从牛莫拉菌中纯化Mbo II甲基化酶（GAAGmA）：在9个和10个碱基对序列处对DNA进行位点特异性切割

McClelland M, Nelson M, Cantor C R

Nucleic Acids Res. 1985 Oct 25;13(20):7171-82. doi: 10.1093/nar/13.20.7171.

The restriction modification methylase M. Mbo II has been purified using a sensitive oligonucleotide linker assay. The enzyme methylates the Mbo II recognition sequence* GAAGA at adenine to produce GAAGmA. M. Mbo II can be used in conjunction with the methylation dependent restriction endonuclease Dpn I (GmATC) to produce cleavage at the 10 base sequence GAAGATCTTC. When M. Mbo II is used in combination with M. Cla I (ATCGATCGAT), cleavage by Dpn I occurs at the four ten base sequences GAAGATCTTC, GAAGATCGAT, ATCGATCTTC and ATCGATCGAT, which is equivalent to a nine base recognition site. The use of combinations of adenine methylases and Dpn I to generate highly selective DNA cleavages at a variety of sequences up to fourteen base pairs is discussed.

限制修饰甲基化酶M. Mbo II已通过灵敏的寡核苷酸连接子分析法进行了纯化。该酶将Mbo II识别序列*GAAGA中的腺嘌呤甲基化，生成GAAGmA。M. Mbo II可与甲基化依赖性限制内切酶Dpn I（GmATC）联合使用，以在10个碱基序列GAAGATCTTC处产生切割。当M. Mbo II与M. Cla I（ATCGATCGAT）联合使用时，Dpn I会在四个10个碱基序列GAAGATCTTC、GAAGATCGAT、ATCGATCTTC和ATCGATCGAT处发生切割，这相当于一个9个碱基的识别位点。本文讨论了使用腺嘌呤甲基化酶和Dpn I的组合在长达14个碱基对的各种序列上产生高度选择性DNA切割的方法。