(2011) Cloning, expression and purification of pwo polymerase from pyrococcus woesei. Iranian journal of microbiology. pp. 118-122. ISSN 20083289 (ISSN)
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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 TM 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.
| Item Type: | Article | ||||||||||||
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| Keywords: | Cloning PCR Pyrococcus woesei bacterial enzyme DNA pwo polymerase unclassified drug article bacterium culture carboxy terminal sequence DNA extraction DNA sequence enzyme activity gene expression molecular cloning molecular weight nonhuman protein purification Pyrococcus furiosus | ||||||||||||
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| Page Range: | pp. 118-122 | ||||||||||||
| Journal or Publication Title: | Iranian journal of microbiology | ||||||||||||
| Journal Index: | Scopus | ||||||||||||
| Volume: | 3 | ||||||||||||
| Number: | 3 | ||||||||||||
| ISSN: | 20083289 (ISSN) | ||||||||||||
| Depositing User: | مهندس مهدی شریفی | ||||||||||||
| URI: | http://eprints.medilam.ac.ir/id/eprint/1616 |
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