An Approach to DNA Crystallization Using the Thermal Reversible Process of DNA Duplexes
An Approach to DNA Crystallization Using the Thermal Reversible Process of DNA Duplexes
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DOI：
10.1021/cg9007075
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发表时间：
2010-03-01
影响因子：
3.8
通讯作者：
Morimoto, Yukio
中科院分区：
文献类型：
--
作者：
Chatake, Toshiyuki;Sazaki, Gen;Morimoto, Yukio
我们提出了一种利用DNA的热可逆过程进行DNA结晶的技术。双链DNA(DsDNA)可以在高温下转化为两个单链DNA(SsDNA)，以改善其水溶性。因此，这种热转化可以用来通过改变溶质物种来增加DNA晶体的溶解度。我们研究了DNA六角体d(CGCGCG)晶体的溶解度及其熔化温度，在该温度下，它的dsDNA与单链DNA发生热转换，反之亦然。在315K附近，van‘t Hoff图显示了两个结晶过程，其溶解度随温度的变化而变化，这一结果与紫外光谱测得的熔融温度(322K)基本一致。这些发现表明，从dsDNA到ssDNA的转换导致了溶解度的增加。利用这种温控技术，可以从少量的DNA样品中获得DNA六角体的单晶；X射线分析表明这些晶体是高品位的。这种易于应用的技术将优于传统的蒸汽扩散技术，因为它允许控制DNA晶体的溶解度，而不需要昂贵的设置。
We propose a technique for DNA crystallization using the thermal reversible process of DNA. A double-stranded DNA (dsDNA) can be converted into two single-stranded DNAs (ssDNAs) at high temperatures to improve its water solubility. Thus, this thermal conversion can be utilized for increasing the solubility of DNA crystals by changing solute species. We investigated the solubility of the crystals of a DNA hexamer d(CGCGCG) and their melting temperature, at which thermal conversion occurs from it dsDNA to ssDNAs or vice versa. A van't Hoff plot showed two crystallization processes with it change in temperature dependency of the solubility at around 315 K. This result almost corresponded to the melting temperature (322 K) measured by UV spectroscopy. These Findings suggest that the conversion from a dsDNA to ssDNAs results in an increase in solubility. Using this temperature-controlled technique, single crystals of the DNA hexamer could be obtained from a small amount of DNA samples; X-ray assessment demonstrated that these crystals were high grade. This easy-to-apply technique would be superior to the conventional vapor diffusion technique in that it allows the solubility of DNA crystals to be controlled with no need for expensive Setups.