插入T3 DNA聚合酶硫氧还蛋白结合结构域可增强Taq DNA聚合酶的持续合成能力和保真度。
Insertion of the T3 DNA polymerase thioredoxin binding domain enhances the processivity and fidelity of Taq DNA polymerase.
作者信息
Davidson John F, Fox Richard, Harris Dawn D, Lyons-Abbott Sally, Loeb Lawrence A
机构信息
Department of Pathology, University of Washington, Seattle, WA 98195, USA.
出版信息
Nucleic Acids Res. 2003 Aug 15;31(16):4702-9. doi: 10.1093/nar/gkg667.
Abstract
Insertion of the T3 DNA polymerase thioredoxin binding domain (TBD) into the distantly related thermostable Taq DNA polymerase at an analogous position in the thumb domain, converts the Taq DNA polymerase from a low processive to a highly processive enzyme. Processivity is dependent on the presence of thioredoxin. The enhancement in processivity is 20-50-fold when compared with the wild-type Taq DNA polymerase or to the recombinant polymerase in the absence of thioredoxin. The recombinant Taq DNA pol/TBD is thermostable, PCR competent and able to copy repetitive deoxynucleotide sequences six to seven times more faithfully than Taq DNA polymerase and makes 2-3-fold fewer AT-->GC transition mutations.
摘要
将T3 DNA聚合酶硫氧还蛋白结合结构域（TBD）插入到亲缘关系较远的耐热Taq DNA聚合酶拇指结构域的类似位置，可将Taq DNA聚合酶从低持续合成能力的酶转变为高持续合成能力的酶。持续合成能力取决于硫氧还蛋白的存在。与野生型Taq DNA聚合酶或不存在硫氧还蛋白时的重组聚合酶相比，持续合成能力提高了20至50倍。重组Taq DNA pol/TBD具有耐热性，具备PCR活性，并且能够比Taq DNA聚合酶更忠实地复制重复脱氧核苷酸序列六到七次，产生的AT→GC转换突变减少2至3倍。
相似文献
1
Insertion of the T3 DNA polymerase thioredoxin binding domain enhances the processivity and fidelity of Taq DNA polymerase.插入T3 DNA聚合酶硫氧还蛋白结合结构域可增强Taq DNA聚合酶的持续合成能力和保真度。
Nucleic Acids Res. 2003 Aug 15;31(16):4702-9. doi: 10.1093/nar/gkg667.
2
A novel strategy to engineer DNA polymerases for enhanced processivity and improved performance in vitro.一种改造DNA聚合酶以提高持续合成能力并改善体外性能的新策略。
Nucleic Acids Res. 2004 Feb 18;32(3):1197-207. doi: 10.1093/nar/gkh271. Print 2004.
3
The thioredoxin binding domain of bacteriophage T7 DNA polymerase confers processivity on Escherichia coli DNA polymerase I.噬菌体T7 DNA聚合酶的硫氧还蛋白结合结构域赋予大肠杆菌DNA聚合酶I持续合成能力。
Proc Natl Acad Sci U S A. 1997 Jan 21;94(2):479-84. doi: 10.1073/pnas.94.2.479.
4
Error-prone replication of repeated DNA sequences by T7 DNA polymerase in the absence of its processivity subunit.在缺乏其持续性亚基的情况下，T7 DNA 聚合酶对重复 DNA 序列进行易错复制。
Proc Natl Acad Sci U S A. 1994 Jul 19;91(15):6830-4. doi: 10.1073/pnas.91.15.6830.
5
Interactions of Escherichia coli thioredoxin, the processivity factor, with bacteriophage T7 DNA polymerase and helicase.大肠杆菌硫氧还蛋白（一种持续性因子）与噬菌体T7 DNA聚合酶及解旋酶的相互作用。
J Biol Chem. 2008 Nov 14;283(46):32077-84. doi: 10.1074/jbc.M805062200. Epub 2008 Aug 30.
