Effective detection and separation method for G-quadruplex DNA based on its specific precipitation with Mg(2+).
Effective detection and separation method for G-quadruplex DNA based on its specific precipitation with Mg(2+).
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DOI：
10.1021/bm100862k
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发表时间：
2010-11
影响因子：
6.2
通讯作者：
Jing Lin;Yi-yong Yan;Tian-Miao Ou;Jia-Heng Tan;Shi-Liang Huang;Ding Li;Zhishu Huang;L. Gu
中科院分区：
文献类型：
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作者：
Jing Lin;Yi-yong Yan;Tian-Miao Ou;Jia-Heng Tan;Shi-Liang Huang;Ding Li;Zhishu Huang;L. Gu
G-四链体是由特定的富含鸟嘌呤的序列形成的一类DNA二级结构。由于其在功能基因组区域的富集及其生物学意义，G-四链体被认为是癌症和其他疾病的重要药物靶标。在这里，我们测试了Mg（2+）对各种DNA寡聚体的沉淀效率，包括单链、双链、三链体、发夹、i基序和一些报道的G-四链体DNA。结果表明，Mg（2+）对具有平行构象的G-四链体DNA具有很高的识别和沉淀效率，可达100%，这为从其他DNA结构中检测和分离G-四链体DNA以及从其他构象的G-四链体中鉴定平行的G-四链体提供了一种廉价、方便的方法。进一步的CD和凝胶电泳实验验证了这种方法的有效性。用透射电子显微镜（TEM）对沉淀物的结构进行了表征，观察到的线性沉淀物表明G-四链体DNA的聚合是通过G-四链体独特的大芳香族表面通过端到端的π-π堆积形成的。
G-quadruplex is a type of DNA secondary structure formed by specific guanine-rich sequences. Because of their enrichment in functional genomic regions and their biological significance, G-quadruplexes are recognized as significant drug targets for cancer and other diseases. Here, we tested the precipitation efficiency of Mg(2+) for various DNA oligomers, including single-stranded, double-stranded, triplex, hairpin, i-motif, and some reported G-quadruplex DNA. It was found that Mg(2+) could specifically recognize and precipitate G-quadruplex DNA with a particularly high efficiency of close to 100% for G-quadruplex structures with parallel conformation, which provided an inexpensive and convenient method for detecting and separating G-quadruplex DNA from other DNA structures as well as identifying parallel G-quadruplex from other conformational G-quadruplexes. Further experiments with both CD and gel electrophoresis validated the effectiveness of this approach. The structure of the precipitate was characterized using transmission electron microscopy (TEM), and the observed linear precipitate suggested that a polymerization of G-quadruplex DNA was formed through π-π stacking of end to end by the unique large aromatic surface of G-quadruplex.