本研究的重点是天然化合物的应用,3,5-二羟基4',来自药用植物Alpinianigra的7-二甲氧基黄酮(DHDM)用于核酸检测和差异细胞染色。发现DHDM与核酸相互作用并形成复合物,针对各种应用进行了调查。它被成功地用于可视化质粒,基因组,和ds线性DNA在琼脂糖凝胶电泳中不影响DNA在凝胶中的迁移率。DHDM的荧光在与dsDNA结合时增加了几倍。评估化合物的光稳定性并显示光漂白效应,其随着时间逐渐降低。化合物的应用进一步扩展到差异细胞染色。当在荧光显微镜下观察时,DHDM对死细胞进行染色,并在细菌的情况下将它们从活细胞中分化出来,酵母,和哺乳动物细胞。发现较高浓度的化合物对癌细胞的细胞毒性较小。核酸染色染料,如溴化乙锭(EtBr),碘化丙啶(PI),等。是致癌物和环境污染物,因此DHDM是一种天然化合物,是一个主要的好处,因此可以作为一个替代目前的染料。
The present
study is focused on application of a natural compound, 3, 5-dihydroxy 4\', 7-dimethoxyflavone (DHDM) from a medicinal plant Alpinia nigra for nucleic acid detection and differential cell staining. DHDM was found to interact with nucleic acid and forms complex, which was investigated for various applications. It was successfully utilized to visualize plasmid, genomic, and ds-linear DNA in agarose gel electrophoresis without affecting the DNA mobility in the gel. Fluorescence of DHDM increased several fold upon binding to dsDNA. Photostability of the compound was assessed and showed photobleaching effect that decreased gradually over time. Application of the compound was further extended to differential cell staining. When observed in fluorescence microscope, DHDM stained the dead cells and differentiated them from live cells in the case of bacterial, yeast, and mammalian cells. Higher concentration of the compound was found to be less cytotoxic to cancerous cells. Nucleic acid staining dyes like Ethidium bromide (EtBr), Propidium iodide (PI), etc. are carcinogens and environmental pollutants and therefore DHDM a natural compound, is a major benefit and thus can serve as an alternative to the current dyes.