6
Domain exchange: chimeras of Thermus aquaticus DNA polymerase, Escherichia coli DNA polymerase I and Thermotoga neapolitana DNA polymerase.结构域交换：嗜热栖热菌DNA聚合酶、大肠杆菌DNA聚合酶I和那不勒斯嗜热栖热菌DNA聚合酶的嵌合体
Protein Eng. 2000 Sep;13(9):645-54. doi: 10.1093/protein/13.9.645.
7
A single highly mutable catalytic site amino acid is critical for DNA polymerase fidelity.单个高度可变的催化位点氨基酸对DNA聚合酶保真度至关重要。
J Biol Chem. 2001 Feb 16;276(7):5044-51. doi: 10.1074/jbc.M008701200. Epub 2000 Nov 7.
8
Inactivation of the 5'-3' exonuclease of Thermus aquaticus DNA polymerase.嗜热水生栖热菌DNA聚合酶5'-3'核酸外切酶的失活
Biochim Biophys Acta. 1995 Nov 7;1264(2):243-8. doi: 10.1016/0167-4781(95)00153-8.
9
Helix-hairpin-helix motifs confer salt resistance and processivity on chimeric DNA polymerases.螺旋-发夹-螺旋基序赋予嵌合DNA聚合酶耐盐性和持续合成能力。
Proc Natl Acad Sci U S A. 2002 Oct 15;99(21):13510-5. doi: 10.1073/pnas.202127199. Epub 2002 Oct 4.
10
Thioredoxin suppresses microscopic hopping of T7 DNA polymerase on duplex DNA.硫氧还蛋白抑制 T7 DNA 聚合酶在双链 DNA 上的微观跳跃。
Proc Natl Acad Sci U S A. 2010 Feb 2;107(5):1900-5. doi: 10.1073/pnas.0912664107. Epub 2010 Jan 11.
引用本文的文献
1
Strategies and procedures to generate chimeric DNA polymerases for improved applications.用于改进应用的嵌合 DNA 聚合酶的生成策略和程序。
Appl Microbiol Biotechnol. 2024 Aug 21;108(1):445. doi: 10.1007/s00253-024-13276-2.
2
DNA Polymerases for Whole Genome Amplification: Considerations and Future Directions.全基因组扩增用 DNA 聚合酶：考虑因素和未来方向。
Int J Mol Sci. 2023 May 26;24(11):9331. doi: 10.3390/ijms24119331.
3
Challenging the proposed causes of the PCR plateau phase.对聚合酶链式反应（PCR）平台期提出的原因提出质疑。
Biomol Detect Quantif. 2019 Mar 2;17:100082. doi: 10.1016/j.bdq.2019.100082. eCollection 2019 Mar.
4
Two Approaches to Enhance the Processivity and Salt Tolerance of Staphylococcus aureus DNA Polymerase.两种提高金黄色葡萄球菌 DNA 聚合酶的持续性和耐盐性的方法。
Protein J. 2019 Apr;38(2):190-198. doi: 10.1007/s10930-019-09818-7.
5
Identification of Thermus aquaticus DNA polymerase variants with increased mismatch discrimination and reverse transcriptase activity from a smart enzyme mutant library.从智能酶突变文库中鉴定具有提高错配识别和逆转录酶活性的嗜热栖热菌 DNA 聚合酶变体。
Sci Rep. 2019 Jan 24;9(1):590. doi: 10.1038/s41598-018-37233-y.
6
Insights into the structural dynamics of the bacteriophage T7 DNA polymerase and its complexes.对噬菌体T7 DNA聚合酶及其复合物结构动力学的见解。
J Mol Model. 2018 Jun 1;24(7):144. doi: 10.1007/s00894-018-3671-2.
7
Localised genetic heterogeneity provides a novel mode of evolution in dsDNA phages.局部遗传异质性为双链 DNA 噬菌体的进化提供了一种新的模式。
Sci Rep. 2017 Oct 23;7(1):13731. doi: 10.1038/s41598-017-14285-0.
