Tehran University of Medical Sciences Iranian Journal of Microbiology 2008-3289 3 3 2011 09 15 118 122 EN Amir Ghasemi Department of Pathobiology, Institute of Public Health, Tehran University of Medical Sciences, Tehran Iran AND Army University of Medical Sciences, Tehran, Iran. Ali Hatef Salmanian National Institute of Genetic Engineering and Biotechnology (NIGEB). Shahrak-e-Pajoohesh, 15th Km, Tehran, Karaj Highway, Tehran, Iran. Nourkhoda Sadeghifard Clinical Microbiology Research Center, Ilam University of Medical Sciences, Ilam, Iran. Amir-Ahmad Salarian Army University of Medical Sciences, Tehran, Iran. Mohammad Khalifeh Gholi Department of Pathobiology, Institute of Public Health, Tehran University of Medical Sciences, Tehran, Iran. 2015 10 01 Background and objectives: Pyrococcus woesei is a hyperthermophilic archaea and produces a heat stable polymerase(Pwo polymerase) that has proofreading activity. Materials and 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. Results and Conclusion: 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. https://ijm.tums.ac.ir/index.php/ijm/article/view/99 https://ijm.tums.ac.ir/index.php/ijm/article/download/99/98