Explanation for the electrophoresis behaviour of DNA condensation induced by pseudopolyrotaxane of different lengths
Explanation for the electrophoresis behaviour of DNA condensation induced by pseudopolyrotaxane of different lengths
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DOI：
10.1002/elps.200800216
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发表时间：
2008-11
期刊：
影响因子：
2.9
通讯作者：
S. Hou;Kun Yang;Xizeng Feng
中科院分区：
文献类型：
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作者：
S. Hou;Kun Yang;Xizeng Feng
DNA缩合作为基因递送载体在活细胞中转运的先决条件受到了广泛关注。电泳分析是研究DNA缩合过程中DNA与基因载体相互作用的常用方法。为了探讨DNA的凝聚状态与电泳行为之间的关系，我们将6-[（6-氨基己基）氨基]-6-脱氧-β-CD二氯化物连接到聚丙二醇上，合成了一系列的假聚轮烷（PPR），并通过电泳分析研究了它们的DNA凝聚效率。在电泳分析中，中等长度的PPR表现出最好的DNA浓缩效率，而较短或较长的同源物仅表现出轻微的DNA浓缩效率。为了解释这一现象，我们研究了这些PPRs/DNA复合物的流体动力学直径，粒径，zeta电位和结合亲和力。我们发现PPRs/DNA复合物的稳定性在决定电泳行为中起重要作用。根据实验数据，我们还指出了电泳法在研究DNA凝聚过程中的不足。本文的研究结果对从事DNA缩合研究的人员有一定的参考价值。
DNA condensation has received much attention as a prerequisite for the transport of gene delivery vectors in living cells. Electrophoresis assay is a common method to investigate the interaction between DNA and gene delivery vectors during the DNA condensation process. In order to probe the relation between DNA condensation status and the corresponding electrophoresis behaviour, we synthesized a series of pseudopolyrotaxanes (PPRs) by threading 6‐[(6‐aminohexyl)amino]‐6‐deoxy‐β‐CD dichloride on the polypropylene glycol and investigated their DNA condensation efficiency by electrophoresis assay. The PPRs of the middle length exhibited the best DNA condensation efficiency in electrophoresis assay, while the shorter or the longer homologues exhibited only slight DNA condensation efficiency. To explain this phenomenon, we investigated the hydrodynamic diameter, the particle size, the zeta potential and the binding affinity of these PPRs/DNA complexes. We found that the stability of PPRs/DNA complexes played an important role in determining the electrophoresis behaviour. Based on the data, we also pointed out the deficiency of electrophoresis assay in investigating the DNA condensation process. Our findings are helpful for researchers engaged in DNA condensation study.