THERMOCOCCUS CELERICRESCENS DNA POLYMERASE AND THERMOCOCCUS CELERICRESCENSE A752K/N213D DNA POLYMERASE AND THEIR USE

摘要

PURPOSE: A Thermococcus celericrescens-derived DNA polymerase having DNA polymerization activity and proofreading activity is provided to enable various nucleic acid polymerization reactions and to improved PCR efficiency and thermal resistance. CONSTITUTION: A Tcel DNA polymerase isolated from Thermococcus celericrescens has an amino acid sequence of sequence number 14. A nucleic acid molecule encoding the polymerase has a base of sequence number 13. A recombinant vector contains the nucleic acid molecule. TcelA752K/N213D DNA polymerase has an optimal activity at pH 8, 70 Deg. C. 100 mM of KCl, and 2 mM or Mg^2+. A recombinant vector, pTCELM vector, contains the nucleic acid molecule encoding the TcelA752K/N213D DNA polymerase. A microorganism transformed by the vector is E.coli Rosetta(DE3)pLysS / pTCELM (deposit number KACC91614P).