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首页> 外文期刊>Biotechnology Progress >Plasmid DNA Purification by Selective Calcium Silicate Adsorption of Closely Related Impurities
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Plasmid DNA Purification by Selective Calcium Silicate Adsorption of Closely Related Impurities

机译：选择性硅酸钙吸附紧密相关杂质的质粒DNA纯化

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摘要

The selective adsorption of supercoiled plasmid,open-circular plasmid,and genomic DNA to gyrolite a compound from the class of crystalline calcium silicate hydrates,is investigated and exploited for purification purposes.Genomic DNA and open-circular plasmid bind to gyrolite adsorbents with greater affinity than the more conforma-tionally constrained supercoiled plasmid.As such the gyrolite adsorbents are an economical and scaleable alternative to chromatographic purification for the removal of DNA impurities from solutons containing sujpercoiled plasmid.The advantage of gyrolite adsorbentd is their lower unit proce and ability to selectively adsorb DNA impurities without binding supercoiled plasmid under ceratin conditions.The effects of ionic strength temperature,chelating agent divalent cation and lyotropic salts on adsorption of highly purified plasmid are studied to understand the forces that bind DNA to gyrolite a structure with hydrophilic and hydrophobic characteristice.The results indicate that DNA binding is governed by hydrogen bonding electrostatic bridging with divalent cations shilding of electrostatic repulsion, hydrophobic asdorption an disruption of integral surface weter layer on gyrolite On the basis of results from a srange of Hofmeister series stalt strongly hydrated anions may enhance DNA adsorption by promoting hydrophobic interations between DNA and gyrolite Conversely the very weakly hydrated chaotrope I~- may enhance adsorption by strongly associating with hydrophbic siloxanes fo gyrolite theredy disrupting an integral water layer which competes for hydrogen bonding sites.
机译：研究和开发了超螺旋质粒，开环质粒和基因组DNA选择性吸附到陀螺石中的一种结晶硅酸钙水合物化合物，用于纯化目的。基因组DNA和开环质粒以更高的亲和力与陀螺石吸附剂结合因此，陀螺吸附剂是色谱纯化的一种经济，可扩展的替代方案，可从含超螺旋质粒的溶液中去除DNA杂质。研究了离子强度温度，螯合剂二价阳离子和溶致盐对高纯度质粒吸附的影响，以了解将DNA结合到具有亲水和疏水特性的陀螺仪上的作用力。有结果表明，DNA结合是通过氢键与二价阳离子的静电键搭桥，静电排斥，疏水性吸附以及对陀螺仪整体表面湿润层的破坏来控制的。根据一系列Hofmeister系列盐的结果，强水合阴离子可以增强DNA吸附相反地​​，非常弱水合的离液剂I--可以通过与疏水性硅氧烷强缔合而增强吸附，从而破坏了完整的水层，后者竞争氢键位。

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