基于色谱法的超螺旋质粒DNA纯化与分析进展

Advances on Purification Process and Analysis of Supercoiled Plasmid DNA Based on Chromatography

质粒DNA含有独立复制的遗传结构,是基因工程的常用工具,广泛应用于分子生物学基础研究、农业转基因检测、医疗诊断与基因治疗等领域。质粒DNA的构型一般分为超螺旋、开口环状及线性3种。初步纯化的质粒DNA溶液中常混有3种构型,并且掺杂一定量的蛋白质、RNA、内毒素以及宿主基因组DNA,这些杂质会影响后续的应用,因此质粒DNA需要进一步精细纯化。该文对质粒DNA的新应用领域、基于色谱的精细纯化技术及产物质量分析体系进行了综述,并展望了高纯度质粒DNA精细纯化及产物分析的发展方向。

Plasmid DNA, a common tool of genetic engineering containing the independent genetic structure of the replication, is widely used in basic research of molecular biology, genetically modified detection, medical diagnosis and gene therapy. Plasmid DNA conformations are generally divided into three types： supercoiled, open circle and linear. Crude plasmid DNA often mixes with three configurations, and the amount of protein, RNA, purities need to be further refined as they affect endotoxin with the host genome DNA. These irasubsequent applications. The new applications of plasmid DNA, refined technologies and products based on chromatography system, and quality analysis of purified plasmid DNA is reviewed in this paper. Finally, the trends of technology of high purity plasmid DNA and analysis of refined product are presented.