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首页> 外文期刊>Separation and Purification Technology >Partition of plasmid DNA in polymer-salt aqueous two-phase systems
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Partition of plasmid DNA in polymer-salt aqueous two-phase systems

机译：质粒DNA在聚合物盐水溶液两相系统中的分配

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摘要

The partition of plasmid DNA (pDNA) in polyethylene glycol (PEG)-phosphate aqueous two-phase systems (ATPS) is presented. A high molecular weight (HMW) and a low molecular weight (LMW) polymer, PEG-1450 and -300, were used in combination with di-potassium hydrogen phosphate. The experimental results demonstrated that the plasmid pTX0161 displays a varied partition behaviour in PEG-phosphate ATPS. In HMW PEG (PEG-1450-phosphate systems), pDNA partitioned to the bottom phase only. In LMW PEG (PEG-300-phosphate systems), pDNA partitioned to all of the phases with respect to the phase composition, system temperature and concentration of lysate used in the ATPS. In systems with volume ratios higher than one, pDNA was mainly recovered in the top phase. For volume ratios between 0.5 and 1, pDNA mainly partitioned to the interface. In systems with volume ratios below 0.5, most of the pDNA was recovered in the bottom phase. For temperatures between 4 and 25 °C, the partition to the top phase decreased whereas partition to the interface steadily increased. At 25 °C, over 80% of pDNA was recovered in the interface. The partition to the bottom phase increased steadily with increasing temperatures up to 40 °C and the partition to the interface decreased. At 20 °C, the recovery of pDNA in the interface gradually increased and reached a maximum at 60% (w/w) lysate with 80% recovery recorded. At 25 °C, over 80% of pDNA was recovered in the interface from lysate concentrations greater than 35% (w/w). At 30 °C, the top phase preference changed to an interface preference between 0 and 20% (w/w) lysate.
机译：介绍了质粒DNA（pDNA）在聚乙二醇（PEG）-磷酸盐水溶液两相系统（ATPS）中的分配。高分子量（HMW）和低分子量（LMW）聚合物PEG-1450和-300与磷酸氢二钾结合使用。实验结果表明，质粒pTX0161在PEG-磷酸盐ATPS中显示出不同的分配行为。在HMW PEG（PEG-1450-磷酸盐系统）中，pDNA仅分配至底部相。在LMW PEG（PEG-300-磷酸盐系统）中，pDNA相对于ATPS中使用的相组成，系统温度和裂解物浓度而言，已划分为所有相。在体积比大于1的系统中，pDNA主要在顶相中回收。对于介于0.5和1之间的体积比，pDNA主要分配到界面上。在体积比低于0.5的系统中，大多数pDNA在底部相中回收。对于4至25°C的温度，至顶层的分配减少，而与界面的分配则稳定增长。在25°C下，界面中回收了超过80％的pDNA。随着温度升高至40°C，对底部相的分配稳定增加，对界面的分配减少。在20°C时，界面中pDNA的回收率逐渐增加，在60％（w / w）裂解液中达到最大值，记录到80％的回收率。在25°C下，从裂解液浓度大于35％（w / w）的界面中回收了超过80％的pDNA。在30°C时，上层优先级更改为0至20％（w / w）裂解物之间的界面优先级。

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