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Literature summary for 2.7.7.7 extracted from

  • Nishioka, M.; Mizuguchi, H.; Fujiwara, S.; Komatsubara, S.; Kitabayashi, M.; Uemura, H.; Takagi, M.; Imanaka, T.
    Long and accurate PCR with a mixture of KOD DNA polymerase and its exonuclease deficient mutant enzyme (2001), J. Biotechnol., 88, 141-149.
    View publication on PubMed

Application

Application Comment Organism
molecular biology long and accurate PCR can be achieved with a mixture of wild type DNA polymerase from Thermococcus kodakaraensis and its exonuclease deficient mutant enzyme N210D is utilized (at the ratio of 1:40) Thermococcus kodakarensis

Cloned(Commentary)

Cloned (Comment) Organism
-
Thermococcus kodakarensis

Protein Variants

Protein Variants Comment Organism
N210D the mutant enzyme shows very weak 3'-5'-exonuclease activity compared to the wild-type enzyme (0.1%). No significant difference in DNA polymerase activity as compared with that of the wild-type enzyme. Mutation frequency in PCR becomes higher as 3'–5' exonuclease activity decreases Thermococcus kodakarensis
Y311F the mutant enzyme shows very weak 3'-5'-exonuclease activity compared to the wild-type enzyme (0.01%). No significant difference in DNA polymerase activity as compared with that of the wild-type enzyme Thermococcus kodakarensis

Organism

Organism UniProt Comment Textmining
Thermococcus kodakarensis P77933
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-

Purification (Commentary)

Purification (Comment) Organism
-
Thermococcus kodakarensis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
deoxynucleoside triphosphate + DNAn the DNA polymerase also shows 3'-5' exonuclease activity. The 3'–5' exonuclease activity of the DNA polymerase is important for DNA elongation with high fidelity Thermococcus kodakarensis diphosphate + DNAn+1
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Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
75
-
3'-5' exonuclease assay at Thermococcus kodakarensis
75
-
DNA polymerase assay at Thermococcus kodakarensis

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
DNA polymerase assay at Thermococcus kodakarensis
8
-
3'-5' exonuclease assay at Thermococcus kodakarensis