title: Cloning, expression and purification of Pwo polymerase from Pyrococcus woesei.
abstract: OBJECTIVE
  Pyrococcus woesei is a hyperthermophilic archaea and produces a heat stable polymerase (Pwo polymerase) that has proofreading activity.
  
  
  METHODS
  In this study, this microorganism was cultured, its DNA was extracted and the pwo gene polymerase was cloned, expressed and purified. The DNA sequence of the cloned gene was verified by sequencing. The pwo polymerase gene consists of 2,328 bps (775 amino acids with about 90 kD molecular weight). Cloning was done by GATEWAY™ Cloning System and for purification of recombinant protein; His6x-Tag was added to the C-terminus of the recombinant protein.
  
  
  CONCLUSIONS
  We could purify Pwo polymerase enzyme by Ni-NTA resin. PCR assay showed that Pwo polymerase activity is comparable to a commercial Pfu polymerase activity.
authors.affiliation: Department of Pathobiology, Institute of Public Health, Tehran University of Medical Sciences, Tehran, Iran.
authors.forename: Amir
authors.initials: A
authors.lastname: Ghasemi
authors.forename: Ali Hatef
authors.initials: AH
authors.lastname: Salmanian
authors.forename: Nourkhoda
authors.initials: N
authors.lastname: Sadeghifard
authors.forename: Amir Ahmad
authors.initials: AA
authors.lastname: Salarian
authors.forename: Mohammad Khalifeh
authors.initials: MK
authors.lastname: Gholi