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KMID : 0578319970070060769
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Molecules and Cells
1997 Volume.7 No. 6 p.769 ~ p.776
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Cloning and Analysis of the DNA Polymerase-encoding Gene from Thermus filiformis
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Jung, Seung Eun
Choi, Jeong Jin/Kim, Hyun Kyu/Kwon, Suk-Tae
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Abstract
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The gene encoding Thermus filformis (Tfi)DNA polymerase was cloned and it nucleotide sequence was determined. The primary structure of Tif DNA polymerase was deduced from its nucleotide sequence. Tfi DNA polymerase comprised of 833 amino acid residues and its molecular mass was determined to be 93,890 Da. The deduced amino acid sequence of Tfi DNA polymerase showed a hogh sequence homology to E. col DNA polymerase I-like DNA polymerases: 78.5% homology to Taq DNA polymerase, 78.4% to Tca DNA polymerase, and 41.8% to E. coli DNA polymerase I. An extremely high sequence identity wa observed in the region containing polymerase activity. The G＋C content of the coding region for the Tfi DNA polymerase gene was 68.5%, which was higher than that of the chromosomal DNA (65%). The G＋C content in the first, second, and third positions of the codons used were 71.8%, 40.9%, and 92.7% respectively. Codon usage in Tfi DNA polymerase was heavily biased towards the use G＋C om the third position. Rare codons with U or A as the third base were sometimes used to avoid using GA(A/T) TC and TCGA sequences, as they are recognition sites for the restriction endonucleases TfiI and TaqI.
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