초고온성 고세균 써모코커스 NA1 균주로부터 새로운 고온성 DNA 중합효소

Title
초고온성 고세균 써모코커스 NA1 균주로부터 새로운 고온성 DNA 중합효소
Alternative Title
A novel DNA Polymerase from a Hyperthermophilic Archaeon Thermococcus sp. NA1
Author(s)
김윤재; 이현숙; 조요나; 강성균; 김상진; 권석태; 이정현
KIOST Author(s)
Lee, Hyun Sook(이현숙)Kang, Sung Gyun(강성균)Lee, Jung Hyun(이정현)
Publication Year
2006-06-24
Abstract
Genomic analysis of Thermococcus sp. NA1 revealed the presence of a family B type DNA polymerase gene (TNA1_pol) consisting of 3,927 bp. Sequence analysis revealed that the gene contained a putative 3'-5' exonuclease domain, a DNA polymerase domain and one in-frame intervening sequence of 1,605 bp (535 amino acids; intein), which is located in the middle of PolIII region. The mature form of the DNA polymerase without intein (TNA1_pol) was overexpressed in Esherichia coli, purified by metal affinity chromatography, and further characterized. Optimal activity of TNA1_pol was observed at pH 7.5 and 75 C. TNA1_pol was highly thermostable with t1/2 of 3.5 h at 100 C and 12.5 h at 95 C, respectively. The optimum condition for PCR amplification was determined, and the PCR parameters (error-rate, processivity and extension rate) of TNA1_pol were measured in comparison with rTaq, Pfu and KOD polymerases. TNA1_pol introduced 1 bp error in 4.45 kb on average in PCR amplification. The processivity of TNA1_pol was determined as 150 nt, and the extension rate of TNA1_pol was measured as 60 bases/sec, comparable to those of rTaq DNA polymerase.
URI
https://sciwatch.kiost.ac.kr/handle/2020.kiost/30955
Bibliographic Citation
10-th Internation Symposium om the Genetics of Industrial Microorganisms, pp.1, 2006
Publisher
GIM
Type
Conference
Language
English
Publisher
GIM
Related Researcher
Research Interests

marine biotechnology,molecular microbiology,해양생명공학,분자미생물학

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