乙醇对鱼精蛋白与DNA相互作用的影响
金仙君，林 莹，宓博凯, 王艳伟*乙醇对鱼精蛋白与DNA相互作用的影响
金仙君，林 莹，宓博凯, 王艳伟*
（温州大学物理与电子信息工程学院, 浙江 温州 325035）
摘 要 鱼精蛋白是近年发展起来的又一新型的天然高分子非病毒载体,可以作为一种良好的基因载体用作输运DNA，而它在不同体积分数乙醇溶液中也会影响其输运效果，作者利用鱼精蛋白这一性质，用单分子荧光电泳技术以及原子力显微镜探究它在不同体积分数的乙醇溶液下对DNA的凝聚效果以及DNA的电荷逆转规律的影响。单分子荧光电泳实验结果表明，DNA的电泳迁移率符号随鱼精蛋白浓度的增大由负变正，即DNA发生电荷逆转现象。溶液中乙醇的浓度越大，DNA的电泳迁移率绝对值越小。DNA的电荷逆转不仅取决于DNA的表面电荷量，可能还取决于DNA的凝聚情况，利用原子力显微镜可观察到DNA凝聚效果随着鱼精蛋白浓度的增加而增强。接着在鱼精蛋白(0.5ng·μL-1)溶液中，加入不同体积分数的乙醇，发现乙醇会促进鱼精蛋白对DNA的凝聚作用，且凝聚效果明显。两组实验结果均表明鱼精蛋白可凝聚DNA并能使DNA发生电荷逆转，且乙醇在鱼精蛋白导致的DNA凝聚与电荷逆转过程中起着一定的作用。
关键词 鱼精蛋白；乙醇；DNA凝聚；电荷逆转；单分子技术
中图分类号：Q6-33 文献标识码：A doi:10.3969/j.issn.1000-6281.2016.06.008
The effect of ethanol on the interaction between protamine and DNA
JIN Xian-jun, LIN-Ying, MI Bo-kai, WANG Yan-wei*
（School of Physics and Electronic Information, Wenzhou University, Wenzhou Zhejiang 325035,China）
Abstract Protamine is used as a new kind of natural non viral vector and a good gene carrier in recent years. It can transport DNA. The transport effect can be affected by the volume fraction of ethanol on solution. We use this characteristic of the protamine to explore DNA condensation and DNA charge reversal rules in different volume fraction of ethanol by using single molecule fluorescence electrophoresis and atomic force microscope technique. The results of single molecule fluorescence electrophoresis experiment show that the symbols of DNA electrophoresis mobility changes from negative to positive with the increasing protamine concentration. The absolute value of electrophoresis mobility is decreased with the increasing volume fraction of ethanol . The charge reversal of DNA depends not only on the surface charge of DNA, may also the morphologies of the condensation of the DNA. We can observe the image of DNA in different concentrations of protamine by atomic force microscope. DNA condensation strengthened with the increasing concentration of protamine. And then we add different volume fraction of ethanol to protamine (0.5ng·μL-1) solution and find that ethanol can promote the interaction between DNA and protamine. Two groups of the experimental results show that protamine condenses DNA and make the DNA occur charge reversal. The ethanol plays a certain role on DNA condensation and charge reversal.
Keywords protamine；ethanol；DNA condensation；charge reversal；single molecule operation
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