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圆二色性和差示扫描量热法研究 DNA 适体二聚体的性质。

The circular dichroism and differential scanning calorimetry study of the properties of DNA aptamer dimers.

机构信息

Department of Nuclear Physics and Biophysics, Faculty of Mathematics, Physics and Informatics, Comenius University, 84248 Bratislava, Slovakia.

出版信息

Biophys Chem. 2011 Apr;155(1):29-35. doi: 10.1016/j.bpc.2011.02.004. Epub 2011 Feb 21.

Abstract

We have applied circular dichroism (CD), temperature-gradient gel electrophoresis (TGGE) and differential scanning calorimetry (DSC) to study the properties of novel bioengineered DNA aptamer dimers sensitive to fibrinogen (F) and heparin (H) binding sites of thrombin and compared them with canonical single stranded aptamer sensitive to fibrinogen binding site of thrombin (Fibri). The homodimer (FF) and heterodimer (FH) aptamers were constructed based on hybridization of their supported parts. CD results showed that both FF and FH dimers form stable guanine quadruplexes in the presence of potassium ions like those in Fibri. The thermal stability of aptamer dimers was slightly lower compared to those of canonical aptamers, but sufficient for practical applications. Both FF and FH aptamer dimers exhibited a potassium-dependent inhibitory effect on thrombin-mediated fibrin gel formation, which was on average two-fold higher than those of canonical single stranded Fibri aptamers.

摘要

我们应用圆二色性(CD)、温度梯度凝胶电泳(TGGE)和差示扫描量热法(DSC)研究新型生物工程 DNA 适体二聚体的性质,这些适体二聚体对凝血酶的纤维蛋白原(F)和肝素(H)结合位点敏感,并将其与对凝血酶纤维蛋白原结合位点敏感的经典单链适体(Fibri)进行了比较。同源二聚体(FF)和异源二聚体(FH)适体基于其支持部分的杂交构建。CD 结果表明,FF 和 FH 二聚体在钾离子存在下像 Fibri 一样形成稳定的鸟嘌呤四链体。与经典适体相比,适体二聚体的热稳定性略低,但足以满足实际应用的需要。FF 和 FH 适体二聚体均表现出对凝血酶介导的纤维蛋白凝胶形成的钾依赖性抑制作用,其抑制作用平均比经典单链 Fibri 适体高两倍。

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