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Literature summary for 2.7.7.7 extracted from

  • Schoenbrunner, N.J.; Fiss, E.H.; Budker, O.; Stoffel, S.; Sigua, C.L.; Gelfand, D.H.; Myers, T.W.
    Chimeric thermostable DNA polymerases with reverse transcriptase and attenuated 3-5 exonuclease activity (2006), Biochemistry, 45, 12786-12795.
    View publication on PubMed
Search for more literature for 2.7.7.7

Application

Application Comment Organism
diagnostics rational design approach to creating modulated proofreading DNA polymerases which can be utilized in a highly sensitive long RT/PCR amenable to the clinical diagnostic setting Thermus sp.
diagnostics rational design approach to creating modulated proofreading DNA polymerases which can be utilized in a highly sensitive long RT/PCR amenable to the clinical diagnostic setting Thermotoga maritima

Protein Variants

Protein Variants Comment Organism
additional information construction of a chimeric DNA polymerase derived from Thermus species Z05 and Thermotoga maritima DNA polymerases. These chimeric DNA polymerases are fashioned using structure-based tools to identify amino acid residues involved in the substrate-binding site of the exonuclease domain of a thermostable DNA polymerase. Mutation of some of these residues results in proteins in which DNA polymerase activity is unaffected, while proofreading activity ranges from 60% of the wild-type level to undetectable levels. Kinetic characterization of the exonuclease activity indicates that the mutations affects catalysis much more than binding. On the basis of their specificity constants (kcat/KM), the mutant enzymes have a 5-15-fold stronger preference for a double-stranded mismatched substrate over a single-stranded substrate than the wild-type DNA polymerase, a desirable attribute for RT/PCR Thermus sp.
additional information construction of a chimeric DNA polymerase derived from Thermus species Z05 and Thermotoga maritima DNA polymerases. These chimeric DNA polymerases are fashioned using structure-based tools to identify amino acid residues involved in the substrate-binding site of the exonuclease domain of a thermostable DNA polymerase. Mutation of some of these residues results in proteins in which DNA polymerase activity is unaffected, while proofreading activity ranges from 60% of the wild-type level to undetectable levels. Kinetic characterization of the exonuclease activity indicates that the mutations affects catalysis much more than binding. On the basis of their specificity constants (kcat/KM), the mutant enzymes have a 5-15-fold stronger preference for a double-stranded mismatched substrate over a single-stranded substrate than the wild-type DNA polymerase, a desirable attribute for RT/PCR Thermotoga maritima

Organism

Organism UniProt Comment Textmining
Thermotoga maritima
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Thermus sp.
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Thermus sp. Z05
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