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首页> 外文期刊>RSC Advances >Cyclization of secondarily structured oligonucleotides to single-stranded rings by using Taq DNA ligase at high temperatures
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Cyclization of secondarily structured oligonucleotides to single-stranded rings by using Taq DNA ligase at high temperatures

机译：通过在高温下使用Taq DNA连接酶将二次结构寡核苷酸的环化对单链环。

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摘要

Single-stranded DNA rings play important roles in nanoarchitectures, molecular machines, DNA detection, etc. Although T4 DNA ligase has been widely employed to cyclize single-stranded oligonucleotides into rings, the cyclization efficiency is very low when the oligonucleotides (l-DNAs) take complicated secondary structures at ambient temperatures. In the present study, this problem has been solved by using Thermus aquaticus DNA ligase (Taq DNA ligase) at higher temperatures (65 and 70 degrees C) where the secondary structures are less stable or completely destroyed. This method is based on our new finding that this ligase successfully functions even when the splint strand is short and forms no stable duplex with l-DNA (at least in the absence of the enzyme). In order to increase the efficiency of cyclization, various operation factors (lengths and sequences of splint, as well as the size of the DNA ring) have been investigated. Based on these results, DNA rings have been successfully synthesized from secondarily structured oligonucleotides in high yields and high selectivity. The present methodology is applicable to the preparation of versatile DNA rings involving complicated secondary structures, which should show novel properties and greatly widen the scope of DNA-based nanotechnology.
机译：单链DNA环在纳米建筑，分子机，DNA检测等中起重要作用。尽管T4 DNA连接酶已被广泛用于将单链寡核苷酸旋转到环中，但是当寡核苷酸（L-DNA）时，环化效率非常低在环境温度下采取复杂的二级结构。在本研究中，通过在较高温度（65和70摄氏度）下使用Thermus Aquaticus DNA连接酶（Taq DNA连接酶）来解决该问题，其中二次结构稳定或完全破坏。该方法基于我们的新发现，即使当夹板链短并且没有与L-DNA的稳定双链体形成（至少在没有酶的情况下）也成功发挥作用。为了提高环化效率，已经研究了各种操作因子（夹板的长度和序列，以及DNA环的尺寸）。基于这些结果，已经成功地以高产率和高选择性从二次结构化寡核苷酸中成功地合成了DNA环。本方法适用于制备涉及复杂的二级结构的通用DNA环，这应该显示新的性质，大大扩大了基于DNA的纳米技术的范围。

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  • 来源
    《RSC Advances》 |2018年第34期|共8页
  • 作者

    Cui Yixiao; Han Xutiange; An Ran; Zhou Guangqing; Komiyama Makoto; Liang Xingguo;

  • 作者单位

    Ocean Univ China Coll Food Sci &

    Engn Qingdao 266003 Peoples R China;

    Ocean Univ China Coll Food Sci &

    Engn Qingdao 266003 Peoples R China;

    Ocean Univ China Coll Food Sci &

    Engn Qingdao 266003 Peoples R China;

    Ocean Univ China Coll Food Sci &

    Engn Qingdao 266003 Peoples R China;

    Ocean Univ China Coll Food Sci &

    Engn Qingdao 266003 Peoples R China;

    Ocean Univ China Coll Food Sci &

    Engn Qingdao 266003 Peoples R China;

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  • 正文语种 eng
  • 中图分类 化学;
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