Cloning, purification, and characterization of a new DNA polymerase from a hyperthermophilic archaeon, Thermococcus sp NA1
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Title
- Cloning, purification, and characterization of a new DNA polymerase from a hyperthermophilic archaeon, Thermococcus sp NA1
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Author(s)
- Kim, Yun Jae; Lee, Hyun Sook; Bae, Seung Seob; Jeon, Jeong Ho; Lim, Jae Kyu; Cho, Yona; Nam, Ki Hoon; Kang, Sung Gyun; Kim, Sang-Jin; Kwon, Suk-Tae; Lee, Jung-Hyun
- KIOST Author(s)
- Kim, Yun Jae(김윤재); Lee, Hyun Sook(이현숙); Lim, Jae Kyu(임재규); Kang, Sung Gyun(강성균); Lee, Jung Hyun(이정현)
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Alternative Author(s)
- 김윤재; 이현숙; 배승섭; 전정호; 임재규; 조요나; 강성균; 김상진; 이정현
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Publication Year
- 2007-07
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Abstract
- Genomic analysis of Thermococcus sp. TNA1 revealed the presence of a 3,927-base-pair (bp) family B-type DNA polymerase gene, TNA1_pol. TNA1_pol, without its intein, was overexpressed in Escherichia coli, purified using metal affinity chromatography, and characterized. TNA1_pol activity was optimal at pH 7.5 and 75 degrees C. TNA1_pol was highly thermostable, with a half-life of 3.5 h at 100 degrees C and 12.5 h at 95 degrees C. Polymerase chain reaction parameters of TNA1_pol such as error-rate, processivity, and extension rate were measured in comparison with rTaq, Pfu, and KOD DNA polymerases. TNA1_pol averaged one incorrect bp every 4.45 kilobases (kb), and had a processivity of 150 nucleotides (m) and an extension rate of 60 bases/s. Thus, TNA1_pol has a much faster elongation rate than Pfu DNA polymerase with 7-fold higher fidelity than that of rTaq.
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ISSN
- 1017-7825
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URI
- https://sciwatch.kiost.ac.kr/handle/2020.kiost/4677
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Bibliographic Citation
- JOURNAL OF MICROBIOLOGY AND BIOTECHNOLOGY, v.17, no.7, pp.1090 - 1097, 2007
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Publisher
- KOREAN SOC MICROBIOLOGY & BIOTECHNOLOGY
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Subject
- HIGH-FIDELITY; PCR; AMPLIFICATION; PROCESSIVITY; SEQUENCES; COMPILATION; EXPRESSION; LITORALIS; ENZYME; KOD1
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Keywords
- hyperthermophile; characterization; family B-type DNA polymerase; processivity; fidelity
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Type
- Article
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Language
- English
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Document Type
- Article
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