Abstract：
:Two DNA polymerase genes have been isolated from Thermococcus strains, Thermococcus zilligii from New Zealand, and the other, Thermococcus 'GT', a fast-growing strain isolated from the Galapagos trench. Both genes were isolated by genomic walking PCR, a technique that does not require expression of the gene product. Phylogenetic analysis of SSU rDNA showed that the two strains were not closely related, as confirmed by an examination of the DNA polymerase sequences. Inteinless versions of each gene were generated by overlap-extension PCR and transferred into plasmid expression vectors. The proteins were produced in an Escherichia coli strain with additional copies of tRNAs corresponding to rarely used codons and purified by standard chromatographic procedures. Both enzymes were able to support PCR, but the Thermococcus 'GT' polymerase required higher concentrations of template than the enzyme from T. zilligii. Both enzymes showed 3' to 5' exonuclease activity, which was abolished in the case of T. zilligii by mutating the aspartic acid at position 141 and the glutamic acid at position 143 to alanine. Both enzymes showed a significant increase in fidelity of replication compared to the family A Thermus aquaticus DNA polymerase, in agreement with other results reported for family B polymerases with proof-reading ability.
journal_name
Protein Expr Purifjournal_title
Protein expression and purificationauthors
Griffiths K,Nayak S,Park K,Mandelman D,Modrell B,Lee J,Ng B,Gibbs MD,Bergquist PLdoi
10.1016/j.pep.2006.07.022subject
Has Abstractpub_date
2007-03-01 00:00:00pages
19-30issue
1eissn
1046-5928issn
1096-0279pii
S1046-5928(06)00248-8journal_volume
52pub_type
杂志文章abstract：:Human D-amino acid oxidase (hDAAO) is a flavoprotein that plays a key role in the pathophysiology of schizophrenia. So far, the biochemical characterization of this enzyme has been hampered by the difficulty of expressing it in a common heterologous host such as Escherichia coli. Increasing amounts of recombinant hDAA...
journal_title：Protein expression and purification
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journal_title：Protein expression and purification
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journal_title：Protein expression and purification
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journal_title：Protein expression and purification
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journal_title：Protein expression and purification
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journal_title：Protein expression and purification
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journal_title：Protein expression and purification
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journal_title：Protein expression and purification
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journal_title：Protein expression and purification
pub_type： 杂志文章
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journal_title：Protein expression and purification
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journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1006/prep.1996.0047
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journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2005.11.027
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journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2014.02.007
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journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2004.06.018
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journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2006.02.019
更新日期：2006-09-01 00:00:00
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journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2018.02.005
更新日期：2018-07-01 00:00:00
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journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2019.105463
更新日期：2019-12-01 00:00:00
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journal_title：Protein expression and purification
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journal_title：Protein expression and purification
pub_type： 杂志文章
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更新日期：2012-01-01 00:00:00
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journal_title：Protein expression and purification
pub_type： 杂志文章
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journal_title：Protein expression and purification
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doi：10.1006/prep.1998.0929
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journal_title：Protein expression and purification
pub_type： 杂志文章
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journal_title：Protein expression and purification
pub_type： 杂志文章
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journal_title：Protein expression and purification
pub_type： 杂志文章
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更新日期：2009-08-01 00:00:00
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journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1006/prep.1999.1120
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journal_title：Protein expression and purification
pub_type： 杂志文章
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journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2004.11.012
更新日期：2005-04-01 00:00:00
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journal_title：Protein expression and purification
pub_type： 杂志文章
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更新日期：2010-10-01 00:00:00
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journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1006/prep.1999.1186
更新日期：2000-04-01 00:00:00
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journal_title：Protein expression and purification
pub_type： 杂志文章
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更新日期：2004-01-01 00:00:00
