Cloning,Sequence Analysis and Three-dimensional Structure Prediction of DNA Pol I from Thermophilic <Emphasis Type="Italic">Geobacillus</Emphasis> sp. <Emphasis Type="Italic">MKK</Emphasis> Isolated from an Iranian Hot Spring |
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Authors: | Mohammad Khalaj-Kondori Majid Sadeghizadeh Khosro Khajeh Hossein Naderi-Manesh Ali Mohammad Ahadi Abdorahman Emamzadeh |
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Institution: | (1) Department of Biology, Faculty of Basic Sciences, Tarbiat Modares University, 14115-175, Tehran, Iran |
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Abstract: | Molecular phylogenetic analysis of a novel thermophilic eubacterium isolated from an Iranian hot spring using 16S rDNA sequence
showed that the new isolate belongs to genera Geobacillus. DNA pol I gene from this isolate was amplified, cloned, sequenced, and the three-dimensional (3D) structure of deduced amino
acid sequence was predicted. Sequence analysis revealed the gene is 2,631 bp long, encodes a protein of 876 amino acids with
a calculated molecular mass of 99 kDa, and belongs to family A DNA polymerases. Comparison of 3′–5′exonuclease domain of Klenow
fragment (KF) with corresponding region of newly identified DNA pol I (MF), the large fragment of Bacillus stearothermophilus DNA pol I (BF) and Klentaq1, revealed not only deletions in three regions compared to KF, but that three of the four critical
metal-binding residues in KF (Asp355, Glu357, Asp424, and Asp501) are altered in MF as well. Predicted 3D structure and sequence
alignments between MF and BF showed that all critical residues in the polymerase active site are conserved. |
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Keywords: | Molecular phylogenetic Thermostable DNA pol I Cloning 3D structure prediction Gsm pol I |
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