Characterization of family IV UDG from Aeropyrum pernix and its application in hot-start PCR by family B DNA polymerase.
Characterization of family IV UDG from Aeropyrum pernix and its application in hot-start PCR by family B DNA polymerase.
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产自 Aeropyrum pernix 的 IV 族 UDG 的表征及其在 B 族 DNA 聚合酶热启动 PCR 中的应用
DOI：
10.1371/journal.pone.0027248
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发表时间：
2011
期刊：
影响因子：
3.7
通讯作者：
Liu JH
中科院分区：
文献类型：
--
作者：
Liu XP;Liu JH
Recombinant uracil-DNA glycosylase (UDG) from Aeropyrum pernix (A. pernix) was expressed in E. coli. The biochemical characteristics of A. pernix UDG (ApeUDG) were studied using oligonucleotides carrying a deoxyuracil (dU) base. The optimal temperature range and pH value for dU removal by ApeUDG were 55–65°C and pH 9.0, respectively. The removal of dU was inhibited by the divalent ions of Zn, Cu, Co, Ni, and Mn, as well as a high concentration of NaCl. The opposite base in the complementary strand affected the dU removal by ApeUDG as follows: U/C≈U/G>U/T≈U/AP≈U/->U/U≈U/I>U/A. The phosphorothioate around dU strongly inhibited dU removal by ApeUDG. Based on the above biochemical characteristics and the conservation of amino acid residues, ApeUDG was determined to belong to the IV UDG family. ApeUDG increased the yield of PCR by Pfu DNA polymerase via the removal of dU in amplified DNA. Using the dU-carrying oligonucleotide as an inhibitor and ApeUDG as an activator of Pfu DNA polymerase, the yield of undesired DNA fragments, such as primer-dimer, was significantly decreased, and the yield of the PCR target fragment was increased. This strategy, which aims to amplify the target gene with high specificity and yield, can be applied to all family B DNA polymerases.
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Aeropyrum pernix 胸腺嘧啶 DNA 糖基化酶的表达和表征。
影响因子：
1.6
作者：
Liu, Xi-Peng;Li, Chun-Peng;Liu, Jian-Hua
通讯作者：
Liu, Jian-Hua
影响因子：
3.3
作者：
Barnes, WM;Rowlyk, KR
通讯作者：
Rowlyk, KR
DOI：
10.1093/dnares/6.2.83
发表时间：
1999-04-30
期刊：
DNA research : an international journal for rapid publication of reports on genes and genomes
影响因子：
--
作者：
Kawarabayasi, Y;Hino, Y;Kikuchi, H
通讯作者：
Kikuchi, H
影响因子：
4.8
作者：
Björnberg, O;Neuhard, J;Nyman, PO
通讯作者：
Nyman, PO
影响因子：
3.5
作者：
LUNDBERG, KS;SHOEMAKER, DD;MATHUR, EJ
通讯作者：
MATHUR, EJ