Cloning, purification, and characterization of a new DNA polymerase from a hyperthermophilic archaeon, Thermococcus sp NA1 SCIE SCOPUS KCI

Cited 35 time in WEB OF SCIENCE Cited 35 time in Scopus
Title
Cloning, purification, and characterization of a new DNA polymerase from a hyperthermophilic archaeon, Thermococcus sp NA1
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)
Lee, Hyun Sook(이현숙)Kang, Sung Gyun(강성균)Lee, Jung Hyun(이정현)
Publication Year
2007-07
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.
ISSN
1017-7825
URI
https://sciwatch.kiost.ac.kr/handle/2020.kiost/4677
Bibliographic Citation
JOURNAL OF MICROBIOLOGY AND BIOTECHNOLOGY, v.17, no.7, pp.1090 - 1097, 2007
Publisher
KOREAN SOC MICROBIOLOGY & BIOTECHNOLOGY
Subject
HIGH-FIDELITY; PCR; AMPLIFICATION; PROCESSIVITY; SEQUENCES; COMPILATION; EXPRESSION; LITORALIS; ENZYME; KOD1
Keywords
hyperthermophile; characterization; family B-type DNA polymerase; processivity; fidelity
Type
Article
Language
English
Document Type
Article
Publisher
KOREAN SOC MICROBIOLOGY & BIOTECHNOLOGY
Related Researcher
Research Interests

marine biotechnology,molecular microbiology,해양생명공학,분자미생물학

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