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首页> 外文期刊>Biochemistry >Substrate Specificity and Biochemical Properties of M3.BstF5I DNA Methyltransferase from the BstF5I Restriction-Modification System
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Substrate Specificity and Biochemical Properties of M3.BstF5I DNA Methyltransferase from the BstF5I Restriction-Modification System

机译:BstF5I限制性修饰系统的M3.BstF5I DNA甲基转移酶的底物特异性和生化特性

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摘要

Optimal conditions for DNA methylation by the M3.BstF5I enzyme from Bacillus stearothermophilus and kinetic parameters of λ phage DNA modification and that of a number of oligonucleotide substrates are established. Comparison of M1.BstF5I and M3.BstF5I kinetic parameters revealed that with similar temperature optima and affinity for DNA, M3.BstF5I has nearly fourfold lower turnover number (0.24 min~(-1)) and modifies the hemimethylated recognition site with lower efficiency under optimal conditions than the unmethylated one. In contrast to another three methylases of the BstF5I restriction-modification system, the M3.BstF5I enzyme is able to optionally modify the noncanonical 5'-GGATC-3' DNA sequence with a rate more than one order of magnitude lower than the methylation rate of the canonical 5'-GGATG-3' recognition site.
机译:建立了嗜热脂肪芽孢杆菌的M3.BstF5I酶进行DNA甲基化的最佳条件,以及λ噬菌体DNA修饰的动力学参数和许多寡核苷酸底物的动力学参数。 M1.BstF5I和M3.BstF5I动力学参数的比较表明,在相似的最佳温度和对DNA的亲和力下,M3.BstF5I的转换数(0.24 min〜(-1))降低了近四倍,并且在较低的效率下修饰了半甲基化的识别位点最佳条件要比未甲基化的条件好。与BstF5I限制性修饰系统的另外三种甲基化酶相比,M3.BstF5I酶能够选择性地修饰非规范性5'-GGATC-3'DNA序列,其降解速率比BstF5I的甲基化速率低一个数量级。规范的5'-GGATG-3'识别站点。

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