CLONING, EXPRESSION AND PURIFICATION OF PWO POLYMERASE FROM PYROCOCCUS WOESEI

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GHASEMI AMIR; SALMANIAN ALI HATEF; SADEGHIFARD NOURKHODA; SALARIAN AMIR AHMAD; KHALIFEH GHOLI MOHAMMAD

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Journal Paper

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Journal: Iranian Journal of Microbiology Year:2011 | Volume:3 | Issue:3 Page(s): 118-122

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Title

CLONING, EXPRESSION AND PURIFICATION OF PWO POLYMERASE FROM PYROCOCCUS WOESEI

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Keywords

PYROCOCCUS WOESEI

CLONING

PCR

Abstract

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.

Cites

EXPRESSION, PURFICATION AND ENDOTOXIN REMOVAL OF BRUCELLA MELITENSIS DNAK AND OMP31 PROTEINS

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APA: Copy

GHASEMI, AMIR, SALMANIAN, ALI HATEF, SADEGHIFARD, NOURKHODA, SALARIAN, AMIR AHMAD, & KHALIFEH GHOLI, MOHAMMAD. (2011). CLONING, EXPRESSION AND PURIFICATION OF PWO POLYMERASE FROM PYROCOCCUS WOESEI. IRANIAN JOURNAL OF MICROBIOLOGY, 3(3), 118-122. SID. https://sid.ir/paper/315286/en

Vancouver: Copy

GHASEMI AMIR, SALMANIAN ALI HATEF, SADEGHIFARD NOURKHODA, SALARIAN AMIR AHMAD, KHALIFEH GHOLI MOHAMMAD. CLONING, EXPRESSION AND PURIFICATION OF PWO POLYMERASE FROM PYROCOCCUS WOESEI. IRANIAN JOURNAL OF MICROBIOLOGY[Internet]. 2011;3(3):118-122. Available from: https://sid.ir/paper/315286/en

IEEE: Copy

AMIR GHASEMI, ALI HATEF SALMANIAN, NOURKHODA SADEGHIFARD, AMIR AHMAD SALARIAN, and MOHAMMAD KHALIFEH GHOLI, “CLONING, EXPRESSION AND PURIFICATION OF PWO POLYMERASE FROM PYROCOCCUS WOESEI,” IRANIAN JOURNAL OF MICROBIOLOGY, vol. 3, no. 3, pp. 118–122, 2011, [Online]. Available: https://sid.ir/paper/315286/en

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