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논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 발행연도
- 2009.1
- 수록면
- 17 - 23 (7page)
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The gene encoding Thermus thermophilus HJ6 DNA polymerase (Tod) was cloned and sequenced. The open reading frame (ORF) of the Tod gene was composed of 2,505 nucleotides and encoded a protein (843 amino acids)
with a predicted molecular weight of 93,795 Da. The deduced amino acid sequence of Tod showed 98% and 86% identities to the Thermus thermophilus HB8 DNA pol and Thermus aquaticus DNA pol, respectively.
The Tod gene was expressed under the control of the bacteriophage λ promoters PR and PL on the expression vector pJLA503 in Escherichia coli strain BL21 (DE3) codon plus. The expressed enzyme was purified by
heat treatment, HiTrapTM Q column, and HiPrepTM Sephacryl S-200 HR 26/60 column chromatographies. The optimal temperature and pH for DNA polymerase activity were found to be 75~80℃ and 9.0, respectively.
The optimal concentrations of Mg2+ and Mn2+ were 2.5 mM and 1 mM, respectively. The enzyme activity was activated by divalent cations, and was inhibited by monovalent cations. The result of the PCR experiment with
Tod DNA polymerase indicates that this enzyme might be useful in DNA amplification and PCR-based applications.
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참고문헌 (14)
참고문헌 신청
Braithwaite, D. K / 1993 / Compilation, alignment, and phylogenetic relationships of DNA polymerases / Nucleic. Acids. Res 21 : 787 ~ 802
CHOI, JEONG JIN / 2004 / Cloning, Expression, and Characterization of DNA Polymerase fromHyperthermophilic Bacterium Aquifex pyrophilus / Journal of Microbiology and Biotechn
Cann, I. K / 1999 / Archaeal DNA replication: identifying the pieces to solve a puzzle / Genetics 152 : 1249 ~ 1267
Chien, A / 1976 / Deoxyribonucleic acid polymerase from the extreme thermophile Thermus aquaticus / J. Bacteriol 127 : 1550 ~ 1557
Choi, J. J / 1999 / Purification and properties of Thermus filiformis DNA polymerase expressed in Escherichia coli / Biotech. Appl. Biochem 30 : 19 ~ 25
CHOI, JEONG JIN / 2004 / Cloning, Expression, and Characterization of DNA Polymerase fromHyperthermophilic Bacterium Aquifex pyrophilus / Journal of Microbiology and Biotechn
Cann, I. K / 1999 / Archaeal DNA replication: identifying the pieces to solve a puzzle / Genetics 152 : 1249 ~ 1267
Chien, A / 1976 / Deoxyribonucleic acid polymerase from the extreme thermophile Thermus aquaticus / J. Bacteriol 127 : 1550 ~ 1557
Choi, J. J / 1999 / Purification and properties of Thermus filiformis DNA polymerase expressed in Escherichia coli / Biotech. Appl. Biochem 30 : 19 ~ 25
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