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Cloning, Purification, and Characterization of a New DNA Polymerase from a Hyperthermophilic Archaeon, Thermococcus sp. NA1
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  • Cloning, Purification, and Characterization of a New DNA Polymerase from a Hyperthermophilic Archaeon, Thermococcus sp. NA1
저자명
Kim. Yun-Jae,Lee. Hyun-Sook,Bae. Seung-Seob,Jeon. Jeong-Ho,Lim. Jae-Kyu,Cho. Yon-A,Nam. Ki-Hoon,Kang. Sung-Gyun,Kim. Sang-Jin,Kw
간행물명
Journal of microbiology and biotechnology
권/호정보
2007년|17권 7호|pp.1090-1097 (8 pages)
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한국미생물생명공학회
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이 논문은 한국과학기술정보연구원과 논문 연계를 통해 무료로 제공되는 원문입니다.
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기타언어초록

Genomic analysis of Thermococcus sp. NA1 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 $75^{circ}C$. TNA1_pol was highly thermostable, with a half-life of 3.5h at $100^{circ}C$ and 12.5h at $95^{circ}C$. 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 (nt) 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.