DNA ligase was compared with previously reported ATP-dependent ligases, and and the result showed high similarity to other archaeal DNA ligases: T. kodakaransis KOD1 (GeneBank accession no. YP184553) (82% identity), T. fumicolans (GeneBank accession no. CAC21199) (81% identity), Pyrococcus abyssi (GeneBank accession no. CAB49465) (75% identity). The ligase was expressed Escherichia coli, and purified using histag affinity chromatography. The recombinant enzyme showed a DNA ligase activity in the presence of ATP and Mg2+.
URI
https://sciwatch.kiost.ac.kr/handle/2020.kiost/31308
Bibliographic Citation
국제효소공학회 프로시딩, pp.95, 2005
Publisher
국제효소공학회
Type
Conference
Language
English
Publisher
국제효소공학회
Related Researcher
Research Interests

marine biotechnology,molecular microbiology,해양생명공학,분자미생물학

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