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首页> 外文期刊>Plasmid: An International Journal Devoted to Extrachromosomal Gene Systems >Tungsten particle-induced nicking of supercoiled plasmid DNA.
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Tungsten particle-induced nicking of supercoiled plasmid DNA.

机译：钨粒子诱导的超螺旋质粒DNA切口。

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摘要

Small particles of metallic tungsten, known also as tungsten microprojectiles, are routinely used for biotechnological purposes. In such applications, tungsten was observed to affect the integrity of plasmid DNA. Here we present evidence that interaction between tungsten particles and intact circular plasmids pU19, pUC119, and ColE1 may result in generation of a limited number of single-strand DNA breaks. As a consequence, supercoiled DNA is converted into its open circular form and no fragmentation products can be detected. The rate of the tungsten-mediated reaction depends on pH but is not influenced by ascorbate, Tris, or EDTA. No DNA nicking can be observed when the tungsten particles are replaced by substances that can be leached out from these particles with water or incubation buffers. Likewise, commercial sodium tungstate, tungsten (VI) oxide, and tungsten (VI) chloride and products of its decomposition remain DNA undamaged. Native plasmid DNA molecules, upon adsorption on the surface of tungsten microparticles, may undergo some nicking without a need for participation of external catalysts. Copyright 2000 Academic Press.
机译：金属钨的小颗粒，也称为钨微粒，通常用于生物技术目的。在这样的应用中，观察到钨影响质粒DNA的完整性。在这里，我们提供的证据表明，钨粒子与完整的环状质粒pU19，pUC119和ColE1之间的相互作用可能会导致产生数量有限的单链DNA断裂。结果，超螺旋DNA被转化成其开放的环状形式，并且没有片段化产物被检测到。钨介导的反应速率取决于pH值，但不受抗坏血酸盐，Tris或EDTA的影响。当钨颗粒被可以用水或温育缓冲液从这些颗粒中浸出的物质代替时，没有观察到DNA切口。同样，商品钨酸钠，氧化钨（VI）和氯化钨（VI）及其分解产物仍保持DNA不变。天然质粒DNA分子吸附在钨微粒的表面上后，可能会发生一些切口，而无需外部催化剂的参与。版权所有2000学术出版社。

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