Biotechnology Letters

, Volume 33, Issue 2, pp 339–346 | Cite as

Cloning, expression, and PCR application of DNA polymerase from the hyperthermophilic archaeon, Thermococcus celer

  • Kee Pum Kim
  • Heejin Bae
  • In Hye Kim
  • Suk-Tae KwonEmail author
Original Research Paper


The family B DNA polymerase gene was amplified from Thermococcus celer genomic DNA by using the degenerate primers and DNA walking PCR. The Tce DNA polymerase gene was cloned and sequenced. The gene contains an ORF of 2,325 bp encoding 774 amino acid residues with a calculated molecular weight of 89,788.9 kDa. The Tce DNA polymerase was purified by heat treatment and heparin column chromatography. The optimal conditions for PCR were determined. Long-range PCR and time-saving PCR were performed using various specific ratios of Taq and Tce DNA polymerases (Tce plus DNA polymerase). Tce plus DNA polymerase surpassed the PCR performance of Tce, Taq and Pfu DNA polymerases in terms of yield and efficiency.


DNA polymerase PCR improvement Thermococcus celer 



This work was supported by the Marine and Extreme Genome Research Center Program of the Ministry of Land, Transportation and Maritime Affairs, Republic of Korea.


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Copyright information

© Springer Science+Business Media B.V. 2010

Authors and Affiliations

  • Kee Pum Kim
    • 1
  • Heejin Bae
    • 1
  • In Hye Kim
    • 1
  • Suk-Tae Kwon
    • 1
    Email author
  1. 1.Department of Genetic EngineeringSungkyunkwan UniversitySuwonRepublic of Korea

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