二氧化氯对脱氧核糖核苷三磷酸和质粒DNA的作用

Effects of Chlorine Dioxide on Deoxyribonucleoside Triphosphate and Plasmid

以75mmol/L的二氧化氯作用于脱氧核糖核苷三磷酸dCTP、dTTP、dGTP、dATP混合物(浓度均为100μmol/L),液相色谱显示它们在260nm波长处的峰面积分别下降了54.23%±2.08%、66.25%±3.32%、55.47%±0.23%、59.59%±3.27%。纯化质粒被二氧化氯作用后电泳条带未出现明显弥散和拖尾现象。45mmol/L以上二氧化氯能彻底抑制质粒DNA的PCR模板活性,150mmol/L的二氧化氯才能使纯化质粒的转化率降至对照组的0.20%±0.20%。实验证明二氧化氯对DNA具有损伤作用,这种损伤可能与嘧啶碱和嘌呤碱的共轭双键被破坏有关。

The mixture of deoxyribonucleoside, dCTP, dTTE dGTP and dATP at concentration of 100 μmol/L for each, was treated by 75 mmol/L of chlorine dioxide for 10 min, high performance liquid chromatographic analyses indicated that all of the four decreased in peak area by 54.23%±2.08%, 66.25%±3.32%, 55.47%±0.23%, 59.59%±3.27% respectively at wavelength of 260 nm. For 0.2μg/μL of purified plasmid DNA, their activity as template in polymerase chain reaction were completely inhibited by chlorine dioxide at concentration of 45 mmol/L or above, and the transformation rate decreased to 0.20%±0.2% when treated by 75 mmol/L of chlorine dioxide. Our result indicated that chlorine dioxide have substantial damages on deoxyribonucleoside and plasmid DNA.