Shukla H, Kobayashi Y, Arenstorf H, Yasukochi Y, Weissman S M
Yale University School of Medicine, Department of Human Genetics, New Haven, CT.
Nucleic Acids Res. 1991 Aug 11;19(15):4233-9. doi: 10.1093/nar/19.15.4233.
We have used a combination of BsuE methyltransferase (M-BsuE) and NotI restriction enzyme to cut genomic DNA at a subset of NotI sites. The usefulness of this system is shown in a re-examination of the restriction map of the human MHC. Combinations of methylases and restriction enzymes can be used to generate cuts at different frequencies in genomic DNA, such that they generate ends complementary to NotI ends, and can be used in conjunction with NotI linking clones in chromosome jumping experiments. These enzyme combinations have the potential to produce cutting sites in genomic DNA spaced at intervals favorable for extensive mapping, fragment enrichment, and cloning efforts.
我们使用了BsuE甲基转移酶(M-BsuE)和NotI限制酶的组合,在NotI位点的一个子集中切割基因组DNA。该系统的实用性在对人类主要组织相容性复合体(MHC)限制酶切图谱的重新检测中得到了体现。甲基化酶和限制酶的组合可用于在基因组DNA中以不同频率产生切割,从而产生与NotI末端互补的末端,并可在染色体跳跃实验中与NotI连接克隆一起使用。这些酶组合有潜力在基因组DNA中产生间隔适宜的切割位点,有利于进行广泛的图谱绘制、片段富集和克隆工作。