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YANG XinLin CHEN Zhe MENG XianMei LI Bo TAN Xin 《科学通报(英文版)》2007,52(13):1802-1806
Activities of trimalonic acid fullerene (TMA C_60) on DNA restrictive enzymatic reaction were investigated by using two restrictive endonucleases Hind III and EcoR I and plasmid pEGFP-N1 with single restric-tive site for both enzymes. Meanwhile, TMA C60 was also tested to clarify its effects on polymerase chain reaction (PCR) with the catalyst of Taq DNA polymerase and the template of plasmid pEGFP-N1. The products from restrictive reactions or PCR were detected by agarose gel electrophoresis. It was found that the product amounts from restrictive reactions or PCR decreased significantly with addition of TMA C60. The inhibition by TMA C60 was dose-dependent and IC50 values for reactions of Hind III, EcoR I and PCR were 16.3, 6.0 and 6.0 μmol/L, respectively. Addition of two scavengers of reactive oxygen species (ROS), L-ascorbic acid-2-phosphate ester magnesium and sodium azide at the con-centrations of 2―10 mmol/L did not antagonize the activities of TMA C60 against PCR and two restrictive reactions. However, increase of Taq DNA polymerase amounts in PCR system antagonized the activities of TMA C60. These data implied that TMA C60 was able to inhibit the activities of the three above-mentioned enzymes involved in DNA metabolism, and that this inhibition probably did not correlate to ROS. 相似文献
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在PCR扩增水平上研究了La^ 3和Ce^ 3对Taq^TMDNA聚合酶的抑制机理,采用动态荧光比色法测定PCR扩增产物浓度,证明了La^ 3和Ce^ 3在10~50μmol/L抑制DNA复制是由于其抑制了Taq^TMDNA聚合酶的活性所致,且该抑制属竞争性的抑制,得到了La^ 3和Ce^ 3对Taq^TMDNA聚合酶的抑制常数分别为12.7μmol/L和14.4μmol/L。 相似文献
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