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Inhibition of Taq DNA polymerase and DNA exonuclease Exo III by an aqueous nanoparticle suspension of a bis-methanophosphonate fullerene

机译:双甲基膦酸酯富勒烯的水性纳米颗粒悬浮液对Taq DNA聚合酶和DNA核酸外切酶Exo III的抑制

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An aqueous nanoparticle suspension of a bis-methanophosphonate fullerene (n-BMPF) was tested to clarify its effects on polymerase chain reaction (PCR) with the catalyst of Taq DNA polymerase and DNA exonuclease Exo Ⅲ and the template of super-coiled plasmid pEGFP-N1. It was found that the product amounts from PCR decreased significantly with addition of the n-BMPF. The inhibition by the n-BMPF was dose-dependent and IC50 values for reactions of PCR were 2.7 μmol/L. Increase of Taq DNA polymerase amounts in PCR system antagonized the activities of the n-BMPF. However, addition of two scavengers of reactive oxygen species (ROS), mannitol and azide at the concentrations of 2~10 mmol/L did not antagonize the activities of the n-BMPF against PCR. These data implied that this inhibition probably did not correlate to ROS. Meanwhile, the inhibition for the DNA exonuclease Exo Ⅲ by the n-BMPF was evident and dose-dependent.
机译:用Taq DNA聚合酶和DNA核酸外切酶ExoⅢ的催化剂以及超螺旋质粒pEGFP- N1发现通过添加n-BMPF,来自PCR的产物量显着降低。 n-BMPF的抑制作用是剂量依赖性的,PCR反应的IC50值为2.7μmol/ L。 PCR系统中Taq DNA聚合酶数量的增加拮抗了n-BMPF的活性。但是,添加2〜10 mmol / L的两种活性氧清除剂(ROS),甘露醇和叠氮化物并不能拮抗n-BMPF对抗PCR的活性。这些数据暗示该抑制作用可能与ROS无关。同时,n-BMPF对DNA核酸外切酶ExoⅢ的抑制作用明显且呈剂量依赖性。

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