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Literature summary for 3.6.1.23 extracted from

  • Dabrowski, S.; Kiaer Ahring, B.
    Cloning, expression, and purification of the His6-tagged hyper-thermostable dUTPase from Pyrococcus woesei in Escherichia coli: application in PCR (2003), Protein Expr. Purif., 31, 72-78.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
the expression of N-terminal His(6)-tagged Pwo dUTPase is performed in Escherichia coli BL21(DE3)pLysS and Eschwerichia coli Rosetta(DE3)pLysS strain that contains plasmid encoding additional copies of rare Escherichia coli tRNAs. Escherichia coli Rosetta(pLysS) strain is found with two times higher expression yield of His(6)-tagged Pwo dUTPase than Escherichia coli BL21(DE3)pLysS. The His(6)-tagged Pwo dUTPase is purified on Ni(2+)-IDA-Sepharose, dialyzed, and the enzyme activity is investigated. THe His(6)-tag domain has no influence on dUTP hydrolytic activity. dUTP is generated during PCR from dCTP, which inhibits the polymerization of DNA catalyzed by DNA polymerase with 3'-5' exonuclease activity Pyrococcus woesei

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
22043
-
x * 22043, His6-tagged enzyme, calculated from sequence Pyrococcus woesei

Organism

Organism UniProt Comment Textmining
Pyrococcus woesei Q6T180
-
-
Pyrococcus woesei DSM 3773 Q6T180
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-

Purification (Commentary)

Purification (Comment) Organism
-
Pyrococcus woesei

Subunits

Subunits Comment Organism
? x * 22043, His6-tagged enzyme, calculated from sequence Pyrococcus woesei

Synonyms

Synonyms Comment Organism
dUTPase
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Pyrococcus woesei

pI Value

Organism Comment pI Value Maximum pI Value
Pyrococcus woesei calculated from sequence, pI: 9.07
-
9.1