【作者】 李朦朦； 杨斌；

【Author】 Mengmeng Li;Bin Yang;College of Chemistry, Xiangtan University;

【机构】 湘潭大学化学学院；

【摘要】 支点介导的DNA链置换策略的研究,引起了研究人员的广泛兴趣,因为它功能强大,在许多DNA电路、DNA机器、DNA生物传感器中的到了应用。研究人员已经开发了多种新的支点活化策略,以实现DNA链置换反应的灵活调控。但是,这些新支点要么采用了共价连接的方式,要么采用双链互补杂交的方式来起作用,而这样的设计策略不够灵活,限制了支点应用的范围和前景。为了解决以上问题,我们构建了一种新型的支点,把它命名为"协同支点",以实现对DNA链置换反应的灵活调控。该协同支点是基于DNA缺口位置,相邻碱基间的碱基堆积效应来起作用的,它对碱基序列没有特殊要求,且相邻两条DNA的序列完全不相关。该协同支点能以不同的浓度,实现对DNA链置换速率的连续调控,并能与可逆支点、支点交换和远程支点联合,构建高级的DNA分子器件。该支点还能构建通用型生物传感器,实现对ATP和RNA的定量测定。它也能与PCR扩增结合,实现对质粒DNA的高灵敏测定,检测限达到a M水平。更多还原

【Abstract】 Toehold-mediated DNA strand displacement has enabled to develop a dreamy set of DNA devices. So far, many new toehold activation mechanisms have been developed. However, almost all those toeholds are inflexible via either a covalently attached manner or a complementary hybridization strategy, which limit the versatility of DNA devices. To solve this problem, we developed a new toehold, named ‘cooperative toehold’, to activate DNA strand displacement. It is based on a base stacking mechanism. The cooperative toehold enabled to continuously tune the rate of DNA strand displacement, as well as more sophisticated strand displacement reactions. The cooperative toehold has also been employed as a universal signal translator for biosensors to qualitatively determine RNA and ATP. Moreover, as a novel specific PCR monitoring system, cooperative toehold-mediated DNA strand displacement can detect the pUC18 plasmid in genomic DNA samples with an aM detection limit.更多还原