Abstract:
:DNA polymerase from the archaeon Pyrococcus abyssi strain Orsay was expressed in Escherichia coli. The recombinant DNA polymerase (Pab) was purified to homogeneity by heat treatment followed by 5 steps of chromatography and characterized for PCR applications. Buffer optimization experiments indicated that Pab PCR performance and fidelity parameters were highest in the presence of 20 mM Tris-HCl, pH 9.0, 1.5 mM MgSO4, 25 mM KCl, 10 mM (NH4)2SO4 and 40 microM of each dNTP. Under these conditions, the error rate was 0.66.10(-6) mutations/nucleotide/duplication. Pab DNA polymerase, having a half life of 5 h at 100 degrees C, was demonstrated to be highly thermostable in PCR conditions compared to commercial Taq and Pfu DNA polymerases. These characteristics enable Pab to be one of the most efficient thermostable DNA polymerases described, exhibiting very high accuracy compared to other available commercial DNA polymerases and robust thermostable activity. This new DNA polymerase is currently on the market under the name Isis DNA Polymerase (Qbiogene Molecular Biology).
journal_name
FEMS Microbiol Lettjournal_title
FEMS microbiology lettersauthors
Dietrich J,Schmitt P,Zieger M,Preve B,Rolland JL,Chaabihi H,Gueguen Ydoi
10.1111/j.1574-6968.2002.tb11460.xkeywords:
subject
Has Abstractpub_date
2002-11-19 00:00:00pages
89-94issue
1eissn
0378-1097issn
1574-6968pii
S0378109702010376journal_volume
217pub_type
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journal_title:FEMS microbiology letters
pub_type: 杂志文章,评审
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