A method to increase the bioactivity of plasmid DNA by heat treatment

Journal of biochemical and biophysical methods Pub Date : 2008-04-24 DOI:10.1016/j.jprot.2008.01.013

Sen Hou, Kun Yang, Ze Liu, Xi-Zeng Feng

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引用次数: 4

Abstract

A method to increase the bioactivity of plasmid DNA by heat treatment has been developed. The structure of the heat treated plasmid DNA was investigated by electrophoresis assay and atomic force microscope (AFM) observation. Electrophoresis assay showed that the heat treated DNA consisted of three components: the supercoiled DNA (component I), the open circular DNA (component II) and the heat denatured DNA component. The bioactivity of the heat treated plasmid DNA was investigated by both DNA condensation experiments and gene transfection experiment with mammal cells. DNA condensation experiments showed that the heat denatured DNA component owned higher sensitivity to spermidine and polyethylenimine (PEI) than component I and component II DNA. Gene transfection experiment with PEI indicated that the heat treated DNA had higher gene transfection efficiency than untreated DNA. Our experiment not only shows an effective approach to increase the bioactivity of plasmid DNA but also leads a way to improve the bioactivity of DNA by physically modifying their structure.

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一种通过热处理提高质粒DNA生物活性的方法

提出了一种通过热处理提高质粒DNA生物活性的方法。用电泳法和原子力显微镜观察热处理后质粒DNA的结构。电泳分析表明，热处理后的DNA由超螺旋DNA(组分I)、开环DNA(组分II)和热变性DNA(组分II)三部分组成。通过DNA浓缩实验和哺乳动物细胞基因转染实验，研究了热处理质粒DNA的生物活性。DNA缩合实验表明，热变性DNA组分对亚精胺和聚乙烯亚胺(PEI)的敏感性高于组分I和组分II DNA。PEI基因转染实验表明，热处理DNA的基因转染效率高于未处理DNA。本实验不仅为提高质粒DNA的生物活性提供了一条有效途径，而且为通过物理修饰DNA的结构来提高DNA的生物活性开辟了一条途径。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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Journal of biochemical and biophysical methods

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