McrBC是一种作用于含有甲基胞嘧啶DNA的特异性核酸内切酶，不作用于非甲基化DNA。 它能够特异性地识别和切割包含两个 (G/A)mC间距在40 bp-2000 bp的DNA序列。 (G/A)mC中的甲基化胞嘧啶可以是5-羟甲基胞嘧啶, 5-甲基胞嘧啶或4-甲基胞嘧啶。 大肠杆菌表达的重组体蛋白质。 在50 μl反应液中，37℃下反应1小时，将0.25 μg的含有单一McrBC酶切位点的线型pBR322 (pBR322-mu-BamH I-M Alu I) 完全分解，所需要的酶量定义为1个活性单位 (U)。 DNA甲基化位点解析 …

2023年9月15日 · McrBC is an endonuclease which cleaves DNA containing methylcytosine* on one or both strands. Sites on the DNA recognized by McrBC consist of two half-sites of the form (G/A)mC. These half-sites can be separated by up to 3 kb, but the optimal separation is 55-103 base pairs. *5-methylcytosine or 5-hydroxymethylcytosine.

2018年6月12日 · McrBC是一种检测基因组小部分区段的DNA甲基化情况的核酸内切酶。 只能降解甲基化的区域，对未甲基化区域不起作用。 酶切体系. 具体操作以及试剂信息相见 McrBC. 发现其中有一个McrBC实验的图，当时还没在意，到了这几天自己要做的时候，才发现是如此的有用. 图片解析. 首先在前文的WGCNA中的M6模块中作者注意到了一个DNA去甲基化酶基因 SlDML2 (已经被证明控制一些成熟相关基因 (RIN 等)的启动子区域的DNA去甲基化)，表明在水果成熟过程 …

McrBC specifically cleaves DNA containing methyl cytosine (mC : 5-hydroxymethylcytosine, 5-methylcytosine, 4-methylcytosine) preceded by a purine nucleotide (Pu : A or G). DNA cleavage by McrBC requires at least two PumC sites within a distance of 40 – 2,000 bp. 1 hour at 37°C in a total reaction volume of 50 μl.

DNA上McrBC的识别位点由两个 (G/A)mC形式的半位点组成，这两个半位点之间的距离可以达到3kb，但是最佳距离为55～103bp。 McrBC的切割需要GTP，当存在不可水解的GTP类似物时，该酶可以特异地结合到甲基化DNA上，但不能进行切割。 McrBC作用于一对PumCG序列元件，以此检测高比例甲基化的CpGs，但是不能识别内部胞嘧啶甲基化了的HpaII/MspI位点 (CCGG)。 注 *：5-甲基胞嘧啶、5-羟甲基胞嘧啶或者N4-甲基胞嘧啶。 测定CpG二核苷酸的甲基化状态。 测 …

Mcr phenotypes reported in [Raleigh, E.A. et al. (1988) Nucl. Acids Res., 16, 1563-1575 PMID: 2831502, References 7 in Restriction of Foreign DNA by E. coli K-12] except those denoted with asterisks. Mrr phenotypes determined at NEB except DH10B (11). EcoKI phenotypes sometimes inferred from genotype. Table Notes:

We offer a range of Escherichia coli bacterial cells made competent with the highest efficiencies by optimized procedure specific to each strain. Choose from 24 new competent cells for a wide variety of applications, including protein expression, routine or difficult cloning, and library generation. Many trial sizes are available. Table 1.

1992年5月20日 · McrBC-mediated restriction of modified DNA has been studied extensively by genetic methods, but little is known of its molecular action. We have used overproducing plasmid constructs to facilitate purification of the McrBL and McrC proteins, and report preliminary characterization of the activity of the complex.

For optimal performance, perform expression in S.O.C. Medium and incubation on antibiotic plates at 30°C. MAX Efficiency® Stbl2TM Competent Cells are a derivative of JM109. The mcrA mutation and the mcrBC-hsdRMS-mrr deletion allow for the cloning of genomic sequences that are methylated (3).

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