8
Derivatives of Bst-like Gss-polymerase with improved processivity and inhibitor tolerance.具有更高持续合成能力和抑制剂耐受性的类Bst Gss聚合酶衍生物。
Nucleic Acids Res. 2017 Sep 19;45(16):9595-9610. doi: 10.1093/nar/gkx645.
9
Engineering human PrimPol into an efficient RNA-dependent-DNA primase/polymerase.将人类 PrimPol 工程改造为一种高效的 RNA 依赖性 DNA 引发酶/聚合酶。
Nucleic Acids Res. 2017 Sep 6;45(15):9046-9058. doi: 10.1093/nar/gkx633.
10
Speeding up biomolecular interactions by molecular sledding.通过分子滑行加速生物分子相互作用。
Chem Sci. 2016 Feb 1;7(2):916-920. doi: 10.1039/C5SC03063C. Epub 2015 Oct 7.
本文引用的文献
1
DNA mismatch repair, microsatellite instability and cancer.DNA错配修复、微卫星不稳定性与癌症
Indian J Exp Biol. 2002 Sep;40(9):989-94.
2
Taq DNA polymerase slippage mutation rates measured by PCR and quasi-likelihood analysis: (CA/GT)n and (A/T)n microsatellites.通过聚合酶链式反应（PCR）和拟似然分析测定的Taq DNA聚合酶滑动突变率：（CA/GT）n和（A/T）n微卫星
Nucleic Acids Res. 2003 Feb 1;31(3):974-80. doi: 10.1093/nar/gkg178.
3
Complete nucleotide sequence and likely recombinatorial origin of bacteriophage T3.噬菌体T3的完整核苷酸序列及可能的重组起源
J Mol Biol. 2002 Jun 21;319(5):1115-32. doi: 10.1016/S0022-2836(02)00384-4.
4
Microsatellite instability in cancer: what problems remain unanswered?癌症中的微卫星不稳定性：哪些问题仍未得到解答？
Surgery. 2002 Jan;131(1 Suppl):S55-62. doi: 10.1067/msy.2002.119305.
5
The fidelity of human DNA polymerase gamma with and without exonucleolytic proofreading and the p55 accessory subunit.具有和不具有核酸外切校正功能的人类DNA聚合酶γ以及p55辅助亚基的保真度
J Biol Chem. 2001 Oct 19;276(42):38555-62. doi: 10.1074/jbc.M105230200. Epub 2001 Aug 14.
6
Prokaryotic DNA polymerase I: evolution, structure, and "base flipping" mechanism for nucleotide selection.原核生物DNA聚合酶I：进化、结构及核苷酸选择的“碱基翻转”机制
J Mol Biol. 2001 May 18;308(5):823-37. doi: 10.1006/jmbi.2001.4619.
7
Induction of microsatellite instability by oxidative DNA damage.氧化性DNA损伤诱导微卫星不稳定性
Proc Natl Acad Sci U S A. 1998 Oct 13;95(21):12468-73. doi: 10.1073/pnas.95.21.12468.
8
Genetic control of microsatellite stability.微卫星稳定性的遗传控制
Mutat Res. 1997 Jan 31;383(1):61-70. doi: 10.1016/s0921-8777(96)00046-8.
9
The thioredoxin binding domain of bacteriophage T7 DNA polymerase confers processivity on Escherichia coli DNA polymerase I.噬菌体T7 DNA聚合酶的硫氧还蛋白结合结构域赋予大肠杆菌DNA聚合酶I持续合成能力。
Proc Natl Acad Sci U S A. 1997 Jan 21;94(2):479-84. doi: 10.1073/pnas.94.2.479.
10
Cancer of the microsatellite mutator phenotype.微卫星突变体表型癌症
Biol Chem. 1996 Nov;377(11):675-84.
Zotero 插件