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研究了一种新型丙烯酸酯(E)-2-氰基-3-[4-(2,4-二氟苯氧基)-苄氨基]-3-甲氧基-丙烯酸2-乙氧基乙酯(MAE)对超螺旋pUC19 DNA的定点切割作用。利用紫外吸收、荧光光谱、凝胶电泳和DNA测序技术研究其核酸切割特异性。紫外吸收光谱的红移和减色效应,以及静态荧光猝灭表明MAE与DNA的作用属于典型的插入模式。不仅如此,凝胶电泳条带的变化以及DNA测序结果表明DNA的定点切割是通过DNA双链的分步水解完成。
A novel acrylate E-2-cyano-3- [4- (2,4-difluorophenoxy) -benzylamino] -3-methoxy- Site-directed cleavage of supercoiled pUC19 DNA with ester (MAE). The specificity of nucleic acid cleavage was studied by ultraviolet absorption, fluorescence spectroscopy, gel electrophoresis and DNA sequencing. The red shift and subtractive effect of UV absorption spectra, as well as the static fluorescence quenching, indicate that the interaction between MAE and DNA belongs to the typical insertion mode. Moreover, changes in gel electrophoresis bands and DNA sequencing results indicate that site-directed DNA cleavage is accomplished by a stepwise hydrolysis of the DNA double-stranded DNA.