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A G-triplex based molecular beacon for label-free fluorescence 'turn-on' detection of bleomycin

机译:基于G-三链接的分子信标,用于无标记的荧光“开启”检测博莱霉素

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摘要

Since bleomycins (BLMs) play a prominent role in the clinical treatment of various cancers, the development of convenient and sensitive detection assays for BLM is of great significance in cancer therapy and related biological mechanism research. Here, taking advantage of the easily controllable and excitation of the G-triplex DNA structure, we reported a facile, label-free G-triplex based functional molecular beacon (G3MB) sensing system for fluorescence "turn-on" detection of BLM based on BLM-Fe(II) mediated DNA strand scission. In the presence of BLM, the stable hairpin structure of G3MB undergoes an irreversible cleavage in the loop region that contains a 5'-GT-3' recognition site for BLM. The released G-tract DNA fragment self-assembles into a G-triplex-ThT complex showing a strong fluorescence. Owing to the effective locking of G-tracts in the stem of the G3MB and the specific DNA strand scission by BLM which is like a key for the release of G-tracts, the assay shows high sensitivity and selectivity with a detection limit of 0.2 nM. In addition, satisfactory results were obtained for the detection of BLM in human serum samples. Critically, the convenient "mix-and-detect" protocol, fast response and no need for modifying DNA offered a potential application of the proposed strategy for BLM assay in biomedical and clinical studies.
机译:由于Bleomycins(Blms)在各种癌症的临床治疗中发挥着突出作用,因此BLM的方便和敏感的检测测定的发展在癌症治疗和相关的生物机制研究方面具有重要意义。在这里,利用G-TripleDNA结构的易于可控和激发,我们报道了一种容易,无标记的G-Tripleclep的功能分子信标(G3MB)感测系统,用于基于的BLM检测荧光“开启”检测BLM-FE(II)介导的DNA链易于进行释放。在BLM存在下,G3MB的稳定发夹结构在含有BLM的5'-GT-3'识别位点的环形区域中经历不可逆的裂解。将释放的G-TRACT DNA片段自组合成G-TRIPLEX-THT复合物,显示出强荧光。由于在G3MB的茎中的G阱和BLM的特异性DNA链裂变中,这是释放G型颈部的键,测定显示出高灵敏度和选择性,检测限为0.2nm 。此外,获得了令人满意的结果,用于检测人血清样品中的BLM。批判性地,方便的“混合和检测”协议,快速响应,不需要改变DNA,提供了在生物医学和临床研究中提出的BLM测定策略的潜在应用。

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