Shandong Provincial Key Laboratory of Chemical Energy Storage and Novel Cell Technology, School of Chemistry and Chemical Engineering, Liaocheng University, Liaocheng, China.
Luminescence. 2021 Aug;36(5):1272-1276. doi: 10.1002/bio.4053. Epub 2021 Apr 27.
A simple, rapid and highly sensitive method for detection of double-stranded DNA (dsDNA) was developed using a novel fluorescence probe composed of a RecA-GFP fusion protein that had specific recognition of ssDNA complexes (RecA-GFP-DNA filament). The RecA-GFP fusion protein not only had strong fluorescence, but could also occur by homologous recombination. In the presence of the target dsDNA, the complementary ssDNA of the RecA-GFP-DNA filaments invaded one end of the dsDNA chain. In addition, the other end of the ssDNA dissociated the RecA-GFP filaments. An assay of the probe showed a linear relationship with dsDNA concentration in the range 1-11 nM, with a correlation coefficient of 0.9923. The limit of detection for dsDNA was determined experimentally to be 0.3 nM (3δ). Compared with conventional methods, this method has the advantages of simple operation, high specificity, and high sensitivity.
一种简单、快速、高灵敏的双链 DNA（dsDNA）检测方法，利用一种新型荧光探针，该探针由 RecA-GFP 融合蛋白组成，对 ssDNA 复合物（RecA-GFP-DNA 丝）具有特异性识别。RecA-GFP 融合蛋白不仅具有强荧光，而且可以通过同源重组发生。在靶 dsDNA 存在下，RecA-GFP-DNA 丝的互补 ssDNA 侵入 dsDNA 链的一端。此外，ssDNA 的另一端使 RecA-GFP 丝解离。该探针的测定显示出与 dsDNA 浓度在 1-11 nM 范围内的线性关系，相关系数为 0.9923。实验确定 dsDNA 的检测限为 0.3 nM（3δ）。与传统方法相比，该方法具有操作简单、特异性高、灵敏度高等优点。
